Krill Prize
Krill Prize
Krill Prize
for Excellence in Scientific Research
in Honor and in Memory of
Benjamin and Gitla Krill Mansbach Shlanger
The Krill Prizes for Excellence in Scientific Research
are awarded in Honor and in Memory of
Benjamin and Gitla Krill Mansbach Shlanger r.i.p .
Mr. Krill Shlanger (born 1912, Germany) was married to Debora Kretzig Stelzer Z’L.
Krill was highly active on behalf of the community in South America, Where he established a fund for the needy. Krill past away in October 2007.
The Krill Prizes for Excellence in Scientific Research, funded by the Krill Family, have been awarded since 2005.
Ten Krill Prizes, are awarded annually, in Exact Sciences, Life Sciences, Medicine, Agriculture and Engineering, to excelling faculty members at the Universities in Israel, who hold the untenured positions of Lecturer, or Senior Lecturer
Recipients are selected by the Wolf Foundation Scholarship Committee, from among outstanding candidates, submitted by all Israeli Universities. The selection is made on the basis of the candidate´s excellence and the importance of his or her field of research
Laureates
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Yoav Shechtman
Krill Prize Laureate 2021
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Moran Shalev Ben-Ami
Krill Prize Laureate 2021
The Weizmann Institute
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Benyamin Rosental
Krill Prize Laureate 2021
Ben-Gurion University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Ido Kaminer
Krill Prize Laureate 2021
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Tamir klein
Krill Prize Laureate 2021
The Weizmann Institute
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Merav Parter
Krill Prize Laureate 2021
The Weizmann Institute
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Guy Katz
Krill Prize Laureate 2021
The Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Naomi Habib
Krill Prize Laureate 2021
The Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Liron Barak
Krill Prize Laureate 2021
Tel-Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Joshua Baraban
Krill Prize Laureate 2021
Ben-Gurion University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Schraga Schwartz
Winner of Krill Prize 2020
Weizmann Institute of Scienc
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Kfir Blum
Winner of Krill Prize 2020
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Tomer Michaeli
Winner of Krill Prize 2020
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yuval Filmus
Winner of Krill Prize 2020
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Meirav Zehavi
Winner of Krill Prize 2020
Ben Gurion University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Idan Hod
Winner of Krill Prize 2020
Ben Gurion University of the Negev
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Adam Teman
Winner of Krill Prize 2020
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yasmine Meroz
Winner of Krill Prize 2020
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yakir Hadad
Winner of Krill Prize 2020
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yonit Hochberg
Winner of Krill Prize 2020
The Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Neta Regev-Rudzki
Krill Prize 2019
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Ofer Firstenberg
Winner of Krill Prize 2019
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Amnon Bar-Shir
Winner of Krill Prize 2019
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Shahar Kvatinsky
Winner of Krill Prize 2019
Technion Institute of Technology
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yaron Fuchs
Winner of Krill Prize 2019
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Baruch Barzel
Winner of Krill Prize 2019
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Malachi Noked
Winner of Krill Prize 2019
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Noga Ron-Zewi
Winner of Krill Prize 2019
University of Haifa
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Dafna Shahaf
Winner of Krill Prize 2019
The Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Ori Katz
Winner of Krill Prize 2019
The Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Itzhak Tamo
Krill Prize Laureate 2018
Tel-Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Amit Sever
Krill Prize Laureate 2018
Tel-Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Meital Landau
Krill Prize Laureate 2018
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Charles E. Diesendruck
Krill Prize 2018
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yakov Babichenko
Krill Prize Laureate 2018
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Ayelet Erez
Krill Prize Laureate 2018
Weismann Institute
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Adi Salomon
Krill Prize Laureate 2018
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Elad Gross
Krill Prize Laureate 2018
The Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Emmanuel Levy
Krill Prize Laureate 2018
Weizmann Institute
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Anat Milo
Krill Prize Laureate 2018
Ben-Gurion University of the Negev
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Avi Shroeder
Krill Prize Laureate 2017
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Carmel Rotchild
Krill Prize Laureate 2017
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Shiri Chechik
Krill Prize Laureate 2017
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yonatan Dubi
Krill Prize Laureate 2017
Ben Gurion University of the Negev
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yoav Goldberg
Krill Prize Laureate 2017
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Zvika Brakerski
Krill Prize Laureate 2017
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Asya Rolls
Krill Prize Laureate 2017
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Nir Bar-Gill
Krill Prize Laureate 2017
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yossi Buganim
