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.

The development, reproduction and P450 enzyme of the white-backed planthopper, Sogatella furcifera (Hemiptera: Delphacidae) under the sublethal concentrations of clothianidin.

Clothianidin is a second-generation neonicotinoid insecticide that can effectively prevent piercing-sucking pests, such as white-backed planthopper (WBPH), Sogatella furcifera (Horváth). In this study, the sublethal effects of clothianidin on the biological traits of S. furcifera were evaluated via the age-stage, two-sex life table procedure. Our results exhibited that the female adult longevity, fecundity and hatchability of F0 generation were significantly decreased after LC10 and (or) LC30 of clothianidin exposure compared to the control. Transgenerational effects showed that the pre-adult period, female adult longevity, total longevity, oviposition days (Od), fecundity and mean generation time (T) of F1 generation were significantly decreased in the LC10 and LC30 groups compared to the control. Moreover, the development times of the third- and fifth-instar nymphs, total preoviposition period (TPOP) and doubling time (DT) were significantly shorter in the LC10 group than in the control and LC30 groups. Furthermore, the intrinsic rate of increase (ri) and finite rate of increase (λ) values of the LC10 group were significantly higher than those of the control group. However, there was no significant difference in the male adult longevity, adult preoviposition period (APOP) and net reproductive rate (R0) between the treated groups and the control. Enzyme activity and gene expression results showed that the P450 enzyme activity and mRNA levels of many P450 genes were significantly increased by clothianidin treatment. In addition, the knockdown of CYP4CE3 and CYP6FJ3, which showed the highest inducing levels, by RNA interference (RNAi) dramatically increased the toxicity of clothianidin against S. furcifera. These results indicated that sublethal concentrations of clothianidin showed a stimulatory effect on the development, but it could adversely affect the survival and reproduction of S. furcifera. Additionally, CYP4CE3 and CYP6FJ3 might play an important role in the detoxification and evolution of clothianidin resistance in S. furcifera.

The Cross-Resistance Pattern and the Metabolic Resistance Mechanism of Acetamiprid in the Brown Planthopper, Nilaparvata lugens (Stål).

Acetamiprid is widely used in paddy fields for controlling Nilaparvata lugens (Stål). However, the risk of resistance development, the cross-resistance pattern and the resistance mechanism of acetamiprid in this pest remain unclear. In this study, an acetamiprid-resistant strain (AC-R) was originated from a field strain (UNSEL) through successive selection with acetamiprid for 30 generations, which reached 60.0-fold resistance when compared with a laboratory susceptible strain (AC-S). The AC-R strain (G30) exhibited cross-resistance to thiamethoxam (25.6-fold), nitenpyram (21.4-fold), imidacloprid (14.6-fold), cycloxaprid (11.8-fold), dinotefuran (8.7-fold), sulfoxaflor (7.6-fold) and isoprocarb (8.22-fold), while there was no cross-resistance to etofenprox, buprofezin and chlorpyrifos. Acetamiprid was synergized by the inhibitor piperonyl butoxide (2.2-fold) and the activity of cytochrome P450 monooxygenase was significantly higher in the AC-R strain compared with the AC-S strain, suggesting the critical role of P450. The gene expression results showed that the P450 gene CYP6ER1 was significantly overexpressed in AC-R compared with the AC-S and UNSEL strains. In addition, the RNA interference (RNAi) of CYP6ER1 significantly increased the susceptibility of AC-R to acetamiprid. Molecular docking predicted that acetamiprid and CYP6ER1 had close binding sites, and the nitrogen atoms had hydrogen bond interactions with CYP6ER1. These results demonstrated that the overexpression of CYP6ER1 contributed to acetamiprid resistance in N. lugens.