Functional orthologs of honeybee CYP6AQ1 in stingless bees degrade the butenolide insecticide flupyradifurone
Flupyradifurone (FPF), a novel butenolide insecticide binding to nicotinic acetylcholine receptors (nAChRs), has been shown to be less acutely toxic to western honey bees (Apis mellifera) than other insecticides such as neonicotinoids sharing the same target-site. A previous study revealed that this...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-12-01
|
| Series: | Ecotoxicology and Environmental Safety |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S014765132301223X |
| _version_ | 1797435811189227520 |
|---|---|
| author | Xingzhi Xiao Julian Haas Ralf Nauen |
| author_facet | Xingzhi Xiao Julian Haas Ralf Nauen |
| author_sort | Xingzhi Xiao |
| collection | DOAJ |
| description | Flupyradifurone (FPF), a novel butenolide insecticide binding to nicotinic acetylcholine receptors (nAChRs), has been shown to be less acutely toxic to western honey bees (Apis mellifera) than other insecticides such as neonicotinoids sharing the same target-site. A previous study revealed that this is due to enhanced oxidative metabolism of FPF, mediated by three cytochrome P450 monooxygenases (P450s), including CYP6AQ1. Therefore, we followed a toxicogenomics approach and investigated the potential role of functional CYP6AQ1 orthologs in FPF metabolism from eight different bee species, including stingless bees (Tribe: Meliponini). We conducted a phylogenetic analysis on four stingless bee species, including Frieseomelitta varia, Heterotrigona itama, Melipona quadrifasciata and Tetragonula carbonaria to identify CYP6AQ1-like functional orthologs. Three non-Meliponini, but tropical bee species, i.e., Ammobates syriacus, Euglossa dilemma and Megalopta genalis were analyzed as well. We identified candidate P450s in all (neo)tropical species with greater than 61% and 67% predicted protein sequence identities when compared to A. mellifera CYP6AQ1 and Bombus terrestris CYP6AQ26, respectively. Heterologous expression in High Five insect cells of these functional orthologs revealed a common coumarin substrate profile and a preference for the O-debenzylation of bulkier substrates. Competition assays using the fluorescent probe substrate 7-benzyloxymethoxy-4-trifluoromethylcoumarin (BOMFC) with these enzymes indicated inhibition of BOMFC metabolism by increasing concentrations of FPF. Furthermore, UPLC-MS/MS analysis revealed the capacity of all CYP6AQ1-like orthologs to metabolize FPF by hydroxylation in vitro at various levels, indicating a conserved FPF detoxification potential in different (neo)tropical bee species including Meliponini. This research, employing a toxicogenomics approach, provides important insights into the potential of stingless and other tropical bee species to detoxify FPF, and highlights the significance of investigating the detoxification mechanisms of insecticides in non-Apis bee species by molecular tools to inform risk assessment and conservation efforts. |
| first_indexed | 2024-03-09T10:53:31Z |
| format | Article |
| id | doaj.art-96766123713e48fbba07cbf25266741a |
| institution | Directory Open Access Journal |
| issn | 0147-6513 |
| language | English |
| last_indexed | 2024-03-09T10:53:31Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Ecotoxicology and Environmental Safety |
| spelling | doaj.art-96766123713e48fbba07cbf25266741a2023-12-01T05:00:35ZengElsevierEcotoxicology and Environmental Safety0147-65132023-12-01268115719Functional orthologs of honeybee CYP6AQ1 in stingless bees degrade the butenolide insecticide flupyradifuroneXingzhi Xiao0Julian Haas1Ralf Nauen2Institute of Crop Science and Resource Conservation, University of Bonn, 53115 Bonn, Germany; Bayer AG, Crop Science Division, R&D, D-40789 Monheim, GermanyBayer AG, Crop Science Division, R&D, D-40789 Monheim, GermanyBayer AG, Crop Science Division, R&D, D-40789 Monheim, Germany; Correspondence to: Bayer AG, Crop Science Division, R&D, Building 