RNAi-Mediated Knockdown of the Sodium Channel Auxiliary Subunit TipE Increases Pyrethroid-Associated Mortality and Reveals Metabolic Transcriptional Responses in Megalurothrips usitatus

Abstract

Background: The bean flower thrips, Megalurothrips usitatus, is a destructive agricultural pest with increasing resistance to pyrethroid insecticides. Most mechanistic studies emphasize mutations in the pore-forming voltage-gated sodium channels α-subunit, whereas the contribution of insect-specific sodium-channel auxiliary subunits remains unclear. Methods: We used dsRNA feeding, pyrethroid bioassays, RNA sequencing, and RT-qPCR validation to evaluate the role of temperature-induced paralytic E (TipE) in M. usitatus. Results: The optimized RNAi condition was 400 μg/mL of dsTipE for 48 h, which reduced TipE expression by 32.55%. TipE knockdown increased adult mortality after exposure to λ-cyhalothrin and permethrin, indicating increased pyrethroid-associated susceptibility under laboratory conditions. Transcriptome sequencing identified 205 differentially expressed genes, with enrichment in metabolism-related pathways. RT-qPCR confirmed reduced expression of CYP307a1, an ecdysteroid-biosynthesis-related P450 gene, and SCD, a lipid-metabolism gene, while Nav α-subunit transcript abundance did not change significantly. Conclusions: These results identify TipE as an auxiliary-subunit factor associated with pyrethroid susceptibility in M. usitatus and provide a transcriptome-based working model linking sodium-channel auxiliary regulation with endocrine- and lipid-metabolism-related responses. This study expands the candidate space for pyrethroid resistance research beyond Nav α-subunit mutations, although the causal roles of CYP307a1 and SCD require further validation.