Phage-bacteria relationships and CRISPR elements revealed by a metagenomic survey of the rumen microbiome.

Viruses are the most abundant biological entities on the planet and play an important role in balancing microbes within an ecosystem and facilitating horizontal gene transfer. Although bacteriophages are abundant in rumen environments, little is known about the types of viruses present or their interaction with the rumen microbiome. We undertook random pyrosequencing of virus-enriched metagenomes (viromes) isolated from bovine rumen fluid and analysed the resulting data using comparative metagenomics. A high level of diversity was observed with up to 28,000 different viral genotypes obtained from each environment. The majority (~78%) of sequences did not match any previously described virus. Prophages outnumbered lytic phages approximately 2:1 with the most abundant bacteriophage and prophage types being associated with members of the dominant rumen phyla (Firmicutes and Proteobacteria). Metabolic profiling based on SEED subsystems revealed an enrichment of sequences with putative functional roles in DNA and protein metabolism, but a surprisingly low proportion of sequences assigned to carbohydrate and amino acid metabolism. We expanded our analysis to include previously described metagenomic data and 14 reference genomes. Clustered regularly interspaced short palindromic repeats (CRISPR) were detected in most of the microbial genomes, suggesting previous interactions between viral and microbial communities.

[1]  B. Boschek,et al.  Endo-N-acetylneuraminidase associated with bacteriophage particles , 1982, Journal of virology.

[2]  A. Klieve,et al.  Morphological diversity of ruminal bacteriophages from sheep and cattle , 1988, Applied and environmental microbiology.

[3]  B Flesher,et al.  Use of phylogenetically based hybridization probes for studies of ruminal microbial ecology , 1988, Applied and environmental microbiology.

[4]  A. Richardson,et al.  The implications of the loss and regain of cotton‐degrading activity for the degradation of straw by Ruminococcus flavefaciens strain 007 , 1990 .

[5]  A. Klieve,et al.  Estimation of ruminal bacteriophage numbers by pulsed-field gel electrophoresis and laser densitometry , 1993, Applied and environmental microbiology.

[6]  J. Nolan,et al.  Natural variability and diurnal fluctuations within the bacteriophage population of the rumen , 1996, Applied and environmental microbiology.

[7]  D. E. Akin,et al.  Bacteria, Fungi, and Protozoa of the Rumen , 1997 .

[8]  K. Novak The complete genome sequence… , 1998, Nature Medicine.

[9]  Kelvin H. Lee,et al.  Genomic analysis. , 2000, Current opinion in biotechnology.

[10]  T. Thingstad Elements of a theory for the mechanisms controlling abundance, diversity, and biogeochemical role of lytic bacterial viruses in aquatic systems , 2000 .

[11]  S. Garcia-Vallvé,et al.  Horizontal gene transfer of glycosyl hydrolases of the rumen fungi. , 2000, Molecular biology and evolution.

[12]  S. Surzycki Preparation of Genomic DNA from Bacteria , 2000 .

[13]  J. Russell,et al.  Factors That Alter Rumen Microbial Ecology , 2001, Science.

[14]  B. White,et al.  Analysis of the rumen bacterial diversity under two different diet conditions using denaturing gradient gel electrophoresis, random sequencing, and statistical ecology approaches , 2001 .

[15]  Hui-Hsien Chou,et al.  DNA sequence quality trimming and vector removal , 2001, Bioinform..

[16]  B. Andresen,et al.  Genomic analysis of uncultured marine viral communities , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[17]  Folker Meyer,et al.  Complete genome sequence and analysis of Wolinella succinogenes , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[18]  M. Clokie,et al.  Marine ecosystems: Bacterial photosynthesis genes in a virus , 2003, Nature.

[19]  R. Mackie,et al.  Opportunities to improve fiber degradation in the rumen: microbiology, ecology, and genomics. , 2003, FEMS microbiology reviews.

[20]  P. Salamon,et al.  Metagenomic Analyses of an Uncultured Viral Community from Human Feces , 2003, Journal of bacteriology.

[21]  C. Buchen-Osmond,et al.  The universal virus database ICTVdB , 2003 .

