Genotyping Wheat Grains to Identify Parental Donation of Alleles v1

In wheat research, KASP (Kompetitive Allele Specific PCR) is widely used to determine the genotypes of progeny from large, segregating populations. However, DNA extraction and genotyping is generally performed on seedling leaf tissue, and thus genotypes that affect seed formation/germination cannot be detected. We have developed a method that relies on DNA extraction from wheat grains to capture these missing genotypes. Furthermore, DNA extraction and genotyping of the endosperm, rather than the embryo, means that we can determine, for each individual, whether an allele is inherited maternally or paternally. This can be used to determine whether an allele is deleterious when inherited either maternally or paternally, or whether particular combinations of alleles are deleterious when inherited from the same parent, but not two separate parents. Moreover, the embryo remains intact throughout this protocol and is capable of germination, allowing adult plants to be recovered from genotypes of interest.