Vertical distribution of picoeukaryotic diversity in the Sargasso Sea.

Eukaryotic molecular diversity within the picoplanktonic size-fraction has primarily been studied in marine surface waters. Here, the vertical distribution of picoeukaryotic diversity was investigated in the Sargasso Sea from euphotic to abyssal waters, using size-fractionated samples (< 2 microm). 18S rRNA gene clone libraries were used to generate sequences from euphotic zone samples (deep chlorophyll maximum to the surface); the permanent thermocline (500 m); and the pelagic deep-sea (3000 m). Euphotic zone and deep-sea data contrasted strongly, the former displaying greater diversity at the first-rank taxon level, based on 232 nearly full-length sequences. Deep-sea sequences belonged almost exclusively to the Alveolata and Radiolaria, while surface samples also contained known and putative photosynthetic groups, such as unique Chlorarachniophyta and Chrysophyceae sequences. Phylogenetic analyses placed most Alveolata and Stramenopile sequences within previously reported 'environmental' clades, i.e. clades within the Novel Alveolate groups I and II (NAI and NAII), or the novel Marine Stramenopiles (MAST). However, some deep-sea NAII formed distinct, bootstrap supported clades. Stramenopiles were recovered from the euphotic zone only, although many MAST are reportedly heterotrophic, making the observed distribution a point for further investigation. An unexpectedly high proportion of radiolarian sequences were recovered. From these, five environmental radiolarian clades, RAD-I to RAD-V, were identified. RAD-IV and RAD-V were composed of Taxopodida-like sequences, with the former solely containing Sargasso Sea sequences, although from all depth zones sampled. Our findings highlight the vast diversity of these protists, most of which remain uncultured and of unknown ecological function.

[1]  D. Vaulot,et al.  Picobiliphytes: A Marine Picoplanktonic Algal Group with Unknown Affinities to Other Eukaryotes , 2007, Science.

[2]  M. D. Keller,et al.  A comparison of HPLC pigment signatures and electron microscopic observations for oligotrophic waters of the North Atlantic and Pacific Oceans , 1996 .

[3]  K. Valentin,et al.  Picoeukaryotic Plankton Diversity at the Helgoland Time Series Site as Assessed by Three Molecular Methods , 2006, Microbial Ecology.

[4]  William A. Siebold,et al.  SAR11 clade dominates ocean surface bacterioplankton communities , 2002, Nature.

[5]  S. Mayama,et al.  Phylogenetic analyses of the polycystine Radiolaria based on the 18s rDNA sequences of the Spumellarida and the Nassellarida , 2005 .

[6]  T. Stoeck,et al.  Novel Eukaryotic Lineages Inferred from Small-Subunit rRNA Analyses of Oxygen-Depleted Marine Environments , 2003, Applied and Environmental Microbiology.

[7]  Hui Zhou,et al.  Genetic diversity of small eukaryotes from the coastal waters of Nansha Islands in China. , 2004, FEMS microbiology letters.

[8]  U. Göbel,et al.  Determination of microbial diversity in environmental samples: pitfalls of PCR-based rRNA analysis. , 1997, FEMS microbiology reviews.

[9]  S. Adl,et al.  The New Higher Level Classification of Eukaryotes with Emphasis on the Taxonomy of Protists , 2005, The Journal of eukaryotic microbiology.

[10]  D. Vaulot,et al.  The Roscoff Culture Collection (RCC): a collection dedicated to marine picoplankton , 2004 .

[11]  Wen-Hsiung Li,et al.  Fundamentals of molecular evolution , 1990 .

[12]  S. Mayama,et al.  Molecular phylogeny of solitary shell-bearing Polycystinea (Radiolaria) , 2004 .

[13]  P. Green,et al.  Base-calling of automated sequencer traces using phred. I. Accuracy assessment. , 1998, Genome research.

[14]  C. Pedrós-Alió,et al.  Unexpected diversity of small eukaryotes in deep-sea Antarctic plankton , 2001, Nature.

[15]  R. Varela,et al.  A multiple PCR-primer approach to access the microeukaryotic diversity in environmental samples. , 2006, Protist.

