论文部分内容阅读
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 study, polymorphism analysis of RN type cytoplasmic male sterile lines and its isonuclear allogeneic maintainer line in soybean by cDNA-AFLP technique was carried out. Total RNA from sterile and maintainer lines was extracted and reverse transcribed into cDNA. Using cDNA as a template, 64 AFLP primers were amplified and screened by PCR, and the specific bands in CMS lines or maintainer lines were recovered and sequenced. 12 homologous sequences were obtained by NCBI-Blast homology analysis Eight sequences are known function genes and four are unknown function genes. Among them, the sequence A1 homologous to the mitochondrial gene, the sequence A1 homologous to the ATP synthase gamma subunit protein gene, and the sequence B1 encoding the homologous meiotic non-cascade protein may be related to infertility. The discovery of these fragments can be helpful for the in-depth study on the mechanism of infertility, and can be used as the basis for rapid identification of CMS and maintainer lines. It is of great significance to shorten the breeding time and accelerate the breeding process.