Development and Validation of Kasp Markers for the CHAT Gene in Winter Wheat.

Wheat Stem Sawfly has long been a pest of spring wheat in the southern regions of Canada and the northern United States. However, it has now become a major pest of winter wheat in Colorado. The Colorado State University Wheat Breeding Program has identified single nucleotide polymorphisms (SNPs) that silence the A and D genomes of a gene responsible for the production of a pheromone that attracts the sawfly to the wheat plant. In the interest of incorporating this SNP into elite varieties, the program needs a higher throughput marker system, such as Kompetitive Allele Specific PCR (KASP). KASP genotyping uses two allele-specific forward primers and one common reverse primer to identify individuals as homozygous or heterozygous for an allele at a certain locus. Our research is focused on the development of a KASP assay system that can identify individuals containing SNPs for the CHAT gene in winter wheat for the CSU Wheat Breeding Program materials. This involves designing genome specific primers for the A and D genomes that amplify the regions surrounding the SNPs, and then using the sequences of that amplicon to develop primers for the KASP assay. This step has been successfully completed and the process of validating the KASP assay is in progress. Once the assay is validated, this system could potentially be used to efficiently screen our breeding materials for the target SNPs that can then be used to determine future crosses and can aid in the incorporation of the desired trait into elite varieties via marker assisted selection to enhance resistance to the wheat stem sawfly pest in winter wheat.