与水稻K型细胞质雄性不育性特异的mtDNA片段的鉴定

Identifying the mtDNA Fragment Which Links to Cytoplasmic Male Sterility in Rice K Type CMS Germplasm

水稻K型雄性不育材料是新发现的一类核质互作不育类型。本研究以水稻K型雄性不育的不育系(K17A)、保持系(K17B)以及杂交F1代(K优17)为研究对象。采用SDS法提取各水稻材料的线粒体DNA(mtDNA),mtDNA经纯化后利用EcoRⅠ和MseⅠ进行双酶切,并进行AFLP分析。利用AFLP分子标记技术从上述材料的mtDNA中找到了K17A、K优17所共有的特异片段,回收并纯化该片段。特异片段经克隆测序后,得知该特异片段序列为442bp所组成。利用GenBank对其进行序列对比分析,表明该序列与水稻其他已知核质互作不育类型特异片段序列不相关,并且该序列与水稻的两种未知功能蛋白的表达相关。

A new type material of male sterility lines named K type was detected recently. The sterility line （K 17A）, maintainer line （D17B） and FI hybrid （Kyoul7） in K type cytoplamic male sterility of rice were studied. The mtDNA of each rice line was extracted by SDS method and recovery, purification and digestion by EcoRⅠand Mse Ⅰ were carried out, and finally they were detected by AFLP approach to find the specific fragment linked to the CMS. The results indicated that the specific fragment of K17A and Kyoul7 should be in common, which is 442 bp length. Sequencing analysis showed the fragment we got is not related to known specific fragment of nucleo-cytoplasmic interaction sterility in rice, but related to two unknown functional protein.