论文部分内容阅读
菜薹(Brassica campestris L.ssp.Chinensis var.utilis Tsenet Lee)属于十字花科芸薹属,原产中国,是华南地区重要的特产蔬菜之一。利用分子标记技术进行菜薹雄性不育相关基因的定位对菜薹品质创新和选育新品种具有重要意义。目前尚未见有利用ISSR分子标记技术进行菜薹雄性不育分析的研究报道。利用ISSR技术对菜薹雄性不育系及其保持系的基因组DNA进行比较分析;结果显示100个ISSR引物中,只有引物UBC843的扩增结果在两系之间表现出了差异性,找到了不育系和保持系间的特异扩增片段。回收该特异扩增片段并进行克隆测序,测序结果表明该片段全长为462bp。与白菜其它育性相关序列比较,同源性均低于50%。根据测序结果设计了一对特异引物,将其转化为更稳定的SCAR标记。经F2群体验证,ISSR引物扩增得到的特异片段很有可能来自核DNA。
Brassica campestris L.ssp.Chinensis var.utilis Tsenet Lee belongs to cruciferous Brassica genus, originating in China. It is one of the important specialty vegetables in southern China. The molecular marker technique for locating male-sterile genes in flowering Chinese cabbage is of great significance to the innovation of quality and the breeding of new varieties. At present, there is no report about the analysis of male sterility of Cauliflower by ISSR molecular marker technology. ISSR technology was used to analyze the genomic DNA of the male sterile line and its maintainer line. The results showed that only the amplification result of the primer UBC843 out of the 100 ISSR primers showed a difference between the two lines, and found no Pedigree and maintainer line-specific amplified fragments. The specific amplified fragment was recovered and cloned and sequenced. The sequencing results showed that the full length of the fragment was 462bp. Compared with other fertility-related sequences in cabbage, homology was less than 50%. According to the sequencing results, a pair of specific primers was designed and transformed into a more stable SCAR marker. The F2 population verified that the specific fragment amplified by ISSR primer is likely to come from nuclear DNA.