Location: Plant Science Research
Project Number: 5062-21000-035-000-D
Project Type: In-House Appropriated
Start Date: Mar 6, 2023
End Date: Mar 5, 2028
Objective 1: Conduct research to optimize gene editing reagents and transformation methods in soybean, alfalfa, and other legumes to increase functional efficiency and improve the screening methods that are used to identify successfully edited plants. Goal 1A: Generate a collection of gene editing reagents comprised of various promoter, Cas9 enzyme, transcriptional/translational enhancers, and terminator combinations and screen their efficiency for mutagenesis of candidate root-trait related targets using a rapid hairy-root transformation assay. Goal 1B: Improve transformation efficiencies of alfalfa and soybean by a 0.5- to 2-fold range and reduce time in tissue culture for plant regeneration in order to generate candidate root-trait related mutant plants. Goal 1C: Develop tissue-culture-free-transgene-free gene editing. Objective 2: Conduct research for functional validation of novel and previously identified genes that contribute to root architecture, nitrogen fixation, and nutrient acquisition in legume species, especially soybean and alfalfa. Goal 2: Use reagents constructed and screened for efficiency in Objective 1 to generate mutant plants for the validation of candidate root-trait related genes in legumes.
Optimize gene editing reagents and the transformation methods of recalcitrant elite, wild, and un-adapted genotypes of soybean, alfalfa, and other legumes to generate mutant plants associated with root system architecture and other root traits. Improve the screening methods used to identify successfully edited plants. Using the generated mutants, validate previously identified root system architecture and other root trait related candidate genes. Demonstrate novel tissue culture-free gene editing of legume using phloem mobile elements.