两种甘蓝Ogura细胞质雄性不育源的分子鉴别

摘要

[目的]比较两种可实用的甘蓝Ogura细胞质雄性不育源(CMSHY和CMSR)在细胞质DNA水平上的区别,以快速、准确鉴别两种不育源.[方法]利用开发的叶绿体SSR(cpSSR)和线粒体SSR(mtSSR)引物,结合聚丙烯酰胺凝胶电泳和测序技术,获得在两种不育源间的分子差异,并对其中有限制性内切酶的位点转化成CAPS标记.[结果]无法在聚丙烯酰胺凝胶电泳上发现32对cpSSR扩增产物的差异,经测序仅发现ACP9引物上SSR重复数的差异.在21对mtSSR引物中,仅mtSSR2引物可以在聚丙烯酰胺凝胶电泳上区分这两种不育源.经测序发现其余5对mtSSR引物上存在9个多态性差异,其中3个为SSR位点的差异.对其中2个可被限制性内切酶MseI酶切的多态性位点转化成CAPS标记,分别命名为m92-143 Mse I、ml-346 Mse I.这些标记均可以用于这两种Ogura细胞质雄性不育源的区分.[结论]利用获得的cpSSR和mtSSR标记,成功区分甘蓝Ogura细胞质雄性不育源CMSHY和CMSR.CAPS标记可快速、简便、准确地区分两种不育源.%[Objective] Two feasible Ogura cytoplasmic male sterility sources (CMS HY and CMSR3) in cabbage were used to identify its cytoplasmic DNA distinction. [Method] With chloroplast SSR (cpSSR) and mitochondrial SSR (mtSSR) primers, the technologies of polyacrylamide gel electrophoresis and DNA sequencing were used to explore the molecular differences between Ogura CMSHY and CMSR3. The markers with restriction enzyme recognition sites were converted into CAPS markers. [Result] cpSSR amplicons could not differ two CMS sources on polyacrylamide gel, but the sequence of amplicon ACP9 differed them on SSR repeat numbers. In 21 mtSSR primers, only amplicon of mtSSR2 could distinguish two CMS sources on polyacrylamide gel. Moreover, nine polymorphic differences were obtained with sequencing in other 5 mtSSR primers. Two polymorphic differences with Mse I restriction enzyme recognition sites were converted into CAPS markers, named as m92-143 Mse I, ml-346 Mse I . [Conclusion] Two cabbage Ogura CMS sources were distinguished with cpSSR and mtSSR markers. CAPS markers could be used as a fast, easy and accurate method to distinguish two CMS sources.