Sharon Ruthstein

Krill Prize 2015
Bar-Ilan University

Sharon Ruthstein

Research Topic:
Exploring the human copper transfer mechanism by electron paramagnetic resonance spectroscopy

Sharon Ruthstein’s research aims to shed light on biological pathways that involve metal ions by using Electron Paramagnetic Resonance Spectroscopy (EPR). More than 30% of all proteins in the cell exploit one or more metals to perform their specific functions, and over 40% of all enzymes contain metals. Metals are commonly found as natural constituents of proteins; however, many metal ions can be toxic when free in biological fluids. Hence, humans and microorganisms have evolved a regulatory machinery to acquire, utilize traffic, detoxify, and otherwise manage the intracellular and extracellular concentrations and types of metal ions. Despite the high regulation of metal ions in the human body, diseases such as Menkes, Wilson, Alzheimer’s, Parkinson’s and Prion’s have been linked with metal binding to proteins. It is therefore vitally important to understand each step of the copper cycle, in order to build a fundamental understanding of sources of the disruption of copper homeostasis that can lead eventually to neurological diseases and disorders. Dr. Ruthstein’s lab is looking into some of the least understood biological processes that are related to metal ion transport and intracellular distribution.

Some of the ongoing projects in her lab are:
(i) Explore metal ion import and export mechanism of the human copper transporter CTR1, from the blood carrier copper protein, albumin, through the CTR1, and to the metallochaperone Atox1.
(ii) Help understand metal binding mechanism of various metal sensors in bacterial cells (CueR and CsoR systems), in order to shed light on the metal regulatory machinery of the bacteria.
(iii) Explore the role of the copper efflux system in E.coli, in particular the Cus system.
(iv) Characterize the role of copper and mutations on the aggregation and folding of proteins, in order to illuminate the microscopic origins of neurological diseases.
To comprehend such processes it is necessary to be sensitive to the structural changes that occur in the protein upon metal binding. The main biophysical tool that is used in the lab of Dr. Ruthstein’s lab is pulsed electron Paramagnetic Resonance (EPR) spectroscopy. The power of EPR lies in the sensitivity to both atomic level changes and nanoscale fluctuations. EPR can characterize properties such as redox state and ligand geometry for different functional states of the protein. The data collected by the EPR experiments is complement by various other biophysical and biochemical approaches, as well as computational methods, such as: CD, NMR, run-off transcription assays, ultra-centrifuge experiments, ITC, and MD simulations, to provide a complete picture on the cellular copper cycle. Dr. Ruthstein’s lab also has several fruitful collaborations in Israel and abroad. One collaboration is with Prof. Hamza (University of Maryland) where the heme’s transfer mechanism by the metallochaperone, HRG-3, in c.elegans is being explored. Additional collaboration is with the lab of Prof. Haim Cohen, Ariel University/BGU. In this collaboration, EPR spectroscopy is used together with other physical methods to characterize the surface radical’s nature in coals. It was revealed that several radical species are observed, and their nature is dependent on the coal rank, and micropores surface area. This work was summarized in three PCCP papers, and the experimental work was published in JoVE. Other collaborations are together with Prof. Lellouche and Dr. Goobes on characterizing the oxidation process of graphene’s surfaces. Additional collaboration is together with Dr. Lior Elbaz (BIU). In this study, Dr. Ruthstein is studying at the molecular level the coordination of heme molecules on graphene surfaces, as well as the interaction of the studied graphene surfaces with oxygen molecules.

Awards and Scholarships

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Nir Shlezinger

Krill Prize 2024
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.

Shai-Lee Horodi

Winner of Kiefer Scholarship in – 2024

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.

Chaya Keller

Krill Prize 2024
Ariel 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.

Raya Sorkin

Krill Prize 2024
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.

Hila Peleg

Krill Prize 2024
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.

Itamar Harel

Krill Prize 2024
The Hebrew 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.

Yaniv Romano

Krill Prize 2024
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.

Renana Gershoni-Poranne

Krill Prize 2024
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.

Neta Shlezinger

Krill Prize 2024
The Hebrew 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.

Mor Nitzan

Krill Prize 2024
The Hebrew 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.

Yoav Livneh

Krill Prize 2024
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.

Yuval Hart

Krill Prize 2023
The Hebrew 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.

Tomer Koren

Krill Prize 2023
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.

Inbal Talgam-Cohen

Krill Prize 2023
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.

Nitzan Gonen

Krill Prize 2023
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.

Viviane Slon

Krill Prize 2023
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.

Yotam Drier

Krill Prize 2023
The Hebrew 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.

Ido Goldstein

Krill Prize 2023
The Hebrew 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.

Shay Moran

Krill Prize 2023
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.

Aviv Tamar

Krill Prize 2023
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.

Leeat Keren

Krill Prize 2023
Weizmann

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.

Ely Kovetz

Krill Prize 2022
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.

Sivan Refaely-Abramson

Krill Prize 2022
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.

Moran Yassour

Krill Prize 2022
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.

Jonathan Ruhman

Krill Prize 2022
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.

Yehonadav Bekenstein

Kril Prize 2022
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.

Ittay Eyal

Krill Prize 2022
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.

Haitham Amal

Krill Prize 2022
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.

Gili Bisker

Krill Prize 2022
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.

Ron Rothblum

Krill Prize 2022
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.

Uri Ben-David

Krill Prize 2022
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.

Yoav Shechtman

Krill Prize 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.

Moran Shalev Ben-Ami

Krill Prize 2021
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.

