Abstract: This paper develops an accurate method to identify six eel species including Anguilla rostrata, A. anguilla, A. japonica, A. mossambica, A. marmorata, and A. bicolor pacifica using polygenic DNA barcoding. Three pairs of universal primers were used to amplify partial fragments of the 16S rRNA, Cyt b and COⅠgenes of the eel species with the total genomic DNA from eel as a template and the amplified products were sequenced. As a result, 16S rDNA (638–643 bp), Cyt b (464–466 bp) and COⅠ(705–707 bp) gene fragments were obtained for each species. Homologous sequences of these genes were found. Three new primer pairs were designed to amplify 638–643, 400 and 609 bp fragments of the 16S rRNA, Cyt b and COⅠ genes by PCR, respectively. The specific 16S rRNA, Cyt b and COⅠfragments of 262, 280 and 300 bp were selected as standard DNA barcodes for the identification of eel species. Homology analysis was performed using DNAMAN software (version 6) and the homology for species discrimination was developed by aligning the DNA barcodes with the GenBank database. Analysis of 30 samples from six eel species by the developed method showed that the species were consistently identified using the DNA barcodes. In conclusion, the method is stable, precise and easy to operate and can be applied to the identification of six species of eels.

Key words: 16S rRNA, Cyt b, COⅠ, polygenic DNA barcode, identification of eel species