Species identification of small fish in Xixuan Island coastal waters of Zhoushan using DNA barcoding

This study aimed to assess the fish species composition in Xixuan Island coastal waters and confirm the applicability of DNA barcoding in fish identification. Fish samples were caught with a gill net in December 2016 and in January, February, April, May and June 2017, and the fish species in these samples were analyzed and identified via both traditional morphological methods and DNA barcoding (mitochondrial 12S rRNA). The sample contained 1,357 individuals, the majority of which were Tridentiger barbatus (829 individuals). A total of 154 individuals were selected for DNA barcode analysis, and 147 sequences were obtained. A total of 35 species, which belonged to six orders and 16 families, were accurately identified. One species was identified to the genus level (Repomucenus), and only one was identified to the family level (Moridae). Most of the species belonged to Gobiidae (12 species). With the exception of those in Moridae, the identified species were commonly detected on the Zhejiang coast. The average Kimura two-parameter (K2P) distances within species, genera, families, and order and between orders were 0.15%, 4.44%, 14.96%, 20.37%, and 26.20%, respectively. The intraspecific K2P distance was markedly greater than the interspecific distance. The A + T content (55.41%) was higher than the G + C content (44.58%). Phylogenetic tree analysis showed that individuals belonging to the same species were clustered together and could be clearly distinguished. Mitochondrial 12S rRNA can be effectively used for juvenile and adult fish identification and has good application prospects in fish species identification. The investigation and species identification of small fish collected monthly from Xixuan Island coastal waters are conducive to understanding the distribution characteristics and species composition of fish in the Zhoushan coastal area.