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Identification of four Donax species by PCR–RFLP analysis of cytochrome c oxidase subunit I (COI)

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Abstract

Four Donax species, D. semistriatus, D. trunculus, D. variegatus and D. vittatus, are found on European coasts. Nevertheless, despite their economic importance there is not a reliable method to differentiate these species independently of their size or condition. Such a method could help to protect consumer rights and avoid commercial fraud due to the replacement of valuable species by less valuable ones with similar morphological traits. In this work, the sequence of the mitochondrial cytochrome oxidase subunit I region was examined in individuals of these species to identify restriction site variation and develop polymerase chain reaction–restriction fragment length polymorphisms assays. Species-specific restriction endonuclease patterns were found with the enzymes AluI, HaeIII and MspI, allowing an exact identification of Donax species. This methodology provides simple, reliable and cost-effective identification of four Donax species and may be useful to prevent commercial fraud and to increase food traceability.

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References

  1. Helm MM, Bourne N, Lovatelli A (comp/ed) (2004) Hatchery culture of bivalves. A practical manual. Fisheries Technical Paper No. 471. FAO, Rome, p 177

  2. Ansell AD (1983) The biology of the genus Donax. Sandy beaches as ecosystems. In: McLanchlan A, Erasmus T (eds) Sandy beaches as ecosystems. Junk, The Hague

    Google Scholar 

  3. Rufino MM, Gaspar MB, Pereira AM, Maynou F, Monteiro CC (2010) Ecology of megabenthic bivalve communities from sandy beaches on the south coast of Portugal. Sci Mar 74:163–178

    Article  Google Scholar 

  4. Salas C, Tirado C, Manjón-Cabeza ME (2001) Sublethal foot-predation on Donacidae (Mollusca: Bivalvia). J Sea Res 46:43–56

    Article  Google Scholar 

  5. Bossier P (1999) Authentication of seafood products by DNA patterns. J Food Sci 64:189–193

    Article  CAS  Google Scholar 

  6. García-Rodríguez FJ, Ponce-Díaz G, Muñoz-García I, González-Armas R, Pérez-Enríquez R (2008) Mitochondrial DNA markers to identify commercial spiny lobster species (Panulirus spp.) from the Pacific coast of Mexico: an application on phyllosoma larvae. Fish Bull 106:204–212

    Google Scholar 

  7. Fernández-Tajes J, Longa A, García-Gil J, Chiu Y-W, Huang Y-S, Méndez J, Lee R-S (2011) Alternative PCR-RFLP methods for mussel Mytilus species identification. Eur Food Res Technol 233:791–796

    Article  Google Scholar 

  8. Klinbunga S, Khamnamtong N, Tassanakajon A, Puanglarp N, Jarayabhand P, Yoosukh W (2003) Molecular genetic identification tools for three commercially cultured oysters (Crassostrea belcheri, Crassostrea iredalei, and Saccostrea cucullata) in Thailand. Mar Biotechnol 5:27–36

    Article  CAS  Google Scholar 

  9. Pereira F, Carneiro J, Amorim A (2008) Identification of species with DNA-based technology: current progress and challenges. Recent Pat DNA Gene Seq 2:187–199

    Article  CAS  Google Scholar 

  10. Mackie IM, Pryde SE, Gonzales-Sotelo C, Medina I, Perez-Martin R, Quinteiro J, Rey-Mendez M, Rehbein H (1999) Challenges in the identification of species of canned fish. Trends Food Sci Technol 10:9–14

    Article  CAS  Google Scholar 

  11. Macedo MC, Macedo MI, Borges JP (1999) Conchas marinhas de Portugal. Editorial Verbo, Lisboa

    Google Scholar 

  12. Tebble N (1966) British bivalve seashells. A handbook for identification. British Museum, Edinburgh

    Google Scholar 

  13. Walsh PS, Metzger DA, Higuchi R (1991) Chelex-100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechniques 10:506–513

    CAS  Google Scholar 

  14. Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol 3:294–299

    CAS  Google Scholar 

  15. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98

    CAS  Google Scholar 

  16. Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics methods and protocols: methods in molecular biology. Humana Press, Totowa, pp 365–386

    Google Scholar 

  17. Fernández A, García T, Asensio L, Rodríguez MA, González I, Hernández PE, Martín R (2001) PCR-RFLP analysis of the internal transcribed spacer (ITS) region for identification of 3 clam species. J Food Sci 66:657–661

    Article  Google Scholar 

  18. Freire R, Insua A, Méndez J (2005) Cerastoderma glaucum 5S ribosomal DNA: characterization of the repeat unit, divergence with respect to Cerastoderma edule, and PCR-RFLPs for the identification of both cockles. Genome 48:427–442

    Article  CAS  Google Scholar 

  19. Freire R, Arias A, Méndez J, Insua A (2011) Identification of European commercial cockles (Cerastoderma edule and C. glaucum) by species-specific PCR amplification of the ribosomal DNA ITS region. Eur Food Res Technol 232:83–86

    Article  CAS  Google Scholar 

  20. Santaclara FJ, Espineira M, Cabado G, Aldasoro A, González-Lavin N, Vieites JM (2006) Development of a method for the genetic identification of mussel species belonging to Mytilus, Perna, Aulacomya, and other genera. J Agric Food Chem 54:8461–8470

