Rapid gene isolation in barley and wheat by mutant chromosome sequencing
Sanchez-Martin J, Steuernagel B, Ghosh S, Herren G, Hurni S, Adamski N, Vrana J, Kubalakova M, Krattinger SG, Wicker T, Dolezel J, Keller B and Wulff BBH (2016) Rapid gene isolation in barley and wheat by mutant chromosome sequencing. Genome Biology17: 221.
Identification of causal mutations in barley and wheat is hampered by
their large genomes and suppressed recombination. To overcome these
obstacles, we have developed MutChromSeq, a complexity reduction
approach based on flow sorting and sequencing of mutant chromosomes, to
identify induced mutations by comparison to parental chromosomes. We
apply MutChromSeq to six mutants each of the barley Eceriferum-q gene and the wheat Pm2
genes. This approach unambiguously identified single candidate genes
that were verified by Sanger sequencing of additional mutants.
MutChromSeq enables reference-free forward genetics in barley and wheat,
thus opening up their pan-genomes to functional genomics.