title: Cloning of Two Acetylcholinesterase Genes and Analysis of Point Mutations Putatively Associated with Triazophos Resistance in Chilo auricilius (Lepidoptera: Pyralidae).
abstract: Acetylcholinesterase (AChE) is the target of organophosphate (OP) and carbamate insecticides. Mutations in the AChE gene (ace) leading to decreased insecticide susceptibility is the main resistance mechanism in insects. In this study, two Chilo auricilius acetylcholinesterase genes, designated as Caace1 and Caace2, were cloned using RT-PCR and RACE. Caace1 cDNA is 2534 bp, with ORF of 2082 bp, and it encodes an acetylcholinesterase 1 (CaAChE1) protein comprising a calculated 693 amino acid (aa) residues. Caace2 cDNA contains 2280 bp, with a full-length ORF of 1917 bp, encoding acetylcholinesterase 2 (CaAChE2) comprising a calculated 638 aa residues. At the aa level, CaAChE1 displays the highest similarity (97%) with the Chilo suppressalis AChE1, and CaAChE2 shows the highest similarity with the C. suppressalis AChE2 (99%). From the restriction fragment length polymorphism (RFLP) PCR (RFLP-PCR) analysis, one mutation in Caace1, similar to the ace1 mutation associated with triazophos resistance in C. suppressalis, was detected. Detailed examination of field populations of C. auricilius indicated this resistance mutation in C. auricilius is still quite infrequent. Based on the assay of AChE activity and RFLP-PCR testing, an individual that contains resistance mutation has lower AChE activities, while the individual that does not contain the resistance mutation has higher AChE activities. This study provides a basis for future investigations into the mechanism of OP resistance in C. auricilius, as well as a guidance for C. auricilius control with reasonable choice of pesticides.
authors.affiliation: Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
authors.forename: Guang-Hua
authors.initials: GH
authors.lastname: Luo
authors.affiliation: Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China.
authors.forename: Xiao-Huan
authors.initials: XH
authors.lastname: Li
authors.affiliation: Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
authors.forename: Zhi-Chun
authors.initials: ZC
authors.lastname: Zhang
authors.affiliation: Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
authors.forename: Bao-Sheng
authors.initials: BS
authors.lastname: Liu
authors.affiliation: Institute of Plant Protection, Jiangxi Academy of Agricultural Sciences, Nanchang 330200, China.
authors.forename: Shui-Jin
authors.initials: SJ
authors.lastname: Huang
authors.affiliation: Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China fangjc@jaas.ac.cn.
authors.forename: Ji-Chao
authors.initials: JC
authors.lastname: Fang
mesh_terms: D000110:Acetylcholinesterase / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002681:China; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D007305:Insecticide Resistance; D007306:Insecticides / Q000494:pharmacology*; D007814:Larva / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D008969:Molecular Sequence Data; D009036:Moths / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D063086:Organothiophosphates / Q000494:pharmacology*; D010802:Phylogeny; D016415:Sequence Alignment; D014230:Triazoles / Q000494:pharmacology*
chemical_list: D018076:DNA, Complementary; D019476:Insect Proteins; D007306:Insecticides; D063086:Organothiophosphates; D014230:Triazoles; C012584:triazophos; D000110:Acetylcholinesterase
keywords: Chilo auricilius; acetylcholinesterase; point mutation; resistance; triazophos