青花菜胞质不育相关基因的SRAP标记筛选  

Identification of the Specific SRAP Marker Associated with Cytoplasmic Male Sterility Gene of Broccoli

作　　者：姚雪琴[1] 李媛[1,2] 谢祝捷[1] 柳李旺[2] 

机构地区：[1]上海市农业科学院园艺研究所 上海市设施园艺技术重点实验室,上海201106 [2]南京农业大学园艺学院,南京210095

出　　处：《分子植物育种》2009年第5期941-947,共7页Molecular Plant Breeding

基　　金：上海市科委科技攻关计划项目(07DZ19602;08DZ1906304);上海市科委重点科技攻关项目(09391911300);上海市农科院青年基金(农青年科技2008(03))共同资助

摘　　要：以青花菜Ogura胞质不育系XY及其保持系XYF基因组DNA为材料进行SRAP分析,共筛选了146对SRAP引物,其中8对引物在两系之间表现差异,经单株验证只有引物对Me6+Em7没有交换单株,找到不育系与保持系间的特异扩增片段M6E7-700,该片段仅在不育系中稳定扩增。测序结果表明该片段全长为689bp,经Blast分析比对,该片段与已报道的所有育性相关基因序列均不同源,而其部分序列与大白菜BAC克隆KBrB042N05和KBrB041L12的部分序列高度同源,表明该片段是一段新发现的与胞质不育育性相关片段,且该片段可能来自核DNA。根据测序结果设计了1对特异引物,将M6E7-700标记转化为更稳定的SCAR标记。本研究首次发现芸薹属作物Ogura-CMS不育系和保持系的核DNA也存在差异,该结果为从质核互作角度解释细胞质雄性不育的分子机制提供了新线索。SRAP （sequence-related amplified polymorphism） was analyzed between Ogura cytoplasmic male sterility line XY and its maintainer line XYF of broccoli. Total 146 pairs of SRAP primers were used. The amplification of eight primers was polymorphic in the two lines. Only the Me6＋Em7 primer had no changed-plant by individual plant testing. And a 689 bp specific band M6E7-700 was detected in male sterility line but not in the maintainer line. Analysis of the sequence showed that this fragment had no homology with all of the male sterility genes which had been reported, and part of this fragment was highly homologous with part of BAC clone KBrB042N05 and KBrB041L12 sequences in Brassica rapa. This suggested that the newly detected fragment related to cytoplasmic male sterility and properly came from nuclear DNA. This study first discovered it is different in nuclear DNA between Ogura-CMS line and its maintainer in the Brassica crop. After cloning and sequencing, specific primers were designed to transform the SRAP marker to more stable SCAR marker, which was named M6E7-700. The results here offer new clues for supporting the molecular mechanism of cytoplasic male sterility by cytoplasm-nuclear interaction.

关 键 词：青花菜 SRAP 细胞质雄性不育 分子标记 

分 类 号：S635.3[农业科学—蔬菜学]