BACKGROUND: Tetranychus urticae is a notorious crop pest with world-wide distribution that has developed resistance to a wide range of acaricides. Here, we investigated the resistance levels of a T. urticae population collected from an ornamental greenhouse in Peloponnese, Greece, and analyzed its resistance mechanisms at the molecular level. RESULTS: Toxicological assays showed resistance levels against compounds with different mode of action, with resistance ratios scaling at: 89-fold for abamectin, >1000-fold for clofentezine, >5000-fold etoxazole, 27-fold for fenpyroximate and pyridaben, 20- and 36-fold for spirodiclofen and spirotetramat, respectively and 116- and >500-fold for cyenopyrafen and cyflumetofen, respectively. Bioassays with synergists indicated the involvement of detoxification enzymes in resistance to abamectin but not to cyflumetofen and spirodiclofen. RNAseq analysis showed significant over-expression of several genes encoding detoxification enzymes such as cytochrome P450 monooxygenases and UDP-glycosyltransferases, which have been previously associated with acaricide resistance. Known target-site resistance mutations were identified in acetyl-choline esterase, chitin synthase 1 and NDUFS7/psst, but also discovered putative novel resistance mutations in targets such as the glutamate-gated chloride channel subunit 3. Interestingly, target site resistance mutations against pyrethroids or bifenazate were not identified possibly indicating a recent reduced selection pressure in Greece, as well as a possible opportunity to rotate these chemistries. CONCLUSION: We identified and characterized a striking case of multiple acaricide resistance in a field population of T. urticae. Exceptionally strong resistance phenotypes, with accumulation of multiple resistance mutations and over-expression of P450s and other detoxification genes in the same field population is reported. This article is protected by copyright. All rights reserved.
