【目的】 本研究旨在克隆韭菜眼蕈蚊Bradysia odoriphaga热休克蛋白Hsp70基因，并对其进行序列和表达模式分析，以及探讨该基因在韭菜迟眼蕈蚊生长发育及响应温度胁迫方面的作用。【方法】 选择韭菜迟眼蕈蚊温度转录组中高温下表达上调的Hsp70序列，设计其基因引物扩增序列，构建qRT-PCR检测体系分析该虫在短时高温热激（30、32、34和36 ℃；1、2、4、6、8、10和12 h）和高温热激后不同恢复时间（25 ℃；1 h、2 h）下的Hsp70表达谱。【结果】 获得韭菜迟眼蕈蚊Hsp70基因cDNA全长序列并命名为BoHsp70（GeneBank登录号：MW250640），包含1 971 bp的开放阅读框，编码656个氨基酸，具有真核生物Hsp70基因家族的3个保守序列，同时在C-末端具有KDEL序列，推测其属于内质网型热休克蛋白。BLAST分析和氨基酸序列系统发育分析结果显示，韭菜迟眼蕈蚊与双翅目蝇类昆虫Hsp70聚类为一个分支。BoHsp70在韭菜迟眼蕈蚊体内不同发育阶段中都有表达，雄成虫体内的表达量高于雌成虫，且在雌雄成虫头部表达量的差异显著。高温胁迫可诱导BoHsp70表达，并在诱导1-2 h内达到最高水平。在30、32和34 ℃热激条件下随热激时间的增加，BoHsp70表达量呈下降趋势，而在36 ℃热激下，BoHsp70表达水平不变。韭菜迟眼蕈蚊在解除高温热激后，BoHsp70表达水平随着恢复时间的增长而下降。【结论】 韭菜迟眼蕈蚊可以通过调节体内Hsp70的表达来应对不良的环境温度。

[Objectives]  To clone the Bradysia odoriphaga heat shock protein Hsp70 gene, analyze its gene sequence and mRNA expression profiles, and investigate its function in development and the thermal stress response. [Methods]  PCR primers were designed to amplify a full-length fragment of the Hsp70 gene based on the nucleotide sequence of B. odoriphaga obtained from transcriptome data. A real-time, fluorescence, quantitative detection system was established to detect Hsp70 expression in B. odoriphaga during exposure to high temperatures for different periods (30, 32, 34 and 36 ℃; 1, 2, 4, 6, 8, 10, 12 h) and after different recovery periods at 25 ℃ (1 h, 2 h). [Results]  The full-length B. odoriphaga Hsp70 gene was obtained and named BoHsp70 (GenBank accession number: MW250640). This gene contained a 1 971 bp open reading frame (ORF) and encoded 656 amino acids. Its amino acid sequence contains three Hsp70 family conserved sequences and a KDEL motif in the C-terminal region, indicating that Hsp70 is a member of the endoplasmic reticulum family of heat shock proteins. BLAST analysis of amino acid sequences indicates that BoHsp70 is very similar to the Hsp70 genes of Dipteran insects, and it clustered on the same branch as Dipteran Hsp70 genes on a phylogenetic tree. BoHsp70 was expressed in different developmental stages of B. odoriphaga. Expression was higher in male adults than in females and was significantly different in the heads of male and female adults. Exposure to a high temperature induced expression of BoHsp70 which peaked after 1-2 h. BoHsp70 expression decreased with the duration of exposure at 30, 32 and 34 ℃, but remained constant over time at 36 ℃. After heat shock treatment, BoHsp70 expression decreased with the duration of recovery time at 25 ℃. [Conclusion]  Collectively, these findings indicate that B. odoriphaga can regulate BoHsp70 expression in response to high temperature stress.