欢迎访问《棉花学报》! 今天是

棉花学报 ›› 2018, Vol. 30 ›› Issue (1): 12-20.doi: 10.11963/1002-7807.llcxcz.20180104

• 研究与进展 • 上一篇    下一篇

棉花细胞质雄性不育恢复基因Rf1候选区域SSCP标记开发与候选基因表达分析

刘利彩,郭立平,戚廷香,王海林,唐会妮,张学贤,乔秀琴,吴建勇*,邢朝柱*   

  1. 棉花生物学国家重点实验室/中国农业科学院棉花研究所,河南 安阳 455000
  • 收稿日期:2015-04-07 出版日期:2018-01-15 发布日期:2018-01-15
  • 通讯作者: 邢朝柱,xingcz@cricaas.com.cn;吴建勇, dr.wujianyong@live.cn
  • 基金资助:
    中央级公益性科研院所基本科研业务费专项资金(SJB1305);农业科研杰出人才基金及其创新团队(棉花亲本创制及杂种优势利用);河南省自然科学基金(162300410328)

SSCP Marker Development and Analysis of Candidate Restorer Gene Rf1 for Cotton Cytoplasmic Male Sterility

Liu Licai, Guo Liping, Qi Tingxiang, Wang Hailin, Tang Huini, Zhang Xuexian, Qiao Xiuqin, Wu Jianyong*, Xing Chaozhu*   

  1. State Key Laboratory of Cotton Biology / Institute of Cotton Research, Chinese Academy of Agricultural Sciences, Anyang, Henan 455000, China
  • Received:2015-04-07 Online:2018-01-15 Published:2018-01-15

摘要: 【目的】棉花细胞质雄性不育恢复基因Rf1的定位对于研究恢复基因的作用机制和改良恢复系具有重要意义。前期研究结果表明在恢复基因Rf1所在Scaffold 333上的目标区间内有9个PPR(Pentatricopeptide repeat)基因成簇分布在160 kb区间内,此区间内还包括另外9个其他类型的基因。为进一步丰富恢复基因目标区域内标记数目,并了解相关基因的表达模式。【方法】利用这18个基因的序列设计出覆盖全部基因的引物共155对,通过单链构象多态性(Single-strand conformation polymorphism,SSCP)技术对可育池和不育池进行筛选,利用荧光定量聚合酶链式反应(Polymerase chain reaction,PCR)对目标区域内的8个非PPR基因在不育系、保持系、恢复系的花蕾不同发育时期的表达模式进行了分析。【结果】共得到15对多态性引物,结合前期的3个简单重复序列标记(Simple sequence repeats,SSR)标记对F2群体进行基因型分析,结果表明这些标记分布在4.8 cM范围内。受恢复基因或不育胞质的影响,在花蕾4个发育时期,大部分基因表达存在明显差异。【结论】本研究获得了与恢复基因紧密连锁的SSCP标记,并初步了解了目标区间内部分基因的表达模式,为进一步开展恢复基因精细定位和候选基因的筛选提供了基础。

关键词: 棉花; 恢复基因Rf1; 遗传定位; PPR基因; SSCP标记

Abstract: [Objective] Locating the cotton cytoplasmic male sterility (CMS) restorer gene Rf1 is important for investigating restorer gene mechanisms and improving restorer lines. In our previous study, a gene cluster, with nine Pentatricopeptide repeat(PPR) genes and nine other genes, was found within the 160-kb Rf1 target region in Scaffold 333. The objective here was to improve the density of Rf1-linked markers in the target region and determine the expression profiles of candidate genes. [Method] Using the sequences of the 18 genes, we designed 155 single-strand conformation polymorphism (SSCP) primers covering all of the gene sequences to identify the polymorphic SSCP markers between the fertile and sterile pools. Additionally, real-time  polymerase chain reaction(PCR) was performed to analyze the expression profiles of eight candidate genes in the four developmental stages of  buds of sterile, maintainer and restoring lines, respectively. [Result] In total, 15 polymorphic primers were identified. A genotype analysis of the F2 population was conducted using the 15 primers and 3 other polymorphic simple sequence repeats (SSR) markers. The markers were distributed in a 4.8 cM range. In addition, owing to the influence of sterile cytoplasm or restorer genes, most of the genes showed different expression patterns in the four developmental stages of the three lines' buds. [Conclusion] SSCP markers tightly linked to Rf1 were identified and the expression profiles of candidate genes were determined. This study provides a basis for the further fine mapping of restorer genes and for candidate gene screening.

Key words: cotton; restorer genes; genetic mapping; PPR gene; SSCP marker

中图分类号: 
  • S562.03:S321.8