遗传 ›› 2005, Vol. 27 ›› Issue (2): 236-240.

• 研究报告 • 上一篇    下一篇

花椰菜细胞质雄性不育基因特异PCR标记的筛选

王春国; 宋文芹WANG Chun-Guo; SONG Wen-Qin   

  1. 南开大学生命科学学院,天津 300071 The College of Life Sciences ,Nankai Uninversity,Tianjin 300071,China
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2005-04-10 发布日期:2005-04-10

Identification of PCR markers associtated with cytoplasmic male sterility in Brassica oleracea var botrytis

  • Received:1900-01-01 Revised:1900-01-01 Online:2005-04-10 Published:2005-04-10

摘要:


基于同源序列的候选基因法(homology-based candidate gene method),通过检索NCBI核酸及蛋白数据库,获得细胞质雄性不育(cytoplasmic male sterility CMS)相关的基因或开放读码框。生物学软件分析,根据保守区设计5对特异引物,PCR扩增,其中引物P9/P10在花椰菜细胞质雄性不育系knxd612中特异扩增出313 bp的片段。单株检测,RT-PCR分析,斑点杂交鉴定,确定此片段为花椰菜细胞质雄性不育系knxd612所特有。序列分析表明该片段与Ogura型胞质不育萝卜,不育相关开放读码框orf138的同源性高达98%。初步结果显示实验所用不育花椰菜胞质亦可能为Ogura型。该结果为进一步从分子水平研究花椰菜细胞质雄性不育打下了坚实的基础。Abstract: The homology-based candidate gene method was used to identified the specific PCR markers linked to cytoplasmic male sterility (CMS) in cauliflower( Brassica oleracea var botrytis.).Searching the DNA and protein data-base of NCBI , correlative genes or open reading frames were indentified .Analysis of biosoft, based on the conservative regions ,five primers were designed . Among them, only primer P9/P10 produced a 313- bp specific fragment. Identified by individual plant testing , analysis of RT-PCR and dot blot ,this fragment was only existed in CMS cauliflower knxd612.Analysis of the sequence indicated it was high homologous(98%) with orf138 of Ogura CMS radish. Primary result suggested that the cytoplasmic type of CMS cauliflower knxd612 may belong to Ogura type. This research offered a good foundation to further investigate the CMS mechanism of cauliflower in molecular level.

关键词: 花椰菜(Brassica oleracea var.Botrytis), 细胞质雄性不育, CMS, cauliflower(Brassica oleracea var.Botrytis), 候选基因
Key words