[22]  Forest Rohwer,et al.  Global Phage Diversity , 2003, Cell.

[23]  Andrew C. Tolonen,et al.  Transfer of photosynthesis genes to and from Prochlorococcus viruses. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Sang Yup Lee,et al.  The genome sequence of the capnophilic rumen bacterium Mannheimia succiniciproducens , 2004, Nature Biotechnology.

[25]  J. García-Martínez,et al.  Intervening Sequences of Regularly Spaced Prokaryotic Repeats Derive from Foreign Genetic Elements , 2005, Journal of Molecular Evolution.

[26]  Peter Salamon,et al.  PHACCS, an online tool for estimating the structure and diversity of uncultured viral communities using metagenomic information , 2005, BMC Bioinformatics.

[27]  Alexander Bolotin,et al.  Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin. , 2005, Microbiology.

[28]  Achim Dickmanns,et al.  Crystal structure of the polysialic acid–degrading endosialidase of bacteriophage K1F , 2005, Nature Structural &Molecular Biology.

[29]  Forest Rohwer,et al.  Here a virus, there a virus, everywhere the same virus? , 2005, Trends in microbiology.

[30]  Laura S. Frost,et al.  Mobile genetic elements: the agents of open source evolution , 2005, Nature Reviews Microbiology.

[31]  S. Ehrlich,et al.  Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin. , 2005, Microbiology.

[32]  Naryttza N. Diaz,et al.  The Subsystems Approach to Genome Annotation and its Use in the Project to Annotate 1000 Genomes , 2005, Nucleic acids research.

[33]  G Vergnaud,et al.  CRISPR elements in Yersinia pestis acquire new repeats by preferential uptake of bacteriophage DNA, and provide additional tools for evolutionary studies. , 2005, Microbiology.

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

[35]  J. S. Godde,et al.  The Repetitive DNA Elements Called CRISPRs and Their Associated Genes: Evidence of Horizontal Transfer Among Prokaryotes , 2006, Journal of Molecular Evolution.

[36]  M. Huynen,et al.  Horizontal gene transfer from Bacteria to rumen Ciliates indicates adaptation to their anaerobic, carbohydrates-rich environment , 2006, BMC Genomics.

[37]  Matthew Berriman,et al.  ACT: the Artemis comparison tool , 2005, Bioinform..

[38]  Jonathan Pevsner,et al.  Basic Local Alignment Search Tool (BLAST) , 2005 .

[39]  R. Edwards,et al.  Viral metagenomics , 2005, Nature Reviews Microbiology.

[40]  Sang Joon Kim,et al.  A Mathematical Theory of Communication , 2006 .

[41]  Florent E. Angly,et al.  The Marine Viromes of Four Oceanic Regions , 2006, PLoS biology.

[42]  M. Rasmussen,et al.  Klebsiella to Salmonella gene transfer within rumen protozoa: implications for antibiotic resistance and rumen defaunation. , 2006, Veterinary microbiology.

[43]  Rick L. Stevens,et al.  The RAST Server: Rapid Annotations using Subsystems Technology , 2008, BMC Genomics.

[44]  M. Fenner,et al.  CRISPR--a widespread system that provides acquired resistance against phages in bacteria and archaea. , 2007 .

[45]  Peter Salamon,et al.  Metagenomic and Small-Subunit rRNA Analyses Reveal the Genetic Diversity of Bacteria, Archaea, Fungi, and Viruses in Soil , 2007, Applied and Environmental Microbiology.

[46]  Ibtissem Grissa,et al.  CRISPRFinder: a web tool to identify clustered regularly interspaced short palindromic repeats , 2007, Nucleic Acids Res..

[47]  Ibtissem Grissa,et al.  The CRISPRdb database and tools to display CRISPRs and to generate dictionaries of spacers and repeats , 2007, BMC Bioinformatics.

[48]  B. White,et al.  Polysaccharide utilization by gut bacteria: potential for new insights from genomic analysis , 2008, Nature Reviews Microbiology.

[49]  Andreas Wilke,et al.  phylogenetic and functional analysis of metagenomes , 2022 .

