胡萝卜atp8基因的克隆表达及细胞质雄性不育的相关性研究

Cloning and Expression of atp8 Gene in Carrot and Its Relationship to Cytoplasmic Male Sterility

为了研究胡萝卜细胞质雄性不育及atp8基因之间的关系,采用同源序列法分离克隆了胡萝卜atp8基因;通过序列分析明确了该基因在不育系和相应保持系的序列差异,并利用半定量RT-PCR和Real-timePCR技术分析了两系该基因的表达情况。结果表明,与不育系相比,胡萝卜保持系atp8基因序列在30～170bp之间核苷酸突变很多,且在154～162bp发生了缺失;Blast分析发现保持系atp8基因与胡萝卜nad6基因的同源性高达100%;表达分析表明,atp8基因在不育系花蕾的前5个时期表达量都明显低于相应的保持系,而在盛花期的表达量与保持系趋于一致。推测atp8基因的异常表达与胡萝卜细胞质雄性不育有着密切关系。

In order to research the relationship between carrot cytoplasmic male sterility and atp8, a mitochondrion gene fragment atp8 in carrot was isolated and cloned using homology-based technique. The sequence differences in carrot sterile line and its corresponding maintainer line were defined according to sequence analysis, and expression analysis of the gene was studied using semi-quantitative RT-PCR and Real-time PCR. The result indicated that compared with sterile line, maintainer line atp8 has a lot of mutations between 30 bp and 170 bp, and the sequence between 154 bp and 162 bp was missed. Blast analysis indicated that maintainer line atp8 shares an identity of 100% with carrot nad6. The expression analysis demonstrates that expression of atp8 in carrot sterile line is significantly lower than in maintainer line besides the flowering stage. These researchers indicated that abnormal expression of atp8 has a close relationship with carrot cytoplasmic male sterility.