TITLE: STRENGTHEN PLANT BIOTECH PROGRAM BY INTEGRATING GENOME EDITING AND ARTIFICIAL INTELLIGENCE TECHNOLOGIES IN TOMATO PROJECTS

The goals of the projectare to use the CRISPR/Cas9 gene editing technology to produce heat tolerant traits for tomatoes; and use artificial intelligence (AI) to develop a high throughput pollen phenotyping program; and tobuild the research/training capacity in gene editing and AI at 1890s.The three specific objectives are: 1).To develop a working system for producing and characterizing CRISPR/Cas9 tomato mutants. Two transcription factor (TF) genes including HeatTF1 (Solyc08g016580.1.1, HS-upregulated in pollen mother cells (PMCs)) and HeatTF2 (Solyc06g053350, HS-down-regulated in PMCs) will be the targeted genes to develop the following modules: a) Experimental procedures for producing CRISPR/Cas9 mutant lines; b). Genotyping on/off target mutations; c) A high throughput phenotyping system for pollen heat tolerance using AI machine learning (ML) and deep learning (DL) technologies; d) Phenotyping for heat tolerance; e). Characterization of cell-type specific transcriptomes and proteomes of male gametophytes in mutant lines; f) Integrated data analysis to determine the function of gene mutation in affecting heat tolerance of pollen and other traits. 2). To generate CRISPR/Cas9 constructs/mutant plants for a list of HS-regulated genes and to select heat tolerant mutants. 3). To enhance 1890 education/training capacity using the project platform.