Krill Prize Laureate 2017
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Eilon Shani
Krill Prize Laureate 2017
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Keren Censor-Hillel
Krill Prize Laureate 2016
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Netanel Lindner
Krill Prize Laureate 2016
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Maya Bar Sadan
Krill Prize Laureate 2016
Ben Gurion University of the Negev
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Jakub Abramson
Krill Prize Laureate 2016
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Itay Halevy
Krill Prize Laureate 2016
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yossi Yovel
Krill Prize Laureate 2016
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Daniel Deutch
Krill Prize Laureate 2016
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Assaf Rinot
Krill Prize Laureate 2016
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Lioz Etgar
Krill Prize Laureate 2016
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Dana Reichmann
Krill Prize Laureate 2016
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Eran Ofek
Krill Prize Laureate 2015
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Ido Amit
Krill Prize Laureate 2015
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Barak Dayan
Krill Prize Laureate 2015
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Natalie Elia Herooty
Krill Prize Laureate 2015
Ben Gurion University of the Negev
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Oded Rechavi
Krill Prize Laureate 2015
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Alex Retzker
Krill Prize Laureate 2015
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Michael Schapira
Krill Prize Laureate 2015
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Sharon Ruthstein
Krill Prize Laureate 2015
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Uri Shapira
Krill Prize Laureate 2015
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Moran Bercovici
Krill Prize Laureate 2015
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Or Dunkelman
Krill Prize Laureate 2014
University of Haifa
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Irit Gat-Viks
Krill Prize Laureate 2014
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Lilach Gilboa
Krill Prize Laureate 2014
The Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Eran Bouchbinder
Krill Prize Laureate 2014
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Shahal Ilani
Krill Prize Laureate 2014
Weizman Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Roie Yerushalmi
Krill Prize Laureate 2014
Hebrew Universuty of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Daniel Podolsky
Krill Prize Laureate 2014
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Amir Yehudayoff
Krill Prize Laureate 2014
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Nathan Keller
Krill Prize Laureate 2014
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Niv Papo
Krill Prize Laureate 2014
Ben Gurion University of Negev
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
David Nicolas Waldmann
Krill Prize Laureate 2013
University of Haifa
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Tomer Volansky
Krill Prize Laureate 2013
Tel – Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Nirit Dudovich
Krill Prize Laureate 2013
Weizmann Institute of Sience
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Anat Levin
Krill Prize Laureate 2013
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Sagiv Shifman
Krill Prize Laureate 2013
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Itai Ynai
Krill Prize Laureate 2013
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Amit Kanigel
Krill Prize Laureate 2013
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Avinoam Zadok
Krill Prize Laureate 2013
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Jacob Hanna
Krill Prize Laureate 2013
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Jacob (kobi) Gal
Krill Prize Laureate 2013
Ben Gurion University of the Negev
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yoel Shkolnisky
Krill Prize Laureate 2012
Tel- Aviv University.
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Alex Bronstein
Krill Prize Laureate 2012
Tel – Aviv University.
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Erez Levanon
Krill Prize Laureate 2012
Bar-Ilan University.
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Amos Tanay
Krill Prize Laureate 2012
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Nachum Ulanovsky
Krill Prize Laureate 2012
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Ido Branslavsky
Krill Prize Laureate 2012
The Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Gil Alexandrowicz
Krill Prize Laureate 2012
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Louisa Meshi
Krill Prize Laureate 2012
Ben-Gurion University of the Negev.
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Amir Amedi
Krill Prize Laureate 2011
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Avishay Gal-Yam
Krill Prize Laureate 2011
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Shai Meiri
Krill Prize Laureate 2011
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Taleb Mokari
Krill Prize Laureate 2011
Ben-Gurion University of the Negev
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Reuven Cohen
Krill Prize Laureate 2011
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Oren Cohen
Krill Prize Laureate 2011
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yuval Shaked
Krill Prize Laureate 2011
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Ehud Altman
Krill Prize Laureate 2010
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Kinneret Keren
Krill Prize Laureate 2010
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Anne Bernheim
Krill Prize Laureate 2010
Ben-Gurion University of the Negev
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Dan Thomas Major
Krill Prize Laureate 2010
Bar Ilan University.
