Acquisition of a reference Tetranychusstrains that are completely susceptible to acaricides and retain identical genetic backgrounds to acaricide-resistant strains is an essential step in elucidating mechanisms of resistance. To establish both completely susceptible and acaricide-resistant strains for this purpose, I collected Tetranychus mite populations from various regions in South Korea including both heavily cultivated and remote regions. Suitability as a susceptible or resistant reference strain was tested by determining species identity as Tetranychus urticae along with baseline susceptibility to common acaricides. The majority of mite populations collected from cultivated areas belonged to a monophyletic group of the previously reported green-type T. urticae as determined by mtCOI- and ITS2-based phylogenetic analysis. These strains showed relatively reduced levels of susceptibility to the acaricides tested, suggestive of the development of resistance. Among them, the AbaR strain was classified as a minor group in the T. urticae complex. The UD strain, originally collected from a remote island region, was found to be susceptible to the acaricides tested and generated an independent phylogenetic branch. The UD strain was also considered a minor group in the T. urticae complex. Phenotypic analysis based on morphological characters confirmed that both the AbaR and UD strains were statistically undistinguishable from the major green-type T. urticae. Taken together, I propose that the UD strain be used as a susceptible reference strain as it provides baseline susceptibility to acaricides and a wild-type genetic background for the resistance studies.