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单核苷酸多态性(single nucleotide polymorphism,SNP)广泛存在于甘薯基因组中,可用于甘薯遗传图谱的构建、关联分析和分子标记辅助选择等重要研究。本研究采用两种特异性扩增的方法:等位基因特异性PCR(AS-PCR)和四引物扩增受阻突变体系PCR(Tetra-primer ARMS-PCR),以及酶切扩增多态性序列(CAPS)酶切的方法,对甘薯徐781和徐薯18之间的2个SNP位点T2-32906(C/G)和T3-30695(G/A)进行检测,并通过比较3种检测方法的检测效果,找出快速高效的甘薯SNP分子标记检测方法。琼脂糖凝胶电泳结果显示:AS-PCR的检测结果在2个样本之间出现假阳性而没有明显的差异条带,Tetra-primer ARMS-PCR和CAPS两种方法的检测结果在2个样本间呈现出明显的差异条带,并且能够根据条带数目区分等位基因是否纯合。三种方法的检测结果表明:相对于其他两种检测方法,Tetra-primer ARMS-PCR呈现出更理想的检测效果。另外通过成本比较,Tetra-primer ARMS-PCR成本明显低于CAPS。因此,Tetra-primer ARMS-PCR是一种可用于甘薯SNP位点快速、高效且费用低廉的分子检测方法。
Single nucleotide polymorphism (SNP) is widely present in the sweet potato genome and can be used for the construction of sweet potato genetic map, association analysis and molecular marker-assisted selection. Two specific amplification methods were used in this study: allele-specific PCR (AS-PCR) and four-primer RT-PCR (Tetra-Primer ARMS-PCR) (C / G) and T3-30695 (G / A) of two SNP sites between sweet potato Xu 781 and Xushu 18 were detected by CAPS digestion method. Method to detect the effect of fast sweet potato to identify SNP molecular marker detection method. The results of agarose gel electrophoresis showed that the results of AS-PCR showed false positives between the two samples without significant difference bands. The results of Tetra-primer ARMS-PCR and CAPS showed that there was no significant difference between the two samples Showing a significant difference in the band, and can be based on the number of bands to distinguish whether the homozygous allele. The results of the three methods showed that the Tetra-primer ARMS-PCR showed a better detection result than the other two detection methods. In addition, through the cost comparison, Tetra-primer ARMS-PCR costs significantly lower than CAPS. Therefore, Tetra-primer ARMS-PCR is a rapid, efficient and inexpensive molecular detection method for sweet potato SNP loci.