大豆RN型细胞质雄性不育系及保持系cDNA-AFLP分析

cDNA-AFLP Analysis of The RN-type Cytoplasmic Male Sterile Line and Its Maintainer Line in Soybean

本研究利用cDNA-AFLP技术对大豆RN型细胞质雄性不育系及其同核异质保持系进行多态性分析。提取不育系和保持系的总RNA,反转录成cDNA。以cDNA为模板,通过PCR对64对AFLP引物进行扩增筛选,对不育系或保持系中特有的差异带进行回收测序,通过NCBI-Blast同源比对分析得到12条同源序列,其中8条序列为已知功能的基因,4条为未知功能基因。其中与线粒体基因同源的序列A4,编码ATP合成酶γ亚基蛋白基因同源的序列A1,编码减数分裂不联会突变蛋白同源的序列B1可能与不育有关。这些片段的发现为不育机制的深入研究提供帮助,同时可作为不育系及保持系快速鉴定的依据,对缩短育种年限,加快育种进程具有重要意义。

In this research c DNA-AFLP technique was carried out to analyze polymorphism between RN-type cytoplasmic male sterile line and its maintainer line in soybean. The total RNA of the male sterile line and the maintainer line were extracted firstly and then transcribed into c DNA. The c DNAs were amplified by PCR with64 pairs of AFLP primers, and then the different displayed bands between the sterile line and the maintainer line were recycled and sequenced. 12 sequences can be found the homology though NCBI-Blast alignment. Among these sequence, 8 sequences had interpretation of gene function annotation, but 4 sequences without interpretation of gene function. Among of them, A4 fragment which encodes mitochondrial gene, A1 fragment which encode ATP synthase gamma chain protein, and B1 fragment which encode meiotic asynaptic mutant protein may be involved in male sterility. The discovery of these sequences provided evidence for research of molecular mechanism on cytoplasmic male sterile. These fragments can distinguished sterile line or maintainer line fastly,which also hoped to shorten the breeding period and promote the breeding process.