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作物学报 ›› 2012, Vol. 38 ›› Issue (02): 360-368.doi: 10.3724/SP.J.1006.2012.00333

• 研究简报 • 上一篇    下一篇

非生物胁迫诱导的GmMYB的克隆与表达分析

孙霞1,**，刘晋跃1,**，袁晓辉1，潘相文1，杜维广2，任海祥2，马永波3，Jun ABE4，邱丽娟5,*，刘宝辉1,*   

  1. 1中国科学院东北地理与农业生态研究所大豆分子育种实验室 / 中国科学院黑土区农业生态院重点实验室, 黑龙江哈尔滨  150081; 2黑龙江省农业科学院牡丹江分院 / 国家大豆改良中心牡丹江试验站, 黑龙江牡丹江 157041; 3辽宁省农业环境保护监测站, 辽宁沈阳 110034; 4日本北海道大学农学院, 日本札幌 060-8589; 5中国农业科学院作物科学研究所, 北京 100081
  • 收稿日期:2011-06-21 修回日期:2011-10-12 出版日期:2012-02-12 网络出版日期:2011-12-01
  • 通讯作者: 刘宝辉, E-mail:liubh@neigaehrb.ac.cn; 邱丽娟, E-mail:qiu_lijuan@263.net
  • 基金资助:

    本研究由中国科学院“百人计划”(KZCX2-YW-BR-11), 国家自然科学基金项目(30971813和31101170), 黑龙江省杰出青年基金(JC200919), 黑龙江省青年基金(QC2011C015)，黑龙江省归国基金(09SRS11), 教育部留学回国人员科研启动基金项目(Y0SQY11001)和所前沿领域项目(2009ZX08009-013B)资助。

Cloning and Expression Analysis of GmMYB Induced by Abiotic Stresses

SUN Xia1,**,LIU Jin-Yue1,**,YUAN Xiao-Hui1,PAN Xiang-Wen1,DU Wei-Guang2,REN Hai-Xiang2,MA Yong-Bo3,Jun ABE 4,QIU Li-Juan5,*,LIU Bao-Hui1,*   

  1. 1 Laboratory of Soybean Molecular Breeding, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences / Key Laboratory of Mollisols Agroecology, Chinese Academy of Sciences, Harbin 150081, China; 2 Mudanjiang Branch of Heilongjiang Academy of Agricultural Sciences, Mudanjiang 157041, China; 3 Liaoning Agricultural Environmental Protection Monitoring Station, Shenyang 110034, China; 4 Research Faculty of Agriculture, Hokkaido University, Sapporo, Hokkaido 060-8589, Japan; 5 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2011-06-21 Revised:2011-10-12 Published:2012-02-12 Published online:2011-12-01
  • Contact: 刘宝辉, E-mail:liubh@neigaehrb.ac.cn; 邱丽娟, E-mail:qiu_lijuan@263.net

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摘要： 本研究室根据一段抗逆的EST序列, 从栽培大豆东农42中克隆到4个开放阅读框均是外显子和内含子间隔构成的R2R3-MYB基因, 其中Gm02g01300、Gm03g38040和Gm10g01340与已公布的Willams 82基因组序列完全一致, Gm19g40650第375位的单核苷酸突变导致多肽链第125位的氨基酸发生置换(GAG375→GAC375, E125→D125)。以人工气候箱内模拟非生物胁迫(盐、碱、干旱和低温)处理栽培大豆东农42芽期, 选择适宜时间点, 采用荧光定量PCR技术, 检测R2R3-MYB基因的表达。结果表明, 4个基因的表达水平都存在明显波动, 呈诱导后短暂上调或下调两种表达模式, 但表达时间、强度和趋势存在明显差异; Gm02g01300受干旱诱导明显, Gm03g38040受多种胁迫条件诱导表达强烈, 推测这些基因在大豆非生物胁迫的调控中起到重要作用; 另外, 在子叶与胚间, 单个基因的表达也存在差异; 多种非生物胁迫条件下, 基因的表达不仅存在时空差异, 可能也具有调控模式的差异。

关键词: 非生物胁迫, GmMYB, 芽期, 表达分析

Abstract: Response to external environment is the outcome of stress-induced gene expression. In this paper, based on one stress-induced EST sequence, we cloned four R2R3-MYB genes from soybean cultivar Dongnong 42, whose genomic sequences consisted of three exons and two introns. Three of them corresponding to Gm02g1300, Gm03g38040, and Gm10g01340 are respectively consistent with the sequences of Willams 82. A mutation at the 375th single nucleotide in the sequence of Gm19g40650 from Dongnong 42 caused a synonymous amino acid substitution (E125–D125). To test the relationship of four MYB genes with stress resistance, we treated the seedlings of cultivar Dongnong 42 with abiotic stresses including salt, alkali, drought and low temperature in the artificial climate chamber. Quantitative PCR analysis indicated that all of the four genes were transient down-regulated or up-regulated when subjected to the stresses, but different in the expression time, level and tendency. Gm02g01300 was induced by drought stress while Gm03g38040 was strongly induced by multiple stresses, indicating that they play important roles in responding to external stresses. There were also differences in the expression of individual gene between cotyledons and embryos. These results under a variety of abiotic stress conditions suggest that the four R2R3-MYB genes are different not only in the expression patterns, but also in the regulation modes.

Key words: Abiotic stress, GmMYB, Bud period, Expression analysis

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