title: Amplification of a cytochrome P450 gene is associated with resistance to neonicotinoid insecticides in the aphid Myzus persicae.
abstract: The aphid Myzus persicae is a globally significant crop pest that has evolved high levels of resistance to almost all classes of insecticide. To date, the neonicotinoids, an economically important class of insecticides that target nicotinic acetylcholine receptors (nAChRs), have remained an effective control measure; however, recent reports of resistance in M. persicae represent a threat to the long-term efficacy of this chemical class. In this study, the mechanisms underlying resistance to the neonicotinoid insecticides were investigated using biological, biochemical, and genomic approaches. Bioassays on a resistant M. persicae clone (5191A) suggested that P450-mediated detoxification plays a primary role in resistance, although additional mechanism(s) may also contribute. Microarray analysis, using an array populated with probes corresponding to all known detoxification genes in M. persicae, revealed constitutive over-expression (22-fold) of a single P450 gene (CYP6CY3); and quantitative PCR showed that the over-expression is due, at least in part, to gene amplification. This is the first report of a P450 gene amplification event associated with insecticide resistance in an agriculturally important insect pest. The microarray analysis also showed over-expression of several gene sequences that encode cuticular proteins (2-16-fold), and artificial feeding assays and in vivo penetration assays using radiolabeled insecticide provided direct evidence of a role for reduced cuticular penetration in neonicotinoid resistance. Conversely, receptor radioligand binding studies and nucleotide sequencing of nAChR subunit genes suggest that target-site changes are unlikely to contribute to resistance to neonicotinoid insecticides in M. persicae.
authors.affiliation: Centre for Sustainable Pest and Disease Management, Rothamsted Research, Harpenden, United Kingdom.
authors.forename: Alin M
authors.initials: AM
authors.lastname: Puinean
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Foster
authors.forename: Linda
authors.initials: L
authors.lastname: Oliphant
authors.forename: Ian
authors.initials: I
authors.lastname: Denholm
authors.forename: Linda M
authors.initials: LM
authors.lastname: Field
authors.forename: Neil S
authors.initials: NS
authors.lastname: Millar
authors.forename: Martin S
authors.initials: MS
authors.lastname: Williamson
authors.forename: Chris
authors.initials: C
authors.lastname: Bass
mesh_terms: D000818:Animals; D001042:Aphids / Q000737:chemistry / Q000187:drug effects / Q000235:genetics*; D001483:Base Sequence; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics*; D005784:Gene Amplification / Q000187:drug effects*; D018628:Gene Dosage; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics*; D007305:Insecticide Resistance; D007306:Insecticides / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009538:Nicotine / Q000494:pharmacology*; D020411:Oligonucleotide Array Sequence Analysis; D016415:Sequence Alignment
chemical_list: D019476:Insect Proteins; D007306:Insecticides; D009538:Nicotine; D003577:Cytochrome P-450 Enzyme System