热激蛋白基因Cf Hsp70-1和Cf Hsp70-2在热驯化增强锈赤扁谷盗高温耐受性中的作用  

The Roles of Heat Shock Protein Genes CfHsp70-1 and CfHsp70-2 in Enhancing the High-Temperature Tolerance after Heat Acclimation in Cryptolestes ferrugineus

作　　者：陈二虎 唐静杰 胡顺杰 唐培安 CHEN ErHu;TANG JingJie;HU ShunJie;TANG PeiAn(College of Food Science and Engineering/Collaborative Innovation Center for Modern Grain Circulation and Safety of Jiangsu Province/Key Laboratory of Grains and Oils Quality Control and Processing of Jiangsu Province,Nanjing University of Finance and Economics,Nanjing 210023)

机构地区：[1]南京财经大学食品科学与工程学院/江苏省现代粮食流通与安全协同创新中心/江苏高校粮油质量安全控制及深加工重点实验室,南京210023

出　　处：《中国农业科学》2025年第5期918-928,共11页Scientia Agricultura Sinica

基　　金：国家重点研发计划(2021YFD2100604);国家自然科学基金(32272388);江苏高校优势学科建设工程资助项目(YXK2103)。

摘　　要：【目的】热激蛋白是生物体重要的分子伴侣,在生物抵御外界不良环境胁迫中具有重要作用。锈赤扁谷盗(Cryptolestes ferrugineus)是一种世界性储粮害虫,拥有极强的环境适应性。论文旨在揭示热激蛋白基因CfHsp70-1和CfHsp70-2在该害虫高温耐受性形成过程中的作用。【方法】分别利用亚致死高温(37和42℃)对锈赤扁谷盗驯化2 h,检测试虫对致死高温(50℃)耐受性变化情况。基于转录组数据鉴定获得两个锈赤扁谷盗关键热激蛋白基因(CfHsp70-1和CfHsp70-2),开展氨基酸序列和系统发育树分析。利用实时荧光定量PCR技术解析CfHsp70-1和CfHsp70-2对亚致死高温胁迫响应的表达模式。使用RNA干扰(RNA interference,RNAi)技术分别沉默CfHsp70-1和CfHsp70-2,分析不同条件下锈赤扁谷盗高温耐受性变化情况。【结果】锈赤扁谷盗经亚致死高温驯化2 h后,不同种群试虫在致死高温(50℃)条件下的半致死时间(LT_(50))显著增加,表明其高温耐受性得到显著提升。序列和进化树分析显示,CfHsp70-1和CfHsp70-2氨基酸序列均拥有3个保守的Hsp70家族特征序列,且与其他鞘翅目昆虫Hsp70蛋白聚为一支。实时荧光定量PCR结果显示,锈赤扁谷盗经37和42℃驯化2 h,热激蛋白基因CfHsp70-1和CfHsp70-2均可被显著诱导表达。通过RNAi技术分别有效沉默CfHsp70-1和CfHsp70-2后,锈赤扁谷盗高温耐受性显著下降,即试虫在50℃条件下的死亡率均显著增加。【结论】热激蛋白基因CfHsp70-1和CfHsp70-2在热驯化增强锈赤扁谷盗高温耐受性中发挥重要作用。【Objective】Heat shock proteins(Hsps)are essential molecular chaperones in organisms and play the crucial roles in resisting adverse environmental stresses.Cryptolestes ferrugineus is a cosmopolitan pest of stored grains with strong environmental adaptability.This study aims to elucidate the crucial roles of heat shock protein genes CfHsp70-1 and CfHsp70-2 in the development of high-temperature tolerance in this pest.【Method】C.ferrugineus was acclimated to sub-lethal temperatures(37 and 42℃)for 2 h to examine the tolerance changes to lethal high temperature(50℃).Two key heat shock protein genes(CfHsp70-1 and CfHsp70-2)were identified based on the transcriptome data of C.ferrugineus,and the amino acid sequences and phylogenetic analysis of Hsp70 proteins were further conducted.The quantitative real-time PCR method was employed to analyze the expression patterns of CfHsp70-1 and CfHsp70-2 in response to sub-lethal heat stress.The RNA interference(RNAi)technology was used to silence CfHsp70-1 and CfHsp70-2,and then the changes in high-temperature tolerance of C.ferrugineus under different conditions were analyzed.【Result】The median lethal time(LT_(50))of different C.ferrugineus populations was significantly increased under lethal heat temperature conditions(50℃)after acclimation of insects to sub-lethal temperatures(37 and 42℃)for 2 h,indicating a substantial enhancement of the high-temperature tolerance.The further sequence and phylogenetic analysis revealed that the amino acid sequences of CfHsp70-1 and CfHsp70-2 contained three conserved Hsp70 family signature motifs,and they clustered together with Hsp70 proteins of other Coleoptera insects.The results of quantitative real-time PCR analysis suggested that the expression levels of heat shock protein genes CfHsp70-1 and CfHsp70-2 were significantly up-regulated after acclimation to 37 and 42℃for 2 h in C.ferrugineus.The gene functional analysis revealed that the high-temperature tolerance of C.ferrugineus was significantly reduced after the ef

关 键 词：锈赤扁谷盗 热驯化 热激蛋白 高温耐受性 RNA干扰 

分 类 号：R73[医药卫生—肿瘤]