甘蓝细胞质雄性不育类型的分子鉴定

Molecular identification of cytoplasmic male sterile types in cabbage

为鉴定几种来源不同的甘蓝细胞质雄性不育材料的类型,获得与其不育性状相关的特异序列,基于同源序列的候选基因法,通过检索NCBI核苷酸及蛋白质数据库获得orf138和orf224开放阅读框序列,依据其保守序列设计2对引物,对甘蓝的5个不育系及其保持系进行PCR扩增。orf138引物在5个甘蓝雄性不育系中均扩增出749 bp的片段,同源性比对发现这5个不育材料的特异片段与萝卜Ogu CMS所具有的Oguorf138基因同源度达100%,表明这几个来源不同的甘蓝细胞质不育材料同属萝卜细胞质不育类型。

In order to identify the cytoplasmic male sterile （CMS） types in cabbage and gain the sterility-related specific sequence, the homology-based candidate gene method was used and the sequences of orf138 and orf224 genes were got by searching the nucleotide and protein databases of NCBI. Based on their conserved sequences,two pair of primers were designed and the DNA amplification of 5 CMS lines of cabbage and their maintainer lines were carried out by PCR. The result indicated that a 749 bp specific fragment was produced in all the 5 CMS lines by amplification with the orf138 primer. The sequencing and comparison showed that the specific fragment from the 5 CMS lines and the Ogu orf138 of Ogu CMS in radish reached 100% in homology,meaning the 5 cabbage CMS lines belong to CMS type of radish.