Location: Vegetable Crops Research
Project Number: 5090-21220-005-028-N
Project Type: Non-Funded Cooperative Agreement
Start Date: Mar 1, 2020
End Date: Jun 30, 2020
Objective:
1. Use enrichment sequencing to identify/verify R genes in late blight resistant germplasm.
2. Develop genetic markers associated with these genes to expedite breeding.
Approach:
Enrichment sequencing will be performed on four different sources of resistance (R) from the ARS and Collaborator breeding programs: Payette Russet; MSR061-1; Palisade Russet; and Defender Russet.
DNA will be extracted from each individual, the R gene sequences will be purified through hybridization with a DNA probe library, and the enriched DNA samples submitted for sequencing. We will utilize single molecule real time sequencing technology. The sequences will then be assembled and compared to a set of sequences from known late blight resistance genes in order to identify which gene is present in the resistant plants. We will utilize the sequences derived from the above approach to target specific genes. These will be candidates for the development of molecular markers. The candidate genes will be used as references to identify DNA sequence variation between resistant and susceptible individuals. This variation will be used to create genotype-specific primers for use in PCR-based screening of populations.
