水稻矮秆基因iga-1的序列变异和表达分析
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金(31200250);国家“863”计划(2012AA101201);广州市对外科技合作项目(2014J4500030);现代农业产业技术体系建设专项资金(CARS-01-12)


Sequence Variation and Expression Analysis of Dwarf Gene iga-1 in Rice (Oryza sativa L.)
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    该研究以水稻矮秆突变体cha-2为材料,对控制其表型性状的iga-1基因进行候选基因筛选,利用基因注释数据库对定位区间进行候选基因预测,通过ORF及其上下游调控区域的测序、序列比对及关键元件分析进行序列变异研究,半定量PCR检测目标基因的表达模式,明确其在基因序列、表达模式的变异,探讨其分子遗传调控机理。结果显示:(1)在隐性核基因iga-1精细定位基础上预测得到3个ORF,其中2个编码dnaJ分子伴侣(含有dnaJ结构域的蛋白),分别是LOC_Os05g26902和LOC_Os05g26926;另外1个为已克隆的水稻矮秆基因RGA1(LOC_Os05g26890)。(2)ORF序列分析表明矮秆突变体cha-2与野生型仅在RGA1基因座存在SNP变异,但未造成氨基酸编码的改变。(3)表达模式分析发现,矮秆突变体cha-2 的RGA1基因在种子萌发期、二叶期、四叶期和分蘖期等4个发育时期均不表达,且在‘中花11’、‘石狩白茅’的遗传背景下稳定遗传,均显现出失活状态,初步确定RGA1为iga-1的候选基因。(4)对RGA1基因上游和下游调控转录关键区进行测序结果表明,突变体cha-2存在865 bp的大片段缺失,包括第1外显子、部分第1内含子和转录起始上游区域。研究推断,突变体cha-2的矮秆基因iga-1正是没有活性的RGA1基因,其转录关键区域的大片段缺失,导致无法正常转录表达。

    Abstract:

    In this study,we screened and confirmed the candidate gene of iga-1 in the fine mapping region,which controls the dwarf phenotype of mutation of cha-2,made clear the sequence variation and expression patterns of the gene,and explored its molecular mechanism of genetic regulation.Candidate gene in the mapping region was predicted using rice annotation database,and sequence variations of the ORF,upstream and downstream were analyzed based on sequencing and sequence alignment.Furthermore,meta-analysis of the crucial motif was performed.And expression pattern was investigated using semi-quantitative PCR.(1)On the basis of early studies of fine mapping of the nuclear genes iga-1,3 ORF were predicted in this study,including 2 dnaJ chaperones LOC_Os05g26902 and LOC_Os05g26926,and another one is the dwarf gene,RGA1( LOC_Os05g26890).(2)It is found that only several SNP were detected in the LOC_Os05g26890 in the mapping region between cha-2 and wild type by sequencing of ORFs,which appear to have no alteration of amino acid coding.(3)Expression analysis results show that RGA1 was unable to express in four developmental stages of cha-2 and was revealed inactivation in genetic background of ‘Zhonghua 11’ or ‘Shisoubaimao’ with genetic stability,which therefore was identified as candidate gene of iga-1.(4)The sequences in upstream and downstream of the RGA1 were examined,and 865 bp deletion was found in the upstream of RGA1 in cha-2,which was induced the whole exon 1,part of intron 1 and upstream region of transcription initiation.We concluded that the iga-1 in mutation of cha-2 was the inactivated RGA1 without normal expression,which was caused by a large fragment deletion in the key region of transcription.

    参考文献
    相似文献
    引证文献
引用本文

陈立凯,卢苏龙,刘永柱,等.水稻矮秆基因iga-1的序列变异和表达分析[J].西北植物学报,2016,36(1):1-7

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2016-02-24
  • 出版日期:

微信公众号二维码

手机版网站二维码