Institute of Molecular Biology & Biotechnology, Foundation for Research & Technology Hellas, 100 N. Plastira Street, GR-700 13, Heraklion, Crete, Greece; Laboratory of Molecular Entomology, Department of Biology, University of Crete, GR-700 13, Heraklion, Crete, Greece.
Institute of Molecular Biology & Biotechnology, Foundation for Research & Technology Hellas, 100 N. Plastira Street, GR-700 13, Heraklion, Crete, Greece.
Pestic Biochem Physiol. 2019 Jun;157:196-203. doi: 10.1016/j.pestbp.2019.03.022. Epub 2019 Apr 1.
Overexpression of the cytochrome P450 monooxygenase CYP6A51 has been previously associated with pyrethroid resistance in the Mediterranean fruit fly (medfly) Ceratitis capitata, an important pest species worldwide; however, this association has not been functionally validated. We expressed CYP6A51 gene in Escherichia coli and produced a functional enzyme with preference for the chemiluminescent substrate Luciferin-ME EGE. In vitro metabolism assays revealed that CYP6A51 is capable of metabolizing two insecticides that share the same mode of action, λ-cyhalothrin and deltamethrin, whereas no metabolism or substrate depletion was observed in the presence of spinosad or malathion. We further expressed CYP6A51 in vivo via a GAL4/UAS system in Drosophila melanogaster flies, driving expression with detoxification tissue-specific drivers. Toxicity bioassays indicated that CYP6A51 confers knock-down resistance to both λ-cyhalothrin and deltamethrin. Detection of CYP6A51 - associated pyrethroid resistance in field populations may be important for efficient Insecticide Resistance Management (IRM) strategies.
细胞色素 P450 单加氧酶 CYP6A51 的过表达先前与地中海实蝇（medfly）Ceratitis capitata 的拟除虫菊酯抗性有关，后者是世界范围内的一种重要害虫；然而，这种关联尚未得到功能验证。我们在大肠杆菌中表达了 CYP6A51 基因，并产生了一种对化学发光底物 Luciferin-ME EGE 具有偏好的功能性酶。体外代谢测定表明，CYP6A51 能够代谢两种作用方式相同的杀虫剂，即氯氟氰菊酯和溴氰菊酯，而在用多杀菌素或马拉硫磷处理时，没有观察到代谢或底物消耗。我们进一步通过 GAL4/UAS 系统在黑腹果蝇中体内表达 CYP6A51，通过解毒组织特异性驱动子驱动表达。毒性生物测定表明，CYP6A51 对氯氟氰菊酯和溴氰菊酯均表现出击倒抗性。在田间种群中检测与 CYP6A51 相关的拟除虫菊酯抗性可能对有效的害虫抗药性管理（IRM）策略很重要。
