Overexpression of Multiple Cytochrome P450 Genes Conferring Clothianidin Resistance in Bradysia odoriphaga

Cytochrome P450 monooxygenases (P450s) play important roles in the detoxification metabolism of xenobiotics and are involved in the resistance of insects to many insecticides. In this study, piperonyl butoxide (PBO), an inhibitor of P450 enzyme activity, significantly increased the toxicity of clothianidin in the clothianidin-resistant (CL-R) population of Bradysia odoriphaga. The enzyme activity of P450 in the CL-R population was significantly higher than that in the SS population. Furthermore, four P450 genes were found to be significantly overexpressed in the CL-R population. Tissue-specific expression analysis indicates that CYP9J57, CYP3828A1, CYP6SX1, and CYP6QE1were most highly expressed in the midgut and/or Malpighian tubules. After exposure to LC30of clothianidin, the expression levels of the four P450 genes were significantly upregulated. The RNAi-mediated knockdown of CYP9J57, CYP3828A1, and CYP6QE1significantly increased the susceptibility of B. odoriphagato clothianidin. These results suggest that P450 genes are involved in clothianidin resistance in B. odoriphaga. This provides a better understanding of P450-mediated clothianidin resistance in B. odoriphagaand will contribute to the management of insect resistance to insecticides.