Identification of flatfish (Pleuronectiforme) species using DNA-based techniques

C G Sotelo, P Calo-Mata, M J Chapela, R I Pérez-Martín, H Rehbein, Georgina Louise Hold, Valerie Joan Stevens, S Pryde, J Quinteiro, M Izquierdo, M Rey-Méndez, C Rosa, Ana T. Santos

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Abstract

Identification of flatfish species using a DNA-based methodology was studied. The polymerase chain reaction was employed to obtain a 464 bp amplicon from mitochondrial cytochrome b gene. The sequences from this fragment belonging to 24 species were analyzed using a genetic distance method, and polymorphic sites were determined. The fragment was found to be highly polymorphic (231 sites), and this permitted the differentiation of most of the species. Phylogenetic tree construction was employed to allow the identification of flatfish species. As a result, each species was grouped in a well-differentiated clade, except for two pairs: Limanda ferruginea and L. limanda, and Solea impar and S. lascaris, which could not be differentiated. On the basis of the sequences obtained, restriction enzymes were selected to provide specific restriction profiles, which allow the differentiation of 21 species of flatfish in a faster and less expensive manner than sequencing. This polymerase chain reaction-restriction fragment length polymorphism methodology (PCR-RFLP) was tested using commercial samples.
Original languageEnglish
Pages (from-to)4562-4569
Number of pages8
JournalJournal of Agricultural and Food Chemistry
Volume49
Issue number10
DOIs
Publication statusPublished - 1 Oct 2001

Keywords

  • polymerase chain reaction
  • authentication
  • flatfish
  • mitochondrial DNA
  • cytochrome b gene
  • sequencing
  • restriction endonucleases
  • fragment length polymorphism
  • capillary zone electrophoresis
  • cytochrome-B gene
  • fish
  • differentiation
  • products

Cite this

Sotelo, C. G., Calo-Mata, P., Chapela, M. J., Pérez-Martín, R. I., Rehbein, H., Hold, G. L., Stevens, V. J., Pryde, S., Quinteiro, J., Izquierdo, M., Rey-Méndez, M., Rosa, C., & Santos, A. T. (2001). Identification of flatfish (Pleuronectiforme) species using DNA-based techniques. Journal of Agricultural and Food Chemistry, 49(10), 4562-4569. https://doi.org/10.1021/jf010452a