Species Identification of ‘Seabream’ (Pagrus major, Evynnis japonica and Dentex tumifrons) by PCR-based DNA Analysis Using the Mitochondrial Cytochrome b Gene

Abstract

The Japan Agricultural Standard (JAS) law revised in 1999 requires correct labeling of species and origin of marine products. We developed methods for identifying three Sparidae species, Red seabream (Pagrus major), Crimson seabream (Evynnis japonica) and Yellowback seabream (Dentex tumifrons) by PCR-restriction fragment length polymorphism (PCR-RFLP) and multiprimer-PCR using species-specific primers. PCR-RFLP analysis of the 498-bp DNA fragment amplified from the mitochondrial cytochrome b gene of three Sparidae species that were digested by two restriction enzymes, SspI or StuI, showed species-specific restriction fragment patterns for these species. To simplify the Sparidae species identification, we also carried out multiprimer-PCR analysis using species-specific primer set based on unique nucleotide substitutions in each species, and showed usefulness of this technique for Sparidae species identification.