Comparative day/night metatranscriptomic analysis of microbial communities in the North Pacific subtropical gyre.

Metatranscriptomic analyses of microbial assemblages (< 5 microm) from surface water at the Hawaiian Ocean Time-Series (HOT) revealed community-wide metabolic activities and day/night patterns of differential gene expression. Pyrosequencing produced 75 558 putative mRNA reads from a day transcriptome and 75 946 from a night transcriptome. Taxonomic binning of annotated mRNAs indicated that Cyanobacteria contributed a greater percentage of the transcripts (54% of annotated sequences) than expected based on abundance (35% of cell counts and 21% 16S rRNA of libraries), and may represent the most actively transcribing cells in this surface ocean community in both the day and night. Major heterotrophic taxa contributing to the community transcriptome included alpha-Proteobacteria (19% of annotated sequences, most of which were SAR11-related) and gamma-Proteobacteria (4%). The composition of transcript pools was consistent with models of prokaryotic gene expression, including operon-based transcription patterns and an abundance of genes predicted to be highly expressed. Metabolic activities that are shared by many microbial taxa (e.g. glycolysis, citric acid cycle, amino acid biosynthesis and transcription and translation machinery) were well represented among the community transcripts. There was an overabundance of transcripts for photosynthesis, C1 metabolism and oxidative phosphorylation in the day compared with night, and evidence that energy acquisition is coordinated with solar radiation levels for both autotrophic and heterotrophic microbes. In contrast, housekeeping activities such as amino acid biosynthesis, membrane synthesis and repair, and vitamin biosynthesis were overrepresented in the night transcriptome. Direct sequencing of these environmental transcripts has provided detailed information on metabolic and biogeochemical responses of a microbial community to solar forcing.

[1]  J. Shendure The beginning of the end for microarrays? , 2008, Nature Methods.

[2]  Tatiana A. Tatusova,et al.  NCBI Reference Sequence (RefSeq): a curated non-redundant sequence database of genomes, transcripts and proteins , 2004, Nucleic Acids Res..

[3]  Ricardo M Letelier,et al.  The role of nitrogen fixation in biogeochemical cycling in the subtropical North Pacific Ocean , 1997, Nature.

[4]  P. Gerhardt,et al.  Methods for general and molecular bacteriology , 1994 .

[5]  M. Wésolowski-Louvel,et al.  Soil eukaryotic functional diversity, a metatranscriptomic approach , 2007, The ISME Journal.

[6]  Sallie W. Chisholm,et al.  Niche Partitioning Among Prochlorococcus Ecotypes Along Ocean-Scale Environmental Gradients , 2006, Science.

[7]  R. Beinart,et al.  In situ transcriptomic analysis of the globally important keystone N2-fixing taxon Crocosphaera watsonii , 2009, The ISME Journal.

[8]  J. Bauer,et al.  14C activity of dissolved organic carbon fractions in the north-central Pacific and Sargasso Sea , 1992, Nature.

[9]  S. Chisholm,et al.  Nutrient gradients in the western North Atlantic Ocean: Relationship to microbial community structure and comparison to patterns in the Pacific Ocean , 2001 .

[10]  Juan Miguel García-Gómez,et al.  BIOINFORMATICS APPLICATIONS NOTE Sequence analysis Manipulation of FASTQ data with Galaxy , 2005 .

[11]  S. Kravitz,et al.  CAMERA: A Community Resource for Metagenomics , 2007, PLoS biology.

[12]  David L. Wheeler,et al.  GenBank , 2015, Nucleic Acids Res..

[13]  Maureen L. Coleman,et al.  Microbial community gene expression in ocean surface waters , 2008, Proceedings of the National Academy of Sciences.

[14]  J. Gilbert,et al.  Detection of Large Numbers of Novel Sequences in the Metatranscriptomes of Complex Marine Microbial Communities , 2008, PloS one.

[15]  David M. Karl,et al.  The Hawaii Ocean Time-series (HOT) program: Background, rationale and field implementation , 1996 .

[16]  Alison Buchan,et al.  Bacterial Taxa That Limit Sulfur Flux from the Ocean , 2006, Science.

[17]  J. Fuhrman,et al.  Extraction from Natural Planktonic Microorganisms of DNA Suitable for Molecular Biological Studies , 1988, Applied and environmental microbiology.

[18]  Nicholas H. Putnam,et al.  The Genome of the Diatom Thalassiosira Pseudonana: Ecology, Evolution, and Metabolism , 2004, Science.

[19]  Robert A. Edwards,et al.  Bacterial carbon processing by generalist species in the coastal ocean , 2008, Nature.

[20]  D. Vaulot,et al.  Photosynthetic picoplankton community structure in the subtropical North Pacific Ocean near Hawaii (station ALOHA) , 1993 .

[21]  C. Bowler,et al.  An ecological and evolutionary context for integrated nitrogen metabolism and related signaling pathways in marine diatoms. , 2006, Current opinion in plant biology.

[22]  A. Halpern,et al.  The Sorcerer II Global Ocean Sampling Expedition: Northwest Atlantic through Eastern Tropical Pacific , 2007, PLoS biology.

[23]  R. Rauhut,et al.  mRNA degradation in bacteria. , 1999, FEMS microbiology reviews.

