Open AccessMolecular identification of biwa trout (Oncorhynchus masou rhodurus) using PCR–RFLP method
Author(s) -
Chihiro Matsumoto,
Yukino Kyota,
Shunya Yamanaka,
Naoki Murakawa,
Ryutaro Kikunaga,
Yoshihiro Yamada,
Hiroyuki Kawachi
Publication year - 2019
Publication title -
journal of food science and technology/journal of food science and technology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.656
H-Index - 68
eISSN - 0975-8402
pISSN - 0022-1155
DOI - 10.1007/s13197-019-03914-3
Subject(s) - oncorhynchus , restriction fragment length polymorphism , trout , identification (biology) , biology , rainbow trout , fishery , zoology , fish <actinopterygii> , polymerase chain reaction , genetics , ecology , gene
Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was used to identify meat from biwa trout ( Oncorhynchus masou rhodurus ), amago trout ( Oncorhynchus masou ishikawae ), yamame trout ( Oncorhynchus masou masou ), and rainbow trout ( Oncorhynchus mykiss ). PCR amplification was conducted using primers flanking conserved regions of NADH dehydrogenase subunits 4 and 5 (ND4-ND5) (2848 bp) and ND1 (1091 bp) genes of mitochondrial DNA following restriction digestion with the enzyme Hae III. Although the segments of ND4-ND5 and ND1 genes showed intraspecies variation, the generation of DNA fragments larger than 300 bp and 160 bp following cleavage by HaeIII of ND4-ND5 and ND1, respectively, was efficient to differentiate the four species. Furthermore, this method was successful in species identification even when using PCR-amplified products obtained from thermally processed biwa trout samples. This sensitive technique can be utilized to reveal commercial fraud, where biwa trout is adulterated with meat from cheaper counterparts.