Molecular identification of the genus Thryssa based on DNA barcoding.

Abstract

DNA barcoding is an effective method for identifying species by analyzing one or a few short standardized DNA sequences. In this study, we examined the utility of mitochondrial cytochrome oxidase subunit I (COI) sequences as a DNA barcode for the identification of six species belonging to the genus Thryssa: T. dussumieri, T. hamiltonii, T. kammalensis, T. mystax, T. setirostris, and T. vitrirostris. We obtained an intraspecific distance of 0.000 for T. vitrirostris and T. hamiltonii, 0.006 for T. mystax, 0.002 for T. dussumieri, and 0.005 for T. kammalensis. The average intraspecific distance was 0.002, while the average interspecific distance was 0.137. Thus, the interspecific genetic distance was approximately 67-fold larger than the intraspecific genetic distance; the average genetic distance among species was greater than the minimum of 0.020 between species suggested elsewhere. The genetic distance between T. vitrirostris and T. mystax was 0.003. A maximum-likelihood phylogenetic tree constructed using best-fitting tree topology showed distinct clusters corresponding to the species (except for T. vitrirostris and T. mystax). The closest relationship was found between T. vitrirostris and T. mystax. These two species clustered together in the phylogenetic tree. This conclusion contradicts the evolutionary relationship based on morphological classification.