    Article  CAS  Google Scholar 

  21. Fernández-Tajes J, Méndez J (2007) Identification of the razor clam species Ensis arcuatus, E. siliqua, E. directus, E. macha, and Solen marginatus using PCR-RFLP analysis of the 5S rDNA region. J Agric Food Chem 55:7278–7282

    Article  Google Scholar 

  22. Freire R, Fernández-Tajes J, Méndez J (2008) Identification of razor clams Ensis arcuatus and Ensis siliqua by PCR-RFLP analysis of ITS1 region. Fish Sci 74:511–515

    Article  CAS  Google Scholar 

  23. López-Piñón MJ, Insua A, Méndez J (2002) Identification of four scallop species using PCR and restriction analysis of the ribosomal DNA internal transcribed spacer region. Mar Biotechnol 4:495–502

    Article  Google Scholar 

  24. Pereira AM, Fernández-Tajes J, Gaspar MB, Méndez J (2012) Identification of the wedge clam Donax trunculus by a simple PCR technique. Food Control 23:268–270

    Article  CAS  Google Scholar 

  25. Hare MP, Palumbi SR, Butman CA (2000) Single-step species identification of bivalve larvae using multiplex polymerase chain reaction. Mar Biol 137(5–6):953–961

    Article  CAS  Google Scholar 

  26. Theologidis I, Fodelianakis S, Gaspar MB, Zouros E (2008) Doubly uniparental inheritance (DUI) of mitochondrial DNA in Donax trunculus (Bivalvia : Donacidae) and the problem of its sporadic detection in Bivalvia. Evolution 62:959–970

    Article  CAS  Google Scholar 

  27. Stewart DT, Saavedra C, Stanwood RR, Ball AO, Zouros E (1995) Male and female mitochondrial-DNA lineages in the blue mussel (Mytilus edulis) species group. Mol Biol Evol 12:735–747

    CAS  Google Scholar 

  28. Sutherland B, Stewart D, Kenchington ER, Zouros E (1998) The fate of paternal mitochondrial DNA in developing female mussels, Mytilus edulis: implications for the mechanism of doubly uniparental inheritance of mitochondrial DNA. Genetics 148:341–347

    CAS  Google Scholar 

  29. Westfall KM, Wimberger PH, Gardner JPA (2010) An RFLP assay to determine if Mytilus galloprovincialis Lmk. (Mytilidae; Bivalvia) is of Northern or Southern hemisphere origin. Mol Ecol Resour 10:573–575

    Article  CAS  Google Scholar 

  30. Hebert PDN, Ratnasingham S, deWaard JR (2003) Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proc R Soc B Biol Sci 270:S96–S99

    Article  CAS  Google Scholar 

  31. Therriault TW, Docker MF, Orlova MI, Heath DD, MacIsaac HJ (2004) Molecular resolution of the family Dreissenidae (Mollusca: Bivalvia) with emphasis on Ponto Caspian species, including first report of Mytilopsis leucophaeata in the Black Sea basin. Mol Phylogenet Evol 30:479–489

    Article  CAS  Google Scholar 

  32. Boudry P, Heurtebise S, Collet B, Cornette F, Gerard A (1998) Differentiation between populations of the Portuguese oyster, Crassostrea angulata (Lamark) and the Pacific oyster, Crassostrea gigas (Thunberg), revealed by mtDNA RFLP analysis. J Experimental Mar Biol Ecol 226:279–291

    Article  Google Scholar 

  33. Yu ZN, Kong XY, Zhang LS, Guo XM, Xiang JH (2003) Taxonomic status of four Crassostrea oysters from China as inferred from mitochondrial DNA sequences. J Shellfish Res 22:31–38

    Google Scholar 

  34. Liu J, Li Q, Kong L, Yu H, Zheng X (2011) Identifying the true oysters (Bivalvia: Ostreidae) with mitochondrial phylogeny and distance-based DNA barcoding. Mol Ecol Resour 1:820–830

    Article  Google Scholar 

  35. Feng Y, Li Q, Kong L, Zheng X (2011) DNA barcoding and phylogenetic analysis of Pectinidae (Mollusca: Bivalvia) based on mitochondrial COI and 16S rRNA genes. Mol Biol Rep 38:291–299

    Article  CAS  Google Scholar 

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Acknowledgments

We thank Jose García for his technical assistance in the laboratory and Luisa Martínez for her constructive and valuable comments. This work was supported by the Consellería de Economía e Industria (Xunta de Galicia) through project 10MMA103013 and by Ministerio de Educación of Spain through a predoctoral FPU fellowship awarded to Ana Nantón.

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This article does not contain any studies with human or animal subjects.

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Correspondence to A. Nantón.

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Nantón, A., Freire, R., Arias-Pérez, A. et al. Identification of four Donax species by PCR–RFLP analysis of cytochrome c oxidase subunit I (COI). Eur Food Res Technol 240, 1129–1133 (2015). https://doi.org/10.1007/s00217-015-2416-z

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  • DOI: https://doi.org/10.1007/s00217-015-2416-z

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