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植物研究 ›› 2022, Vol. 42 ›› Issue (6): 1023-1032.doi: 10.7525/j.issn.1673-5102.2022.06.012

• 分子生物学 • 上一篇    下一篇

橡胶树胶乳高表达热激蛋白HbHSP90.4基因抗逆功能分析

陈华峰1, 代龙军2, 刘明洋2, 郭冰冰2, 杨洪2, 王立丰2()   

  1. 1.东北林业大学,哈尔滨 150040
    2.农业农村部橡胶树生物学与遗传资源利用重点实验室,省部共建国家重点实验室培育基地-海南省热带作物栽培生理学重点实验室,农业农村部儋州热带作物科学观测实验站,海南省高性能天然橡胶材料工程重点实验室,中国热带农业科学院橡胶研究所,海口 571101
  • 收稿日期:2022-06-17 出版日期:2022-11-20 发布日期:2022-11-22
  • 通讯作者: 王立丰 E-mail:lfwang@catas.cn
  • 作者简介:陈华峰(1978—),男,硕士,主要从事植物代谢及生理生态学研究。
  • 基金资助:
    国家自然科学基金面上项目(31870577)

Stress Tolerance Functional Analysis of the High Expression Heat Shock Protein HbHSP90.4 Gene from the Latex of Hevea brasiliensis

Huafeng CHEN1, Longjun DAI2, Mingyang LIU2, Bingbing GUO2, Hong YANG2, Lifeng WANG2()   

  1. 1.Northeast Forestry University,Harbin 150040
    2.Key Laboratory of Biology and Genetic Resources of Rubber Tree,Ministry of Agriculture and Rural Affairs,P. R. China;State Key Laboratory Incubation Base for Cultivation & Physiology of Tropical Crops;Danzhou Investigation & Experiment Station of Tropical Crops,Ministry of Agriculture and Rural Affairs,P. R. China;Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101
  • Received:2022-06-17 Online:2022-11-20 Published:2022-11-22
  • Contact: Lifeng WANG E-mail:lfwang@catas.cn
  • About author:CHEN Huafeng(1978—),male,postgraduate,major in plant metabolism and physiological ecology.
  • Supported by:
    National Natural Science Foundation of China(31870577)

摘要:

为了分析热激蛋白90(HSP90)基因在橡胶树(Hevea brasiliensis)逆境胁迫和激素转导中的作用,利用PCR技术从橡胶树品种热研73397胶乳中克隆得到HbHSP90.4基因全长cDNA序列,该基因含有1个2 451 bp开放阅读框(ORF),编码816个氨基酸。生物信息学分析结果表明,HbHSP90.4含有HSP90 superfamily和HATPase superfamily结构域,属于HSP90家族成员。系统进化分析发现该蛋白与木薯MeHSP90具有较近的亲缘关系。亚细胞定位预测显示HbHSP90.4基因定位在内质网。qRT-PCR结果表明HbHSP90.4基因主要在橡胶树胶乳中表达。干旱、冷胁迫、橡胶树白粉菌侵染、H2O2和MeJA处理均可促进胶乳HbHSP90.4基因上调表达,而其在ETH、SA和ABA处理中均呈现显著下调表达。构建植物表达载体HbHSP90.4-mScarlet,为进一步的转基因植物的做成准备了材料。本研究为阐明胶乳HbHSP90.4基因响应橡胶树逆境胁迫过程和植物激素信号传导途径分子调控机制奠定坚实基础。

关键词: 橡胶树, HbHSP90.4, 生物信息学, 表达分析, 载体构建

Abstract:

In order to analyze the role of Heat Shock Protein 90(HSP90) in stress and phytohormone signaling transduction in rubber tree, the full-length cDNA sequence of the HbHSP90.4 gene was cloned from rubber tree Reyan73397 latex by PCR, and HbHSP90.4 gene contained a 2 451 bp open reading frame(ORF) and encoded 816 amino acids. The results of bioinformatics analysis showed that HbHSP90.4 contained the HSP90 superfamily and HATPase superfamily domains, which was a member of the HSP90 family. Phylogenetic analysis revealed that this protein was closely related to cassava MeHSP90. Subcellular localization predictions showed that the HbHSP90.4 gene was localized in endoplasmic reticulum. The qRT-PCR results showed that the HbHSP90.4 gene was mainly expressed in rubber latex. Drought, cold, powdery mildew infection, H2O2 and MeJA treatments promoted the up-regulation of the HbHSP90.4 gene in latex respectively, while its expression was significantly down-regulated in ETH, SA and ABA treatments respectively. The plant expression vector HbHSP90.4-mScarlet was also constructed to prepare the material for further transgenic plants. This study laid a solid foundation for the elucidation of the molecular regulation of the latex HbHSP90.4 gene in response to rubber tree stress and the phytohormone signaling pathway.

Key words: Rubber tree, HbHSP90.4, bioinformatics, expression analysis, expression vector construction

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