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High-Resolution Melting of 18S rDNA sequences (18S-HRM) for discrimination of bivalve’s species at early juvenile stage: application to a spat survey

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Abstract

It is particularly difficult to identify bivalve species at early development stages (larvae and spat) by morphological examination. Targeting genetic marker commonly used for species identification, we developed a cheap and user friendly tool for bivalve identification through High-Resolution Melting (HRM) analysis. Based on the polymorphism of the nuclear 18S ribosomal RNA gene among 11 orders of bivalve, the 18S-HRM method allowed discriminating species of the Ostreida, Mytilida and Pectinida orders. In this study, this method proved to be a very sensitive tool able to authenticate one species that differed by only one mutation from another one. We applied the method by identifying bivalve species in spat collections from different benthic substrates moored at subtidal and intertidal areas in euhaline conditions. This field survey showed that the settlement of the invasive species Crassostrea gigas was limited vertically to intertidal substrates. In conclusion, the 18S-HRM analysis is a fast, closed-tube method with high resolution and cost effectiveness, and is thus a good alternative method to identify bivalve species. This HRM assay can by applied for species identification of bivalves in the context of ecological management as well as food traceability for bivalves with economical interests.

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Acknowledgements

Sincere thanks are due to P.E. Galand for his helpful suggestions and for correcting the English and also to the two anonymous reviewers who carefully edited our manuscript.

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Authors and Affiliations

  1. Laboratoire des Sciences de l’Environnement Marin, UMR CNRS 6539, Institut Universitaire Européen de la Mer, Université de Bretagne Occidentale, 29280, Plouzané, France

    Anne-Leila Meistertzheim & Morgane Lejart

  2. Centre de Recherches Insulaires et Observatoire de l’Environnement de Polynésie Française-CRIOBE, USR 3278 CNRS/EPHE/UPVD, Laboratoire d’Excellence “CORAIL”, Université de Perpignan, Bâtiment CBETM, 52 Avenue Paul Alduy, 66860, Perpignan Cedex, France

    Anne-Leila Meistertzheim

  3. Univ. Perpignan Via Domitia, Centre de Formation et de Recherche sur les Environnements Méditerranéens, UMR 5110, 66860, Perpignan, France

    Laurent Héritier

  4. France Energies Marines, 15 rue Johanes Kepler, 29200, Brest, France

    Morgane Lejart

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  1. Anne-Leila Meistertzheim
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  2. Laurent Héritier
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Correspondence to Anne-Leila Meistertzheim.

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This research program was financially supported by the French national program PROGIG (Prolifération de Crassostrea gigas, LITEAU II, contract number CV 05000131).

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The authors declare no conflict of interest.

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All applicable international, national and/or institutional guidelines for the care and use of animals were followed.

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Meistertzheim, AL., Héritier, L. & Lejart, M. High-Resolution Melting of 18S rDNA sequences (18S-HRM) for discrimination of bivalve’s species at early juvenile stage: application to a spat survey. Mar Biol 164, 133 (2017). https://doi.org/10.1007/s00227-017-3162-5

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