A male sterile mutant, S201, was identified in Brassica napus. Genetic analysis revealed that the male sterility trait was controlled by a recessive nuclear gene, male sterility (MS), which was stably inherited. The results of microscopy showed that the main reason for male sterility was a defect in microspore development, resulting in the absence of typical exine and mature microspores. Bulked segregant analysis (BSA) and genotyping of an F2 population showed that the MS gene was located in a 1.4 Mb region. Sequence analysis showed that the CYP704B1 gene in this region contained two non-synonymous SNPs, leading to substitutions of two amino acids. A high-throughput KASP marker was characterized to detect the presence of the ms gene in the breeding population. The data presented here indicate that the male sterile mutant S201 can be applied in rapeseed breeding by producing the male sterile line and that the KASP marker developed for male sterility will be useful in marker-assisted selection of male sterile individuals in rapeseed-breeding programs.
