杜仲泛应激蛋白基因EuUSP1的克隆和功能分析
Cloning and functional analysis of universal stress protein gene EuUSP1 in Eucommia ulmoides
胡盼 1赵德刚2
作者信息
- 1. 贵州大学生命科学学院,农业生物工程研究院,山地植物资源保护与种质创新教育部重点实验室,贵阳 550025
- 2. 贵州大学生命科学学院,农业生物工程研究院,山地植物资源保护与种质创新教育部重点实验室,贵阳 550025;贵州省农业科学院,贵州省农业生物技术重点实验室,植物保育技术应用工程研究中心,贵阳 550006
- 折叠
摘要
泛应激蛋白(universal stress protein,USP)是植物抵抗胁迫的保护蛋白.为探究杜仲泛应激蛋白基因EuUSP1的分子结构和功能,本研究以杜仲组培苗为材料,通过RT-PCR和RACE克隆EuUSP1 cDNA,进行生物信息学分析,构建了过表达载体遗传转化三星烟草进行功能分析,并利用qPCR技术进行时空表达特征分析.结果表明,该基因转录cDNA全长782 bp,ORF长495 bp,5'UTR 149 bp,3'UTR 138 bp,编码蛋白164个氨基酸残基.EuUSP1在杜仲幼茎的表达量高于幼叶和幼根,且在7月表达显著高于其他时期.过表达EuUSP1对烟草植株形态没有明显影响,但在干旱胁迫下,转EuUSP1基因植株缓解干旱引起的萎蔫.在干旱胁迫10 d时转基因烟草植株叶片的CAT、SOD和POD活性显著高于野生型,分别为237.3、533.65和8 800 U·g-1.干旱胁迫前转基因植株MDA含量显著低于野生型,为6.99 nmol·g-1.综上说明EuUSP1有助于增强抗旱性.
Abstract
Universal stress proteins(USPs)play a crucial role in protecting plants against various stressors.In this study,we aimed to investigate the molecular structure and function of Eucommia ulmoides univer-sal stress protein gene,EuUSP1.Tissue culture seedlings of E.ulmoides were used as materials,and Eu-USP1 cDNA was cloned through RT-PCR and RACE techniques.Bioinformatics analysis was conducted to gain insights into the gene.An overexpression vector was constructed and genetically transformed into Samsung tobacco for functional analysis.Spatiotemporal expression patterns were analyzed using qPCR technology.The results revealed that the transcriptional cDNA of EuUSP1 has a total length of 782 bp,with an open reading frame(ORF)length of 495 bp,a 5'untranslated region(UTR)of 149 bp,and a 3'UTR of 138 bp.It encodes a protein consisting of 164 amino acid residues.The expression level of EuUSP1 in young stems of E.ulmoides was higher than those in young leaves and roots.Furthermore,its expression showed a significant increase in July compared to other months.The transgenic overexpression of EuUSP1 did not significantly affect the morphology of tobacco plants.However,under drought stress conditions,the transgenic plants expressing EuUSP1 gene exhibited reduced wilting.After 10 days of drought stress,the activities of catalase(CAT),superoxide dismutase(SOD),and peroxidase(POD)enzymes in the leaves of the transgenic tobacco plants were significantly higher than those in the wild-type plants,and the respective enzyme activity values were 237.3,533.65,and 8 800 U·g-1.Additionally,the malondialdehyde(MDA)content before drought stress was significantly lower than that in the transgenic plants,with a value of 6.99 nmol·g-1.Overall,our findings suggest that EuUSP1 enhances drought resistance in E.ulmoides.
关键词
杜仲/EuUSP1基因/抗逆性Key words
Eucommia ulmoides/stress resistance/EuUSP1 gene引用本文复制引用
基金项目
国家高技术研究发展计划("863"计划)子课题(2013AA102605)
贵州省农业科学院人才专项(2022-02)
贵州省农业科学院人才专项(2023-02)
贵州省人才办(RCJD2018-14)
出版年
2024
NETL
NSTL
万方数据