[16]  John Bunge,et al.  Predicting microbial species richness. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[17]  D. Vaulot,et al.  Genetic diversity and habitats of two enigmatic marine alveolate lineages , 2006 .

[18]  F. Rodríguez-Valera,et al.  Toward the monophyly of Haeckel's radiolaria: 18S rRNA environmental data support the sisterhood of polycystinea and acantharea. , 2002, Molecular biology and evolution.

[19]  Rodrigo Lopez,et al.  Multiple sequence alignment with the Clustal series of programs , 2003, Nucleic Acids Res..

[20]  A. Rooney Mechanisms underlying the evolution and maintenance of functionally heterogeneous 18S rRNA genes in Apicomplexans. , 2004, Molecular biology and evolution.

[21]  E. Delong,et al.  Community Genomics Among Stratified Microbial Assemblages in the Ocean's Interior , 2006, Science.

[22]  H. Ducklow,et al.  Annual flux of dissolved organic carbon from the euphotic zone in the northwestern Sargasso Sea , 1994, Nature.

[23]  C. Pedrós-Alió,et al.  Marine microbial diversity: can it be determined? , 2006, Trends in microbiology.

[24]  D. Vaulot,et al.  Florenciella parvula gen. et sp. nov. (Dictyochophyceae, Heterokontophyta), a small flagellate isolated from the English Channel , 2004 .

[25]  J. Vallino,et al.  Efficient export of carbon to the deep ocean through dissolved organic matter , 2005, Nature.

[26]  A. Worden Picoeukaryote diversity in coastal waters of the Pacific Ocean , 2006 .

[27]  D. Vaulot,et al.  Composition and temporal variability of picoeukaryote communities at a coastal site of the English Channel from 18S rDNA sequences , 2004 .

[28]  H. Claustre,et al.  Abundance and diversity of prymnesiophytes in the picoplankton coμmunity from the equatorial Pacific Ocean inferred from 18S rDNA sequences , 2000 .

[29]  F. Azam,et al.  Microbes, Molecules, and Marine Ecosystems , 2004, Science.

[30]  T. Cavalier-smith,et al.  Phylum-specific environmental DNA analysis reveals remarkably high global biodiversity of Cercozoa (Protozoa). , 2004, International journal of systematic and evolutionary microbiology.

[31]  T. A. Hall,et al.  BIOEDIT: A USER-FRIENDLY BIOLOGICAL SEQUENCE ALIGNMENT EDITOR AND ANALYSIS PROGRAM FOR WINDOWS 95/98/ NT , 1999 .

[32]  A. Yamaguchi,et al.  Structure and size distribution of plankton communities down to the greater depths in the western North Pacific Ocean , 2002 .

[33]  Ramon Massana,et al.  Study of Genetic Diversity of Eukaryotic Picoplankton in Different Oceanic Regions by Small-Subunit rRNA Gene Cloning and Sequencing , 2001, Applied and Environmental Microbiology.

[34]  I. Sarashina,et al.  Molecular phylogeny of acantharian and polycystine radiolarians based on ribosomal DNA sequences, and some comparisons with data from the fossil record. , 2006, European journal of protistology.

[35]  C. Berney,et al.  Small-subunit ribosomal RNA gene sequences of Phaeodarea challenge the monophyly of Haeckel's Radiolaria. , 2004, Protist.

[36]  C. Pedrós-Alió,et al.  Diversity and Distribution of Marine Microbial Eukaryotes in the Arctic Ocean and Adjacent Seas , 2006, Applied and Environmental Microbiology.

[37]  C. Pedrós-Alió,et al.  Picoeukaryotic diversity in an oligotrophic coastal site studied by molecular and culturing approaches. , 2004, FEMS microbiology ecology.

[38]  Thomas Huber,et al.  Bellerophon: a program to detect chimeric sequences in multiple sequence alignments , 2004, Bioinform..

[39]  C. Pedrós-Alió,et al.  Phylogenetic and Ecological Analysis of Novel Marine Stramenopiles , 2004, Applied and Environmental Microbiology.

