title: A mutation in the PSST homologue of complex I (NADH:ubiquinone oxidoreductase) from Tetranychus urticae is associated with resistance to METI acaricides.
abstract: The acaricidal compounds pyridaben, tebufenpyrad and fenpyroximate are frequently used in the control of phytophagous mites such as Tetranychus urticae, and are referred to as Mitochondrial Electron Transport Inhibitors, acting at the quinone binding pocket of complex I (METI-I acaricides). Because of their very frequent use, resistance evolved fast more than 20 years ago, and is currently wide-spread. Increased activity of P450 monooxygenases has been often associated with resistance, but target-site based resistance mechanisms were never reported. Here, we report the discovery of a mutation (H92R) in the PSST homologue of complex I in METI-I resistant T. urticae strains. The position of the mutation was studied using the high-resolution crystal structure of Thermus thermophilus, and was located in a stretch of amino acids previously photo-affinity labeled by fenpyroximate. Selection experiments with a strain segregating for the mutant allele, together with marker-assisted back-crossing of the mutation in a susceptible background, confirmed the involvement of the mutation in METI-I resistance. Additionally, an independent genetic mapping approach; QTL analysis identified the genomic region of pyridaben resistance, which included the PSST gene. Last, we used CRISPR-Cas9 genome editing tools to introduce the mutation in the Drosophila PSST homologue.
authors.affiliation: Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam, P.O. Box 9424, 1090 GE Amsterdam, The Netherlands.
authors.forename: Sabina
authors.initials: S
authors.lastname: Bajda
authors.affiliation: Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience Engineering, Coupure Links 653, Ghent University, B-9000 Ghent, Belgium.
authors.forename: Wannes
authors.initials: W
authors.lastname: Dermauw
authors.affiliation: Institute of Molecular Biology & Biotechnology, Foundation for Research & Technology Hellas, 100 N. Plastira Street, GR-700 13 Heraklion, Crete, Greece.
authors.forename: Rafaela
authors.initials: R
authors.lastname: Panteleri
authors.affiliation: Kyoto University, Graduate School of Agriculture, Laboratory of Ecological Information, Kyoto 606-8502, Japan.
authors.forename: Naoya
authors.initials: N
authors.lastname: Sugimoto
authors.affiliation: Institute of Molecular Biology & Biotechnology, Foundation for Research & Technology Hellas, 100 N. Plastira Street, GR-700 13 Heraklion, Crete, Greece; Department of Biology, University of Crete, 71409 Heraklion, Greece.
authors.forename: Vassilis
authors.initials: V
authors.lastname: Douris
authors.affiliation: Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience Engineering, Coupure Links 653, Ghent University, B-9000 Ghent, Belgium.
authors.forename: Luc
authors.initials: L
authors.lastname: Tirry
authors.affiliation: Kyoto University, Graduate School of Agriculture, Laboratory of Ecological Information, Kyoto 606-8502, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Osakabe
authors.affiliation: Institute of Molecular Biology & Biotechnology, Foundation for Research & Technology Hellas, 100 N. Plastira Street, GR-700 13 Heraklion, Crete, Greece; Laboratory of Pesticide Science, Department of Crop Science, Agricultural University of Athens, 75 Iera Odos Street, GR-11855 Athens, Greece.
authors.forename: John
authors.initials: J
authors.lastname: Vontas
authors.affiliation: Institute for Biodiversity and Ecosystem Dynamics, University of Amsterdam, P.O. Box 9424, 1090 GE Amsterdam, The Netherlands; Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience Engineering, Coupure Links 653, Ghent University, B-9000 Ghent, Belgium. Electronic address: thomas.vanleeuwen@ugent.be.
authors.forename: Thomas
authors.initials: T
authors.lastname: Van Leeuwen
mesh_terms: D056810:Acaricides; D000595:Amino Acid Sequence; D000818:Animals; D060829:Arthropod Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004351:Drug Resistance / Q000235:genetics*; D042967:Electron Transport Complex I / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012313:RNA / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D040102:Tetranychidae / Q000254:growth & development / Q000502:physiology*
chemical_list: D056810:Acaricides; D060829:Arthropod Proteins; D018076:DNA, Complementary; D012313:RNA; D042967:Electron Transport Complex I
keywords: 20 kDa subunit; Acari; Drosophila; METI-I; NUKM; Rotenone