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作物学报 ›› 2011, Vol. 37 ›› Issue (12): 2152-2157.doi: 10.3724/SP.J.1006.2011.02152

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

大豆逆境诱导基因GmPRP的克隆与表达

翟莹,雷婷婷,闫帆,黄开猛,李晓薇,张庆林,张海军,苏连泰,孙昕,王英,李景文*,王庆钰*   

  1. 吉林大学植物科学学院, 吉林长春 130062
  • 收稿日期:2011-05-10 修回日期:2011-07-25 出版日期:2011-12-12 网络出版日期:2011-09-29
  • 通讯作者: 王庆钰, E-mail: wqy414cn@yahoo.com.cn; 李景文, E-mail: ljwk9@163.com
  • 基金资助:

    本研究由转基因生物新品种培育重大专项子课题(2008ZX08004-003)，国家自然科学基金面上项目(30971808)，吉林省科技发展计划重点项目(20080204)，长春市科技局国际科技合作项目(08GH10)和“211”三期建设项目资助。

Cloning and Expression of a Stress-induced GmPRP Gene in Soybean (Glycine max)

ZHAI Ying, LEI Ting-Ting,YAN Fan, HUANG Kai-Meng, LI Xiao-Wei, ZHANG Qing-Lin, ZHANG Hai-Jun, SU Lian-Tai, SUN Xin, WANG Ying, LI Jing-Wen*,WANG Qing-Yu*   

  1. College of Plant Sciences, Jilin University, Changchun 130062, China
  • Received:2011-05-10 Revised:2011-07-25 Published:2011-12-12 Published online:2011-09-29
  • Contact: 王庆钰, E-mail: wqy414cn@yahoo.com.cn; 李景文, E-mail: ljwk9@163.com

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1277

被引次数

9

摘要： 通过对大豆吉林32未成熟胚表达谱的分析，利用RT-PCR技术从大豆中克隆了一个新的脯氨酸富集蛋白基因，命名为GmPRP。GmPRP的开放阅读框长396 bp, 其分子量13.79 kD，具有131个氨基酸残基，等电点8.96，其DNA序列无内含子。GmPRP蛋白序列N端含有一段信号肽，中间为脯氨酸富集区，C端为半胱氨酸富集区。该蛋白与四季豆和木豆的PRP同源性最高, 具有较近的亲缘关系。GmPRP的683 bp启动子序列含有10种与逆境相关的顺式作用元件，分别为ABRE-like、G-box、W-box、GT-1、MYB、MYC、BIHD10s、DPBF、SEBF和WRKY。实时荧光定量PCR分析表明, 该基因表达量在大豆的根和叶中最高，在茎和胚中其次，在花中最低，且受干旱、高盐、低温、机械伤害及SA(水杨酸)、ETH(乙烯)、ABA(脱落酸)、MeJA (茉莉酸甲酯)的诱导上调表达。

关键词: GmPRP, 大豆, 逆境胁迫, 启动子, 表达分析

Abstract: Plant proline-rich proteins (PRPs) are putative cell wall proteins, which are usually associated with different abiotic and biotic stress conditions. A soybean mRNA sequence encoding a proline-rich protein (PRP) was cloned and designated as GmPRP from Jilin 32 immature embryo gene expression profiles using RT-PCR. The GmPRP consisted of an ORF with a length of 396 bp, and encoded 131 amino acids (13.79 kD) with an isoelectric point of 8.96. There was no intron in the DNA sequence of GmPRP. Except a repetitive proline-rich domain, GmPRP also contained a signal peptide in the N-terminal domain and a conserved eight cysteine motif in the C-terminal domain. The amino acid sequences of GmPRP, PvPRP and CcHyPRP shared high homology through phylogenetic analysis. The length of the promoter was 683 bp, containing several stress-induced elements: ABRE-like,G-box,W-box,GT-1,MYB,MYC,BIHD10s,DPBF,SEBF, and WRKY. Real-time quantitative PCR (qPCR) analysis revealed that GmPRP expressed highly in root and leaf and low in flower. qPCR was also performed to investigate the expression profiles of the GmPRP under different stresses such as drought, high salt, low temperature, wound, SA (salicylic acid), ETH (ethane), ABA (abscisic acid) and MeJA (methyl jasmonate). Under these stresses GmPRP showed up-regulated expression patterns. These results revealed that GmPRP might be involved in multiple pathways of plants responding to the different environmental conditions.

Key words: GmPRP, Soybean, Adversity stresses, Promoter, Expression analysis

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