Project Number: 2090-21000-034-021-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Sep 30, 2021
End Date: Sep 29, 2024
The objectives are to investigate pathogenicity mechanisms of grain legume pathogens and to develop management practices for controlling grain legume diseases including developing biological control using mycovirus-harboring stains and identifying new resistance sources.
A. Approaches for pathogenicity mechanisms: A1. Investigate up-regulated genes of Sclerotinia sclerotiorum during plant infection and at acidic environment by performing RNA-seq, transcriptome analysis, and differentially expressed gene (DEG) analysis. A2. Predict presence of signal peptide using SignalP 4.1 server and identify potential secreted proteins (effectors) genes. A3. Conduct targeted gene deletions on selected potential effector genes using the split-marker technique, confirm putative deletion mutants and purify the confirmed mutants to homokaryotic state with respect to the deletion alleles, using repeated hyphal tipping and/or through ascospore purification. A4. Determining phenotypic difference in morphology (growth, sclerotial production) and pathogenicity between the deletion mutants and the wild type strains. A5. Conduct experiments to demonstrate secretion, and protein-protein interactions of the effector proteins with plant receptors in modulating disease development, using modern molecular techniques such as yeast-two-hybrid (Y2H), bimolecular fluorescence complementation (BiFC) and co-immunoprecipitation (Co-IP). A6. Prepare official reports and publish the research findings in peer-reviewed journals. B. Approaches to management of grain legume diseases: B1. Identify resistance sources in wild relatives of pea, chickpea and lentil to fungal and oomycete diseases commonly found in the US Pacific Northwest through experimentation in the field, greenhouse and laboratory. B2. Determine the genetics of resistance and resistance mechanisms using classical and modern molecular techniques. B3. Identify microbial agents in indigenous and introduced microbes, especially mycovirus-harboring strains, that are suitable for managing fungal diseases through testing in the laboratory, greenhouse and, with permission from the Animal and Plant Health Inspection Service (APHIS), in the field. B4. Investigate the interactions of plant genotypes with microbial biocontrol agents in disease development and develop strategies for practical applications. B5. Demonstrate potential utility of the developed disease management strategies. B6. Prepare official reports and publish the research findings.