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Species identification method for Scombrops boops and Scombrops gilberti based on polymerase chain reaction-restriction fragment length polymorphism analysis of mitochondrial DNA
Authors:Shiro Itoi  Noriyuki Takai  Satomi Naya  Keitaro Dairiki  Akira Yamada  Seiji Akimoto  Kiyoshi Yoshihara  Haruo Sugita
Affiliation:Department of Marine Science and Resources, Nihon University, Fujisawa, Kanagawa 252-8510, and;Kanagawa Prefectural Fisheries Research Institute, Miura, Kanagawa 238-0237, Japan
Abstract:ABSTRACT:   Gnomefish Scombrops boops and Scombrops gilberti are commercially important fishes in Japan, but these species are often confused in the markets because of their morphological similarity. To identify these two species, we performed nucleotide sequencing and restriction fragment length polymorphism (RFLP) analysis on 16S ribosomal RNA (rRNA) gene and the control region in mitochondrial DNA. Five and 12 nucleotide substitutions were observed between species in the 777-bp 16S rRNA gene and 471-bp control region, respectively. Diagnostic restriction sites for discriminating between S. boops and S. gilberti were found in the 16S rRNA gene, but not in the control region. Polymerase chain reaction (PCR)–RFLP analysis using two enzymes, Eco NI and Mva I, clearly discriminated between S. boops and S. gilberti identified by meristic characters. The PCR–RFLP analysis identified most of the 168 Scombrops young caught in the coastal waters of the Izu and Miura peninsulas as S. boops , suggesting that S. gilberti juveniles are rare in this area.
Keywords:16S rRNA    control region    mtDNA    PCR–RFLP    Scombropidae    Scombrops boops    Scombrops gilberti
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