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Functional characterization of cytochrome P450s associated with pyrethroid resistance in the olive fruit fly Bactrocera oleae

View ORCID ProfileAnastasia Kampouraki, View ORCID ProfileDimitra Tsakireli, View ORCID ProfileVenetia Koidou, View ORCID ProfileMarianna Stavrakaki, Stavroula Kaili, Yannis Livadaras, View ORCID ProfileLinda Grigoraki, View ORCID ProfilePanagiotis Ioannidis, View ORCID ProfileEmmanouil Roditakis, View ORCID ProfileJohn Vontas
doi: https://doi.org/10.1101/2022.12.07.519177
Anastasia Kampouraki
aPesticide Science Lab, Agricultural University of Athens, 75 Iera Odos, 118 55 Athens, Greece
bInstitute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, GR-700 13 Heraklion, Crete, Greece
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  • For correspondence: vontas@imbb.forth.gr kampouraki.an@gmail.com
Dimitra Tsakireli
aPesticide Science Lab, Agricultural University of Athens, 75 Iera Odos, 118 55 Athens, Greece
bInstitute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, GR-700 13 Heraklion, Crete, Greece
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Venetia Koidou
bInstitute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, GR-700 13 Heraklion, Crete, Greece
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Marianna Stavrakaki
aPesticide Science Lab, Agricultural University of Athens, 75 Iera Odos, 118 55 Athens, Greece
cInstitute of Olive Tree, Subtropical Crops and Viticulture, Hellenic Agricultural Organization – Demeter, 71307, Heraklion, Greece
dDepartment of Agriculture, School of Agricultural Sciences, Hellenic Mediterranean University, Estavromenos, 71410 Heraklion, Greece
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Stavroula Kaili
bInstitute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, GR-700 13 Heraklion, Crete, Greece
eDepartment of Biology, University of Crete, Vassilika Vouton, 71409 Heraklion, Crete, Greece
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Yannis Livadaras
bInstitute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, GR-700 13 Heraklion, Crete, Greece
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Linda Grigoraki
bInstitute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, GR-700 13 Heraklion, Crete, Greece
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Panagiotis Ioannidis
bInstitute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, GR-700 13 Heraklion, Crete, Greece
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Emmanouil Roditakis
cInstitute of Olive Tree, Subtropical Crops and Viticulture, Hellenic Agricultural Organization – Demeter, 71307, Heraklion, Greece
dDepartment of Agriculture, School of Agricultural Sciences, Hellenic Mediterranean University, Estavromenos, 71410 Heraklion, Greece
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John Vontas
aPesticide Science Lab, Agricultural University of Athens, 75 Iera Odos, 118 55 Athens, Greece
bInstitute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, GR-700 13 Heraklion, Crete, Greece
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  • For correspondence: vontas@imbb.forth.gr kampouraki.an@gmail.com
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Abstract

Resistance to pyrethroid insecticides has evolved in Bactrocera oleae populations in Greece, threatening the efficacy of control interventions based on this insecticide class. Here we report the collection of populations from Crete with resistance levels reaching up to 331-folds, compared to susceptible laboratory strains and show that pyrethroid resistance is substantially suppressed by the PBO synergist, suggesting the involvement of detoxification enzymes. To identify specific candidate genes implicated in resistance, we performed comparative transcriptomic analysis, between the pyrethroid resistant populations from Crete and the susceptible laboratory strains, using both whole bodies and Malpighian tubules. Several genes were found differentially transcribed between resistant and susceptible flies in each comparison, with P450s being among the most highly over-expressed detoxification genes in pyrethroid resistant populations. Four of the over-expressed P450s (Cyp6A61, Cyp6G6, Cyp4P6 and Cyp6G28) were recombinantly expressed in Escherichia coli and in vitro metabolism assays revealed that CYP6A61 is capable of metabolizing alpha-cypermethrin, while CYP6G6, CYP4P6 and CYP6G28 are capable of metabolizing deltamethrin. No metabolism of neonicotinoid insecticides was recorded. We further silenced CYP6G6 in vivo, via RNAi, which led to a small, but significant increase in deltamethrin toxicity. The study provides valuable information towards the development of molecular diagnostics and evidence-based insecticide resistance management strategies.

Competing Interest Statement

The authors have declared no competing interest.

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Posted December 08, 2022.
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Functional characterization of cytochrome P450s associated with pyrethroid resistance in the olive fruit fly Bactrocera oleae
Anastasia Kampouraki, Dimitra Tsakireli, Venetia Koidou, Marianna Stavrakaki, Stavroula Kaili, Yannis Livadaras, Linda Grigoraki, Panagiotis Ioannidis, Emmanouil Roditakis, John Vontas
bioRxiv 2022.12.07.519177; doi: https://doi.org/10.1101/2022.12.07.519177
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Functional characterization of cytochrome P450s associated with pyrethroid resistance in the olive fruit fly Bactrocera oleae
Anastasia Kampouraki, Dimitra Tsakireli, Venetia Koidou, Marianna Stavrakaki, Stavroula Kaili, Yannis Livadaras, Linda Grigoraki, Panagiotis Ioannidis, Emmanouil Roditakis, John Vontas
bioRxiv 2022.12.07.519177; doi: https://doi.org/10.1101/2022.12.07.519177

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