中国农业科学 ›› 2025, Vol. 58 ›› Issue (5): 918-928.doi: 10.3864/j.issn.0578-1752.2025.05.008

• 植物保护 • 上一篇    下一篇

热激蛋白基因CfHsp70-1CfHsp70-2在热驯化增强锈赤扁谷盗高温耐受性中的作用

陈二虎(), 唐静杰, 胡顺杰, 唐培安()   

  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
  • 收稿日期:2024-10-31 接受日期:2024-12-05 出版日期:2025-03-07 发布日期:2025-03-07
  • 通信作者:
    唐培安,E-mail:
  • 联系方式: 陈二虎,E-mail:erhuchen1104@163.com。
  • 基金资助:
    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)

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

CHEN ErHu(), TANG JingJie, HU ShunJie, TANG PeiAn()   

  1. 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
  • Received:2024-10-31 Accepted:2024-12-05 Published:2025-03-07 Online:2025-03-07

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CfHsp70-1CfHsp70-2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(CfHsp70-1CfHsp70-2),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CfHsp70-1CfHsp70-2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CfHsp70-1CfHsp70-2,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(LT50)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CfHsp70-1CfHsp70-2均可被显著诱导表达。通过RNAi技术分别有效沉默CfHsp70-1CfHsp70-2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CfHsp70-1CfHsp70-2在热驯化增强锈赤扁谷盗高温耐受性中发挥重要作用。

关键词: 锈赤扁谷盗, 热驯化, 热激蛋白, 高温耐受性, RNA干扰

Abstract:

【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 (LT50) 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 effective silencing of CfHsp70-1 and CfHsp70-2 via RNAi, that is, the mortality of the test insects at 50 ℃ increased significantly.【Conclusion】The heat shock protein genes CfHsp70-1 and CfHsp70-2 are involved in enhancing the high-temperature tolerance after heat acclimation of C. ferrugineus.

Key words: Cryptolestes ferrugineus, heat acclimation, heat shock protein (Hsp), high-temperature tolerance, RNA interference (RNAi)