抗SCN位点rhg1与Rhg4在种质资源中的单倍型分析及分子标记开发  

Haplotype Analysis and Molecular Marker Development of Soybean Nematode Cyst Resistance Loci rhg1 and Rhg4 in Soybean Germplasm

作　　者：盛碧涵 刘兵[1] 陈秀兰[1] 朱治佳 谭云夫 李文滨[1] 赵雪[1] 韩英鹏[1] 

机构地区：[1]东北农业大学大豆生物学教育部重点实验室/东北农业大学农业部东北大豆生物学与遗传育种重点实验室,黑龙江哈尔滨150030

出　　处：《大豆科学》2017年第3期345-350,共6页Soybean Science

基　　金：黑龙江省自然科学基金面上项目(C2015011);国家自然科学基金(31671717);东北农业大学学术骨干计划项目(15XG04);黑龙江省博士后项目(LBH-Z15017)

摘　　要：rhg1和Rhg4是抗大豆胞囊线虫病种质所具有的主要的抗性基因,利用与rhg1和Rhg4位点紧密连锁的SSR标记和SNP标记对15份抗感病大豆种质进行基因分型。结果表明:rhg1位点连锁SSR标记Satt309对抗病种质的检出率为55.56%,Rhg4位点连锁SSR标记Sat_162对抗病种质的检出率为66.67%;rhg1位点序列引物PCR产物630bp位置存在腺嘌呤与胞嘧啶(A/C)突变,感病等位为A碱基,抗病等位为C碱基,在Rhg4位点标记引物SHMT的PCR产物上2 749 bp位置存在胞嘧啶和鸟嘌呤(C/G)突变,感病等位为C碱基,抗病等位为G碱基。rhg1和Rhg4位点内SNP对抗病种质的检出率分别为77.78%和100%。利用高分辨率溶解曲线法设计rhg1和Rhg4位点SNP标记,在扫描温度为65℃起始,60℃保持,94℃终止时可得到理想的分型结果。Soybean cyst nematode （SCN）, also named verticillium wilt, is a kind of soil spread parasitic disease. SCN dis- ease is caused by soybean cyst nematode infection on soybean roots and seriously threats to the soybean production, rhg1 and Rhg4 were the main loci for resistance to soybean cyst nematode （SCN）. SSR and SNP markers tightly linked with rhg1 and Rhg4 loci were used to genotype fifteen soybean germplasm. The positive rates of resistance germplasm for Satt309 which was linked with rhg1 loci was 55.56% and that for Sat_162 which was linked with rhg1 loci was 66. 67%. The sequencing result showed that the adenine/cytosine （A/C） mutation existed at 630 bp location of the key fragment at rhg1 loci. The disease- susceptible allele was base ‘ A’while the disease-resistant allele was ‘ C’. The cytosine and guanine （C/A） mutation existed at 2 749 bp location of the key gene SHMT of Rhg4 loci. The disease-susceptible allele was base ‘C’while the disease-resist- ant allele was ‘G’. The positive rates of resistance germplasm for SNP markers in rhg1 and Rhg4 loci were 77.78% and 100%, respectively. The genotyping method based on high resolution melting curve （HRM） for SNP markers of rhg1 and Rhg4 loci was established. The ideal genotyping results were obtained when the condition of scanning was starting at 65℃, holding at 60℃ and ending at 94℃.

关 键 词：大豆胞囊线虫病 rhg1 Rhg4 微卫星标记 单核苷酸多态性 

分 类 号：S435.651[农业科学—农业昆虫与害虫防治]