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作物学报 ›› 2013, Vol. 39 ›› Issue (03): 563-569.doi: 10.3724/SP.J.1006.2013.00563

• 研究简报 • 上一篇    

异源表达棉花GhPRP5基因增强了拟南芥对盐和ABA的敏感性

张德静,秦丽霞,李龙,饶玥,李学宝,许文亮*   

  1. 华中师范大学生命科学学院 / 遗传调控与整合生物学湖北省重点实验室,湖北武汉 430079
  • 收稿日期:2012-07-10 修回日期:2012-10-09 出版日期:2013-03-12 网络出版日期:2012-12-11
  • 通讯作者: 许文亮, E-mail: wenliangxu@yahoo.com.cn, Tel: 027-67867814
  • 基金资助:

    本研究由国家自然科学基金项目(30870142和30400022)和华中师范大学中央高校基本科研业务费(120002040249)资助。

Expression of Cotton GhPRP5 Gene in Arabidopsis Enhances Susceptibility to ABA and Salt Stresses

ZHANG De-Jing,QIN Li-Xia,LI Long,RAO Yue,LI Xue-Bao,XU Wen-Liang*   

  1. Hubei Key Laboratory of Genetic Regulation and Integrative Biology / College of Life Sciences, Central China Normal University, Wuhan 430079, China
  • Received:2012-07-10 Revised:2012-10-09 Published:2013-03-12 Published online:2012-12-11
  • Contact: 许文亮, E-mail: wenliangxu@yahoo.com.cn, Tel: 027-67867814

摘要:

富含脯氨酸的蛋白(proline-rich proteins, PRPs)代表一类富含脯氨酸和羟脯氨酸的细胞壁结构蛋白质,最先在伤害诱导的胡萝卜贮藏根中被发现。越来越多的证据显示这类蛋白在应答多种生物和非生物胁迫中起作用。我们之前从棉花cDNA文库中分离了一个命名为GhPRP5的编码富含脯氨酸蛋白的基因,为研究其功能,构建了GhPRP5的过量表达载体,转化拟南芥,获得GhPRP5高表达的8个株系的纯合体。在正常培养条件下,转基因株系和野生型种子的萌发率一致,但盐胁迫和ABA处理显著抑制了转基因拟南芥株系种子的萌发。盐胁迫条件下野生型的绿苗率明显高于转基因拟南芥株系,与正常生长条件相比,ABA处理抑制转基因拟南芥主根伸长的程度更大。利用Quantitative RT-PCR技术分析几个胁迫相关标记基因的表达情况表明,盐和ABA诱导了RD29ARD29BKIN1的表达,但诱导水平在转基因株系和野生型中不一样,说明GhPRP5参与调控拟南芥胁迫相关标记基因的表达,但具体参与的胁迫应答信号传导途径仍需进一步研究。

关键词: 棉花GhPRP5, 转基因拟南芥, 盐胁迫, ABA

Abstract:

Pro-rich proteins (PRPs) represent one family of Pro- and Hyp-rich structural cell wall proteins that are initially identified as wound-induced gene products in carrot storage roots. Accumulated evidences demonstrate that PRP genes are regulated by various abiotic and biotic stresses and may play a role in plant responses to changes in living conditions. In our previous study, a gene encoding a proline-rich protein designated as GhPRP5 was isolated from cotton cDNA libraries. To validate its function, in this study, we introduced the coding region of GhPRP5into the vector pBI121 under the control of the CaMV 35S promoter and then transformed the vector into Arabidopsis thaliana. Eight independent T4 homozygous lines with high expression of GhPRP5 were obtained. Germination rate of transgenic lines overexpressing GhPRP5 was not affected under normal conditions; however, salt stress and ABA significantly inhibited the germination of the transgenic lines. When growing on media with NaCl, the GhPRP5-overexpressed plants displayed much less cotyledon greening rate compared with the wild type. In contrast to the normal growth conditions, ABA inhibited the elongation of primary root more severely in the transgenic lines. Quantitative RT-PCR technique was used to analyze the transcription of several stress gene markers (RD29A, RD29B, KIN1,and ABI1) in the transgenic lines and the wild type plants under salt stress and ABA treatments. Expressions of RD29A, RD29B, and KIN1 were induced by ABA and NaCl in the transgenic and the wild type plants, though the induction levels in the transgenic lines were different from those in the wild type. This finding suggests that GhPRP5 is implicated in the regulation of stress gene expression in Arabidopsis. The plant stress signal transduction pathway in which GhPRP5 may be involved needs to be further studied.

Key words: GhPRP5, Transgenic Arabidopsis, Salt stress, ABA

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