Overexpression of ace2 compensating for the acetylcholinesterase activity loss from ace1 mutations accelerated the development of eggs and early nymphs in Nilaparvata lugens.

Acetylcholinesterase (AChE) guarantees the acetylcholine signal in insect central nervous system, and is the target of organophosphorus and carbamate insecticides, towards which resistance was often reported due to AChE mutations. In Nilaparvata lugens, a major pest on rice, two mutations (G119S and F331C) were detected in AChE1 in a chlorpyrifos-resistant (CHL) strain. The double mutations in AChE1 reduced total AChE activity in metabolizing acetylcholine, such as low protein stability, substrate affinity and catalytic efficiency, which needed compensation in a special way. In CHL strain, the transcriptional level of ace2 encoding AChE2 was systematically elevated, such as over 30-fold overexpression in brain. The ace2 overexpression not only compensated for AChE activity loss in brain due to AChE1 mutations, but also accelerated the development of eggs and early nymphs in CHL strain. When performing ace2 RNAi in CHL eggs, the egg and early nymph duration were recovered. In CHL eggs, the transcriptional levels of three basic helix-loop-helix transcription factors (Ase2, Ato1 and SCL), which were closely related to neural development, were significantly upregulated. Their respective RNAi in CHL strain also significantly recovered the egg duration, as RNAi towards ace2, which partially explained the reason for the accelerated development of eggs and early nymphs. The results revealed AChE2 non-canonical function in insect embryonic development, and uncovered a physiological effect caused by ace2 overexpression as a compensation action for resistance mechanism due to AChE1 mutations, providing a base for insecticide resistance management in insects.