摘要
【目的】热激蛋白基因Hsps在生物体抗逆性过程中具有重要的作用,本文旨在通过阐释瓜实蝇Bactrocera cucurbitae (Coquillett)热激蛋白基因Hsp90在其对环境温度适应性及抗药性过程中的功能作用,为瓜实蝇综合治理提供理论基础。【方法】采用RACE-PCR技术克隆瓜实蝇的Hsp90基因cDNA序列,利用生物信息学工具分析序列特征,并通过实时定量PCR技术分析高温胁迫、阿维菌素诱导和抗性及敏感品系中该基因的表达情况。【结果】克隆获得的瓜实蝇Hsp90基因cDNA全长2 654 bp,命名为Bc-Hsp90,GenBank登录号为KP864677,含2 145 bp的开放阅读框,编码715个氨基酸蛋白,具有HSP90蛋白家族共有模式和1个基序标签,C-末端具有MEEVD基序,属于胞质型热激蛋白。系统发育分析显示,Bc-Hsp90基因高度保守。不同高温(32-40℃)胁迫处理1 h和2 h后,瓜实蝇成虫体内Bc-Hsp90的相对表达量均显著高于对照组(26℃),并在38℃和40℃表达量最高;阿维菌素长期筛选的RS品系,Bc-Hsp90表达量是对照SS品系的2.13倍,但RS品系以LC90剂量诱导后,Bc-Hsp90表达量显著下调,随着恢复时间延长其表达量逐渐升高,96 h后恢复到对照水平。【结论】推测Bc-Hsp90在瓜实蝇耐热性和抗阿维菌素过程中可能起着重要作用。
[Objectives] To investigate the function of the heat shock protein Hsp90 gene of Bactrocera cucurbitae(Coquillett)(Bc-Hsp90) under different temperatures and pesticide stress in order to improve control of this pest.[Methods]The cDNA full-length fragment of the Bc-Hsp90 was cloned using the RACE-PCR method. Bioinformatics tools were used to analyze the sequence characteristics, and real-time quantitative PCR was used to detect the expression levels of the gene under heat stress, and after exposure to the pesticide abamectin, in abamectin-resistant and susceptible strains.[Results] The complete sequence of 2 654 bp was obtained(Accession number: KP864677) and was found to contain a 2 145 bp open reading frame(ORF) encoding 715 amino acids. Family signatures of the HSP90 protein and the cytoplasm of the Hsp90 gene C-terminal region MEEVD motif were detected. Phylogenetic analysis indicates that Bc-Hsp90 is highly conserved. Real-time quantitative PCR results showed that 1 h and 2 h of heat stress(32-40℃)up-regulated the expression of Bc-Hsp90. Expression peaked at 38℃ and 40℃ and expression in an abamectin resistant strain(RS) was 2.13 times higher than in a sensitive strain(SS). The expression of the Bc-Hsp90 gene was down-regulated after exposure to an LC90 dose of abamectin and returned to normal levels after 96 h.[Conclusion] The expression level of Bc-Hsp90 could play an important role in heat resistance and pesticide tolerance in B. cucurbitae.
作者
姜建军
黄立飞
黎柳锋
陈红松
王凤英
杨朗
JIANG Jian-Jun;HUANG Li-Fei;LI Liu-Feng;CHEN Hong-Song;WANG Feng-Ying;YANG Lang(Guangxi Key Laboratory of Biology for Crop Diseases and Insect Pests,Plant Protection Research Institute,Guangxi Academy of Agricultural Sciences,Nanning 530007,China)
出处
《应用昆虫学报》
CAS
CSCD
北大核心
2019年第3期444-453,共10页
Chinese Journal of Applied Entomology
基金
广西自然科学基金(2018GXNSFAA138068
2015GXNSFBA139072)
广西科技重大专项(AA17204041)
广西作物病虫害生物学重点实验室项目(2016-ST-4)
关键词
瓜实蝇
高温胁迫
抗药性
HSP90
表达
Bactrocera cucurbitae
heat stress
resistance
Hsp90
gene expression
