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植物研究 ›› 2017, Vol. 37 ›› Issue (5): 751-760.doi: 10.7525/j.issn.1673-5102.2017.05.016

• 研究报告 • 上一篇    下一篇

油菜RbohB基因的克隆及逆境响应表达分析

张腾国, 罗丹瑜, 郭艳峰, 郑晟, 王娟   

  1. 西北师范大学生命科学学院, 兰州 730070
  • 收稿日期:2016-12-19 出版日期:2017-09-15 发布日期:2017-09-15
  • 作者简介:张腾国(1971-),男,博士,教授,主要从事抗逆生理及分子生物学研究。
  • 基金资助:
    国家自然科学基金(31460099)

Cloning and Expression Analysis of RbohB Gene in Brassica campestris L.

ZHANG Teng-Guo, LUO Dan-Yu, GUO Yan-Feng, ZHENG Sheng, WANG Juan   

  1. School of Life Sciences, Northwest Normal University, Lanzhou 730070
  • Received:2016-12-19 Online:2017-09-15 Published:2017-09-15
  • Supported by:
    The National Natural Science Foundation of China(31460099)

摘要: 利用RACE技术从‘陇油6号’油菜中克隆得到一个新的RbohB基因,全长2 694 bp,开放阅读框2 541 bp,编码846个氨基酸。实时荧光定量PCR分析表明RbohB基因表达受低温、高盐、H2O2诱导,MAPKK抑制剂U0126预处理12 h再经低温、高盐、H2O2诱导,与单独处理结果相比,RbohB基因表达明显降低,表明该基因在油菜适应低温、高盐、H2O2胁迫过程中发挥作用,U0126对该基因的转录有抑制作用。NADPH氧化酶活性受H2O2处理的诱导,U0126预处理12 h再经H2O2处理,与单独H2O2处理结果相比,NADPH氧化酶活性明显降低,表明MAPKK抑制剂U0126对该酶活性有抑制作用。

关键词: 油菜, RbohB基因, 分子克隆, 表达分析

Abstract: A novel RbohB gene was isolated from Brassica campestris L. Longyou 6 by RACE. The full-length cDNA of RbohB is 2 694 bp, with open reading frame (ORF) 2 541 bp, encoding a 846 amino acid protein. By RT-PCR analysis, transcript level of RbohB was increased in response to cold, high salt and H2O2 stress. However, MAPKK inhibitor U0126 pretreatment 12 h before cold, high salt and H2O2 stress, compared with the single stress, the expression level of RbohB was significantly decreasing. The results suggested that RbohB played an important role during cold, high salt and H2O2 stress in B.campestris L. and U0126 has inhibitory effect to RbohB gene transcription. The activity level of NADPH oxidase was increased in response to H2O2 stress. However, U0126 pretreatment 12 h before H2O2 stress, compared with the single H2O2 stress, NADPH oxidase activity was significantly decreasing. The result showed that MAPKK inhibitor U0126 has inhibitory effect to NADPH oxidase activity.

Key words: Brassica campestris L., RbohB gene, molecular cloning, expression analysis

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