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雄性不育是农作物利用杂种优势、进行轮回选择和群体改良的重要手段,在农作物生产中具有巨大的利用价值。该研究为了鉴定青花菜细胞质雄性不育材料的不育胞质类型,以期今后为青花菜种质资源的收集、利用及分子标记辅助育种提供新的不育标记。根据Gen Bank中orf138基因保守序列设计特异引物,对20个青花菜种质资源基因组DNA进行PCR扩增。结果表明:特异引物P1/P2在12个青花菜雄性不育基因型中均扩增出392 bp的片段,在8个可育基因型中未扩增出条带,与田间育性鉴定结果相符。获得青花菜Ogu胞质雄性不育的特异基因orf138序列,Gen Bank中的登录号为HQ149728;用Blastn在Gen Bank中进行同源性比对分析,发现12个不育材料的特异片段与已报道的萝卜Ogu CMS所具有的Ogu orf138基因(Genbank登录号:Z18896.1)同源度高达100%。序列同源比对发现orf138基因存在变异位点。研究结果可为青花菜雄性不育细胞质的分子鉴定、进一步阐明胞质雄性不育败育机理,以及指导青花菜新型不育系的创建和杂种优势高效利用提供理论依据。
Male sterility is an important means for crop utilization heterosis, reincarnation selection and group improvement, and has great value in crop production. In order to identify the types of sterile cytoplasm of broccoli cytoplasmic male sterile (CMS) materials, this study aims to provide new markers of sterility for the collection, utilization and molecular marker assisted breeding of broccoli germplasm resources in the future. Genomic DNA from 20 Broccoli germplasms was amplified by PCR using specific primers designed according to the conserved sequence of orf138 in Gen Bank. The results showed that the specific primers P1 / P2 amplified 392 bp fragments in 12 genotypes of broccoli and no bands were amplified in 8 fertile genotypes, which was consistent with the results of field fertility identification . Genomic accession number of HQ149728 was obtained from the orf138 sequence of cytoplasmic male sterility of Ogura. Blastn was used for homology alignment analysis in Gen Bank. It was found that the specific fragments of 12 inbred lines were similar to those reported The Ogu CMS gene has the homology of up to 100% with the Ogu orf138 gene (GenBank accession number: Z18896.1). Sequence homology alignment revealed that the orf138 gene has a mutation site. The results could provide a theoretical basis for the molecular identification of male sterile cytoplasm of broccoli and further elucidate the mechanism of abortion of cytoplasmic male sterility, as well as guide the creation of novel male sterile lines of broccoli and the efficient utilization of heterosis.