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植物研究 ›› 2022, Vol. 42 ›› Issue (5): 811-820.doi: 10.7525/j.issn.1673-5102.2022.05.012

• 分子生物学 • 上一篇    下一篇

橡胶树热激蛋白HbHSP90.8-1基因的克隆与功能分析

刘明洋1, 肖化兴1, 王立丰2, 梁晓宇1, 张宇1, 王萌1()   

  1. 1.海南大学植物保护学院,热带农林生物灾害绿色防控教育部重点实验室,海口 570228
    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,中国热带农业科学院橡胶研究所,海口 571101
  • 收稿日期:2021-05-07 出版日期:2022-09-20 发布日期:2022-09-15
  • 通讯作者: 王萌 E-mail:wangmeng@hainanu.edu.cn
  • 作者简介:刘明洋(1996—),女,硕士研究生,主要从事植物分子病理学研究。
  • 基金资助:
    国家自然科学基金(32160370);现代农业产业技术体系建设专项资金(CARS-33-BC1);海南省高层次人才基金(2019RC164)

Cloning and Functional Analysis of Heat Shock Protein HbHSP90.8-1 from Hevea brasiliensis Müll. Arg.

Mingyang LIU1, Huaxing XIAO1, Lifeng WANG2, Xiaoxu LIANG1, Yu ZHANG1, Meng WANG1()   

  1. 1.School of Plant Protection,Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests(Hainan University),Ministry of Education,Hainan University,Haikou 570228
    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:2021-05-07 Online:2022-09-20 Published:2022-09-15
  • Contact: Meng WANG E-mail:wangmeng@hainanu.edu.cn
  • About author:LIU Mingyang(1996—),female,master,mainly engaged in the research on plant molecular pathology.
  • Supported by:
    National Natural Science Foundation of China(32160370);Earmarked fund for China Agriculture Research System(CARS-33-BC1);High-level Talent Fund of Hainan Province(2019RC164)

摘要:

白粉病是橡胶树生长发育过程中主要叶部病害之一。热激蛋白90(HSP90)分子伴侣在植物逆境胁迫抗性中起着重要作用。为研究HSP90家族成员结构及其在橡胶树抗白粉病中的功能,利用PCR技术从橡胶树品种热研73397叶片中克隆HbHSP90.8-1并采用生物信息学等技术对其结构和功能进行分析。结果表明,HbHSP90.8-1 cDNA序列全长2 844 bp,开放阅读框(ORF)2 454 bp,编码817个氨基酸。HbHSP90.8-1编码一个有信号肽、无跨膜结构且定位预测在内质网上的稳定亲水蛋白,存在HSP90 superfamily和HATPase superfamily结构域。系统进化分析结果表明橡胶树HbHSP90.8-1与木薯MeHSP90的亲缘关系最近,与麻风树JcHSP90聚为一类。通过qRT-PCR分析结果表明HbHSP90.8-1在橡胶树不同组织中均有表达,其在胶乳中的表达量最高;HbHSP90.8-1基因在白粉菌侵染、H2O2、ABA和ETH处理叶片中,表达量均呈现显著上调趋势;在MeJA和SA激素处理下,HbHSP90.8-1表达量呈现下调的趋势。说明HbHSP90.8-1参与橡胶树对白粉菌响应过程以及植物抗病相关激素信号转导途径。

关键词: 橡胶树, 白粉病, HbHSP90.8-1, 生物信息学, 表达分析

Abstract:

Powdery mildew is one of the major leaf diseases in growth of rubber tree. Heat shock protein 90(HSP90) molecular chaperone plays an important role in plant stress resistance. To study the structure of the rubber tree HSP90 family members and their function in powdery mildew resistance, HbHSP90.8-1 was cloned from rubber tree variety Reyan73397 leaf using PCR technology and its structure and function were analyzed by bioinformatics method. The results showed that the cDNA sequence of HbHSP90.8-1 was 2 844 bp in full length, 2 454 bp open reading frame(ORF) and encoded 817 amino acids. HbHSP90.8-1 encoded a stable hydrophilic protein with signal peptide, no transmembrane structure and predicted positioning on endoplasmic reticulum, contained HSP90 superfamily and HATPase superfamily domains. Phylogenetic analysis showed that HbHSP90.8-1 was closely related to MeHSP90, and classified into Jatropha JcHSP90 and Castor RcHSP90. The qRT-PCR analysis showed that HbHSP90.8-1 was expressed in different tissues of rubber tree, and its expression level was the highest in latex. The expression of HbHSP90.8-1 was significantly expressed and up-regulated in leaves treated with Oidium heveae infection, H2O2, ABA and ETH respectively. However, under the treatment of MeJA and SA hormone, the expression of HbHSP90.8-1 showed a downward trend. The study suggested that HbHSP90.8-1 related to the powdery mildew resistance and the hormone signal transduction pathway related to plant disease resistance, and provided a technical guidance for the subsequent study on the molecular regulation mechanism of HbHSP90.8-1 in the powdery mildew resistance in rubber tree.

Key words: rubber tree, powdery mildew, HbHSP90.8-1, bioinformatics, expression analysis

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