Benyamin Rosental

Krill Prize 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.

Ido Kaminer

Krill Prize 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.

Tamir klein

Krill Prize 2021
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.

Merav Parter

Krill Prize 2021
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.

Guy Katz

Krill Prize 2021
The Hebrew 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.

Naomi Habib

Krill Prize 2021
The Hebrew 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.

Liron Barak

Krill Prize 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.

Joshua Baraban

Krill Prize 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.

Karam Natour

Winner of Kiefer Scholarship in – 2020

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.

Schraga Schwartz

Krill Prize 2020
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.

Kfir Blum

Krill Prize 2020
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.

Tomer Michaeli

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.

Yuval Filmus

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.

Meirav Zehavi

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.

Idan Hod

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.

Adam Teman

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.

Yasmine Meroz

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.

Yakir Hadad

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.

Yonit Hochberg

Krill Prize 2020
Hebrew Univ.

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.

Neta Regev-Rudzki

Krill Prize 2019
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.

Ofer Firstenberg

Krill Prize 2019
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.

Amnon Bar-Shir

Krill Prize 2019
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.

Shahar Kvatinsky

Krill Prize 2019
Technion 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.

Yaron Fuchs

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.

Baruch Barzel

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.

Malachi Noked

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.

Noga Ron-Zewi

Krill Prize 2019
Haifa 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.

Dafna Shahaf

Krill Prize 2019
Hebrew 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.

Ori Katz

Krill Prize 2019
Hebrew 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.

Itzhak Tamo

Krill Prize 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.

Amit Sever

Krill Prize 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.

Meital Landau

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.

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.

Yakov Babichenko

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.

Ayelet Erez

Krill Prize 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.

Adi Salomon

Krill Prize 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.

Elad Gross

Krill Prize 2018
Hebrew 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.

Emmanuel Levy

Krill Prize 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.

Anat Milo

Krill Prize 2018
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.

Avi Shroeder

Krill Prize 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.

Carmel Rotchild

Krill Prize 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.

Shiri Chechik

Krill Prize 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.

Yonatan Dubi

Krill Prize 2017
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.

Yoav Goldberg

Krill Prize 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.

Zvika Brakerski

Krill Prize 2017
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.

Asya Rolls

Krill Prize 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.

Nir Bar-Gill

Krill Prize 2017
Hebrew 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.

Yossi Buganim

Krill Prize 2017
Hebrew 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.

Yael Frank

Winner of Kiefer Scholarship in – 2017

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.

Eilon Shani

Krill Prize 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.

Keren Censor-Hillel

Krill Prize 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.

Netanel Lindner

Krill Prize 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.

Maya Bar Sadan

Krill Prize 2016
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.

Jakub Abramson

Krill Prize 2016
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.

Itay Halevy

Krill Prize 2016
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.

Yossi Yovel

Krill Prize 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.

Daniel Deutch

Krill Prize 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.

Assaf Rinot 

Krill Prize 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.

Lioz Etgar

Krill Prize 2016
Hebrew 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.

Dana Reichmann

Krill Prize 2016
Hebrew Univ.

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.

Eran Ofek

Krill Prize 2015
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.

Ido Amit 

Krill Prize 2015
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.

Barak Dayan

Krill Prize 2015
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.

Natalie Elia Herooty

Krill Prize 2015
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.

Oded Rechavi

Krill Prize 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.

Alex Retzker

Krill Prize 2015
Hebrew 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.

Michael Schapira

Krill Prize 2015
Hebrew Univ.

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.

Sharon Ruthstein

Krill Prize 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.

Uri Shapira

Krill Prize 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.

Moran Bercovici

Krill Prize 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.

Naama Arad

Winner of Kiefer Scholarship 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.

Or Dunkelman

Krill Prize 2014
Haifa 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.

Irit Gat-Viks

Krill Prize 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.

Lilach Gilboa

Krill Prize 2014
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.

Eran Bouchbinder

Krill Prize 2014
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.

Shahal Ilani

Krill Prize 2014
Weizman 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.

Roie Yerushalmi

Krill Prize 2014
Hebrew Univ.

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.

Daniel Podolsky

Krill Prize 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.

Amir Yehudayoff

Krill Prize 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.

Nathan Keller

Krill Prize 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.

Niv Papo

Krill Prize 2014
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.

David Nicolas Waldmann

Krill Prize 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.

Tomer Volansky

Krill Prize 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.

Nirit Dudovich

Krill Prize 2013
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.

Anat Levin

Krill Prize 2013
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.

Sagiv Shifman

Krill Prize 2013
Hebrew 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.

Itai Ynai

Krill Prize 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.

Amit Kanigel

Krill Prize 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.

Avinoam Zadok

Krill Prize 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.

Jacob Hanna

Krill Prize 2013
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.

Jacob (kobi) Gal

Krill Prize 2013
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.

Tamar Harpaz

Winner of Kiefer Scholarship 2013

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.

Yoel Shkolnisky

Krill Prize 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.

Alex Bronstein

Krill Prize 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.

Erez Levanon

Krill Prize 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.

Amos Tanay

Krill Prize 2012
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.

Nachum Ulanovsky

Krill Prize 2012
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.

Ido Branslavsky

Krill Prize 2012
Hebrew 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.

Gil Alexandrowicz

Krill Prize 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.

Louisa Meshi

Krill Prize 2012
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.

Amir Amedi

Krill Prize 2011
Hebrew 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.

Avishay Gal-Yam

Krill Prize 2011
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