[50]  E. Koonin,et al.  Genomics of bacteria and archaea: the emerging dynamic view of the prokaryotic world , 2008, Nucleic acids research.

[51]  Rick L. Stevens,et al.  Functional metagenomic profiling of nine biomes , 2008, Nature.

[52]  Forest Rohwer,et al.  Roles of viruses in the environment. , 2009, Environmental microbiology.

[53]  Forest Rohwer,et al.  The GAAS Metagenomic Tool and Its Estimations of Viral and Microbial Average Genome Size in Four Major Biomes , 2009, PLoS Comput. Biol..

[54]  D. Bolton,et al.  Transfer of Antibiotic Resistance Marker Genes between Lactic Acid Bacteria in Model Rumen and Plant Environments , 2009, Applied and Environmental Microbiology.

[55]  F. Rohwer,et al.  Viruses manipulate the marine environment , 2009, Nature.

[56]  F. Rohwer,et al.  Explaining microbial population genomics through phage predation , 2009, Nature Reviews Microbiology.

[57]  K. Nelson,et al.  Gene-centric metagenomics of the fiber-adherent bovine rumen microbiome reveals forage specific glycoside hydrolases , 2009, Proceedings of the National Academy of Sciences.

[58]  Alvaro G. Hernandez,et al.  Diversity and Strain Specificity of Plant Cell Wall Degrading Enzymes Revealed by the Draft Genome of Ruminococcus flavefaciens FD-1 , 2009, PloS one.

[59]  Brandi L. Cantarel,et al.  The Carbohydrate-Active EnZymes database (CAZy): an expert resource for Glycogenomics , 2008, Nucleic Acids Res..

[60]  Jaysheel D. Bhavsar,et al.  Census of the Viral Metagenome within an Activated Sludge Microbial Assemblage , 2010, Applied and Environmental Microbiology.

[61]  M. Weinbauer,et al.  Trade-Offs between Competition and Defense Specialists among Unicellular Planktonic Organisms: the “Killing the Winner” Hypothesis Revisited , 2010, Microbiology and Molecular Biology Reviews.

[62]  William J. Kelly,et al.  The Genome Sequence of the Rumen Methanogen Methanobrevibacter ruminantium Reveals New Possibilities for Controlling Ruminant Methane Emissions , 2010, PloS one.

[63]  K. Nelson,et al.  Comparative Genome Analysis of Prevotella ruminicola and Prevotella bryantii: Insights into Their Environmental Niche , 2010, Microbial Ecology.

[64]  Tal Pupko,et al.  Inference and Characterization of Horizontally Transferred Gene Families Using Stochastic Mapping , 2009, Molecular biology and evolution.

[65]  H. Deveau,et al.  CRISPR/Cas system and its role in phage-bacteria interactions. , 2010, Annual review of microbiology.

[66]  S. Abedon,et al.  Bacteriophage host range and bacterial resistance. , 2010, Advances in applied microbiology.

[67]  H. Cadillo-Quiroz,et al.  CRISPR Associated Diversity within a Population of Sulfolobus islandicus , 2010, PloS one.

[68]  Forest Rohwer,et al.  Viruses in the fecal microbiota of monozygotic twins and their mothers , 2010, Nature.

[69]  Peter Salamon,et al.  Viral and microbial community dynamics in four aquatic environments , 2010, The ISME Journal.

[70]  L. Marraffini,et al.  CRISPR interference: RNA-directed adaptive immunity in bacteria and archaea , 2010, Nature Reviews Genetics.

[71]  Dong Li,et al.  The Glycobiome of the Rumen Bacterium Butyrivibrio proteoclasticus B316T Highlights Adaptation to a Polysaccharide-Rich Environment , 2010, PloS one.

[72]  S. Tringe,et al.  Metagenomic Discovery of Biomass-Degrading Genes and Genomes from Cow Rumen , 2011, Science.

[73]  R. Aminov Horizontal Gene Exchange in Environmental Microbiota , 2011, Front. Microbio..

[74]  Jing Chen,et al.  Community cyberinfrastructure for Advanced Microbial Ecology Research and Analysis: the CAMERA resource , 2010, Nucleic Acids Res..