青花菜 Ogu 不育胞质分子鉴定和序列分析

摘要

Cytoplasmic male sterility (CMS)is an important tool forheterosis utilization,recurrent selection and popu-lation improvement in crops,and has great utility value in crop production.In order to identify the cytoplasmic male sterile (CMS)types and obtaln the sterility-related sequences in broccoli (Brassica oleracea var.italica ),to provide a new Ogu marker and reference for broccoli germplasm collection,utilization and molecular marker assisting selection, specific primers P1/P2 were designed to amplify twenty broccoli germplasm genomic DNA according to the conserved sequences of orf 138 gene.A 392 bp specific fragment was detected in twelve infertile broccoli genotypes by PCR am-plification,eight fertile genotypes with no band,harmonized with morphology in field.The sequencing and comparison showed the sequence (Genbank accession code:HQ149728)as high as 100% homologous to the reported orf 138 (Genbank accession code:Z18896.1)in radish.Sequence homology comparison detected orf 138 gene contalned some variation sites.In this study,the molecular identification of broccoli CMS,was helpful to further elucidate the mecha-nism of CMS abortion,to guide creation new CMS lines,and to provide a theoretical basis for heterosis efficient utili-zation.%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 GenBank 中 orf 138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 orf 138序列,GenBank 中的登录号为 HQ149728；用 Blastn 在 GenBank 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 orf 138基因(Genbank 登录号：Z18896．1)同源度高达100％.序列同源比对发现 orf 138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.