Fecha de publicación: -- Fuente: PubMed "pollination" Mol Breed. 2026 Nov 3;46(11):115. doi: 10.1007/s11032-026-01737-7. eCollection 2026 Nov.ABSTRACTCytoplasmic male sterility (CMS) is a crucial pollination control system for heterosis utilization in rapeseed (Brassica napus) worldwide, with Polima CMS (Pol CMS) and Ogura CMS (Ogu CMS) being the most widely utilized systems. To enable high-throughput identification of cytoplasm types, we developed 4 Kompetitive Allele-Specific PCR (KASP) markers targeting or flanking the mitochondrial sterility genes orf224/atp6 (Pol CMS) and orf138 (Ogu CMS). Genotyping of 101 rapeseed accessions with diverse genetic backgrounds demonstrated that these markers accurately discriminated among cytoplasm types​. By combining these cytoplasmic markers with markers for the nuclear fertility restorer gene (Rf), we established a dual-marker system capable of simultaneously determining cytoplasmic type (S/N) and nuclear genotype (RfRf/Rfrf/rfrf). This system enables systematic tracing of the specific causes of purity decline in male-sterile lines, restorer lines, and hybrids, with results fully consistent with field phenotyping. Furthermore, using this dual-marker-assisted selection strategy, we efficiently identified 24 isoplasmic restorer lines from 187 germplasm accessions. In conclusion, the KASP markers developed in this study provide an efficient and precise tool for identifying mitochondrial sterility genes in the Pol CMS and Ogu CMS systems, thereby substantially improving the efficiency of parental line identification and purity assessment.SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at https://doi.org/10.1007/s11032-026-01737-7.PMID:42985285 | PMC:PMC13731633 | DOI:10.1007/s11032-026-01737-7