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BACKGROUND Acaricide application remains an integral component of IPM for the two-spotted spider mite Tetranychus urticae. Species and strains of phytoseiid predatory mites vary significantly in their response to acaricides. For the success of IPM, it is imperative to identify the determinants of selectivity and molecular stress responses of acaricides in predatory mites. RESULTS The three classical acaricides bifenazate, cyflumetofen, and fenbutatin oxide, did not affect survival and fecundity of Phytoseiulus persimilis regardless of the route of exposure. Selectivity of the orange oil- and terpenoid blend-based botanical acaricides was low via a combination of direct exposure, acaricide-laced diet, and residual exposure but improved when limiting exposure only to the diet. To gain insights into the molecular stress responses, the transcriptome of P. persimilis was assembled. Subsequent gene expression analysis of predatory mites orally exposed to fenbutatin oxide and orange oil yielded only a limited xenobiotic stress response. In contrast, P. persimilis exhibited target-site resistance mutations, including I260M in SdhB, I1017M in CHS1, and kdr and super-kdr in VGSC. Extending the screen using available Phytoseiidae sequences uncovered I136T, S141F in cytb, G119S in AChE, and A2083V in ACC, well-known target-sites of acaricides. CONCLUSION Selectivity of the tested botanical acaricides to P. persimilis was low but could be enhanced by restricting exposure to a single route. Differential gene expression analysis did not show a robust induced stress response after sub-lethal exposure. In contrast, this study uncovered target-site mutations that may help explain the physiological selectivity of several classical acaricides to phytoseiid predators. This article is protected by copyright. All rights reserved.