Grapholita molesta is one of the most important fruit pests worldwide. Abamectin is a biological pesticide frequently used to control fruit borers like G. molesta in part owing to its translaminar properties. In this study, we characterized the toxicity of abamectin to G. molesta larvae using the diet incorporation method. The sublethal effects of abamectin on the development, reproduction, detoxification enzyme activity, and related gene expression of G. molesta were assessed. The results showed that the LC20 and LC50 values of the insecticide against G. molesta 72 h post-treatment were 1.17 mg L–1 and 5.85 mg L–1, whereas the LC20 and LC50 values 96 h post-treatment were 0.34 mg L–1 and 3.63 mg L–1. When compared to the control, sublethal concentrations of abamectin 1) significantly increased the mortality of the larvae, prepupae, and pupae of G. molesta, 2) prolonged the duration of 3rd to 5th instar larva, prepupal and pupal periods, 3) shortened the longevity of adults, and 4) reduced female fecundity. The enzymatic activity of glutathione S-transferase (GST) varied significantly after exposure to sublethal concentrations of abamectin, but the cytochrome P450 monooxygenases and carboxylesterase activity were not significantly affected. Thirteen of the 25 GST genes were significantly upregulated under different sublethal concentrations of abamectin. The combined findings increase our understanding of the effects of abamectin on G. molesta and the potential role of GSTs in the metabolic interactions of abamectin in this pest, and have applications for more rational and effective use of abamectin to control G. molesta.