Generation and analysis of base editor (BE) CRISPR/Cas9-based SlSBP tomato mutants

• Grant number: 19/24101-7
• Support type: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): December 01, 2019
• Effective date (End): November 30, 2022
• Field of knowledge: Biological Sciences - Genetics - Plant Genetics
• Principal researcher: Fabio Tebaldi Silveira Nogueira
• Grantee: Carlos Hernan Barrera Rojas
• Home Institution: Escola Superior de Agricultura Luiz de Queiroz (ESALQ). Universidade de São Paulo (USP). Piracicaba , SP, Brazil
• Associated research grant: 18/17441-3 - Molecular control of shoot architecture: interplay between microRNAs, transcription factors and phytohormones, AP.TEM

Abstract

Fundamental questions in Plant Biology include how cells and tissues maintain their identity over time and how they switch from one stable state to the next. Understanding such basic questions may help us to better predict how plants modulate their shoot architecture in response to endogenous cues and the environment. Shoot architecture is one of the main developmental factors affecting plant productivity. Predictable, plants have evolved intricate regulatory networks to module their shoot architecture, which include phytohormones, transcription factor-regulated transcriptional programming, and epigenetic factors like microRNAs. However, how these different factors integrate to control shoot architecture at molecular and cellular levels is still unclear in most species. Following our findings that microRNAs (miRNAs) can regulate different aspects of sugarcane and tomato development by interacting with phytohormones, the characterization of miRNA-based patterning mechanisms and their association with phytohormones is a central aspect of our work. For example, we are studying the interaction between the gibberellin-negative regulators DELLA with miRNA-controlled pathways during flower and shoot development. Moreover, we are evaluating the interactions between phytohormones and microRNA modules in the control of axillary shoot branching, which directly influence shoot architecture. For each of the research areas, we intend to employ developmental genetics, genome editing (via CRISPR/Cas9-based technologies), next generation sequencing (NGS), imaging, and bioinformatics approaches. Therefore, the main objective of this project is to generate and characterize CRISPR/Cas9-based SlSBP mutants, which will enormously help to unravel the molecular mechanisms underlying the establishment of tomato shoot architecture. (AU)