title: A nicotinic acetylcholine receptor mutation conferring target-site resistance to imidacloprid in Nilaparvata lugens (brown planthopper).
abstract: Neonicotinoids, such as imidacloprid, are nicotinic acetylcholine receptor (nAChR) agonists with potent insecticidal activity. Since its introduction in the early 1990s, imidacloprid has become one of the most extensively used insecticides for both crop protection and animal health applications. As with other classes of insecticides, resistance to neonicotinoids is a significant threat and has been identified in several pest species, including the brown planthopper, Nilaparvata lugens, a major rice pest in many parts of Asia. In this study, radioligand binding experiments have been conducted with whole-body membranes prepared from imidacloprid-susceptible and imidacloprid-resistant strains of N. lugens. The results reveal a much higher level of [3H]imidacloprid-specific binding to the susceptible strain than to the resistant strain (16.7 +/- 1.0 and 0.34 +/- 0.21 fmol/mg of protein, respectively). With the aim of understanding the molecular basis of imidacloprid resistance, five nAChR subunits (Nlalpha1-Nlalpha4 and Nlbeta1) have been cloned from N. lugens.A comparison of nAChR subunit genes from imidacloprid-sensitive and imidacloprid-resistant populations has identified a single point mutation at a conserved position (Y151S) in two nAChR subunits, Nlalpha1 and Nlalpha3. A strong correlation between the frequency of the Y151S point mutation and the level of resistance to imidacloprid has been demonstrated by allele-specific PCR. By expression of hybrid nAChRs containing N. lugens alpha and rat beta2 subunits, evidence was obtained that demonstrates that mutation Y151S is responsible for a substantial reduction in specific [3H]imidacloprid binding. This study provides direct evidence for the occurrence of target-site resistance to a neonicotinoid insecticide.
authors.affiliation: Key Laboratory of Monitoring and Management of Plant Diseases and Insects, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Zewen
authors.initials: Z
authors.lastname: Liu
authors.forename: Martin S
authors.initials: MS
authors.lastname: Williamson
authors.forename: Stuart J
authors.initials: SJ
authors.lastname: Lansdell
authors.forename: Ian
authors.initials: I
authors.lastname: Denholm
authors.forename: Zhaojun
authors.initials: Z
authors.lastname: Han
authors.forename: Neil S
authors.initials: NS
authors.lastname: Millar
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D002681:China; D003001:Cloning, Molecular; D017931:DNA Primers; D004330:Drosophila; D006430:Hemiptera / Q000187:drug effects / Q000235:genetics*; D007093:Imidazoles / Q000633:toxicity*; D007305:Insecticide Resistance / Q000235:genetics; D008969:Molecular Sequence Data; D000073943:Neonicotinoids; D009574:Nitro Compounds; D017354:Point Mutation / Q000235:genetics*; D011869:Radioligand Assay; D011978:Receptors, Nicotinic / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014316:Tritium
chemical_list: D017931:DNA Primers; D007093:Imidazoles; D000073943:Neonicotinoids; D009574:Nitro Compounds; D011978:Receptors, Nicotinic; D014316:Tritium; C082359:imidacloprid