Abstract
Temperature is a key ecological factor influencing insect development and survival. Temperature stress triggers insect cell apoptosis. However, factors surrounding the response of insects to various temperature stresses at different developmental stages remain unclear. The molecular mechanisms by which these factors reduce apoptosis are also not well understood. In this study, transcriptome sequencing and differential expression analysis were conducted on the W[1118] strain of Drosophila melanogaster at various developmental stages under different temperature treatments (6 °C, 26 °C, 35 °C/37 °C). The analysis revealed that DmenHSP68 is a differentially expressed gene for different developmental stages and under different temperature stresses. The RNA interference (RNAi) suppression of DmenDNAJA1 (HSP40 family), DmenHSP68 (HSP70 family), and DmenHSP83 (HSP90 family) significantly decreased adult survival rates under temperature stress. RT-PCR results showed a significant upregulation of apoptosis-related genes. The levels of apoptosis markers, such as reactive oxygen species (ROS), cytochrome c (Cytc) levels, and Caspase-3 activity significantly increased, while adenosine triphosphate (ATP) levels significantly decreased. This study provides a theoretical foundation for further elucidation of the molecular mechanisms underlying apoptosis in Drosophila under different temperature stresses.