Periodic Reporting for period 2 - RobustHormoneTrans (Robustness and specialization among hormone transporters: Redundant and unique roles)
Reporting period: 2019-07-01 to 2020-12-31
The goal of this proposal is to reveal the robust and specialized function of the NPF and ABC plant hormone transporter families. The project places key technological challenges that require multi-disciplinary expertise to examine how plants balance redundancy and specialization to tightly regulate hormone localization. Broad and targeted transportome screens using multi-targeted artificial miRNAs and CRISPR technology in Arabidopsis and tomato, respectively, are designed to unveil novel plant hormone transporters, with an emphasis on subcellular localized transporters and the missing GA exporters. Specialization and robustness of candidate transporters are evaluated by integrating in vitro transport assays with in vivo growth and development experiments.
2) We have engineered CRISPR-based multiplex populations to identify redundant hormone transporter families in Arabidopsis and tomato.
3) We have dissected the contributions of specialization and robustness to hormone transporter functions.
4) We are developing a genetic system that will allow to reveal the hormone subcellular map
In addition, we have created hormone subcellular maps. Since multiple lines of evidence demonstrate that bioactive hormones, their intermediates, and conjugates are regulated at the subcellular level, we constructed a genetically based method that allows the isolation of different plant organelles. We utilize our unique method to draw the first hormone subcellular maps in plants. Results from this project provide researches worldwide with detailed quantitative information regarding subcellular localization of active hormones, their intermediates, and conjugates. Beyond the scientific knowledge obtained in this project, our proposed method to isolate different plant organelles enables subcellular transcriptomics, metabolomics, and proteomics research.