玉米C型细胞质雄性不育特异片段的分离及验证

Separation and verification of a specific AFLP fragment from the C-type cytoplasmic male sterility in maize

对一套玉米同核异质不育系及保持系（包括N48-2、T48-2、C48-2、S48-2）的线粒体DNA进行了扩增片断长度多态性（AFLP）分析比较。从选择性扩增产物中筛选到了一条C48-2所特有的差异条带C48-223。对C48-223进行克隆、测序分析表明,它是玉米C型细胞质雄性不育线粒体atp6基因编码序列的一部分。Southern、Northern杂交分析证实了C48-223在C48-2线粒体上的特异性。这说明该差异片段有可能同C型细胞质雄性不育性状的表现有关。

The analyses of mitochondrial DNA extracted from one set of isonuclear alloplasm materials including N48-2, T48- 2, C48-2 and S48-2 in maize and the comparison between them were carried out with AFLP technique. C48-223, one differential DNA segment of C48-2, was found when the mtDNA fragments were amplified with selective primers. The band was isolated, purified and sequenced, and the results showed that C48-223 was a part of atp6-C, which is a chimeric gene uniquely associated with the C male-sterile cytoplasm of maize. The results of Southern blot indicated that C48-223 was a real band. Via Northern blot, the signals can only be detected at the tetrad stage and uninucleate stage of another RNA extracted from C48-2. All these indicate that the fragment probably has strong relationship with the C male-sterile cytoplasm of maize.