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棉花学报 ›› 2017, Vol. 29 ›› Issue (1): 1-8.doi: 10.11963/issn.1002-7807.201701001

• 研究与进展 •    下一篇

采用基于高分辨率熔解曲线技术的SNP标记定位徐州142无絮的短绒控制基因

吴春珲(),马启峰,李海晶,王文魁,裴文锋,李兴丽,吴嫚,张金发,于霁雯(),喻树迅()   

  1. 中国农业科学院棉花研究所/棉花生物学国家重点实验室,河南 安阳 455000
  • 收稿日期:2015-12-31 出版日期:2017-01-15 发布日期:2017-01-15
  • 通讯作者: 于霁雯,喻树迅 E-mail:wchd163@163.com;yujw@cricaas.com.cn;yu@cricaas.com.cn
  • 作者简介:吴春珲(1991―),男,硕士研究生,wchd163@163.com
  • 基金资助:
    国家863计划(2013AA102601-01-14);国家科技支撑计划(2014BAD03B01)

Genetic Mapping of a Fuzzless Mutant Gene in Upland Cotton (Gossypium hirsutum L.) Using High Resolution Melting-Based Single Nucleotide Polymorphism Markers

Wu Chunhui(), Ma Qifeng, LI Haijing, Wang Wenkui, Pei Wenfeng, Li Xingli, Wu Man, Zhang Jinfa, Yu Jiwen(), Yu Shuxun()   

  1. Institute of Cotton Research, Chinese Academy of Agricultural Sciences / State Key Laboratory of Cotton Biology, Anyang, Henan 455000, China
  • Received:2015-12-31 Online:2017-01-15 Published:2017-01-15
  • Contact: Jiwen Yu,Shuxun Yu E-mail:wchd163@163.com;yujw@cricaas.com.cn;yu@cricaas.com.cn

摘要: 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20.2 cM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 cM,n2与最近的SNP标记Cricaas20158遗传距离为5.5 cM,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。

关键词: 陆地棉; 高分辨率熔解曲线; 单核苷酸多态性; 简单重复序列; 棉纤维; 基因定位

Abstract:

[Objective] This study aimed to map the fuzzless mutant (i.e., naked seed) gene n2 in XZ142w cotton plants. [Method] An F2 population of upland cotton (Gossypium hirsutum L.) was generated from a cross between XZ142 and the lintless-fuzzless mutant XZ142w. First, 108 simple sequence repeat (SSR) markers were used for mapping. High resolution melting (HRM) technology was then applied to screen 50 single nucleotide polymorphism (SNP) primer pairs designed based on a comparative transcriptomic analysis of the two parents. The SNP markers were used for genotyping. [Results] The n2 gene was first mapped to a 20.2-cM region on chromosome 26 based on 108 SSR markers. Nine pairs of SNP markers from the HRM screening were used to genotype the F2 progeny. Using the linkage map based on SSR markers, the genetic interval of n2 was finally narrowed to a 19.5-cM region. At a genetic distance of 5.5 cM, the closest marker was Cricaas20158 (an SNP marker). The linkage map was mostly consistent with an in silico physical map based on a sequenced upland cotton genome. [Conclusion] The application of HRM technology is useful for detecting cotton SNPs and mapping the n2 gene.

Key words: upland cotton; high resolution melting; single nucleotide polymorphism; simple sequence repeat; cotton fiber; gene mapping