Periodic Reporting for period 4 - RobustHormoneTrans (Robustness and specialization among hormone transporters: Redundant and unique roles)
Reporting period: 2022-07-01 to 2023-12-31
ERC Project Summary Report: Deciphering Plant Hormone Transport
Throughout the project's duration, we embarked on a journey to unravel the intricate mechanisms governing plant hormone transport, recognizing the pivotal role these molecules play in orchestrating various aspects of plant development. Leveraging state-of-the-art genetic tools such as CRISPR and artificial miRNAs, our aim was to shed light on how plants regulate hormone localization within tissues and cells, ultimately impacting their growth and responses to environmental cues.
Our research delved deep into the transportome of Arabidopsis and tomato, employing multi-disciplinary approaches to unearth novel hormone transporters. By dissecting the roles of these transporters at subcellular levels, we sought to decipher the delicate balance between redundancy and specialization in hormone localization regulation.
During the course of our investigations, significant breakthroughs were achieved. We identified the first subcellular transporters for crucial plant hormones, unveiling their critical roles in root endodermal suberization and other developmental processes. Furthermore, our findings challenged conventional understanding by revealing nuanced interactions between different hormones, shedding light on the complexity of hormone signaling networks.
Looking back, our research endeavors expanded beyond mere exploration. We delved into the realm of stress responses, uncovering novel hormone transporters vital for plant resilience under adverse conditions. The innovative genetic toolboxes developed, such as the Multi-Knock CRISPR libraries, empowered us to uncover hidden traits, revolutionizing plant genetics studies in the process.
As the project concludes, our comprehensive approach bridging genetics, biochemistry, and plant physiology, and genetic tools has yielded invaluable insights into hormone transport mechanisms. These findings hold promise for advancements in agriculture, environmental resilience, and sustainable food production, marking a significant contribution to the scientific community's understanding of the fundamental processes governing plant biology.
Throughout the project's duration, we embarked on a journey to unravel the intricate mechanisms governing plant hormone transport, recognizing the pivotal role these molecules play in orchestrating various aspects of plant development. Leveraging state-of-the-art genetic tools such as CRISPR and artificial miRNAs, our aim was to shed light on how plants regulate hormone localization within tissues and cells, ultimately impacting their growth and responses to environmental cues.
Our research delved deep into the transportome of Arabidopsis and tomato, employing multi-disciplinary approaches to unearth novel hormone transporters. By dissecting the roles of these transporters at subcellular levels, we sought to decipher the delicate balance between redundancy and specialization in hormone localization regulation.
During the course of our investigations, significant breakthroughs were achieved. We identified the first subcellular transporters for crucial plant hormones, unveiling their critical roles in root endodermal suberization and other developmental processes. Furthermore, our findings challenged conventional understanding by revealing nuanced interactions between different hormones, shedding light on the complexity of hormone signaling networks.
Looking back, our research endeavors expanded beyond mere exploration. We delved into the realm of stress responses, uncovering novel hormone transporters vital for plant resilience under adverse conditions. The innovative genetic toolboxes developed, such as the Multi-Knock CRISPR libraries, empowered us to uncover hidden traits, revolutionizing plant genetics studies in the process.
As the project concludes, our comprehensive approach bridging genetics, biochemistry, and plant physiology, and genetic tools has yielded invaluable insights into hormone transport mechanisms. These findings hold promise for advancements in agriculture, environmental resilience, and sustainable food production, marking a significant contribution to the scientific community's understanding of the fundamental processes governing plant biology.
The project spans several primary research topics aimed at understanding plant hormone transport mechanisms, uncovering hidden traits through next-generation genetic tools, and developing innovative approaches to overcome functional redundancy in plants. Summary of the work performed and the main results achieved:
Transport Mechanisms of Plant Hormones: The project delved into understanding the intricate mechanisms underlying the transport of plant hormones, particularly gibberellins (GA) and abscisic acid (ABA). Through rigorous experimentation and modeling, the team identified NPF2.14 as a pivotal subcellular transporter responsible for the accumulation of GA and ABA in the root endodermis. This groundbreaking discovery shed light on the crucial role of these hormones in endodermal suberization and highlighted their non-antagonistic actions. Additionally, the study unveiled the mechanism driving long-distance movement of GA12 from shoot to root, providing insights into tissue-specific hormone storage and release dynamics.
Single-Cell Kinetics of Auxin Transport and Activity: Employing cutting-edge technologies, researchers investigated the dynamics of auxin transport and activity at the single-cell level within Arabidopsis roots. By developing an inducible auxin biosynthesis system and innovative image-analysis tools, we quantitatively characterized auxin movement patterns and their impact on root growth kinetics. The study uncovered novel aspects of auxin flux, including its directionality, dependency on specific transport proteins, and role in root twisting and skewing. These findings significantly advanced our understanding of auxin signaling and root development processes.
ABCB-Mediated Auxin Transport in Root Tissues: Collaborating with VIB Ghent, the project identified a cluster of ABCB transporters crucial for regulating lateral root spacing in Arabidopsis. Through gene silencing and CRISPR-based approaches, we elucidated the role of these transporters in modulating auxin oscillations and orchestrating lateral root formation. The study highlighted the significance of auxin transport dynamics in outer root tissues, offering valuable insights into root architecture and nutrient uptake strategies.
ABA Homeostasis and Long-Distance Translocation: In a bid to unravel the mechanisms governing abscisic acid (ABA) homeostasis, we uncovered the role of ABCG17 and ABCG18 as key ABA importers localized in the shoot. Their study unveiled a sophisticated mechanism whereby these transporters regulate ABA levels in different cellular compartments, thereby influencing stomatal conductance and lateral root emergence. By elucidating the shoot-to-root ABA translocation pathway, the research provided critical insights into plant adaptation to abiotic stresses and paved the way for targeted strategies to enhance drought tolerance in crops.