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Eran Halperin
Krill Prize Laureate 2010
Tel-Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Masha Niv
Krill Prize Laureate 2010
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Julia Kempe
Krill Prize Laureate 2009
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Nathalie Questembert-Balaban
Krill Prize Laureate 2009
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Ilan Koren
Krill Prize Laureate 2009
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Boaz Tsaban
Krill Prize Laureate 2009
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Debbie Lindell
Krill Prize Laureate 2009
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Eli C. Lewis
Krill Prize Laureate 2009
Ben-Gurion University of Negev
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Shiri Artstein-Avidan
Krill Prize Laureate 2008
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Ido Dagan
Krill Prize Laureate 2008
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Roy Bar-Ziv
Krill Prize Laureate 2008
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Eli Berger
Krill Prize Laureate 2008
University of Haifa
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Yuval Dor
Krill Prize Laureate 2008
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Jeff Steinhauer
Krill Prize Laureate 2008
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Zeev Zalevsky
Krill Prize Laureate 2007
Bar-Ilan University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Tal Alexander
Krill Prize Laureate 2007
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Roded Sharan
Krill Prize Laureate 2007
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Dr. Yoav Tsori
Krill Prize Laureate 2007
Ben-Gurion University of the Negev
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Oren Froy
Krill Prize Laureate 2007
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Amir Orian
Krill Prize Laureate 2007
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Leeor Kronik
Krill Prize Laureate 2006
Weizmann Institute of Science
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Tal Pupko
Krill Prize Laureate 2006
Tel Aviv University
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Itamar Simon
Krill Prize Laureate 2006
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Shulamit Levenberg
Krill Prize Laureate 2006
Technion
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
Dorit Aharonov
Krill Prize Laureate 2006
Hebrew University of Jerusalem
Dr. Eilon Shani
Scientific Overview:
Plant growth and development is mediated to a large extent by hormones. Plants regulate hormone response pathways at multiple levels including biosynthesis, metabolism, perception, and signaling (1-3). In addition, plants exhibit the unique ability to spatially regulate hormone distribution ( 4, 5). This ability is illustrated most clearly in the case of auxin (!AA). The combined activity of auxin influx and efflux carrier proteins generates auxin maxima and local gradients that inform developmental patterning. The regulation of the cellular localization of PIN-FORMED (PIN) efflux transporters determines the direction of auxin flow from one cell to another ( 6, 7). Until recently, little was known about the transport mechanisms and the distribution patterns of hormones other than auxin. These are exciting days for the plant hormone community as novel gibberellin (GA), abscisic acid (ABA), and cytokinin (CK) transporters have recently been identified (8-14) joining earlier findings on auxin transporters (15, 16), all are members ofNPF, ABC, or PUP families. My research group and others identified the NPFs as the first GA transporter in plants 14• Although our studies suggest that individual NPF and ABC genes have specialized functions, genetic analyses of the family have been limited by the scarcity of loss-of-function phenotypes. The major reason for this is that plant genomes are highly redundant. There are huge numbers of genes encoding NPF and ABC transporters (53 and 120, respectively, in Arabidopsis) that exhibit high redundancy despite diverse substrate specificity of the encoded proteins. This redundancy, with over 80% of all protein-coding genes belonging to families, illustrates the huge challenge to the field of plant genetics in general and to the study of hormone transport specifically. On top of that, certain transporters, such as some NPFs, can transport multiple hormone substrates (GA, ABA) generating a highly robust system that is unique and interesting but puzzling (J 2, 14, 17). Furthermore, transporters with high specificity for conjugated hormones, which must balance the subcellular active hormone pools, have not been identified.
We plan to address the burning questions that emerge from this recent progress in the field, all of which are challenging due to the functional redundancy of plant hormone transporters: 1) Do closely related NPF and ABC members transport multiple substrates and what is the biological importance of such activity? 2) Why have GA exporters, which must exist to enable the movement of the hormone, not been identified? 3) Are there specialized hormone transporters for conjugated hormones and what is their contribution to the rapid hormone response at tissue and subcellular levels? 4) Do subcellularly localized hormone transporters affect the hormone response and what is subcellular hormone map?
The major goal of the lab goals is to investigate the robust and specialized functions of plant hormone transporter families. In our lab we will are using multiple-targeted artificial microRNAs and CRISPR technology to establish Arabidopsis and tomato populations, respectively, deficient in multiple redundant hormone transporters. First stage, we generated a unique targeted forward genetic approach that bypasses functional redundancy in plants with a dynamic screening range. This allows us to first target the NPF and ABC families in Arabidopsis and tomato. At a second stage, we are using targeted high-throughput screens to identify developmental redundant phenotypes. These loss-of-function phenotypes combined with hormone-mediated physiological assays, analyses of transporter expression patterns and localization, and biochemical transport assays allows us to study NPF and ABC hormone transport mechanisms, identify missing exporters, and evaluate subcellular localization of hormone transporters. Our work in the past three years has identified novel putative GA, CK, IAA and ABA hormone transporters, including CRISPR application in tomato that recovered striking GA-mediated growth defects when six novel GA transporters were knocked out in tomato, thus emphasizing the outstanding strength of our screens. Having the novel CRISPR multiple-targeted platform at our disposal is a significant achievement, since it not only contribute to our understanding of hormone transport mechanisms but will also serve as a scientific breakthrough to overcome functional redundancy in all fields of plant biology and agricultural breeding.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.
In addition, there is growing amount of evidence suggesting that hormone transporters localized on intracellular compartments actively regulate the subcellular distributions of hormones and hormone intermediate (J 8-20). One of the labs aims is to obtain a deeper understanding of hormones distribution in snbcellular resolution, and address the question of quantitative endogenous hormone levels in different compartments. To profile cellular hormone metabolites within discrete organelles, we are using an innovative strategy to isolate distinct cellular compartments and reveal the complete profiles of active hormone and hormone precursors and conjugates in organelles. This approach will generate the first subcellular hormone localization map and will enable the quantification and characterization of the in vivo activities of known and unidentified subcellular hormone transporters. We believe that our studies will lead to a fuller understanding of hormone transporter functions and specialized subcellular activities.