The diamondback moth (DBM), Plutella xylostella, is a globally distributed and important economic pest. Chemical control is the primary approach to regulate populations of this pest. Chlorantraniliprole is the first commercial insecticide that belongs to the new chemical class of diamide insecticides. In this study, the resistant strain was observed 1578-fold resistance to chlorantraniliprole. Point mutation (G4946E) in ryanodine receptor (RyR) showed a high frequency. Enzyme assays indicated that glutathione S-transferase (GST) activity in the resistant strain was 2.4 times higher compared with the susceptible strain, whereas no difference was seen for P450 and esterase. In addition, the expression of two GSTs genes was up-regulated. These findings pave the way for the complete understanding of the mechanisms of diamide insecticides resistance in insects.