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UDP-glycosyltransferases (UGTs) are a diverse superfamily of enzymes. Insects utilize uridine diphosphate-glucose (UDP-glucose) as a glycosyl donor for glycosylation in vivo, involved in the glycosylation of lipophilic endosymbionts and xenobiotics, including phytotoxins. UGTs act as second-stage detoxification metabolizing enzymes, which are essential for the detoxification metabolism of insecticides and benzoxazine compounds. However, the UGT genes responsible for specific glycosylation functions in <i>S. frugiperda</i> are unclear at present. In this study, we utilized CRISPR/Cas9 to produce a <i>SfUGT50A15-KO</i> strain to explore its possible function in governing sensitivity to chemical insecticides or benzoxazinoids. The bioassay results suggested that the <i>SfUGT50A15-KO</i> strain was significantly more sensitive to chlorantraniliprole, emamectin benzoate, and benzoxazinoids than the wild-type strains. This finding suggests that the overexpression of the <i>SfUGT50A15</i> gene may be linked to <i>S. frugiperda</i> resistance to pesticides (chlorantraniliprole and emamectin benzoate) as well as benzoxazinoids (BXDs).