甜菜细胞质雄性不育相关基因的克隆表达及CMS的相关性研究

已有的研究表明，线粒体atpA基因和orf187片段与作物雄性不育关系密切。为了研究atpA基因和orf187片段与甜菜细胞质雄性不育之间的关系，采用同源序列法分离克隆了甜菜体内atpA和orf187序列；序列分析揭示这2个片段序列在不育系和相应保持系间存在差异，进一步利用半定量RT-PCR和Real-time PCR技术分析了两系的atpA基因和orf187片段的表达情况。结果表明，与保持系相比，atpA在不育系中发生了碱基的置换及插入；orf187在不育系中缺失6个碱基，而在保持系中多了6 bp的重复序列；基因表达分析也显示，不育系atpA在花蕾发育的前2个时期表达量明显低于相应的保持系，而在大花蕾期其表达量又高于相应的保持系；orf187在不育系小花蕾时期表达量明显高于相应的保持系，随着花蕾的发育，该序列在不育系中的表达逐渐减弱并明显低于对应保持系中的表达量。推测atpA和orf187的结构变异及异常表达与甜菜细胞质雄性不育有着密切关系。

Cloning and Expression of Cytoplasmic Male Sterility Associated Genes in Sugar Beet and Its Relationship to CMS

The results indicated that Mitochondrial atpA gene and orf187 fragment were closely related to Male Sterility in Crops. In order to research the relationship with atpA gene and orf187 fragment, cytoplasmic male sterility in sugarbeet, atpA gene and orf187 fragment in sugarbeet were isolated and cloned using homology-based technique. The sequence differences in sugarbeet sterile line and its corresponding maintainer line were defined according to sequence analysis, expression analysis of atpA gene and orf187 fragment was studied further use of the semi-quantitative RT-PCR and Real-time PCR. The results indicated that compared with maintainer line, sterile line atpA had the replacement of bases and insertion;6 base pairs were missing in sterile line orf187. The expression analysis demonstrates that expression of atpA in sugarbeet sterile line is significantly lower than in maintainer line besides the big flower bud period, while expression of orf187 in sugarbeet sterile line is apparently higher than in maintainer line besides the big flower bud period. These results indicated that structural variation and abnormal expression pattern of atpA and orf187 has a close relationship with sugarbeet cytoplasmic male sterility.