Alternative Splicing and Expression Reduction of P450 Genes Mediating the Oxidation of Chlorpyrifos Revealed a Novel Resistance Mechanism in Nilaparvata lugens

Cytochrome P450 enzymes metabolize various xenobiotics in insects. Compared to numerous P450s associated with insecticide detoxification and resistance, fewer have been identified to bioactivate proinsecticides in insects. Here we reported that two P450s, CYP4C62 and CYP6BD12, in Nilaparvata lugens could bioactivate chlorpyrifos, an organophosphorus insecticide, into its active ingredient chlorpyrifos-oxon in vivo and in vitro. RNAi knockdown of these two genes significantly reduced the sensitivity to chlorpyrifos and the formation of chlorpyrifos-oxon in N. lugens. Chlorpyrifos-oxon was generated when chlorpyrifos was incubated with the crude P450 enzyme prepared from N. lugens or recombinant CYP4C62 and CYP6BD12 enzymes. The expression reduction of CYP4C62 and CYP6BD12 and alternative splicing in CYP4C62 reduced the oxidation of chlorpyrifos into chlorpyrifos-oxon, which contributed importantly to chlorpyrifos resistance in N. lugens. This study revealed a novel mechanism of insecticide resistance due to the bioactivation reduction, which would be common for all currently used proinsecticides.