MutChromSeq to identify the resistance gene Pm2 in hexaploid wheat

The gene Pm2 confers resistance to Blumeria graminis in wheat. In order to determine the sequence of Pm2 we sequenced the wildtype and 6 susceptible EMS mutants that have lost the function of Pm2. The loss is likely to be due to a non-synonymous mutation in the coding region of Pm2. We flow-sorted 5D chromosomes from wildtype and suscrptible mutants and sequenced them. Wildtype data was denovo assembled and data from individual mutants mapped against the assembly. A subsequent SNP calling revealed two candidate contigs with a mutation in each mutant line. In one of the contigs all mutations fall within an open reading frame associated with an NLR-type resistance gene.