Distinguishment of rodentia species has been difficult due to the high similarities in morphology between species.As a useful tool for rapid species assignment and cryptic species discovery,DNA barcoding can be employed to resolve the puzzle.In this research,a 648bp of mitochondrial COI gene from Murinae species was used as DNA barcode to evaluate the effectiveness of DNA barcode for species identification and to enrich the DNA barcode reference sequence library for rodentia.The COI gene from 79 individuals belonging to 12 speices of Murinae was determined combined with additional 154 sequences downloaded from the genbank.In total,233 individuals belonging to 29 Murinae species were investigated.Several methods were employed to analyze the dataset including the pairwise intra- and interspecific distances,tree-based methods,as well as the character-based methods.The mean intraspecific distance of Murinae was 2.01%,while the mean interspecific distance was 12.39%.And most species exhibited much lower intraspecific distances than the mean interspecific distances.The successful identification rate by distance-based methods exceeded 90.6%.The neighbour-joining analysis could unambiguously discriminate 96.4% of the surveyed species with high bootstrap values,and the ML and Bayesian analysis showed the similar results.Character-based methods,including CAOS and BLOG analyses,successfully authenticated about 98% of the Murinae individuals.Character-based methods performed extraordinarily better than other analytic methods in identifying closely related species such as Niviventer niviventer & N.Fulvescens,which is difficult to distinguish by morphological traits,distance-based methods or tree-based methods.The present study indicated that when proper analytic methods were applied or combined,DNA barcoding could effectively identify species of Murinae in china.