生物技术通报 ›› 2022, Vol. 38 ›› Issue (12): 214-222.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0290

• 研究报告 • 上一篇    下一篇

橡胶树硫氧还蛋白基因HbCXXS1的克隆及表达分析

王帅1,2(), 袁坤2, 何其光2, 胡义钰2, 冯成天2, 王真辉2, 刘进平1, 刘辉2()   

  1. 1.海南大学热带作物学院,海口 570228
    2.中国热带农业科学院橡胶研究所 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,海口 571101
  • 收稿日期:2022-03-30 出版日期:2022-12-26 发布日期:2022-12-29
  • 作者简介:王帅,男,硕士研究生,研究方向:植物分子生物学;E-mail:791647230@qq.com
  • 基金资助:
    海南省自然科学基金项目(320RC732);海南省重大科技计划项目(ZDKJ2021004);现代农业产业技术体系建设专项资金(CARS-33)

Cloning and Expression Analysis of HbCXXS1,a Thioredoxin Gene in Hevea brasiliensis

WANG Shuai1,2(), YUAN Kun2, HE Qi-guang2, HU Yi-yu2, FENG Cheng-tian2, WANG Zhen-hui2, LIU Jin-ping1, LIU Hui2()   

  1. 1. College of Tropical Crops,Hainan University,Haikou 570228
    2. Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences Key Laboratory of Biology and Genetic Resources of Rubber Tree,Ministry of Agriculture and Rural Affairs,State Key Laboratory Incubation Base for Cultivation & Physiology of Tropical Crops,Haikou 571101
  • Received:2022-03-30 Published:2022-12-26 Online:2022-12-29

摘要:

硫氧还蛋白通过调控细胞内的氧化还原状态在植物生长发育及非生物胁迫应答等过程中发挥重要作用。研究硫氧还蛋白基因HbCXXS1在橡胶树各组织以及激素和非生物胁迫处理下的表达,为解析其生物学功能奠定基础。采用RT-PCR克隆HbCXXS1,对其进行生物信息学分析,并采用实时荧光定量PCR分析该基因在橡胶树不同组织及不同激素和非生物胁迫条件下的表达模式。结果表明,HbCXXS1开放阅读框为372 bp,编码123个氨基酸。HbCXXS1蛋白为亲水性蛋白,具有硫氧还蛋白保守结构域,活性中心为CXXS,属于h型硫氧还蛋白的第III组。HbCXXS1在橡胶树胶乳中的表达显著高于其他组织,且在死皮植株胶乳中的表达显著低于健康植株。脱落酸、茉莉酸甲酯、干旱和盐胁迫处理抑制了HbCXXS1的表达,乙烯利和低温处理上调了HbCXXS1的表达,而水杨酸和过氧化氢处理的不同时间点,HbCXXS1的表达既有上调也有下调。HbCXXS1可能在橡胶树产排胶和非生物胁迫应答中发挥重要调控作用。

关键词: 硫氧还蛋白, 橡胶树, 克隆, 表达分析, 非生物胁迫

Abstract:

Thioredoxin plays an important role in plant growth and development and abiotic stress response by regulating cellular redox homeostasis. This study aims to investigate the expressions of thioredoxin gene HbCXXS1 in various tissues of rubber tree(Hevea brasiliensis)and under different hormones and abiotic stresses,for laying a foundation for analyzing its biological function. The HbCXXS1 gene was cloned by RT-PCR and analyzed by bioinformatics. Quantitative real-time PCR was applied to analyze the expression patterns of HbCXXS1 in different tissues of rubber tree and in response to various hormones and abiotic stresses. The results showed that HbCXXS1 contained an open reading frame of 372 bp encoding 123 amino acids. HbCXXS1 was a hydrophilic protein,and contained a conserved thioredoxin domain with a CXXS active site,which belonged to group III of thioredoxin h. HbCXXS1 in the latex was expressed significantly higher than that in other tissues. In addition,its expression was significantly decreased in latex of tapping panel dryness trees as compared with that of healthy trees. The expression of HbCXXS1 was inhibited by abscisic acid,methyl jasmonate,drought and salt stresses but was up-regulated by ethephon and cold stress. Moreover,HbCXXS1 expression was both up-regulated and down-regulated at different time points after salicylic acid and hydrogen peroxide treatments. These results suggested that HbCXXS1 might play an important regulatory role in latex production and latex flow,as well as abiotic stress responses in rubber tree.

Key words: thioredoxin, rubber tree, cloning, expression analysis, abiotic stress