[40]  P. Kemp,et al.  Estimating prokaryotic diversity: When are 16S rDNA libraries large enough? , 2004 .

[41]  P. Green,et al.  Consed: a graphical tool for sequence finishing. , 1998, Genome research.

[42]  David Posada,et al.  MODELTEST: testing the model of DNA substitution , 1998, Bioinform..

[43]  E. Delong,et al.  Archaeal dominance in the mesopelagic zone of the Pacific Ocean , 2001, Nature.

[44]  Sangcheol Na,et al.  Active ingestion of fluorescently labeled bacteria by mesopelagic heterotrophic nanoflagellates in the East Sea, Korea , 2000 .

[45]  B. Díez,et al.  Unveiling the Organisms behind Novel Eukaryotic Ribosomal DNA Sequences from the Ocean , 2002, Applied and Environmental Microbiology.

[46]  A. Worden,et al.  In-depth analyses of marine microbial community genomics. , 2006, Trends in microbiology.

[47]  D. Caron,et al.  TOWARDS A MOLECULAR PHYLOGENY OF COLONIAL SPUMELLARIAN RADIOLARIA , 1999 .

[48]  D. Vaulot,et al.  A Single Species, Micromonas pusilla (Prasinophyceae), Dominates the Eukaryotic Picoplankton in the Western English Channel , 2004, Applied and Environmental Microbiology.

[49]  D. Caron,et al.  Phylogenetic relationships between the Acantharea and the Polycystinea: a molecular perspective on Haeckel's Radiolaria. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[50]  T. Cavalier-smith Only six kingdoms of life , 2004, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[51]  E. Delong,et al.  Genomic perspectives in microbial oceanography , 2005, Nature.

[52]  Wei Qian,et al.  Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis. , 2000, Molecular biology and evolution.

[53]  N. B. Petrov,et al.  The twilight of Heliozoa and rise of Rhizaria, an emerging supergroup of amoeboid eukaryotes. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[54]  M. Sogin,et al.  Benthic eukaryotic diversity in the Guaymas Basin hydrothermal vent environment , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[55]  L. Guillou,et al.  Oligonucleotide Probes for the Identification of Three Algal Groups by Dot Blot and Fluorescent Whole-Cell Hybridization , 2000, The Journal of eukaryotic microbiology.

[56]  D. Vaulot,et al.  Eukaryotic picoplankton communities of the Mediterranean Sea in summer assessed by molecular approaches (DGGE, TTGE, QPCR). , 2006, FEMS microbiology ecology.

[57]  D. Caron,et al.  Protistan Diversity Estimates Based on 18S rDNA from Seawater Incubations in the Western North Atlantic 1 , 2005, The Journal of eukaryotic microbiology.

[58]  Purificación López-García,et al.  Autochthonous eukaryotic diversity in hydrothermal sediment and experimental microcolonizers at the Mid-Atlantic Ridge , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[59]  D. W. Coats,et al.  PARASITISM OF PHOTOSYNTHETIC DINOFLAGELLATES BY THREE STRAINS OF AMOEBOPHRYA (DINOPHYTA): PARASITE SURVIVAL, INFECTIVITY, GENERATION TIME, AND HOST SPECIFICITY1 , 2002 .

[60]  C. Pedrós-Alió,et al.  Distribution and abundance of uncultured heterotrophic flagellates in the world oceans. , 2006, Environmental microbiology.

[61]  C. Berney,et al.  How many novel eukaryotic 'kingdoms'? Pitfalls and limitations of environmental DNA surveys , 2004, BMC Biology.

[62]  R. Wachter,et al.  Oceanic 18S rDNA sequences from picoplankton reveal unsuspected eukaryotic diversity , 2001, Nature.

[63]  Purificación López-García,et al.  The molecular ecology of microbial eukaryotes unveils a hidden world. , 2002, Trends in microbiology.

[64]  David J. Allen,et al.  Analysis of photosynthetic picoeukaryote diversity at open ocean sites in the Arabian Sea using a PCR biased towards marine algal plastids , 2006 .

[65]  P Green,et al.  Base-calling of automated sequencer traces using phred. II. Error probabilities. , 1998, Genome research.