辣椒核质互作雄性不育系线粒体不育基因CoxⅡ和atp6的克隆与序列分析

Clone and Sequence Analysis of the Specific Mitochondrial Genes CoxⅡ and atp6 Related to Cytoplasmic Male Sterile in Pepper

以辣椒核质互作雄性不育系9704A、保持系9704B和恢复系9701(简称"三系")为材料,根据GenBank报道的茄科作物线粒体CoxⅡ和atp6基因编码序列分别设计引物,PCR扩增辣椒"三系"线粒体DNA目的基因片段,研究辣椒"三系"线粒体DNACoxⅡ和atp6基因的差异及与雄性不育的关系。结果表明:从辣椒核质互作雄性不育系中扩增到两个基因的部分序列atp6-706和CoxⅡ-708,GenBank登录号分别为FJ986191和FJ986190,生物信息学分析发现分离的atp6-706和CoxⅡ-708片段与GenBank报道的茄科作物线粒体CoxⅡ和atp6基因的相似性高达95%以上;在保持系和恢复系中均未能扩增到任何序列,说明辣椒雄性不育系的CoxⅡ和atp6基因与保持系和恢复系在线粒体DNA水平上存在差异,这种结构上的变化暗示可能与辣椒雄性不育相关。

In order to study the relation between cytoplasmic male sterile（CMS） and the variation in DNA sequence of CoxⅡ and atp6 gene in the cytoplasmic male-sterile（CMS） line,pepper CMS line 9704A,its maintainer 9704B,and restorer lines 9701,were used as experimental materials.The primers were designed according to the reserved sequences in the encoding region of mitochondrial genes CoxⅡ and atp6,associated with cytoplasmic male sterility（CMS）,in some Solanaceae species from GenBank.The CoxⅡ and atp6 gene fragments,CoxⅡ-708 and atp6-706（GenBank accession numbers：FJ986190 and FJ986191）,associated with cytoplasmic male sterility（CMS） were cloned from the mitochondrial DNA of a pepper（Capsicum annuum L.） cytoplasmic male sterile line using PCR.These fragments hadn＇t complete encoding region but showed the homology of 95% with CoxⅡ and atp6 genes from the referred Solanaceae in GenBank.No band was cloned from the mitochondrial DNA of maintainer and restorer lines.The result suggested a close relationship of pepper CMS with the variation in DNA sequence of CoxⅡ and atp6 gene in CMS line.