Location: Plant, Soil and Nutrition Research
Project Number: 8062-21000-050-030-A
Project Type: Cooperative Agreement
Start Date: Apr 22, 2024
End Date: Sep 30, 2025
Objective:
The ARS PI is developing a genetic diversity panel of tomato and its near wild relatives to rapidly associate nutrient quality, ripening, shelf-life and related fleshy fruit traits with naturally occurring genetic variants found in the diversity of cultivated and inter-fertile tomato wild relatives. This project has and will continue to point to numerous candidate allelic variants likely to underpin the manifestation of important plant characteristics. It is necessary to functionally test candidate variants to both validate the discovery pipeline and to confirm the value and importance of key allelic variants for use in breeding or biotechnology based crop improvement. Cooperator has efficient and low cost infrastructure and protocols for developing gene-edited, targeted gene suppression and gene over-expression genetically modified plants that can be returned to the ARS PI for evaluation. Agreement objectives are to cooperatively identify candidate genes for evaluation in the ARS PI lab, create tomato lines altered in said target genes through the Cooperator and evaluate resulting genetically modified lines back in the ARS PI lab.
Approach:
The ARS PI lab will identify candidate genes for testing and use standard biotechnology approaches routine in the lab to develop DNA plant transformation constructs for targeted CRISPR/Cas9-based gene editing, over-expression through direction of constitutively expressed or fruit-specific promoters, and/or repression via RNAi. These approaches are routine in the ARS PI lab. Cooperator will transfer ARS PI supplied DNA constructs into disarmed Agrobacterium tumefaciens, validate their sequence integrity and deploy them for plant transformation (tomato) using efficient techniques routinely used by the Cooperator. Cooperator will recover transgenic callus, induce plant regeneration and supply 5 - 10 independently transformed lines per DNA construct back to the ARS PI who will confirm genetic modification and evaluate any trait changes in the modified plants.
