百子莲脱水素基因ApY2SK2逆境应答模式及启动子调控功能研究
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金(31670693,31971705);


Stress Response Pattern of Agapanthus praecox Dehydrin Gene ApY2SK2 and Its Promoter Functional Analysis
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:

    在百子莲胚性细胞中筛选到对超低温保存复合逆境具有积极响应的保护类蛋白脱水素(ApY2SK2),为探明ApY2SK2基因在复合逆境中的应答模式,该研究采用染色体步移技术克隆并分析了ApY2SK2编码基因上游1 200 bp的启动子序列。结果表明:(1)序列分析显示,该启动子含有多个与逆境和激素诱导相关的顺式调控元件;实时荧光定量PCR结果表明,ApY2SK2基因的表达具有组织特异性,在百子莲的叶和果中表达量较高,且在多种胁迫处理与ABA激素诱导下,其表达量显著升高。(2)成功构建了5个ApY2SK2启动子不同缺失片段驱动GUS基因的融合表达载体,经农杆菌转化、抗性筛选和PCR检测鉴定,获得T3代纯和转基因拟南芥株系。(3) GUS组织化学染色结果显示,GUS基因在拟南芥幼苗全株、成年苗的叶、花和成熟果实中表达活性较强,但在未成熟果实中无明显表达;烟草瞬时表达结果显示,与对照组相比,在脱水胁迫和ABA处理下的ApY2SK2启动子不同缺失片段驱动GUS基因表达具有显著差异。(4)转基因拟南芥GUS活性测定结果显示,ApY2SK2启动子MBS元件和ABRE元件可响应干旱与渗透胁迫信号;ApY2SK2启动子LTR元件参与低温响应;ApY2SK2启动子-1 199~-262 bp区域包含多个串联的ABRE顺式调控元件(-373~-211 bp)对响应ABA信号具有主要调控作用。该研究结果揭示了ApY2SK2启动子的组织特异性,且启动子上的关键顺式调控元件对不同的胁迫和激素信号响应具有决定性调控作用。

    Abstract:

    ApY2SK2 a dehydrin gene from Agapanthus praecox was identified to have the positive protective effect on embryonic cells during cryopreservation. In order to reveal the regulatory mechanism of ApY2SK2 response to complex stresses of cryopreservation, a 1 200 bp promoter sequence of ApY2SK2 was cloned and analyzed by using Chromosome Walking technique. The results showed that: (1) ApY2SK2 promoter contained multiple stress, hormones response and plant embryo developmentrelated cisacting elements. Quantitative Realtime PCR assay suggested that the ApY2SK2 promoter had tissuespecific expression pattern, and ApY2SK2 gene had the highest expression level in the leaves and fruits of A. praecox. This gene was significantly upregulated response to various stress and ABA treatments. (2) Five different deletion fragments of ApY2SK2 promoter were constructed into pBI121 expression vectors, and using Agrobacterium mediated transformation and PCR identification to obtain the A. thaliana T3 generation transgenic lines. (3) GUS histochemical staining results showed that GUS staining signal were mainly distribute at the whole seedling, and the leaves, flowers and mature fruits of adult transgenic plants. Furthermore, gene transient transformation in leaf epidermis of tobacco test showed that the driver activity of 5 different deletion promoter fragments were significantly different under PEG and ABA treatments. (4) Quantitative determination of GUS enzyme activity suggested that MBS element and ABRE element of the ApY2SK2 promoter involved in response to drought and osmotic signals, respectively. The LTR element participates in the low temperature response. Additionally, -1 199 to -262 bp of ApY2SK2 promoter contained multiple tandem ABRE cisacting regulators (locate at -373 to -211 bp) that play an important role in response to ABA signaling. This study finally confirmed that ApY2SK2 promoter has tissue specific regulatory functions, and some core cisacting elements play a decisive role in regulating the ApY2SK2 response to different stresses and hormone signals.

    参考文献
    相似文献
    引证文献
引用本文

陈晓童,吕 可,刘 涛,等.百子莲脱水素基因ApY2SK2逆境应答模式及启动子调控功能研究[J].西北植物学报,2021,41(8):1267-1278

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:
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2021-09-10
  • 出版日期:

微信公众号二维码

手机版网站二维码