南京林业大学学报（自然科学版）2025，Vol.49Issue(1)：28-36,9.DOI:10.12302/j.issn.1000-2006.202304018
松墨天牛GST基因克隆及高温胁迫下的表达特性分析
Cloning of glutathione S-transferases gene from Monochamus alternatus and its expression characteristics under heat stress
摘要
Abstract
[Objective]To clone the glutathione S-transferases(GST)genes of Monochamus alternatus(Coleoptera:Cerambycidae)and to determine the expression levels of these genes under heat stress in adult males and females,as well as at different developmental stages of M.alternatus,we used molecular biology and bioinformatics methods.[Method]Three GST cDNA sequences were cloned,and their structural characteristics were analyzed using DNAMAN 9.0 and I-TASSER software.qRT-PCR was employed to measure the relative expression levels of GST genes in adult and fourth instar larvae of M.alternatus under varying temperatures and treatment durations.The role of these GST genes in protecting the organism from oxidative stress was assessed through disc diffusion assays.[Result]Three GST cDNA sequences were cloned and named MaltGSTe1,MaltGSTe2,and MaltGSTt1.MaltGSTe1 and MaltGSTe2 belong to the Epsilon family,while MaltGSTt1 belongs to the Theta family.Analysis of three-dimensional protein structures indicated that these GST are cytoplasmic.The relative expression levels of MaltGSTe1,MaltGSTe2 and MaltGSTt1 in fourth instar larvae significantly changed under heat stress,with MaltGSTe2 showing the greatest change.In males,the expression level of MaltGSTt1 was significantly down-regulated.Escherichia coli expressing the GST genes demonstrated enhanced antioxidant capacity,with MaltGSTe2 exhibiting the strongest activity.[Conclusion]We successfully cloned three GST genes and investigated their expression characteristics under high temperature stress in M.alternatus.High temperature stress was found to induce up-regulation of GST gene expression.Disc diffusion assays confirmed that heterogeneously expressed GST proteins have antioxidant capabilities.These findings suggest that GST genes play a role in the response mechanism to high temperature stress by protecting against oxidative stress,providing a theoretical foundation for exploring the heat resistance mechanisms of M.alternatus in subtropical regions.关键词
松墨天牛/高温胁迫/谷胱甘肽S转移酶/GST基因/氧化应激/基因克隆/表达分析/生物信息学Key words
Monochamus alternatus/heat stress/glutathione S-transferases/GST gene/oxidative stress/gene clone/express analysis/bioinformatics分类
农业科技引用本文复制引用
李子纯,郝德君,李慧,李长燕,许丹雯仪,杨华磊,赵培渊..松墨天牛GST基因克隆及高温胁迫下的表达特性分析[J].南京林业大学学报（自然科学版）,2025,49(1):28-36,9.基金项目
国家自然科学基金项目(32201562) （32201562）
江苏省自然科学基金项目(BK20220412) （BK20220412）
江苏省大学生创新训练计划重点项目(202310298002Z). （202310298002Z）
