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Applied and Environmental Microbiology, July 2001, p. 2932-2941, Vol. 67, No. 7
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.7.2932-2941.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Study of Genetic Diversity of Eukaryotic Picoplankton in Different Oceanic Regions by Small-Subunit rRNA Gene Cloning and Sequencing

Beatriz Díez, Carlos Pedrós-Alió,* and Ramon Massana

Departament de Biologia Marina, Institut de Ciències del Mar, CSIC, E-08039 Barcelona, Catalunya, Spain

Received 10 January 2001/Accepted 11 April 2001

Very small eukaryotic organisms (picoeukaryotes) are fundamental components of marine planktonic systems, often accounting for a significant fraction of the biomass and activity in a system. Their identity, however, has remained elusive, since the small cells lack morphological features for identification. We determined the diversity of marine picoeukaryotes by sequencing cloned 18S rRNA genes in five genetic libraries from North Atlantic, Southern Ocean, and Mediterranean Sea surface waters. Picoplankton were obtained by filter size fractionation, a step that excluded most large eukaryotes and recovered most picoeukaryotes. Genetic libraries of eukaryotic ribosomal DNA were screened by restriction fragment length polymorphism analysis, and at least one clone of each operational taxonomic unit (OTU) was partially sequenced. In general, the phylogenetic diversity in each library was rather great, and each library included many different OTUs and members of very distantly related phylogenetic groups. Of 225 eukaryotic clones, 126 were affiliated with algal classes, especially the Prasinophyceae, the Prymnesiophyceae, the Bacillariophyceae, and the Dinophyceae. A minor fraction (27 clones) was affiliated with clearly heterotrophic organisms, such as ciliates, the chrysomonad Paraphysomonas, cercomonads, and fungi. There were two relatively abundant novel lineages, novel stramenopiles (53 clones) and novel alveolates (19 clones). These lineages are very different from any organism that has been isolated, suggesting that there are previously unknown picoeukaryotes. Prasinophytes and novel stramenopile clones were very abundant in all of the libraries analyzed. These findings underscore the importance of attempts to grow the small eukaryotic plankton in pure culture.

* Corresponding author. Mailing address: Departament de Biologia Marina, Institut de Ciències del Mar, CSIC, Passeig Joan de Borbó s/n, E-08039 Barcelona, Catalunya, Spain. Phone: 34-932216416. Fax: 34-932217340. E-mail: cpedros{at}icm.csic.es.

Applied and Environmental Microbiology, July 2001, p. 2932-2941, Vol. 67, No. 7
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.7.2932-2941.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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