6260, Alfred Nobel Str. 50, D40789 Monheim, Germany.Flupyradifurone (FPF), a novel butenolide insecticide binding to nicotinic acetylcholine receptors (nAChRs), has been shown to be less acutely toxic to western honey bees (Apis mellifera) than other insecticides such as neonicotinoids sharing the same target-site. A previous study revealed that this is due to enhanced oxidative metabolism of FPF, mediated by three cytochrome P450 monooxygenases (P450s), including CYP6AQ1. Therefore, we followed a toxicogenomics approach and investigated the potential role of functional CYP6AQ1 orthologs in FPF metabolism from eight different bee species, including stingless bees (Tribe: Meliponini). We conducted a phylogenetic analysis on four stingless bee species, including Frieseomelitta varia, Heterotrigona itama, Melipona quadrifasciata and Tetragonula carbonaria to identify CYP6AQ1-like functional orthologs. Three non-Meliponini, but tropical bee species, i.e., Ammobates syriacus, Euglossa dilemma and Megalopta genalis were analyzed as well. We identified candidate P450s in all (neo)tropical species with greater than 61% and 67% predicted protein sequence identities when compared to A. mellifera CYP6AQ1 and Bombus terrestris CYP6AQ26, respectively. Heterologous expression in High Five insect cells of these functional orthologs revealed a common coumarin substrate profile and a preference for the O-debenzylation of bulkier substrates. Competition assays using the fluorescent probe substrate 7-benzyloxymethoxy-4-trifluoromethylcoumarin (BOMFC) with these enzymes indicated inhibition of BOMFC metabolism by increasing concentrations of FPF. Furthermore, UPLC-MS/MS analysis revealed the capacity of all CYP6AQ1-like orthologs to metabolize FPF by hydroxylation in vitro at various levels, indicating a conserved FPF detoxification potential in different (neo)tropical bee species including Meliponini. This research, employing a toxicogenomics approach, provides important insights into the potential of stingless and other tropical bee species to detoxify FPF, and highlights the significance of investigating the detoxification mechanisms of insecticides in non-Apis bee species by molecular tools to inform risk assessment and conservation efforts.http://www.sciencedirect.com/science/article/pii/S014765132301223XCytochrome P450DetoxificationFlupyradifuroneMeliponiniPollinatorsToxicogenomics |
| spellingShingle | Xingzhi Xiao Julian Haas Ralf Nauen Functional orthologs of honeybee CYP6AQ1 in stingless bees degrade the butenolide insecticide flupyradifurone Ecotoxicology and Environmental Safety Cytochrome P450 Detoxification Flupyradifurone Meliponini Pollinators Toxicogenomics |
| title | Functional orthologs of honeybee CYP6AQ1 in stingless bees degrade the butenolide insecticide flupyradifurone |
| title_full | Functional orthologs of honeybee CYP6AQ1 in stingless bees degrade the butenolide insecticide flupyradifurone |
| title_fullStr | Functional orthologs of honeybee CYP6AQ1 in stingless bees degrade the butenolide insecticide flupyradifurone |
| title_full_unstemmed | Functional orthologs of honeybee CYP6AQ1 in stingless bees degrade the butenolide insecticide flupyradifurone |
| title_short | Functional orthologs of honeybee CYP6AQ1 in stingless bees degrade the butenolide insecticide flupyradifurone |
| title_sort | functional orthologs of honeybee cyp6aq1 in stingless bees degrade the butenolide insecticide flupyradifurone |
| topic | Cytochrome P450 Detoxification Flupyradifurone Meliponini Pollinators Toxicogenomics |
| url | http://www.sciencedirect.com/science/article/pii/S014765132301223X |
| work_keys_str_mv | AT xingzhixiao functionalorthologsofhoneybeecyp6aq1instinglessbeesdegradethebutenolideinsecticideflupyradifurone AT julianhaas functionalorthologsofhoneybeecyp6aq1instinglessbeesdegradethebutenolideinsecticideflupyradifurone AT ralfnauen functionalorthologsofhoneybeecyp6aq1instinglessbeesdegradethebutenolideinsecticideflupyradifurone |