[24]  Jizhong Zhou Microarrays for bacterial detection and microbial community analysis. , 2003, Current opinion in microbiology.

[25]  Michael Y. Galperin,et al.  The COG database: a tool for genome-scale analysis of protein functions and evolution , 2000, Nucleic Acids Res..

[26]  James R. Knight,et al.  Genome sequencing in microfabricated high-density picolitre reactors , 2005, Nature.

[27]  K. Goodwin,et al.  Leisingera methylohalidivorans gen. nov., sp. nov., a marine methylotroph that grows on methyl bromide. , 2002, International journal of systematic and evolutionary microbiology.

[28]  Daniel H. Huson,et al.  Simultaneous Assessment of Soil Microbial Community Structure and Function through Analysis of the Meta-Transcriptome , 2008, PloS one.

[29]  B. Heikes,et al.  Atmospheric methanol budget and ocean implication , 2002 .

[30]  A. Pardee,et al.  Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. , 1992, Science.

[31]  R. Kiene,et al.  New and important roles for DMSP in marine microbial communities , 2000 .

[32]  H. Bürgmann,et al.  mRNA Extraction and Reverse Transcription-PCR Protocol for Detection of nifH Gene Expression by Azotobacter vinelandii in Soil , 2003, Applied and Environmental Microbiology.

[33]  Andrew Hansen,et al.  Unicellular cyanobacteria fix N2 in the subtropical North Pacific Ocean , 2001, Nature.

[34]  Tatiana Tatusova,et al.  NCBI Reference Sequence (RefSeq): a curated non-redundant sequence database of genomes, transcripts and proteins , 2004, Nucleic Acids Res..

[35]  D. Scanlan,et al.  Diel rhythmicity in amino acid uptake by Prochlorococcus. , 2008, Environmental microbiology.

[36]  T. Urich,et al.  Archaea predominate among ammonia-oxidizing prokaryotes in soils , 2006, Nature.

[37]  M. Moran,et al.  Transcriptional response of Silicibacter pomeroyi DSS-3 to dimethylsulfoniopropionate (DMSP). , 2007, Environmental microbiology.

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

[39]  中尾 光輝,et al.  KEGG(Kyoto Encyclopedia of Genes and Genomes)〔和文〕 (特集 ゲノム医学の現在と未来--基礎と臨床) -- (データベース) , 2000 .

[40]  Forest Rohwer,et al.  An application of statistics to comparative metagenomics , 2006, BMC Bioinformatics.

[41]  Mary Ann Moran,et al.  Transporter genes expressed by coastal bacterioplankton in response to dissolved organic carbon , 2010, Environmental microbiology.

[42]  J. Belasco,et al.  1 – mRNA Degradation in Prokaryotic Cells: An Overview , 1993 .

[43]  M. Moran,et al.  Analysis of Microbial Gene Transcripts in Environmental Samples , 2005, Applied and Environmental Microbiology.

[44]  J. Belasco,et al.  Control of messenger RNA stability. , 1993 .

[45]  D. Scanlan,et al.  Genetic diversity of eukaryotic ultraphytoplankton in the Gulf of Naples during an annual cycle , 2007 .

[46]  P. Novelli,et al.  Methane oxidation and methane fluxes in the ocean surface layer and deep anoxic waters , 1987, Nature.

[47]  J. Paul,et al.  Real-Time PCR Quantification of rbcL (Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase) mRNA in Diatoms and Pelagophytes , 2002, Applied and Environmental Microbiology.

[48]  Martin Gallagher,et al.  Uptake of methanol to the North Atlantic Ocean surface , 2004 .

[49]  S. Karlin,et al.  Predicted Highly Expressed Genes of Diverse Prokaryotic Genomes , 2000, Journal of bacteriology.

[50]  J. Eberwine,et al.  Amplified RNA synthesized from limited quantities of heterogeneous cDNA. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[51]  R. Overbeek,et al.  The use of gene clusters to infer functional coupling. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[52]  Adam Godzik,et al.  Cd-hit: a fast program for clustering and comparing large sets of protein or nucleotide sequences , 2006, Bioinform..

[53]  J. D. Elsas,et al.  Molecular Microbial Ecology Manual , 2013, Springer Netherlands.

[54]  S. Giovannoni,et al.  The small genome of an abundant coastal ocean methylotroph. , 2008, Environmental microbiology.

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

[56]  E. Lander,et al.  Genomic mapping by fingerprinting random clones: a mathematical analysis. , 1988, Genomics.

[57]  F. Neidhardt,et al.  Growth of the bacterial cell , 1983 .

[58]  M. Moran,et al.  Section 8 Update - Environmental transcriptomics: a method to access expressed genes in complex microbial communities , 2004 .

[59]  R. Oremland,et al.  Identification of Methyl Halide-Utilizing Genes in the Methyl Bromide-Utilizing Bacterial Strain IMB-1 Suggests a High Degree of Conservation of Methyl Halide-Specific Genes in Gram-Negative Bacteria , 2001, Applied and Environmental Microbiology.

[60]  Alexander F. Auch,et al.  MEGAN analysis of metagenomic data. , 2007, Genome research.

[61]  B. De Baets,et al.  Genome analysis of the smallest free-living eukaryote Ostreococcus tauri unveils many unique features. , 2006, Proceedings of the National Academy of Sciences of the United States of America.