ARMS-PCR对棉花SNP分型的研究  

Study on SNP Genetic Typing of Sea-island Cotton Genome Group

作　　者：郑炜佳[1] 曲延英[1] 谢元元[1] 陈全家[1] 冯文林[1] 马顺尧[1] 柴颜军[1] 梁维维[1] 

机构地区：[1]新疆农业大学农学院,乌鲁木齐830052

出　　处：《新疆农业科学》2013年第12期2182-2188,共7页Xinjiang Agricultural Sciences

基　　金：国家"863"项目"高产优质多抗棉花分子育种与品种创制"(2012AA101108)

摘　　要：【目的】在已知SNP的情况下,找到适合棉花SNP的简便验证方法。【方法】利用四引物扩增突变体系(tetra-primer amplification refractory mutationsystem PCR,Tetra-primer ARMS-PCR)对棉花SNP分型进行验证。【结果】根据深圳华大基因已组装的棉花转录本做为参考,分别将新海21号和新陆中36号的转录组测序样品比对到参考基因,通过SOAPsnp技术检测样品的单核苷酸多态性。棉花海陆杂交亲本的38个单核苷酸多态性位点设计出的66组SNP引物进行验证。通过优化PCR反应体系,反应条件,调整内外引物浓度和PCR体系的优化,从66组引物中扩增出11组引物,得到了良好的分型效果。【结论】四引物扩增受阻突变体系聚合酶链式反应是一种简单快捷而有效的SNP基因分形方法。【Objective】Under the condition of the known SNPS to find a simple validation SNP method of cotton. 【Method】Four primer PCR amplification blocked systems( tetra- primer amplification refractory mutation system PCR,Tetra- primer ARMS- PCR) technology were used to validate SNP genetic typing of sea- island cotton. 【Result】Cotton transcript was used as reference according to the Shenzhen Huada genomics, sequence transcript sample of Xinhai 21 and Xinluzhong 36 were respectively compared to the reference gene to test the single nucleotide polymorphism of the sample through SOAPsnp technology. From the 38 single nucleotide polymorphism of the sea- island cotton hybrid parent,66 pairs SNP primers were designed for authentication. Through the optimized PCR reaction system,reaction conditions,the concentration of inside and outside primers were adjusted and PCR system was further optimized. 11 set of primers were amplified from 66 pairs primers,and good classification effect was obtained.【Conclusion】The result showed that the four primer polymerase chain reaction amplification blocked mutation system is a kind of simple,quick and effective method of SNP genetic typing.

关 键 词：棉花 单核苷酸多态性 基因分型 ARMS-PCR 

分 类 号：S562[农业科学—作物学] S188