Contribution of multiple overexpressed carboxylesterase genes to indoxacarb resistance in Spodoptera litura

As an important family of detoxification enzymes, carboxylesterases (CarEs) have important roles in the development of insecticide resistance in almost all agricultural pests. Previous studies have suggested that enhancement of CarE activity is an important mechanism mediating indoxacarb resistance in Spodoptera litura, and several CarE genes have been found to be overexpressed in indoxacarb-resistant strains. However, the functions of these CarE genes in indoxacarb resistance needs to be further investigated.The synergist triphenyl phosphate effectively reduced the resistance of S. litura to indoxacarb, suggesting an involvement of CarEs in indoxacarb resistance. Among seven identified S. litura CarE genes (hereafter SlituCOE), six were overexpressed in two indoxacarb-resistant strains, but there were no significant differences in gene copy number. Knockdown of SlituCOE009 and SlituCOE050 enhanced indoxacarb sensitivity in both susceptible and resistant strains, whereas knockdown of SlituCOE090, SlituCOE093 and SlituCOE074 enhanced indoxacarb sensitivity in only the resistant strain. Knockdown of the sixth gene, SlituCOE073, did not have any effect. Furthermore, simultaneous knockdown of the five SlituCOE genes had a greater effect on increasing indoxacarb sensitivity than silencing them individually. By contrast, overexpression of the five SlituCOE genes individually in Drosophila melanogaster significantly decreased the toxicity of indoxacarb to transgenic fruit flies. Furthermore, modeling and docking analysis indicated that the catalytic pockets of SlituCOE009 and SlituCOE074 were ideally shaped for indoxacarb and N-decarbomethoxylated metabolite (DCJW), but the binding affinity for DCJW was stronger than for indoxacarb.This study reveals that multiple overexpressed CarE genes are involved in indoxacarb resistance in S. litura.