Identification and Characterization of CYP6 Family Genes from the Oriental Fruit Moth (Grapholita molesta) and Their Responses to Insecticides

Abstract

Cytochrome P450 (CYP) monooxygenases comprise a superfamily of proteins that detoxify xenobiotics and plant secondary metabolites in insects. The CYP6 family is unique to the class Insecta, and its members participate in the metabolism of exogenous substances. In this study, we sequenced and characterized the full-length cDNAs of eight CYP6 family genes from Grapholita molesta (Busck), a global pest of pome fruits. P450 genes with the exception of CYP6AN35, which was most highly expressed in adults, consistently showed high expression in third- or fourth-instar larvae. The analysis of different tissues of adults showed that most of these genes were predominantly expressed in the midgut, Malpighian tubules, and/or fat body. The expression of these eight CYP6 genes was differentially affected by three representative insecticides: malathion (organophosphate), deltamethrin (pyrethroid), and chlorantraniliprole (carbamate). All eight CYP6 genes responded to malathion treatment. Only three CYP6 genes were highly expressed in deltamethrin-treated individuals. Chlorantraniliprole treatment exerted weak effects on gene expression. Interestingly, CYP6AN35 was a highly expression level in the adult head and its expression was induced by all three insecticides. CYP6AN35 may be a key gene in the metabolism of insecticides. This study provides a fundamental understanding of the functions of the CYP6 gene family in insecticide metabolism in G. molesta.

Inducible Cytochrome P450s in the Fat Body and Malpighian Tubules of the Polyphagous Pests of Spodoptera litura Confer Xenobiotic Tolerance

“…In addition to investigating the structures of P450s, the overexpression of P450s, especially those of the CYP3 and 4 clades, has been explored, and the results indicated that these enzymes are related to insecticide resistance or tolerance. For example, the expression levels of AgCYP6CY7, AgCYP6CY21, AgCYP4CJ1, and AgCYP380C6 in a cyantraniliprole-resistant A. gossypii strain, 45 GmCYP6AB112 and GmCYP6AB113 of Grapholita molesta, 46 and PxCYP6BG1 of Plutella xylostella 47 were increased in response to diamide treatments. Moreover, upregulated P450 expression levels were found to be associated with plant allelochemical exposure.…”

“…For example, the expression levels of AgCYP6CY7 , AgCYP6CY21 , AgCYP4CJ1, and AgCYP380C6 in a cyantraniliprole-resistant A. gossypii strain, GmCYP6AB112 and GmCYP6AB113 of Grapholita molesta , and PxCYP6BG1 of Plutella xylostella were increased in response to diamide treatments. Moreover, upregulated P450 expression levels were found to be associated with plant allelochemical exposure. ,, DaCYP6CR2 and DaCYP6DE5 expression in D.…”

RNA interference in cytochrome P450 monooxygenase (CYP) gene results in reduced insecticide resistance in Megalurothrips usitatus Bagnall

“…Studies have shown that the P450 genes related to insecticide resistance are mainly concentrated in the CYP3 (including CYP6 and CYP9) and CYP4 families ( Feyereisen, 2012 ). In this study, the MusiDN2722 sequence cloned from M. usitatus was used to construct a phylogenetic tree and was found to belong to the CYP6 clade, the family members of which have also been shown to play important roles in the detoxification and metabolism of toxic substances ( Pan et al, 2018 ; Han et al, 2022 ). Through comparison with CYP genes of the same family in other insects, it was found that MusiDN2722 shared the characteristic CYP450 sequence with other model insects, containing highly conserved hydrogen-bonding regions, including Helix-C (WxxxR), Helix-I (AGxxT), Helix-K (ExxR), Meander (FxxGxRxxxG), and the heme-binding domain (PxxFxPxxF) ( Feyereisen, 2005 ).…”

Identification and Functional Characterization of CYP4D2 Putatively Associated with β-Cypermethrin Detoxification in Phortica okadai

“…melanogaster dietary exposure to permethrin and cyp4e3 knockdown caused a significant elevation of oxidative stress-associated markers in the Malpighian tubules, which included lipid peroxidation based on the production of 4-hydroxynonenal; these findings have increased our understanding of the molecular mechanisms of permethrin detoxification in the Malpighian tubules [ 36 ]. These observations indicate that the fat body, midgut, and Malpighian tubules are essential tissues for detoxification [ 37 , 38 ]. In particular, the expression of Pocyp4d2 was higher in the midgut than in other tissues.…”