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本试验旨在开发一种新的设计共显性SNP标记的方法,以提高SNP标记检验的效率,并将其成功应用于番茄抗黄化曲叶病毒基因Ty-1的SNP标记开发中,提高了种质资源鉴定和育种分离世代材料筛选的效率。通过对抗病基因Ty-1和等位感病基因ty-1的cDNA序列进行比对,挑选了两个稳定扩增的多元非同义突变位点,分别用来设计Ty-1和ty-1的SNP标记NL3和NL2,将两个标记进行混合得到双重SNP标记NL2-3。通过用NL2-3检验已知的抗病和感病番茄材料,验证了其多态性和稳定性。并用NL2-3对抗感病材料的F3代杂交分离群体和普通自交系进行了Ty-1的筛选,并经过田间观察进一步验证了该标记的可靠性。此方法开发得到的双重SNP标记只需要进行一次PCR反应和一次凝胶电泳就可以区分纯合抗、杂合抗、纯合感3种基因型,提高了育种选择的效率。
The aim of this study was to develop a new method for designing co-dominant SNP markers to improve the efficiency of SNP marker detection. The experiment was successfully applied to the development of SNP markers in tomato TYV gene Ty-1 The identification of germplasm resources and the separation efficiency of breeding materials. By aligning the cDNA sequences of the Ty-1 gene and the allelic gene Ty-1, two stable non-synonymous mutational sites were selected for Ty-1 and Tyr- 1 SNP markers NL3 and NL2, the two markers are mixed to obtain a double SNP marker NL2-3. The polymorphism and stability of the tomato material were tested by NL2-3 test against the disease-resistant and susceptible tomato materials. The screening of Ty-1 was carried out by using the F3 progeny of NL2-3 to combat susceptible materials and normal inbred lines. The reliability of the marker was further verified by field observations. The double SNP marker developed by this method only needs one PCR reaction and one gel electrophoresis to distinguish the three genotypes of homozygous, heterozygous and homozygous, and improve the efficiency of breeding selection.