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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。
rhg1 and rhg4 were the major resistance genes against soybean cyst nematode germplasm. 15 resistant soybean germplasms were genotyped by SSR markers and SNP markers closely linked to rhg1 and Rhg4 loci. The results showed that the detection rate of the disease-resistant germplasm was 55.56% with the SSR marker Satt309 of rhg1 locus, and 66.67% with the SSR marker Sat_162 of Rhg4 locus; the 630 bp of the PCR product of rhg1 locus A / C mutation existed in Adenine and cytosine (A / C), the susceptible allele was A base and the disease resistance allele was C base. Cytosine was found at 2 749 bp on PCR product of Rhg4 locus primer SHMT and Guanine (C / G) mutation, susceptible allele C base, resistance allele G base. The detection rates of SNPs against disease-resistant germplasm in rhg1 and Rhg4 loci were 77.78% and 100% respectively. The rhp1 and Rhg4 SNPs were designed by high resolution lysing curve. The ideal typing results were obtained when the scanning temperature was 65 ° C, 60 ° C and terminated at 94 ° C.