The Roles of Four Novel P450 Genes in Pesticides Resistance in Apis cerana cerana Fabricius: Expression Levels and Detoxification Efficiency

Abstract

Cytochrome P450 monooxygenases (P450s) are widely distributed multifunctional enzymes that play crucial roles in insecticide detoxification or activation. In this study, to ascertain the molecular mechanisms of P450s in the detoxification of Chinese honeybees, Apis cerana cerana Fabricius (A. c. cerana), we isolated and characterized four new P450 genes (Acc301A1, Acc303A1, Acc306A1, and Acc315A1). The open reading frames of the four genes are 1263 to 1608 bp in length and encode four predicted polypeptides of 499 to 517 amino acids in length. Real-time quantitative PCR (RT-qPCR) results showed that expression of all four genes was observed in all developmental stages. In addition, Western blot assays further indicated the RT-qPCR results that showed that the four genes were induced by pesticide (thiamethoxam, deltamethrin, dichlorovos, and paraquat) treatments. Furthermore, we also used double-stranded RNA-mediated RNA interference to investigate the functions of Acc301A1, Acc303A1,and Acc306A1 in the antioxidant defense of honeybees. RNA interference targeting Acc301A1, Acc303A1, and Acc306A1 significantly increased the mortality rate of A. c. cerana upon pesticide treatment. These results provide important evidence about the role of the four P450 genes involved in detoxification.

Evidence-based insecticide resistance in South American tomato leaf miner,Phthorimaea absoluta(Meyrick) under laboratory selection

“…CYP450s are the major genes associated with resistance to most of the chemical insecticides (Nahlen et al ., 2003; Brooke et al ., 2008; Zhang et al ., 2019). We attempted for the first time to study the CYP450 genes in P. absoluta and identified four novel genes viz., CYP248f (flubendiamide), CYP272c, CYP724c (cyantraniliprole), and CYP648i.…”

Low toxicity crop fungicide (fenbuconazole) impacts reproductive male quality signals leading to a reduction of mating success in a wild solitary bee

“…It is possible that the synthetic pathways of some of the key compounds that are used by females to assess male fitness were altered after fungicide exposure and that this leads to the observed changes in chemical profile. There is evidence of an indirect regulation of cuticular hydrocarbons through detoxification and metabolic resistance to pesticides in insects that are mediated by cytochrome P450 monooxygenases (Balabanidou et al, 2016; Zhang et al, 2019). P450 genes are modulators of biosynthetic production of hydrocarbons in insects (Chen et al, 2016) and subfamilies of these genes are associated with metabolic resistance to systemic pesticides in honeybees (Gong & Diao, 2017) and in Osmia bees (Beadle et al, 2019).…”

“…It is possible that the synthetic pathways of some of the key compounds that are used by females to assess male fitness were altered after fungicide exposure and that this leads to the observed changes in chemical profile. There is evidence of an indirect regulation of cuticular hydrocarbons through detoxification and metabolic resistance to pesticides in insects that are mediated by cytochrome P450 monooxygenases (Balabanidou et al, 2016;Zhang et al, 2019).…”

Functionally characterized arthropod pest and pollinator cytochrome P450s associated with xenobiotic metabolism

“…P450 functional validation seem to rely more on the in vitro systems, and also in vivo, mostly using genome modification set of techniques. Three P450s have been validated only using RNAi (A. cerana cerana CYP301A1, CYP303A1, CYP306A1 (Zhang et al, 2019))(Figure 3E and Table 2D). Thirteen P450s were validated using only in vitro systems.…”