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2025年4月6日 星期日
  2017, Vol. 25 Issue (4): 558-566    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
葡萄温度胁迫相关基因VvMIPS的克隆及功能分析
郭扬1,侯丽霞1,朱丹2,刘新1
1. 青岛农业大学
2.
Cloning and Functional Analysis of VvMIPS Relevant to Temperature Stress in Grape (Vitis vinifera)
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摘要 肌醇-1-磷酸合成酶(L-myo-inositol-1-phosphate synthase, MIPS)是合成肌醇等多糖的关键酶,在植物的生长发育及非生物胁迫应答过程中发挥重要功能。为了探究葡萄(Vitis vinifera)中VvMIPS的功能,本研究以抗性品种左优红组培苗为材料克隆得到MIPS基因(GenBank登录号: 1984676)全长,序列分析结果显示,VvMIPS基因片段长度为1 533 bp,编码510个氨基酸,蛋白分子量约为56.3 kD,等电点为5.37,与大豆(Glycine max)、苜蓿(Medicago sativa)及车轴草(Trifolium pratense)的MIPS家族蛋白同源关系较近。qRT-PCR分析结果显示,VvMIPS基因在葡萄花芽和茎中表达量最高,根中的表达量最少;高温和低温均能诱导VvMIPS基因的表达;同时,VvMIPS基因受胁迫信号分子如H2S、脱落酸(abscisic acid, ABA)、H2O2诱导上调表达;通过构建重组质粒pET28a-VvMIPS,利用大肠杆菌(Escherichia coli)BL21表达系统发现,VvMIPS基因的表达能够改善大肠杆菌对低温和高温胁迫的耐受性。研究结果为葡萄耐寒和耐高温品种的遗传改良提供了基因资源。
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郭扬
侯丽霞
朱丹
刘新
关键词 葡萄肌醇-1-磷酸合成酶基因(MIPS)基因克隆温度胁迫功能分析    
Abstract:Myo-inositol-1-phosphate synthase (MIPS) is a key rate-limiting enzyme in inositol biosynthetic, and plays important roles in plant growth regulation and abiotic stress response. To investigate fuction of MIPS in grape (Vitis vinifera), the full-length cDNA of VvMIPS was cloned from cultivar Zuoyouhong. Sequence analysis showed that the size of VvMIPS was 1 533 bp (GenBank No. 1984676), encoding a protein of 510 amino acids with molecular weight of 56.3 kD, and isoelectric point of 5.37. VvMIPS presented high similarity with MIPS proteins from Glycine max, Medicago sativa and Trifolium pratense. qRT-PCR analysis showed the expression of VvMIPS was the highest in flower buds, stems, and the lowest in root. In addition, VvMIPS was highly induced by low and high temperature stress, and induced by H2S, abscisic acid (ABA) and H2O2. The recombinant plasmid pET28a-VvMIPS was transformed into Escherichia coli BL21, and expression of VvMIPS protein was tested. The results showed that VvMIPS protein could enhance the resistance of E.coli to low and high temperature stress. This study provides the genetic resources for grape cultivars improvement with lower or higher temperature stress tolerance.
Key wordsVitis vinifera    L-myo-inositol-1-phosphate synthase gene (MIPS)    Gene clone    Temperature stress    Functional analysis
收稿日期: 2016-12-02      出版日期: 2017-03-31
ZTFLH:  S663  
基金资助:国家自然科学基因;国家自然科学基金;国家自然科学基金;青岛农业大学高层次人才科研基金
通讯作者: 刘新     E-mail: liuxin6080@126.com
引用本文:   
郭扬 侯丽霞 朱丹 刘新. 葡萄温度胁迫相关基因VvMIPS的克隆及功能分析[J]. , 2017, 25(4): 558-566.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2017/V25/I4/558
 
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