[1]李大培,等.核桃V-ATPase c亚基基因(JrVHAc4)的克隆和抗旱功能分析[J].南京林业大学学报(自然科学版),2019,43(02):079-85.[doi:10.3969/j.issn.1000-2006.201811014]
 LI Dapei,WANG Yi,ZHANG Shangkun,et al.Drought resistance analysis of a V-ATPase c subunit(JrVHAc4)gene from Juglans regia[J].Journal of Nanjing Forestry University(Natural Science Edition),2019,43(02):079-85.[doi:10.3969/j.issn.1000-2006.201811014]
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核桃V-ATPase c亚基基因(JrVHAc4)的克隆和抗旱功能分析
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《南京林业大学学报(自然科学版)》[ISSN:1000-2006/CN:32-1161/S]

卷:
43
期数:
2019年02期
页码:
079-85
栏目:
研究论文
出版日期:
2019-03-30

文章信息/Info

Title:
Drought resistance analysis of a V-ATPase c subunit(JrVHAc4)gene from Juglans regia
文章编号:
1000-2006(2019)02-0079-07
作者:
李大培1 2 王 艺1 张尚昆1 赵 翔1 2 赵焕元1 2 刘玉梅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,陕西 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,陕西 杨凌 712100)
Author(s):
LI Dapei12WANG Yi 1ZHANG Shangkun 1ZHAO Xiang 12ZHAO Huanyuan 12LIU Yumei 12YANG Guiyan 12 *
(1. Walnut and Chestnut Experiment Station of Shanyang, College of Forestry, Northwest A & F University,Yangling 712100, China; 2. Key Laboratory of Economic Plant Resources Development and Utilization in Shaanxi Province, College of Forestry, Northwest A & F University, Yangling 712100, China)
关键词:
核桃 干旱胁迫 V-ATPase 表达分析 酵母表达系统
Keywords:
Juglans regia drought stress V-ATPase expression analysis yeast expression system
分类号:
Q786
DOI:
10.3969/j.issn.1000-2006.201811014
文献标志码:
A
摘要:
【目的】V-ATPase是植物逆境响应的重要酶,有必要了解V-ATPase相关亚基参与核桃响应逆境胁迫的功能机制。【方法】从‘香玲’核桃转录组中克隆获得1条V-ATPase c 亚基基因(命名为JrVHAc4),通过分析启动子预测其逆境响应功能。对JrVHAc4基因进行干旱胁迫下的表达分析,同时构建JrVHAc4酵母表达载体转入酵母表达系统研究其抗旱功能。【结果】JrVHAc4基因开放读码框(ORF)全长495 bp,拟推导的蛋白分子量为16 527.61 u,包含164个氨基酸,理论等电点为8.62; 其上游1 047 bp启动子包含MYC、DOF、MYB等与干旱响应相关的顺式作用元件。干旱胁迫下,JrVHAc4基因在核桃叶和根中均被显著诱导,并存在差异。对JrVHAc4转基因酵母进行不同浓度甘露醇胁迫,与对照酵母相比,发现JrVHAc4基因的表达能显著提高转基因酵母的生长活性。【结论】JrVHAc4基因能响应干旱胁迫,并能提高酵母的抗旱能力,JrVHAc4可作为核桃抗逆重要候选基因。
Abstract:
【Objective】 V-ATPase is an important enzyme in plant stress response. This study is under stunll the functional mechanism of V-ATPase subunits in response to stress in walnuts. 【Method】 A V-ATPase c subunit gene(named JrVHAc4)was cloned from the ‘Xiangling’ walnut transcriptome. The stress response function was predicted by analyzing its promoter and expression under drought stress. Meanwhile, JrVHAc4 was inserted into a yeast expression vector pYES2 and transformed into yeast INVSC1 to study its drought resistance function. 【Result】 The open reading frame(ORF)of JrVHAc4 was 495 bp in length, the deduced protein was 16 527.61 u and contained 164 amino acids, whose theoretical isoelectric point(pI)was 8.62. An upstream 1 047 bp promoter segment was identified and found to contain abundant cis-elements, among which some are related to drought response, such as MYC, DOF and MYB. JrVHAc4 was significantly induced in leaves and roots by drought stress, and the induction in leaves and roots were different. Compared with control yeast, we found that expression of JrVHAc4 increased the growth activity of transgenic yeast significantly under different concentrations of mannitol stress. 【Conclusion】 JrVHAc4 can respond to drought stress and improve the drought resistance of transgenic yeasts, JrVHAc4 can be used as an important candidate gene for walnut resistance.

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备注/Memo

备注/Memo:
收稿日期:2018-11-06 修回日期:2018-12-22
基金项目:国家自然科学基金项目(31800510); 陕西省自然科学基础研究计划项目(2018JQ3066); 大学生创新创业训练计划项目(201803072)。
第一作者:李大培(deplee1021@gmail.com)。
*通信作者:杨桂燕(yangguiyan@yahoo.com),讲师,ORCID(0000-0002-7669-6951)。
更新日期/Last Update: 2019-03-30