The brown planthopper (BPH), Nilaparvata lugens (Stål), is one of the most destructive pests that seriously threatens the high-quality and safe production of rice. However, due to the unscientific use of chemical insecticides, N. lugens has developed varying levels of resistance to insecticides, including nitenpyram and clothianidin. The ATP-binding cassette (ABC) transporter plays a nonnegligible role in phase III of the detoxification process, which may play an important role in insecticide resistance. In the present study, NlABCG3 was significantly overexpressed in both the NR and CR populations compared with susceptible populations. Silencing NlABCG3 significantly increased the susceptibility of BPH to nitenpyram and clothianidin. In addition, RNAi-mediated knockdown of three key genes in the miRNA biogenesis pathway altered the level of NlABCG3. Subsequently, the luciferase reporter assays demonstrated that novel_268 binds to the NlABCG3 coding region and downregulates its expression. Furthermore, injection of miRNA inhibitors or mimics of novel_268 significantly altered the susceptibility of N. lugens to nitenpyram and clothianidin. These results suggest that miRNA novel_268 targeting NlABCG3 is involved in nitenpyram and clothianidin resistance in N. lugens. These findings may help to enhance our knowledge of the transcriptional regulation of the ABC transporter that mediate insecticide resistance in N. lugens.
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