Abamectin is metabolized by CYP392A16, a cytochrome P450 associated with high levels of acaricide resistance in Tetranychus urticae.
Abamectin is one of the most important insecticides worldwide. It is used against major agricultural pests and insects of public health importance, as well as against endoparasites in animal health. Abamectin has been used successfully for the control of the spider mite Tetranychus urticae, a major agricultural pest with global distribution, an extremely diverse host range, and a remarkable ability to develop resistance against insecticides including abamectin. Target site resistance mutations may explain a large part of resistance, although genetic evidence and transcriptomic data indicated that additional mechanisms may also be implicated in the abamectin resistant phenotype. To investigate a functional link between cytochrome P450-mediated metabolism and abamectin resistance, we recombinantly expressed three cytochrome P450s (CYP392A16, CYP392D8 and CYP392D10) that have been associated with high levels of abamectin resistance in a resistant T. urticae strain isolated from Greece. CYP392A16 was expressed predominately in its P450 form however, both CYP392D8 and CYP392D10 were expressed predominately as P420, despite optimization efforts on expression conditions. CYP392A16 catalyses the hydroxylation of abamectin (Kcat=0.54 pmol/min/pmol P450; Km=45.9 μM), resulting in a substantially less toxic compound as confirmed by bioassays with the partially purified metabolite. However, CYP392A16 did not metabolize hexythiazox, clofentezine and bifenthrin, active ingredients that also showed reduced toxicity in the abamectin resistant strain. Among a number of fluorescent and luminescent substrates screened, Luciferin-ME EGE was preferentially metabolized by CYP392A16, and it may be a potential diagnostic probe for metabolic resistance detection and monitoring.
Faculty of Applied Biotechnology and Biology, Department of Biology, University of Crete, 71409 Heraklion, Greece.
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M
Riga
Faculty of Applied Biotechnology and Biology, Department of Biology, University of Crete, 71409 Heraklion, Greece.
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D
Tsakireli
Faculty of Applied Biotechnology and Biology, Department of Biology, University of Crete, 71409 Heraklion, Greece.
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A
Ilias
Faculty of Applied Biotechnology and Biology, Department of Biology, University of Crete, 71409 Heraklion, Greece.
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E
Morou
Faculty of Environmental & Analytical Chemistry, Department of Chemistry, University of Crete, 71409 Heraklion, Greece.
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A
Myridakis
Faculty of Environmental & Analytical Chemistry, Department of Chemistry, University of Crete, 71409 Heraklion, Greece.
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EG
Stephanou
Bayer CropScience AG, RD-SMR Pest Control Biology, Alfred Nobel Str. 50, D-40789 Monheim, Germany.
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Nauen
Laboratory of Agrozoology, Department of Crop Protection, Faculty of Agricultural and Applied, Biological Sciences, Ghent University, B-9000 Ghent, Belgium.
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Dermauw
Laboratory of Agrozoology, Department of Crop Protection, Faculty of Agricultural and Applied, Biological Sciences, Ghent University, B-9000 Ghent, Belgium; Institute for Biodiversity and Ecosystem Dynamics (IBED), University of Amsterdam (UvA), Science Park 904, 1098 XH Amsterdam, The Netherlands.
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Van Leeuwen
Department of Vector Biology, Liverpool School Tropical Medicine, UK.
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M
Paine
Faculty of Applied Biotechnology and Biology, Department of Biology, University of Crete, 71409 Heraklion, Greece. Electronic address: vontas@biology.uoc.gr.
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Vontas
D056810:Acaricides / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D060829:Arthropod Proteins / Q000235:genetics / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D004351:Drug Resistance / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D007559:Ivermectin / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D040102:Tetranychidae / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism
D056810:Acaricides; D060829:Arthropod Proteins; D011994:Recombinant Proteins; C048324:abamectin; D007559:Ivermectin; D003577:Cytochrome P-450 Enzyme System
Avermectin; Detoxification; Diagnostic; Multiple resistance