High-throughput detection of knockdown resistance in Myzus persicae using allelic discriminating quantitative PCR.
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.
Anstead
James A
JA
Department of Plant and Invertebrate Ecology, Rothamsted Research, Harpenden AL5 2JQ, UK. ansteadj@bbsrc.ac.uk
Williamson
Martin S
MS
Eleftherianos
Ioannis
I
Denholm
Ian
I
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
D005456:Fluorescent Dyes; D007306:Insecticides; D011722:Pyrethrins; D015222:Sodium Channels