Kashi is an important production area of Punica granatum L., investigation found that Tenuipalpus hornotinus is the dominant pest mite that harms P. granatum in the region. Upon literature review, it was found that its identification only relies on morphology, and there was no report on the DNA barcode of the mite. Due to its small size and numerous similar species, morphological identification of this mite is difficult. Therefore, this study utilized morphology combined with DNA barcode technology to achieve rapid and accurate identification of this mite. This study conducted on-site observations, collected samples, and made specimens, observing under the microscope combined with morphological data to compare and identify morphological features of this mite; meanwhile, genes such as COⅠ, ITS2 and 28S-rRNA were amplified using DNA barcode technology and sequenced, the obtained sequences were analyzed and a phylogenetic tree was constructed using MEGA7.0 software. The pest mite was identified as T. hornotinus by morphological features and the barcode sequence of the pest mite was obtained using DNA barcode technology. Rapid and accurate identification of T. hornotinus has achieved by morphology and DNA barcode technology, providing a species basis for the subsequent prevention and control of this pest mite.