A simple and innovative method for species identification of phytoplankton cells on minute quantities of DNA

Authors

  • Estelle Masseret,

    Corresponding author
    1. Université Montpellier II, CNRS, Ifremer, IRD, UMR 5119 Ecosystèmes Lagunaires, cc093, place E. Bataillon, 34095 Montpellier, France.
    Search for more papers by this author
  • Marjorie Enquebecq,

    1. Université Montpellier II, CNRS, Ifremer, IRD, UMR 5119 Ecosystèmes Lagunaires, cc093, place E. Bataillon, 34095 Montpellier, France.
    Search for more papers by this author
  • Mohamed Laabir,

    1. Université Montpellier II, CNRS, Ifremer, IRD, UMR 5119 Ecosystèmes Lagunaires, cc093, place E. Bataillon, 34095 Montpellier, France.
    Search for more papers by this author
  • Benjamin Genovesi,

    1. Université Montpellier II, CNRS, Ifremer, IRD, UMR 5119 Ecosystèmes Lagunaires, cc093, place E. Bataillon, 34095 Montpellier, France.
    Search for more papers by this author
  • André Vaquer,

    1. Université Montpellier II, CNRS, Ifremer, IRD, UMR 5119 Ecosystèmes Lagunaires, cc093, place E. Bataillon, 34095 Montpellier, France.
    Search for more papers by this author
  • Jean-Christophe Avarre

    1. IRD, UR175 CAVIAR, 361 Rue Jean-François Breton, BP5095, 34196 Montpellier cedex 05, France.
    Search for more papers by this author

E-mail estelle.masseret@univ-montp2.fr; Tel. (+33) 467144762; Fax (+33) 467143719.

Summary

Dinoflagellates belonging to the genus Alexandrium are often involved in harmful algal blooms. Their ecological exploration is thus essential to increase our knowledge on these toxic events. Yet, population genetic studies, taxonomic identification and environmental monitoring are hampered by major constraints: the necessity to establish monoclonal cultures from environmental samples and the sensitivity of available molecular tools. The present work describes a very simple and sensitive method for extraction and amplification of DNA at the infra-single-cell level. Its on-slide format allows for easy visual control of both quality and quantity of the templates. Combined with a semi-multiplex PCR protocol designed on the 18S-28S rDNA-ITS region of Alexandrium catenella and Alexandrium tamarense, this procedure allowed the identification and discrimination of these species from both monoclonal cultures and natural samples.

Ancillary