Abstract
The two-spotted spider mite (Tetranychus urticae) is a highly destructive and economically significant pest in agricultural, horticultural, and ornamental agroecosystems worldwide, including hop (Humulus lupulus) and mint (Mentha spp.) fields in the Pacific Northwest (PNW) region of the United States. Repeated acaricide applications and rotations have led to widespread resistance, resulting in control failures. In this study, we investigated the mechanisms of resistance to four different acaricides (bifenthrin, bifenazate, etoxazole, and abamectin) across 23 field-collected TSSM populations by integrating diagnostic bioassays, genetic screening for resistance-associated mutations, structural modeling, and molecular docking. Several kdr mutations and mutation combinations were detected in TuVGSC across all tested populations. The G132A in Tucytb was identified in 68.75% of hop and 40% of mint TSSM populations, while the I1017F in TuCHS 1 was found in 94% of hop and 100% of mint populations. Structural analysis revealed key interactions between acaricides and target proteins in both wild-type and mutant variants, providing novel insights into the functional impacts of these mutations. Our findings enhance the understanding of TSSM adaptation to acaricides among different crops, supporting the development of more effective resistance management strategies to mitigate economic losses in hops, mint, and other crop production.