Using a combination of the cytochrome oxidase subunit I gene (COI) and internal transcribed spacer (ITS) sequence information, cross-breeding, and morphological criteria, the taxonomic status of Tetranychus takafujii was reassessed. Described from Japan and very close to Tetranychus evansi, only the male empodium II separated the two species. Six T. evansi samples collected in Brazil, France, Kenya, Spain (including the Canary Islands), Taiwan, and two samples from Japan previously species-identified as T. takafujii, were examined. DNA sequences consistently divide the samples into two groups: group I held the mites from Brazil and France and group II contained the remaining samples of T. evansi together with the two mites collected in Japan. Likewise, crossbreeding experiments detected the same two groups that were reproductively partially incompatible. However, the Japanese samples were completely compatible with the rest of the group II T. evansi samples. The empodium II shape failed to distinguish consistently between the T. evansi and T. takafujii samples, but rather highlighted its variability, questioning its utility for separating these taxa. Congruent results support the co-specificity of the two taxa. It is concluded that T. takafuji is a junior synonym of T. evansi.