AME 28:157-166 (2002)  -  doi:10.3354/ame028157

Application of fluorescent in situ hybridization coupled with tyramide signal amplification (FISH-TSA) to assess eukaryotic picoplankton composition

Fabrice Not*, Nathalie Simon, Isabelle C. Biegala, Daniel Vaulot

Station Biologique, UMR 7127, CNRS, INSU et Université Pierre et Marie Curie, Place George Teissier, 29680 Roscoff, France

ABSTRACT: Photosynthetic picoeukaryotes (phytoplankton cells with a diameter smaller than 2 to 3 μm) contribute significantly to both biomass and primary production in the oligotrophic open ocean and coastal waters, at certain times of the year. The identification of these organisms is difficult because of their small size and simple morphology, therefore hindering detailed ecological studies of their distribution and role. In this paper, we demonstrate the use of oligonucleotide probes specific to algal classes or to lower order taxa in combination with fluorescent in situ hybridization and tyramide signal amplification (FISH-TSA) to determine eukaryotic picophytoplankton diversity. Target cells were detected and enumerated using epifluorescence microscopy. The sensitivity of the technique and the specificity of the probes were tested on pure and mixed picoplanktonic strains, as well as on natural samples from the English Channel. In these samples, the community was dominated by cells belonging to the division Chlorophyta. Haptophyta, Bolidophyceae and Pelagophyceae were also detected at low abundance. The FISH-TSA method is readily applicable to the study of picoplankton diversity in natural communities.

KEY WORDS: Fluorescent in situ hybridization · Tyramide signal amplification · Picoplankton · Eukaryotes · Coastal waters · Diversity

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