笃斯越橘E3连接酶基因VuARI2的克隆及其抗寒功能分析
作者:
作者单位:

大连大学生命健康学院

作者简介:

通讯作者:

中图分类号:

S663.2

基金项目:

辽宁省教育厅面上项目(LJKZ1194)


Cloning and Cold Resistance Functional Analysis of Bog Bilberry E3 Ligases VuARI2 Gene
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    目的】E3泛素连接酶基因参与多种植物对低温胁迫响应的调控,是否参与笃斯越橘低温胁迫响应尚未进行深入研究。因此,本研究通过对笃斯越橘E3泛素连接酶基因VuARI2的克隆及在拟南芥中异源表达,探索笃斯越橘VuARI2基因在低温胁迫响应中的功能。【方法】以笃斯越橘为试验材料,用RT-PCR和RACE技术从茎中克隆获得VuARI2 基因;并对其进行系统的生物信息学分析;利用qRT-PCR技术分析该基因在不同组织中以及低温胁迫处理下的表达模式;在拟南芥中异源表达VuARI2并对其进行冷驯化后表达水平、生理指标及冰冻处理后的抗寒性进行分析。【结果】VuARI2基因编码区长1770 bp,编码589个氨基酸。VuARI2与野山茶、咖啡树、葡萄和河岸葡萄的ARI2同源关系较近。qRT-PCR结果显示,VuARI2基因表达具有组织特异性,在叶和花芽中VuARI2的表达量显著高于根和茎;茎和花芽VuARI2表达显著受低温诱导。转基因植物的VuARI2基因表达量在冷驯化后显著高于野生型;叶绿素含量降幅显著低于野生型;脯氨酸和可溶性糖含量显著高于野生型;超氧化物歧化酶、过氧化物酶和过氧化氢酶活性均显著高于野生型;而相对电导率和丙二醛含量显著低于野生型。冷驯化后转基因植物在冰冻低温下生存率显著高于野生型,相对电导率显著低于野生型,提高了植物抗寒性。【结论】克隆获得了笃斯越橘VuARI2基因,过表达VuARI2基因可以提高植物抗寒性。

    Abstract:

    [Objectives] plant E3 ligases family has been recognized for its involvement in regulating plant responses to low temperature stress,its role in bog bilberry under low temperature stress remains largely unexplored. In this study,we cloned VuARI2 gene,transformation model plants Arabidopsis thaliana to explore the function of VuARI2 gene in responses to low temperature stress. [Methods] VuARI2 gene was cloned from stem of bog bilberry by RACE and RT-PCR and conducted a comprehensive bioinformatics analysis. Expression of VuARI2 in different tissues and stems and flower buds after cold acclimation were analyzed by qRT-PCR. Furthermore,Heterologously express the VuARI2 in Arabidopsi Thaliana,and the transgenic plants were analyzed for the expression levels of VuARI2,physiological indexes,and freezing resistance after cold acclimation. [Results] VuARI2 was successfully cloned,and its open reading frame of 1770 bp,which encoded 589 amino acids. The multi-sequence alignment and phylogenetic tree analysis of VuARI2 amino acids indicated that VuARI2 protein was closely related to ARI2 protein in wild camellia,coffee tree,grape and riverside grape. RT qPCR showed that the expression of VuARI2 in leaves and flower buds of bog bilberry was significantly higher than that in roots and stems,and the expression of VuARI2 in stems and flower buds was significantly induced by low temperature. After cold acclimation,expression levels of VuARI2 gene in transgenic plants was significantly higher in wild type,chlorophyll content decreased significantly lower than that of the wild type;proline content and soluble sugar content were significantly higher than that of the wild type;superoxide dismutase activity;peroxidase activity and catalase activity were significantly higher than that of the wild type;while relative conductivity and malondialdehyde content were significantly lower than that of the wild type. After cold acclimation,the survival rate of transgenic plants was significantly higher than that of the wild type,and the relative conductivity was significantly lower than that of the wild type at freezing temperature,significantly enhance the freezing resistance of plants. [Conclusions] Cloning and obtaining VuARI2-overexpressed A. thaliana plants,functional analysis showed that overexpress of VuARI2 gene could significantly enhance the cold resistance of plants.

    参考文献
    相似文献
    引证文献
引用本文

张润梅,乌凤章.笃斯越橘E3连接酶基因VuARI2的克隆及其抗寒功能分析[J].西北植物学报,2024,44(9):1420-1432

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2024-03-19
  • 最后修改日期:2024-04-20
  • 录用日期:2024-04-22
  • 在线发布日期: 2024-07-15
  • 出版日期: 2024-09-05
文章二维码