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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (2): 99-103.

所属专题： 生物技术； 油料作物； 园艺

• 林学 园艺 园林 • 上一篇    下一篇

白菜型油菜WRKY基因片段的克隆与表达分析

宋琴 赵福宽 孙凊鹏 杨爱珍   

  • 收稿日期:2010-05-15 修回日期:2010-08-25 出版日期:2011-01-14 发布日期:2011-01-14
  • 基金资助:

    北京市教委“植物逆境生物学科研平台建设”资助项目

Cloning & Expression Analysis of WRKY Gene Segment in Brassica campestris

  • Received:2010-05-15 Revised:2010-08-25 Online:2011-01-14 Published:2011-01-14

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摘要：

WRKY转录因子能广泛地参与植物的生物与非生物胁迫反应。为了研究WRKY转录因子在ABA诱导下的表达调控，首先利用同源克隆法在白菜型油菜中克隆WRKY基因和Actin基因片段, 用BLAST和DNAMAN软件对核酸及氨基酸序列进行分析；通过荧光定量PCR技术检测白菜型油菜WRKY基因在ABA处理条件下的相对表达趋势；然后利用相对定量PCR技术（real-time relative quantification PCR，以下简称RT-qPCR）检测WRKY基因在ABA(100 μM)诱导不同时段的差异表达。在白菜型油菜中克隆到一段中克隆到一段长度为680 bp的WRKY基因片段和一段长度为933 bp的β-actin基因片段。RT-qPCR实验结果表明，BcWRKY能被ABA诱导表达，且在诱导1 h后表达量最高。本实验文成功地克隆了白菜型油菜的WRKY转录因子基因和Actin基因片段，并证明了白菜型油菜WRKY基因的表达受ABA的影响。

关键词: 广腹螳螂, 广腹螳螂, 龙眼长跗萤叶甲, 密度, 功能反应

Abstract:

The WRKY transcription factor is considered to be involved in biotic and abiotic stress responses widely in plants. The purpose of the paper is to study the relative expression trends of WRKY transcription factor gene under ABA treatment in Brassica campestris. Actin gene and WRKY gene were cloned by using the homology cloning method. The sequences of nucleic acid and amino acid were analyzed using BLAST and DNAMAN software. Relative expression trends of WRKY gene were detected by applying real-time relative quantification PCR (RT-qPCR) under ABA (100 μM) treatment; A 680 bp NAC gene segment and a 933 bp Actin gene segment were acquired in Brassica campestris. The result of RT-qPCR analysis revealed that the relative expression of WRKY gene reaches the peak at 1h with ABA treatment. In this paper, Actin and WRKY gene in Brassica campestris were cloned successfully, which was proved to play an important role in ABA signal pathway

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