DNA Barcoding of the Genus Discogobio (Teleostei, Cyprinidae) in China

Discogobio is a genus of small, economically important freshwater fishes that are widely distributed in Southwestern China. The species of the genus are morphologically very similar, which makes their taxonomic identification quite challenging. DNA barcoding technology can identify species at the molecular level, thus overcoming the limitations of morphological classification. In this study, we collected 16 morphological species of Discogobio from China, analyzed the mitochondrial cytochrome oxidase I subunit (COI) gene sequences of 206 samples, and applied DNA barcoding to identify the species. The COI amplicon was 651 bp in length, and the mean base contents were: (T) 28.83%, (C) 27.63%, (A) 25.97%, (G) 17.57%. The AT content (54.8%) was higher, and the base composition was biased. The intraspecific differences in the genus Discogobio were not significant, and the genetic distances were all less than 2%. The average interspecific genetic distances (3.94%) were about 18.8 times the average intraspecific genetic distances (0.21%), suggesting that there are barcode gaps among the species of the genus Discogobio. Five different species definition methods, Automatic Barcode Gap Discovery (ABGD), Assemble Species by Automatic Partitioning (ASAP), Bayesian Poisson Tree Process (bPTP), Generalized Mixed Yule Combination (GMYC), and Refined Single Linkage (RESL), were used to infer molecular operational taxonomic units (MOTU). The number of MOTUs ranged from 9 to 18. Phylogenetic analysis based on COI gene haplotypes showed that most species formed well-evolved branches on the phylogenetic tree, and the clustering among species was obvious without mixing. The results of this study provide reliable DNA barcoding information for species identification within the genus Discogobio, which is of great significance for taxonomic identification.