Comprehensive analysis of the overexpressed cytochrome P450-based insecticide resistance mechanism in Spodoptera litura.

Cytochrome P450s play critical roles in the metabolic resistance of insecticides in insects. Previous findings showed that enhanced P450 activity was an important mechanism mediating indoxacarb resistance, and multiple P450 genes were upregulated in indoxacarb resistant strains of Spodoptera litura. However, the functions of these P450 genes in insecticide resistance remain unknown. Here, the P450 inhibitor PBO effectively decreased the resistance of S. litura to indoxacarb. Ten upregulated P450 genes were characterized, all of which were overexpressed in response to indoxacarb induction. Knockdown of nine P450 genes decreased cell viability against indoxacarb, and further silencing of three genes (CYP339A1 , CYP340G2 , CYP321A19) in larvae enhanced the sensitivity to indoxacarb. Transgenic overexpression of these three genes increased resistance to indoxacarb in Drosophila melanogaster. Moreover, molecular modeling and docking predicted that these three P450 proteins could bind tightly to indoxacarb and N -decarbomethoxylated metabolite (DCJW). Interestingly, these three P450 genes may also mediate cross-resistance to chlorantraniliprole, λ-cyhalothrin and imidacloprid. Additionally, heterologous expression and metabolic assays confirmed that three recombinant P450s could effectively metabolize indoxacarb and DCJW. This study strongly demonstrates that multiple overexpressed mitochondrial and microsomal P450 genes were involved in insecticide resistance in S. litura. [Display omitted] • Synergist bioassay confirmed the important roles of P450s in indoxacarb resistance. • Ten inducible and constitutively overexpressed P450 genes were identified and analyzed. • Several overexpressed P450 genes were involved in indoxacarb resistance. • Multiple resistant P450 genes may mediate cross-resistance to other insecticides. • This study provides direct evidence for P450s-mediated insecticides resistance. [ABSTRACT FROM AUTHOR]