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【目的】定位徐州142无絮(XZ142w)突变体的短绒控制基因n2。【方法】以陆地棉(Gossypium hirsutum L.)徐州142(XZ142)×XZ142w的F2群体为研究对象,利用108个简单重复序列(Simple sequence repeat,SSR)标记对n2进行初步定位,再根据2个亲本材料中有单核苷酸多态性(Single nucleotide polymorphic,SNP)的差异基因设计50对SNP引物,用高分辨率熔解曲线(High resolution melting,HRM)技术从中筛选在亲本间有多态性的SNP引物,并用于后代的基因分型。【结果】利用108个SSR标记将n2初步定位在26号染色体的20.2c M的遗传区间内;用HRM技术筛选到9对亲本间有多态性的SNP引物,成功实现基因分型;并结合以SSR构建的连锁图谱,将n2的遗传区间缩小为19.5 c M,n2与最近的SNP标记Cricaas20158遗传距离为5.5 c M,且遗传图谱上的标记与四倍体陆地棉测序物理图谱基本一致。【结论】HRM技术可用于棉花中的SNP检测和n2基因的定位。
【Objective】 The objective of this study was to locate the flue-controlling gene n2 of Xuzhou 142 non-wilt (XZ142w) mutant. 【Method】 The F2 population from Xuzhou 142 (XZ142) × XZ142w of Gossypium hirsutum L. was used as the research object, and 108 simple sequence repeat (SSR) markers were used to locate n2. According to 2 Fifty pairs of SNP primers were designed based on the single nucleotide polymorphic (SNP) difference genes in parental material. High-resolution melting (HRM) technique was used to screen for polymorphisms among parents Of SNP primers and used for genotyping of offspring. 【Result】 The results showed that n2 was initially located in the genetic interval of 20.2cM on chromosome 26 using 108 SSR markers. Nine SNP primers with polymorphisms among parents were screened by HRM and the genotypes were successfully combined. The linkage map constructed by SSR narrowed the genetic interval of n2 to 19.5 cM, the genetic distance between n2 and the nearest SNP marker Cricaas20158 was 5.5 cM, and the genetic map was basically the same as the tetraploid upland cotton sequencing physical map. 【Conclusion】 HRM can be used to detect SNP and n2 gene in cotton.