橡胶树HbMC2在酵母中的表达和抗逆性分析

doi:10.13560/j.cnki.biotech.bull.1985.2018-0298

生物技术通报 ›› 2018, Vol. 34 ›› Issue (9): 202-208.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0298

橡胶树HbMC2在酵母中的表达和抗逆性分析

刘辉, 邓治, 杨洪, 代龙军, 李德军

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省热带作物栽培生理学重点实验室,儋州 571737

收稿日期:2018-03-30 出版日期:2018-09-26 发布日期:2018-10-10
作者简介:刘辉,男,副研究员,研究方向：植物分子生物学;E-mail：liuhui@catas.cn
基金资助:
中国热带农业科学院基本科研业务费专项资金（1630022017028,1630022014006）

Expression and Stress Tolerance Analysis of HbMC2 Gene from Hevea brasliensis in Yeast

LIU Hui, DENG Zhi, YANG Hong, DAI Long-jun, LI De-jun

Hainan Provincial Key Laboratory of Tropical Crops Cultivation and Physiology,Key Laboratory of Biology and Genetic Resources of Rubber Tree,Ministry of Agriculture,Rubber Research Institute,Chinese Academy of Tropical Agricultural Sciences,Danzhou 571737

Received:2018-03-30 Published:2018-09-26 Online:2018-10-10

摘要/Abstract

摘要： Metacaspase介导的程序性细胞死亡在植物生长发育、抵御生物和非生物胁迫过程中发挥重要作用。前期研究发现橡胶树metacaspase基因HbMC2的表达受干旱和高盐等非生物胁迫调控。为明确HbMC2的功能,采用RT-PCR从橡胶树中扩增HbMC2编码区序列并将其插入到pYES2载体中,构建酵母表达载体pYES2-HbMC2。将pYES2-HbMC2质粒转化到酿酒酵母INVSc1菌株中,获得重组酵母INVSc1（pYES2-HbMC2）。PCR及RT-PCR检测结果表明,HbMC2已成功转入重组酵母并能在半乳糖诱导下表达。与对照酵母INVSc1（pYES2）相比,重组酵母INVSc1（pYES2-HbMC2）在氧化、干旱和盐胁迫下的生长受到抑制、细胞死亡率增加。结果表明酵母中表达HbMC2降低了对非生物胁迫的抗性,HbMC2是非生物胁迫抗性的负调节因子。

关键词: 橡胶树, metacaspase, 非生物胁迫, 酵母表达系统, 程序性细胞死亡

Abstract: Metacaspase-mediated programmed cell death plays an essential role in plant growth and development as well as resistances to biotic and abiotic stresses. Our previous study revealed that the rubber tree（Hevea brasliensis）metacaspase gene HbMC2 was regulated by abiotic stresses,such as drought,high salinity,etc. To investigate the function of HbMC2,the coding sequence of HbMC2 was amplified from rubber tree by RT-PCR and inserted into the pYES2 vector,and its yeast expression vector pYES2-HbMC2 was constructed. The plasmid of pYES2-HbMC2 was transformed into the yeast strain INVSc1 and the recombinant yeast INVSc1（pYES2-HbMC2）was obtained. PCR and RT-PCR analysis showed that HbMC2 gene was successfully introduced and expressed in the recombinant yeast when induced by galactose. Compared with the control yeast INVSc1（pYES2）,the recombinant yeast INVSc1（pYES2-HbMC2）demonstrated growth inhibition and its cell death rate increased under oxidative,drought,and salt stresses. These results indicate that expression of HbMC2 gene in yeast reduces tolerance to abiotic stress,i.e.,HbMC2 is a negative regulator of abiotic stress tolerance.

Key words: rubber tree, metacaspase, abiotic stress, yeast expression system, programmed cell death

刘辉, 邓治, 杨洪, 代龙军, 李德军. 橡胶树HbMC2在酵母中的表达和抗逆性分析[J]. 生物技术通报, 2018, 34(9): 202-208.

LIU Hui, DENG Zhi, YANG Hong, DAI Long-jun, LI De-jun. Expression and Stress Tolerance Analysis of HbMC2 Gene from Hevea brasliensis in Yeast[J]. Biotechnology Bulletin, 2018, 34(9): 202-208.

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橡胶树HbMC2在酵母中的表达和抗逆性分析

Expression and Stress Tolerance Analysis of HbMC2 Gene from Hevea brasliensis in Yeast

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