Although a large number of laboratory bioassay methods have been developed for detecting and characterizing resistance, most of these are limited to defining phenotypes and provide little or no information on the underlying genes or mechanisms. Thus, although bioassays remain the indispensable mainstay of most large-scale resistance monitoring programs, much attention is being paid to developing more incisive techniques that not only offer greater precision and turnover rates, but also diagnose the type of mechanism(s) present and, whenever possible, the genotypes of resistant insects. A variety of approaches are being adopted for this purpose, including electrophoretic or immunological detection of resistance-causing enzymes, kinetic and end-point assays for quantifying the activity of enzymes or their inhibition by insecticides, and DNA-based diagnostics for mutant resistance alleles.
The sensitivity of these techniques is exemplified by work on the green peach aphid, Myzus persicae. In northern Europe, this insect possesses three coexisting resistance mechanisms: an overproduced carboxylesterase conferring resistance to organophosphates, an altered acetylcholinesterase conferring resistance to certain carbamates, and target-site resistance (i.e., knockdown resistance, kdr) to pyrethroids. These mechanisms collectively confer strong resistance in this species to virtually all available aphicides. Fortunately, it is now possible to diagnose all three mechanisms in individual aphids by using an immunoassay for the overproduced esterase, a kinetic microplate assay for the mutant AChE, and a molecular diagnostic for the kdr allele. The combined use of these techniques against field populations provides up-to-date information on the incidence of the mechanisms and serves to inform growers of potential control problems and in the development of optimal strategies for the management of M. persicae.
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