油菜RbohB基因的克隆及逆境响应表达分析

doi:10.7525/j.issn.1673-5102.2017.05.016

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

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

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

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/Abstract

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

中图分类号:

S565.4

张腾国, 罗丹瑜, 郭艳峰, 郑晟, 王娟. 油菜RbohB基因的克隆及逆境响应表达分析[J]. 植物研究, 2017, 37(5): 751-760.

ZHANG Teng-Guo, LUO Dan-Yu, GUO Yan-Feng, ZHENG Sheng, WANG Juan. Cloning and Expression Analysis of RbohB Gene in Brassica campestris L.[J]. Bulletin of Botanical Research, 2017, 37(5): 751-760.

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油菜RbohB基因的克隆及逆境响应表达分析

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

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