1a. Objectives (from AD-416)
To functionally characterize soybean response to pathogens, with focus on Sclerotinia sclerotiorum. Candidate defense-related genes will be identified from soybean response to the pathogen or products of the pathogen. Functional characterization will include repression and/or enhancement of expression of the candidate genes in soybean and/or Arabidopsis.
1b. Approach (from AD-416)
The ARS scientist will identify candidate defense genes, whereas the collaborator's lab will assist with construct design for over and under expression of genes in host plants and will generate transgenic soybean material. The ARS scientist will assist in propagation of the soybean transgenics and will conduct disease assays on this material to determine effect on defense.
3. Progress Report
Conducted oxalate leaf infiltration experiments with transgenic and parent soybean lines to collect final biological rep for each line from three different treatments (total of 3 reps for statistical purposes). Extracted and purified RNA from leaf samples, and shipped our lab for microarray analysis, where statistical analysis was performed. Completed sets 2 and 3 biological reps for mist chamber Sclerotinia infection assays for qRT-PCR analysis by our lab. Improved on Arabidopsis inoculation method. Ordered nine Arabidopsis knock-out lines from ABRC–Salk Institute and are testing these to verify homozygousity of the T-DNA insertion. Examining which gene(s) to focus on for RNAi studies and methods for generating constructs. Bombarded soybean tissue for transformation using the RNAi construct design by the lab to knock-out a G-protein related gene in soybean. Activities of this project were monitored through personal meetings, phone calls and e-mails.