《植物生理学报》 2015, 51(11): 1955-1962

巴西橡胶树sHSP23.8基因的克隆、生物信息学及表达分析

李德军^1,*,**, 郭会娜^2,3,*, 邓治¹, 刘辉¹, 陈江淑^1,2, 姜达², 夏立琼², 夏志辉^2,**

¹中国热带农业科学院橡胶研究所, 农业部橡胶树生物学与遗传资源利用重点实验室, 海南儋州571737; ²海南大学农学院, 海口570228; ³右江民族医学院, 广西百色533000

通信作者：李德军；E-mail: zhxia-111@163.com, dragonldj@163.com；Tel: 0898-23301174, 0898-66279271

摘　要：

本文采用RACE技术, 从巴西橡胶树中克隆到一个小热激蛋白基因。该基因全长cDNA为1 002 bp, 最长开放阅读框645 bp, 预测编码蛋白包含214个氨基酸残基, 与植物sHSP23.8蛋白具高度相似, 将该基因命名为HbsHSP23.8。生物信息学分析显示, HbsHSP23.8具有α-晶体蛋白保守结构域, 可能定位在叶绿体, HbsHSP23.8蛋白中预测到19个磷酸化位点。二级结构预测结果显示HbsHSP23.8由α螺旋、β转角、延伸链和随机卷曲组成。实时定量RT-PCR分析结果表明, HbsHSP23.8在巴西橡胶树胶乳、叶片、树皮、雄花、雌花、花药中均有表达。在橡胶树叶片不同发育时期, HbsHSP23.8表达存在明显变化。此外, HbsHSP23.8表达受高盐、干旱、低温、乙烯和茉莉酸处理调控, 表明HbsHSP23.8可能在巴西橡胶树逆境胁迫应答、乙烯和茉莉酸信号途径中发挥作用。

关键词：巴西橡胶树; 小热激蛋白23.8; 逆境反应; 基因克隆; 表达分析

收稿：2015-07-18   修定：2015-09-30

资助：国家自然科学基金(31270651和31200514)和中国热带农业科学院橡胶研究所基本科研业务费专项(16300-22015003)。

Cloning, Bioinformatics and Expression Analysis of sHSP23.8 Gene in Hevea brasiliensis

LI De-Jun^1,*,**, GUO Hui-Na^2,3,*, DENG Zhi¹, LIU Hui¹, CHEN Jiang-Shu^1,2, JIANG Da², XIA Li-Qiong², XIA Zhi-Hui^2,**

¹Key Laboratory of Biology and Genetic Resources of Rubber Tree, Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou, Hainan 571737, China; ²College of Agriculture, Hainan University, Haikou 570228, China; ³Youjiang Medical University for Nationalities, Baise, Guangxi 533000, China

Corresponding author: LI De-Jun; E-mail: zhxia-111@163.com, dragonldj@163.com; Tel: 0898-23301174, 0898-66279271

Abstract:

In this study, a full-length cDNA sequence of a small heat shock protein (sHSP) gene was cloned from Hevea brasiliensis with the RACE method. The full-length cDNA of HbsHSP23.8 was 1 002 bp in size with a 645 bp open reading frame, encoding a deduced polypeptide of 214 amino acids. The deduced protein showed high identity to plant sHSP23.8 proteins, therefore we named this genes as HbsHSP23.8. Bioinformatic analyses indicated that HbsHSP23.8 contained a conserved alpha-crystallin domains (ACD) domain, and was likely located in chloroplast. 19 phosphorylation sites were predicted within HbsHSP23.8, and the secondary structure of HbsHSP23.8 contained α-helix, β-turn, extended strand and random coil. Real time RT-PCR analysis indicated that HbsHSP23.8 was expressed in latex, barks, leaves, barks, male flowers, female flowers and anthers. With the development of leaves, the HbsHSP23.8 expression showed a significant change. In addition, the expression of HbsHSP23.8 was regulated by NaCl, drought, low temperature, ET and JA treatments, suggesting that HbsHSP23.8 might play important roles in stresses responses as well as ET and JA signals in rubber tree.

Key words: Hevea brasiliensis; small heat shock protein 23.8; stress response; gene cloning; expression analysis