Molecular identification and distinctness of NSa male sterile cytoplasm in Brassica napus.
Molecular identification and distinctness of NSa male sterile cytoplasm in Brassica napus.
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DOI：
10.3724/sp.j.1006.2008.01946
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发表时间：
2008-01-01
期刊：
影响因子：
--
通讯作者：
Wang, W. M.
中科院分区：
文献类型：
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作者：
Cheng JiHua, Cheng JiHua;Li YunChang, Li YunChang;Wang, W. M.
细胞质雄性不育作为一种有效的授粉控制系统在油菜杂交生产中得到了广泛应用。随着十字花科野生植物的开发和生物技术的应用，近年来报道了几种油菜细胞质雄性不育系统。传统的细胞质鉴定方法耗时费力，效率低。因此，建立一种高效、准确的细胞质分类鉴定方法是十分必要的。基于重复序列的聚合酶链反应（rep-PCR）已被证明是鉴定基因组中含有丰富重复序列的线粒体和原核生物的一种很好的方法。采用rep-PCR技术对Nsa、Pol、nap、No、Ogu和Ath 6种细胞质雄性不育材料进行指纹图谱分析。根据两对引物的PCR扩增带型，可以明确地区分出6个油菜雄性不育细胞质。其中，ERIC750和ERIC4000是甘油三酯与葡萄球菌体细胞杂交的NSa CMS所特有的条带。利用基因特异性引物的PCR结果也表明，NSa是油菜细胞质雄性不育的新系统。从而建立了一种油菜细胞质分类方法。这两个特异标记不仅在分子水平上证实了NSa细胞质与其他雄性不育细胞质的差异性，也为这一潜在CMS材料在油菜育种和生产中的知识产权保护提供了有用的工具。
Cytoplasmic male sterility (CMS) has been widely used as an efficient pollination control system in rapeseed (Brassica napus) hybrid production. With the exploitation of wild species of Crucifer and the application of biotechnology, several cytoplasmic male sterile systems of oilseed rape have been reported in recent years. Traditional methods used for the identification of cytoplasm, however, are time and labor consuming with low efficiency. Thus, it is highly desirable to develop an efficient and accurate identification method for cytoplasm classification. Repetitive sequence based polymerase chain reaction (rep-PCR) has proved to be a good method for the identification of mitochondrial and procaryotic organisms which contain abundant repetitive sequences in genome. Fingerprints of six cytoplasmic male sterility materials including Nsa, Pol, nap, No, Ogu, and Ath were mapped using rep-PCR. Based on the PCR amplification band patterns by two primer pairs, six male sterile cytoplasms in rapeseed could be distinguished unambiguously. Two bands, ERIC750 and ERIC4000, were unique to NSa CMS which was derived from somatic hybrids between B. napus and S. arvensis. The PCR results using gene specific primers also showed that NSa is a novel cytoplasmic male sterility system in oilseed rape. Thus, a method for cytoplasm classification in oilseed rape was established. The two specific markers for NSa cytoplasm not only confirmed the distinctness of NSa cytoplasm from other male sterile cytoplasm at molecular level, but also provided useful tools for the protection of intellectual property right of this potential CMS material in rapeseed breeding and production.