Next-Generation Multi-Targeted CRISPR Genetic Toolboxes: The project pioneered the development of innovative genetic toolboxes, namely Multi-Knock and mTACT, aimed at overcoming functional redundancy in plant genetics. Leveraging CRISPR technology, we designed genome-scale libraries targeting multiple gene family members, thereby uncovering hidden traits and dissecting gene functions. These toolboxes, equipped with cell type-specific promoters and optimized target selection algorithms, offered unprecedented opportunities for forward-genetic screens and tissue-specific genetic manipulation. With transformative potential in plant genetics research and crop breeding, these advancements hold promise for addressing global agricultural challenges and enhancing food security.
The results obtained from these studies provide a comprehensive understanding of hormone transport mechanisms, auxin dynamics at the single-cell level, and the role of transporters in plant growth, development, and stress responses. The developed genetic toolboxes offer novel approaches to uncover hidden traits and dissect gene functions, promising transformative impacts on plant genetics research and breeding efforts. Future perspectives include further exploration of ABA transporters, validation in different plant species, and application in addressing global agricultural challenges.
Transport Mechanisms of Plant Hormones: The project delved into understanding the intricate mechanisms underlying the transport of plant hormones, particularly gibberellins (GA) and abscisic acid (ABA). Through rigorous experimentation and modeling, the team identified NPF2.14 as a pivotal subcellular transporter responsible for the accumulation of GA and ABA in the root endodermis. This groundbreaking discovery shed light on the crucial role of these hormones in endodermal suberization and highlighted their non-antagonistic actions. Additionally, the study unveiled the mechanism driving long-distance movement of GA12 from shoot to root, providing insights into tissue-specific hormone storage and release dynamics.
Single-Cell Kinetics of Auxin Transport and Activity: Employing cutting-edge technologies, researchers investigated the dynamics of auxin transport and activity at the single-cell level within Arabidopsis roots. By developing an inducible auxin biosynthesis system and innovative image-analysis tools, we quantitatively characterized auxin movement patterns and their impact on root growth kinetics. The study uncovered novel aspects of auxin flux, including its directionality, dependency on specific transport proteins, and role in root twisting and skewing. These findings significantly advanced our understanding of auxin signaling and root development processes.
ABCB-Mediated Auxin Transport in Root Tissues: Collaborating with VIB Ghent, the project identified a cluster of ABCB transporters crucial for regulating lateral root spacing in Arabidopsis. Through gene silencing and CRISPR-based approaches, we elucidated the role of these transporters in modulating auxin oscillations and orchestrating lateral root formation. The study highlighted the significance of auxin transport dynamics in outer root tissues, offering valuable insights into root architecture and nutrient uptake strategies.
ABA Homeostasis and Long-Distance Translocation: In a bid to unravel the mechanisms governing abscisic acid (ABA) homeostasis, we uncovered the role of ABCG17 and ABCG18 as key ABA importers localized in the shoot. Their study unveiled a sophisticated mechanism whereby these transporters regulate ABA levels in different cellular compartments, thereby influencing stomatal conductance and lateral root emergence. By elucidating the shoot-to-root ABA translocation pathway, the research provided critical insights into plant adaptation to abiotic stresses and paved the way for targeted strategies to enhance drought tolerance in crops.
Next-Generation Multi-Targeted CRISPR Genetic Toolboxes: The project pioneered the development of innovative genetic toolboxes, namely Multi-Knock and mTACT, aimed at overcoming functional redundancy in plant genetics. Leveraging CRISPR technology, we designed genome-scale libraries targeting multiple gene family members, thereby uncovering hidden traits and dissecting gene functions. These toolboxes, equipped with cell type-specific promoters and optimized target selection algorithms, offered unprecedented opportunities for forward-genetic screens and tissue-specific genetic manipulation. With transformative potential in plant genetics research and crop breeding, these advancements hold promise for addressing global agricultural challenges and enhancing food security.
The results obtained from these studies provide a comprehensive understanding of hormone transport mechanisms, auxin dynamics at the single-cell level, and the role of transporters in plant growth, development, and stress responses. The developed genetic toolboxes offer novel approaches to uncover hidden traits and dissect gene functions, promising transformative impacts on plant genetics research and breeding efforts. Future perspectives include further exploration of ABA transporters, validation in different plant species, and application in addressing global agricultural challenges.
The ERC project aimed to deepen our understanding of hormone transport mechanisms, develop advanced genetic tools, and tackle the challenge of functional redundancy in plants. Along this journey, we have made significant strides that push the boundaries of current knowledge.
The outcomes of our endeavors extend beyond scientific discovery. They hold the potential to revolutionize agricultural practices, accelerate crop breeding programs, and bolster global food security. Moving forward, we envision further exploration of hormone transport mechanisms, validation across diverse plant species, and the application of genetic toolboxes to address pressing challenges in agriculture. In essence, our pioneering efforts are poised to reshape the landscape of plant genetics and crop improvement for years to come.
The outcomes of our endeavors extend beyond scientific discovery. They hold the potential to revolutionize agricultural practices, accelerate crop breeding programs, and bolster global food security. Moving forward, we envision further exploration of hormone transport mechanisms, validation across diverse plant species, and the application of genetic toolboxes to address pressing challenges in agriculture. In essence, our pioneering efforts are poised to reshape the landscape of plant genetics and crop improvement for years to come.