大豆类病变皱叶突变体NT301遗传分析和2对基因定位

Genetic analysis and two pairs of genes mapping in soybean mutant NT301 with disease-like rugose leaf

通过研究类病变突变体,挖掘抗病基因,利用分子设计育种的方法快速培育优良抗病大豆新品种,可减轻化学农药对环境的污染,降低病害的抗药性。本研究以^(60)Coγ诱变获得的类病变皱叶突变体NT301为父本,分别与W82、KF1和KF35进行杂交,构建了F_(2)和F_(2:3)分离群体,通过SSR标记和SNP分析,将目标基因1(rl1)缩小到18号染色体937 kb区间内,包含66个候选基因,将目标基因2(rl2)缩小到8号染色体130 kb区间内,包含15个候选基因。接着,本研究利用基因芯片技术对近等基因系进行了基因表达谱研究,得到了差异表达基因参与的KEGG调控通路。另外对8号染色体的15个候选基因进行半定量与荧光定量RT-PCR表达分析发现,只有基因Glyma.08G332500在突变体NT301与野生型中的差异达到了4倍,推测Glyma.08G332500基因是突变体NT301的候选基因。

Research on lesion mimic mutant,mining resistance genes,and developing superior disease-resistant new soybean varieties by molecular design breeding methods can contribute to the alleviating the environmental pollution caused by chemical pesticides and drug resistance to disease.In this study,the disease-like rugose leaf mutant NT301 obtained by^(60)Coγmutagenesis as the male parent was crossed with W82,KF1,and KF35,respectively,to construct F_(2) and F(2:3) segregating populations.Using SSR and SNP markers,target gene 1(rl1)was narrowed to 937 kb on chromosome 18 with 66 genes and target gene 2(rl2)was narrowed to 130 kb on chromosome 8 with 15 genes.The gene expression patterns of the wild type and NT301 were compared using gene chip technology,and the KEGG pathways of the differentially expressed genes were assessed.Moreover,semi quantitative and quantitative RT-PCR methods were used to analyze the relative expression levels of candidate genes on chromosome 8.The results showed that the relative expression level of Glyma.08G332500 in NT301 was four times higher than the wild type.In contrast,the expression levels of other genes showed no more than double difference.Therefore,we suggest that Glyma.08G332500 may be a candidate gene for NT301.