甘蓝型油菜细胞质雄性不育材料1575A的分子鉴定

Molecular Identification of Cytoplasmic Male Sterility Line 1575A of Brassica napus

根据已报道的油菜育性相关基因orf224、orf138和orf222设计引物对1575A、陕3A、Ogura和nap等4份甘蓝型油菜的mtDNA进行PCR扩增。结果显示:引物对Syworf1382/Xyworf1382在1575A和Ogura的mtDNA中均有1 000 bp左右的扩增片段;测序结果表明,在1575A中的该扩增片段包含已报道基因orf138(登录号为AB055435)的全部编码序列;该引物对在陕3A和nap中都没有相应扩增片段。引物对Syworf222/Xyworf222在除Ogura外的其他3个材料的mtDNA中都有196 bp左右的扩增片段;测序结果显示,扩增产物与已报道基因orf222(登录号为DQ872162)同源性为100%。初步推测1575A属于Ogu CMS的改良类型。

In this study,primers were designed according to the reported fertility-related gene orf224,orf138 and orf222 and the mtDNA amplification of 4 CMS lines of Brassica napus were carried out by PCR.The result indicated that about 1 000 bp specific fragment was produced in the 1575A and Ogura lines by amplification with the primer pair Syworf1382/Xyworf1382.Sequencing results indicated that amplified bands in the 1575A contained the entire coding sequence of orf138（Accession numbers：AB055435）.And no band was amplified in Shaan3A and nap CMS with the primer pair Syworf1382/Xyworf1382.In addition to outside Ogura mtDNA,other three materials amplified a 196 bp specific fragment with the primer pair Syworf222/ Xyworf222.Sequencing results indicated that the specific fragment and the sequence of reported gene orf222（Accession numbers：DQ872162） reached 100% in homology.The result concluded that 1575A belong to an improved Ogu CMS.