Marker-assisted selection in the creation of rapeseed maitainer lines

The article presents the results of a study on the application of marker-assisted selection (MAS) for developing sterility maintainer lines in the breeding of F1 hybrids of spring rapeseed (Brassica napus L.). Fifteen lines and hybrids from the collection of LLC “N.N. Timofeev Breeding Station” were analyzed using multiplex PCR to identify five types of cytoplasm (Ogura, Ogura-NWSUAF, Polima, Cam, Nap) based on markers of the orf138, orf222 and orf224 genes. This method enabled a detailed analysis of the genetic material and identified plants with Cam (RYa016) and Polima (DDShf) cytoplasm types. Specifically, molecular genotyping revealed the absence of samples with normal cytoplasm, two samples with Ogura-type cytoplasm, and ten with Ogura-NWSUAF, confirming the prevalence of these variants in the studied collection. Using four pairs of molecular markers and phenotypic evaluation, hybridological analysis and molecular screening of progeny from crosses with the CMS tester line M8ms (Ogura-NWSUAF type) were conducted to assess the presence/absence of the Rfo fertility restorer gene, which restores fertility only for the Ogura cytoplasm type. All plants in the M8ms × RYa016 combination formed sterile flowers with underdeveloped anthers and a complete absence of viable pollen, whereas fertile flowers of the RYa016 line exhibited well-formed anthers with viable pollen (fertility 98%). In other crossing combinations, a 1:1 segregation of fertile and sterile plants was observed, as expected in a typical test cross of a heterozygote Rfrf with a recessive homozygote rfrf. The analysis included evaluation of the genotype and phenotype of the obtained hybrids, revealing that the RYa016 line has Cam-type cytoplasm and lacks the Rfo gene, making it suitable as a sterility maintainer for Ogura-type cytoplasm. The results demonstrate the high efficiency of marker-assisted selection, which significantly accelerates and simplifies the process of creating F1 rapeseed hybrids.

яровой рапс, закрепитель стерильности, молекулярные маркеры, мультиплексная ПЦР, ген-восстановитель фертильности