番茄Mi-1基因的SNP分型

SNP genotyping of tomato Mi-1 gene

SNP分型是SNP分子标记辅助育种,以及分子遗传作图等研究的重要基础技术。试验利用在番茄Mi-1基因中开发出的一个A/T型SNP位点,应用等位基因聚合酶链式反应技术,成功的对438个F2群体进行了SNP分型。结果表明,这一F2群体中有72个纯合T/T基因型,175个杂合A/T基因型,70个纯合A/A基因型,以及121个不明原因缺失的F2单株,已得基因型经卡方测验符合1:2:1分离比例;同时,试验通过在下游引物3′末端第二、三位点引入错配碱基,以及优化PCR反应体系为Mg2+1.875 mmol.L-1,dNTP 100μmol.L-1,提高了反应的准确性,为番茄抗根结线虫辅助育种打下了基础。

SNP genotyping is an important research for SNP molecular marker-assisted breeding and molecular genetic mapping techniques,etc.The study successfully carried out 438 F2 groups SNP genotypes with SNPs detected in tomato Mi-1 gene,and the allele specific polymerase chain reaction technology.The results showed that 72 of them were T/T homozygous genotype,175 of them were A/T heterozygous genotype,70 of them were A/A homozygous genotype,and 121 of them unexplained lost.The separation of genotypes accorded with the ratio of 1： 2： 1 by χ2 test;The method of introducing proper mismatched based at the second or the third sites of the 3＇-end of the anti-primer and optimizing the reaction system to be Mg2＋ 1.875 mmol.L-1,and dNTP 100 μmol.L-1 by gradient PCR which had improved the accuracy of the reaction.It laid the foundation for assisted-breeding of the tomato root-knot nematode resistance.