Overexpression of NADPH-cytochrome P450 reductase is associated with sulfoxaflor resistance and neonicotinoid cross-resistance in Nilaparvata lugens (Stål)

Nicotinamide adenine dinucleotide phosphate (NADPH)–cytochrome P450 reductase (CPR), a crucial electron-transfer partner of P450 systems, is required for various biological reactions catalyzed by P450 monooxygenase. Our previous study indicated that enhanced P450 enzyme detoxification and CYP6ER1 overexpression contributed to sulfoxaflor resistance in Nilaparvata lugens. However, the association between CPR, sulfoxaflor resistance, and neonicotinoid cross-resistance in N. lugens remains unclear. In this study, the sulfoxaflor-resistant (SFX-SEL) (RR = 254.04-fold), resistance-decline (DESEL) (RR = 18.99-fold), and susceptible unselected (UNSEL) strains of N. lugens with the same genetic background were established. Real-time quantitative polymerase chain reaction (RT-qPCR) revealed that the N. lugens CPR (NlCPR) expression level in the SFX-SEL strain was 6.85-fold and 6.07-fold higher than in UNSEL and DESEL strains, respectively. NlCPR expression was significantly higher in the abdomens of UNSEL, DESEL, and SFX-SEL fourth-instar nymphs than in other tissues (thoraxes, heads, and legs). Additionally, sulfoxaflor stress significantly increased NlCPR mRNA levels in the UNSEL, SFX-SEL and DESEL strains. NlCPR silencing by RNA interference (RNAi) dramatically increased the susceptibility of the UNSEL, DESEL, and SFX-SEL strains to sulfoxaflor, but the recovery of SFX-SEL was more obvious. Furthermore, NlCPR silencing led to a significant recovery in susceptibility to nitenpyram, dinotefuran, clothianidin, and thiamethoxam across all strains (UNSEL, DESEL, and SFX-SEL), with the greatest degree of recovery in the sulfoxaflor-resistant strain (SFX-SEL). Our findings suggest that NlCPR overexpression contributes to sulfoxaflor resistance and neonicotinoid cross-resistance in N. lugens. This will aid in elucidating the significance of CPR in the evolution of P450-mediated metabolic resistance in N. lugens.