Transcription factor CREB/ATF regulates overexpression of CYP6CY14 conferring resistance to cycloxaprid in Aphis gossypii.

Aphis gossypii Glover as a destructive agricultural pest has evolved resistance to various insecticides. Cycloxaprid is a novel structure neonicotinoid insecticide with excellent toxicity against A. gossypii. However, the resistance mechanism of A. gossypii to cycloxaprid was unclear. In the present study, a cycloxaprid-resistant (Cpd-R) strain (80.1-fold) of A. gossypii was obtained by continuous selection. Bioassay results showed that piperonyl butoxide significantly increased the toxicity of cycloxaprid by 10.5-fold to the Cpd-R strain. The activity of P450s was significantly higher in Cpd-R strain than in susceptible (Cpd-S) strain. The transcriptomic and qRT-PCR results showed that CYP6CY14, CYP380C44 and CYP303A1 were significantly upregulated in Cpd-R strain compared with Cpd-S strain. Furthermore, knockdown of CYP6CY14, CYP380C44 and CYP303A1 via RNA interference (RNAi) significantly increased the sensitivity of Cpd-R strain to cycloxaprid. Based on the higher expression of CYP6CY14 and RNAi results, transgenic Drosophila assay was conducted to further clarify the role of CYP6CY14 in cycloxaprid resistance, and results showed a significant increase in resistance to cycloxaprid in D. melanogaster. Additionally, the results of RNAi, dual-luciferase reporter and yeast one-hybrid (Y1H) indicated that CREB/ATF directly regulates CYP6CY14 expression. These findings provide necessary basis for clarifying the resistance mechanism of cycloxaprid in A. gossypii.