Quick Genotyping for an ACCase Herbicide Resistance Gene in Wheat Using KASP Assay

Through a forward genetics screen using an induced mutagenesis method, mutant lines of wheat resistant to the ACCase inhibitor quizalofop p-ethyl were previously characterized, and further crosses were performed to integrate the trait into advanced breeding lines for a commercial release. Quick and effective genotyping of these lines to tell whether they carry the resistance gene on one, two, or all three copies of the genome will be essential for diagnostic issues that may arise with the technology in a field setting. A method for genotyping these lines was previously accomplished via the KASP assay using a pre-amplification step for purification of the ACCase gene from each of the three genome copies. We have developed a different method for genotyping the ACCase gene in wheat using the KASP assay without the need for a pre-amplification step. This allows for quicker and less reagent intensive calls of whether a line is a one-gene, two-gene, or three-gene quizalofop p-ethyl resistant wheat, forgoing the accuracy of knowing the specific genome(s) that carry the trait.