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为了研究V型细胞质雄性不育小麦的不育机理,以V型细胞质雄性不育小麦的一套近等基因系为材料,利用cDNA-RAPD方法对不育系、保持系和杂种F1线粒体RNA进行了比较分析。共用了284个引物,有104个引物在不育系和保持系间存在扩增差异,22个引物在不育系和杂种F1间存在扩增差异,找到了一个在不育系中特异表达的差异片段S238999,Northern杂交证实该片段在不育系、保持系和杂种F1的转录的确有差异,它的转录本丰度随着恢复基因的导入明显降低,表明该片段与V型小麦细胞质雄性不育具有一定的相关性。通过NCBI数据库的同源性比对,发现该片段仅与一个Hordeum vulgare基因组DNA片段有84 bp的同源序列,表明这是一个新的可能与V型小麦细胞质雄性不育相关的序列。
In order to study the mechanism of sterility in V-type CMS, a set of near-isogenic lines of V-type CMS was used as materials to perform CMS-line and hybrid F1 mitochondrial RNA by cDNA-RAPD A comparative analysis. A total of 284 primers were used, of which 104 primers had amplification differences between male sterile line and maintainer line. There were 22 amplification differences between male sterile lines and F1 hybrids, The differential fragment S238999 and Northern blot showed that the transcript of this fragment was indeed different in the male sterile line, maintainer line and hybrid F1. The transcript abundance of the fragment was significantly decreased with the introduction of the restorer gene, indicating that this fragment was not associated with the cytoplasmic male sterility of V-type wheat Education has a certain relevance. According to the homology comparison of NCBI database, it was found that the fragment was only 84 bp homologous to a Hordeum vulgare genomic DNA fragment, indicating that it is a new sequence possibly related to the cytoplasmic male sterility of V-type wheat.