核桃JrGRAS2基因响应热胁迫的表达及功能分析

doi:10.7525/j.issn.1673-5102.2018.01.015

植物研究 ›› 2018, Vol. 38 ›› Issue (1): 125-131.doi: 10.7525/j.issn.1673-5102.2018.01.015

核桃JrGRAS2基因响应热胁迫的表达及功能分析

粟莉圆^1,2, 李孝哲¹, 陈淑雯^1,2, 张芳芳^1,2, 赵婷婷^1,2, 杨桂燕^1,2

1. 西北农林科技大学林学院山阳核桃板栗试验示范站, 杨凌 712100;
2. 西北农林科技大学林学院陕西省经济植物资源开发利用重点实验室, 杨凌 712100

收稿日期:2017-05-24 出版日期:2018-01-15 发布日期:2018-01-06
通讯作者: 杨桂燕,E-mail:yangguiyan@yahoo.com E-mail:yangguiyan@yahoo.com
作者简介:粟莉圆(1996—),女,本科生,主要从事林木遗传育种研究.
基金资助:
中央高校基本科研业务费（2452016057/2452015171）；中国博士后科学基金面上资助项目（2016M590979）；西北农林科技大学校重点大学生创新创业训练计划项目（1201610712031）

Expression and Function Analysis of Walnut JrGRAS2 Gene under Heat Stress

SU Li-Yuan^1,2, LI Xiao-Zhe¹, CHEN Shu-Wen^1,2, ZHANG Fang-Fang^1,2, ZHAO Ting-Ting^1,2, YANG Gui-Yan^1,2

1. Walnut and Chestnut Experiment Station of Shanyang, College of Forestry, Northwest A&F University, Yangling 712100;
2. Key Laboratory of Economic Plant Resources Development and Utilization in Shaanxi Province, College of Forestry, Northwest A&F University, Yangling 712100

Received:2017-05-24 Online:2018-01-15 Published:2018-01-06
Supported by:
Fundamental Research Funds for the Central Universities(2452016057/2452015171);China Postdoctoral Science Foundation Project(2016M590979);The Program of Innovative Entrepreneurship Training Programs for Northwest Agriculture & Forestry University Students(1201610712031)

摘要/Abstract

摘要： GRAS转录因子在植物响应逆境中起重要作用。为更好的了解核桃（Juglans regia）在逆境胁迫下的适应机制，本研究从‘香玲’核桃转录组中克隆获得一条GRAS基因（命名为JrGRAS2），对其在不同高温胁迫下的表达进行分析，并将该基因插入酵母表达载体pYES2中构建重组载体pYES2-JrGRAS2，将pYES2-JrGRAS2转入酿酒酵母（Saccharomyces cerevisiae）INVSCI，同时以转化pYES2的重组酵母作为阴性对照，在酵母表达系统中研究该基因的抗热胁迫功能。结果显示，该基因开放读码框（ORF）全长1296bp，拟推导的蛋白分子量为47405.83Da，含有氨基酸数为431，理论等电点为5.66。在热胁迫下，JrGRAS2基因被显著诱导，特别是在36℃胁迫0.5h的茎内，其表达相对于对照被上调了335.5倍。对两种酵母进行热胁迫，发现转JrGRAS2基因酵母表现出较对照更高的生存活性。表明JrGRAS2基因具有响应热胁迫的能力，且能提高酵母的抗性，JrGRAS2基因可作为核桃逆境应答的重要候选基因。

关键词: 核桃, 热胁迫, GRAS转录因子, 表达分析, 酵母表达系统

Abstract: The GRAS transcription factor is important for plant response to abiotic stress. To well understand the adaptive mechanism to adverse environment of walnut tree, a GRAS gene was cloned from the transcriptome of Juglans regia(Named as JrGRAS2). The expression of JrGRAS2 was analyzed under heat stress, and JrGRAS2 was inserted into the yeast expression vector pYES2 for constructing recombinant plasmid pYES2-JrGRAS2, which was transformed into a Saccharomyces cerevisiae strain(INVSCI). The yeast transformed with empty pYES2 was used as a negative control. The yeast expression system was used for analysis the heat stress tolerance. The full length open reading frame(ORF) of JrGRAS2 was 1 296 bp, the deduced protein was 47 405.83 Da with 431 amino acids, and the theoretical isoelectric point(pI) was 5.66. Under heat stress, JrGRAS2 was highly induced, especially exposed to 36℃ for 0.5 h in the stems, it was induced to 335.5-fold of control. When both recombinant yeasts were treated with 53℃, the JrGRAS2 expressed yeast displayed higher vitality and survival rate than the control yeast. Therefore, JrGRAS2 gene could effectively response to heat stress and improve heat tolerance of transgenic yeasts, JrGRAS2 may be an important candidate gene for walnut response to adverse stimulus.

Key words: Juglans regia, heat stress, GRAS transcription factor, expression analysis, yeast expression system

中图分类号:

Q943.2

粟莉圆, 李孝哲, 陈淑雯, 张芳芳, 赵婷婷, 杨桂燕. 核桃JrGRAS2基因响应热胁迫的表达及功能分析[J]. 植物研究, 2018, 38(1): 125-131.

SU Li-Yuan, LI Xiao-Zhe, CHEN Shu-Wen, ZHANG Fang-Fang, ZHAO Ting-Ting, YANG Gui-Yan. Expression and Function Analysis of Walnut JrGRAS2 Gene under Heat Stress[J]. Bulletin of Botanical Research, 2018, 38(1): 125-131.

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核桃JrGRAS2基因响应热胁迫的表达及功能分析

Expression and Function Analysis of Walnut JrGRAS2 Gene under Heat Stress

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