Chemosensory Proteins Protect Nilaparvata lugens from Imidacloprid by Sequestering the Insecticide and Facilitating Metabolic Detoxification

The involvement of chemosensory proteins (CSPs) in binding to insecticides has been implicated. However, our understanding of CSP-mediated insecticide resistance remains limited. Herein, 15 CSP genes were identified and characterized from Nilaparvata lugens. Expression analysis identified six CSPs with overexpression in the imidacloprid-resistant strain, whose involvement in imidacloprid resistance was validated by RNA interference. Among them, four CSPs were successfully expressed using a prokaryotic expression system, and their binding affinities to imidacloprid were confirmed through fluorescence competitive binding assays. Knockdown of them impaired the capacity of N. lugens to metabolize imidacloprid and inhibited the activity of metabolic detoxification pathways, while their overexpression in Escherichia coli enhanced bacterial metabolic efficiency toward imidacloprid. Furthermore, the transcriptional regulation of CSP2 and CSP15 was found to be mediated by AhR/ARNT and CncC/MafK. These findings suggest that the overexpression of CSPs in N. lugens promotes imidacloprid resistance by sequestering the insecticide and enhancing metabolic detoxification.