title: High-throughput detection of knockdown resistance in Myzus persicae using allelic discriminating quantitative PCR.
abstract: The peach-potato aphid Myzus persicae (Sulzer) has developed resistance to pyrethroid insecticides as a result of a mechanism conferring reduced nervous system sensitivity, termed knockdown resistance (kdr). This reduced sensitivity is caused by two mutations, L1014F (kdr) and M918T (super-kdr), in the para-type voltage-gated sodium channel. We have developed a diagnostic dose bioassay to detect kdr and provide preliminary information on the genotype present. We also developed two allelic discrimination PCR assays to determine precisely the genotypes of the two mutations (L1014F and M918T) in individual M. persicae using fluorescent Taqman MGB probes. In combination with assays for elevated carboxylesterase levels and modified acetylcholinesterase (MACE), this suite of assays allows for rapid high-throughput diagnosis, in individual aphids, of the three main resistance mechanisms of practical importance in the UK.
authors.affiliation: Department of Plant and Invertebrate Ecology, Rothamsted Research, Harpenden AL5 2JQ, UK. ansteadj@bbsrc.ac.uk
authors.forename: James A
authors.initials: JA
authors.lastname: Anstead
authors.forename: Martin S
authors.initials: MS
authors.lastname: Williamson
authors.forename: Ioannis
authors.initials: I
authors.lastname: Eleftherianos
authors.forename: Ian
authors.initials: I
authors.lastname: Denholm
mesh_terms: D000483:Alleles; D000818:Animals; D001042:Aphids / Q000235:genetics*; D005456:Fluorescent Dyes; D005820:Genetic Testing; D007305:Insecticide Resistance / Q000235:genetics*; D007306:Insecticides / Q000494:pharmacology* / Q000633:toxicity; D017354:Point Mutation; D016133:Polymerase Chain Reaction / Q000379:methods*; D011722:Pyrethrins / Q000633:toxicity*; D012680:Sensitivity and Specificity; D015222:Sodium Channels / Q000235:genetics
chemical_list: D005456:Fluorescent Dyes; D007306:Insecticides; D011722:Pyrethrins; D015222:Sodium Channels