四川农业大学学报 ›› 2012, Vol. 30 ›› Issue (04): 373-378.doi: 10.3969/j.issn.1000-2650.2012.04.001

• •    下一篇

75K γ-黑麦碱基因的分子鉴定与转移

王庆1,2, 袁中伟2, 王延谦2, 刘登才2   

  1. 1. 长江师范学院, 重庆 涪陵 408000;
    2. 四川农业大学小麦研究所, 四川 温江 611130
  • 收稿日期:2012-08-04 出版日期:2012-12-31 发布日期:2017-02-28
  • 作者简介:王庆,硕士。
  • 基金资助:
    国家科技计划863项目(2011AA100103);国家自然科学基金项目(30700495)

Molecular Identifying and Transferring of 75K γ-secalin in Rye into Wheat

WANG Qing1,2, YUAN Zhong-wei2, WANG Yan-qian2, LIU Deng-cai2   

  1. 1. Yangtze Normal University, Fuling 408100, Chongqing, China;
    2. Triticeae Research Institute, Sichuan Agricultural University, Wenjiang 611130, Sichuan, China
  • Received:2012-08-04 Online:2012-12-31 Published:2017-02-28
  • Contact: 刘登才,研究员,博士生导师,主要从事麦类遗传育种与种质资源研究,E-mail:dcliu7@yahoo.com。 E-mail:dcliu7@yahoo.com

摘要: [目的] 研究75K γ-黑麦碱基因的系统进化关系,并将秦岭黑麦75K γ-黑麦碱基因导入普通小麦遗传背景中。[方法] 利用75K γ-secalin亚基的特异引物secF/secR对5种黑麦进行扩增、克隆、测序及序列分析。以普通小麦(中国春)为母本,黑麦为父本,获得杂种。自杂种后代中,采SDS-PAGE技术分析杂交后代的75K γ-secalins蛋白亚基。用根尖制片进行体细胞染色体数目观察和原位杂交分析。[结果] 序列分析结果表明,我国秦岭黑麦(AS156)与美国Wrens黑麦(CIse35)、土耳其4041黑麦(PI168199)、南非的Polko黑麦(PI412949)、智利278黑麦(PI436188)的栽培品系均具有75K γ-黑麦碱典型的一级结构,但是在重复区中序列差异较大(包括缺失/插入和SNP)。聚类分析表明,我国的秦岭黑麦与同在北纬的土耳其和美国黑麦亲缘关系近,而与位于南纬的南非、智利的两种黑麦关系较远。以中国春为遗传背景,将位于秦岭黑麦2R染色体上的75K γ-黑麦碱基因导入六倍体小麦,获得结实正常,农艺性状稳定的品系,编号WYQ122。通过SDS-PAGE技术和基因测序分析,已确认WYQ122中携带75K γ-黑麦碱基因,同时原位杂交也验证黑麦染色体的存在。细胞学鉴定表明,WYQ122植株间染色体数目有41条和42条两种类型。[结论] 黑麦的75K γ-黑麦碱基因存在差异。以染色体代换系的方式将该基因转入普通小麦中。

关键词: 黑麦, 75K γ-黑麦碱, 分子鉴定, 转移

Abstract: [Objective] In this study, we aimed to study the genetic phylogenetic relationship of 75K γ-secalin gene in rye(Secale cereale L) and transfer the gene in Chinese rye line Qinling into common wheat. [Method] The experiments were contained of amplification, cloning, sequencing, and sequence analysis of five kinds rye using specific primers secF/secR of 75K γ-secalin subunit. We obtained hybrids of the ordinary wheat(Chinese Spring) as female parent and the rye as male parent. We analyzed 75K γ-secalins protein subunits of hybrids by the SDS-PAGE from the progenies. We observed somatic chromosome number and analyzed in situ hybridization with apical producer. [Results] Sequence analysis showed that the cultivar had typical primary structure of 75K γ-secalin in Qinling(AS156) from China, Wrens(CIse35) from USA, 4041(PI168199) from Turkey, Polko(PI412949) from South Africa, and 278(PI436188) from Chile, respectively. However, they showed a lot variations, including insertion/delection and SNP in repeat region. Moreover, the sequence from AS156 had a deletion of Cys in C-terminal. A clade was clustered by the rye sequences from north latitude of China, USA, and Turkey and the other clade from south latitude of South Africa and Chile. The 75K γ-secalin gene on chromosome 2R in Chinese rye line Qinling was transferred in common wheat line Chinese Spring. A F8 line, WYQ122, with this gene shows good fertility and consistent character in agronomy. This gene in WYQ122 was verified by SDS-PAGE analysis and sequencing data of 75K γ-secalin gene. Cytological observation indicated that WYQ122 has 41 or 42 chromosomes. GISH proved this line containing rye chromosomes. [Conclusion] There was genetic diversity for 75K γ-secalin gene among rye lines. This gene was transferred into common wheat via chromosomal substitution.

Key words: Secale cereale L, 75K γ-secalin, molecular identification, transfer

中图分类号: 

  • S512.1