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中国农学通报 ›› 2012, Vol. 28 ›› Issue (4): 104-109.

所属专题: 生物技术 园艺

• 林学 园艺 园林 • 上一篇    下一篇

甘蓝细胞质雄性不育相关基因orf138的分子特性分析

朱琴 康宗利 简元才 丁云花 康俊根   

  • 收稿日期:2011-09-18 修回日期:2011-11-22 出版日期:2012-02-05 发布日期:2012-02-05

The Molecular Characteristics of Ogura Cytoplasmic Male Sterility Related Gene orf138 in Cabbage (Brassica oleracea var. capitata)

  • Received:2011-09-18 Revised:2011-11-22 Online:2012-02-05 Published:2012-02-05

摘要:

为了研究甘蓝雄性不育机制,根据萝卜CMS相关基因orf138的序列信息,设计特异引物,并在甘蓝不同类型不育系和保持系中鉴定PCR产物的稳定性。随后利用Tail-PCR技术,扩增获得此基因的侧翼序列并进行了生物信息学分析。结果表明在甘蓝不育型材料中,能够稳定扩增出300 bp左右的单一条带,而在其他细胞质不育类型和可育材料中均未扩出条带,经多次验证结果稳定可靠。甘蓝中orf138的上下游侧翼序列有效碱基1789 bp,通过生物信息学分析,获得包括起始密码和终止密码的orf138的完整序列共417 bp。同源性比对结果显示:与甘蓝型油菜﹑白菜和萝卜的orf138片段具有高度保守性。分析侧翼序列表明甘蓝orf138的3’端是由ORF83、trnfM、ORF125等基因片段构成的一个复杂序列。获得了甘蓝OguCMS特异的分子鉴定标记,明确了orf138在甘蓝线粒体中的位置,以上结果为甘蓝雄性不育的进一步研究奠定良好的基础。

关键词: 油茶饼粕, 油茶饼粕, 蒸制, 残油榨取, 二榨油品质

Abstract:

The objective was to study the mechanism of cytoplasmic male sterile (CMS) of cabbage. The orf138 gene fragment in cabbage CMS line was amplified based on specific primers designed according to radish orf138 gene sequence information. The upstream and downstream mitochondrion sequences of orf138 sterile line were identified by use of Tail-PCR technology. The results showed that a single PCR product about 300 bp was obtained in OguCMS cabbage line, while never in fertile maintainer lines and other types of cytoplasmic male sterility lines. Upstream and downstream flanking sequence of orf138 were aligned and resulted in a 1789 bp sequence. Bioinformatics analysis showed that this sequence was 417 bp orf138 including the complete sequence. Homology comparison showed that orf138 gene of cabbage, rapeseed and radish were highly conserved. The 3' flanking mitochondrion sequence of the orf138 in cabbage showed a complex sequence including trnfM, ORF125, ORF83. The research obtained an effective specific DNA marker Bo138300BF/R and illustrated the genome location of orf138 in cabbage mitochondrion. The results lay a good foundation for further study of cabbage OguCMS.