Location: Grain Legume Genetics Physiology Research
Project Number: 2090-21000-034-008-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Sep 30, 2017
End Date: Sep 29, 2022
Objective:
Identify and characterize pathogenicity effectors of fungal pathogens including Sclerotinia sclerotiorum that are preferentially expressed during plant infection.
Approach:
1) Isolate mRNAs from fungal pathogen strains grown at different pH conditions, and isolate mRNA from legume crops infected by wild type and mutant strains of the pathogens.
2) Construct mRNA libraries and sequence the libraries using Illumina sequencing technology. Assemble the raw reads into unigenes and map the unigenes onto the genomes of pathogens.
3) Perform differential gene expression (DEG) analyses and determine genes that are up-regulated under acidic environment and during plant infection.
4) Predict presence of signal peptide using SignalP 4.1 server and identify potential secreted proteins (effectors) genes.
5) Conduct targeted gene deletions on selected potential effector genes and purify and confirm putative deletion mutants.
6) Determining phenotypic difference in growth, sclerotial production and pathogenicity between the deletion mutants and wild type strains.
7) Conduct experiments to demonstrate secretion, and protein-protein interactions of the effector protein with plant receptors in modulating disease development.
8) Publish the research findings.
