MutChromSeq to identify the resistance gene Pm2 in hexaploid wheat. MutChromSeq_Pm2

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.