Location: Plant, Soil and Nutrition Research
Project Number: 8062-21000-050-020-A
Project Type: Cooperative Agreement
Start Date: Apr 15, 2023
End Date: Apr 14, 2025
Objective:
Understanding the genetic basis of fruit ripening control is essential in accurately targeting loci and genes useful for enhancing fruit quality, shelf-life and nutrition. Cooperator has facilities and expertise to efficiently generate gene-edited tomato lines that will be provided to ARS for phenotypic assessment of candidate gene function. The cooperator also has excellent plant growth facilities and imaging equipment essential for aspects of phenotyping necessary in determining gene expression patterns and activities. BTI additionally houses the primary international database of tomato genome, gene expression, genetic diversity and phenotype information (solgenomics.net) that will be easily accessed and ultimately used as a distribution point for data resulting from agreement activities.
Approach:
ARS will identify target ripening genes for analysis and develop gene-editing constructs for deployment in transgenic tomato plants. ARS will provide DNA of isolated and sequence-validated gene-editing constructs and seed for generation of cotyledon material for transformation. Cooperator will perform plant transformation and target the generation of 10 independent tomato transformation events (T0) per DNA construct. Cooperator will isolate leaf DNA from each plant and provide to ARS for validation of transforamtion and identification of any target gene edits. Cooperator will grow edited plants to maturity and collect T1 seed. Cooperator will grow gene-edited T1 plants to maturity for phenotyping (days to breaker, days to red-ripe, ethylene evolution, texture analysis by penetrometer, carotenoid profiling) by ARS. Cooperator will provide access to confocal microscopy for image analysis of fruit tissues. Cooperator will provide T2 generation seed to ARS for any lines deemed by ARS to be of interest for further analysis. Cooperator will house resulting data in the Solgenomics database.
