Abstract: The aim of the study was to identify a kind of common pollination insect in alfalfa field in Xinjiang based on DNA barcoding and morphological characteristics. Genomic DNA of the insect was extracted by Genomic DNA Mini Preparation Kit, mitochondrial cytochrome c oxidase subunit Ⅰ (COⅠ) gene fragment was amplified with a pair of universal primers of Lepidoptera insect. PCR product was purified by Gel Extraction Kit, cloned by the pUCm-T Vector Cloning Kit and then sent to a sequencing company. Finally the bee was identified based on morphological characteristics and the sequences in the BOLD database. The results showed that the sequences of gene fragments had the highest similarity (99%) with two unidentified specimens of Anthidium in Kyrgyzstan. Then were the two specimens of Anthidium florentinum in Greece and Russia, with the similarity of more than 96%. Sequences of the gene fragments had low similarity with other leaf cutting specimens in the BOLD database. Morphological characteristics also showed that this insect was the same with the description of A. florentinum. Therefore, this kind of leafcutting bee in alfalfa field in Alar, Xinjiang was identified as A. florentinum.