New Perspective on Uncultured Bacterial Phylogenetic Division OP11

ABSTRACT Organisms belonging to the OP11 candidate phylogenetic division of Bacteria have been detected only in rRNA-based sequence surveys of environmental samples. Preliminary studies indicated that such organisms represented by the sequences are abundant and widespread in nature and highly diverse phylogenetically. In order to document more thoroughly the phylogenetic breadth and environmental distribution of this diverse group of organisms, we conducted further molecular analyses on environmental DNAs. Using PCR techniques and primers directed toward each of the five described subdivisions of OP11, we surveyed 17 environmental DNAs and analyzed rRNA gene sequences in 27 clonal libraries from 14 environments. Ninety-nine new and unique sequences were determined completely, and approximately 200 additional clones were subjected to partial sequencing. Extensive phylogenetic comparisons of the new sequences to those representing other bacterial divisions further resolved the phylogeny of the bacterial candidate division OP11 and identified two new candidate bacterial divisions, OP11-derived 1 (OD1) and Sulphur River 1 (SR1). The widespread environmental distribution of representatives of the bacterial divisions OD1, OP11, and SR1 suggests potentially conspicuous biogeochemical roles for these organisms in their respective environments. The information on environmental distribution offers clues for attempts to culture landmark representatives of these novel bacterial divisions, and the sequences are specific molecular signatures that provide for their identification in other contexts.

[1]  E. Stackebrandt,et al.  Nucleic acid techniques in bacterial systematics , 1991 .

[2]  D. Hillis,et al.  Taxonomic sampling, phylogenetic accuracy, and investigator bias. , 1998, Systematic biology.

[3]  C. Kuske,et al.  Diverse uncultivated bacterial groups from soils of the arid southwestern United States that are present in many geographic regions , 1997, Applied and environmental microbiology.

[4]  L. Shimkets,et al.  Bacterial diversity of a Carolina bay as determined by 16S rRNA gene analysis: confirmation of novel taxa , 1997, Applied and environmental microbiology.

[5]  J. Bull,et al.  Experimental phylogenetics: generation of a known phylogeny. , 1992, Science.

[6]  N. Pace,et al.  Expanding the Known Diversity and Environmental Distribution of an Uncultured Phylogenetic Division of Bacteria , 2000, Applied and Environmental Microbiology.

[7]  D. Lane 16S/23S rRNA sequencing , 1991 .

[8]  T. Ueda,et al.  Molecular phylogenetic analysis of a soil microbial community in a soybean field , 1995 .

[9]  F. Dewhirst,et al.  Bacterial Diversity in Human Subgingival Plaque , 2001, Journal of bacteriology.

[10]  A. Graybeal,et al.  Is it better to add taxa or characters to a difficult phylogenetic problem? , 1998, Systematic biology.

[11]  N. Pace,et al.  Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[12]  P. Hugenholtz,et al.  A multiple-outgroup approach to resolving division-level phylogenetic relationships using 16S rDNA data. , 2001, International journal of systematic and evolutionary microbiology.

[13]  J. Borneman,et al.  Molecular microbial diversity in soils from eastern Amazonia: evidence for unusual microorganisms and microbial population shifts associated with deforestation , 1997, Applied and environmental microbiology.

[14]  Philip Hugenholtz,et al.  Impact of Culture-Independent Studies on the Emerging Phylogenetic View of Bacterial Diversity , 1998, Journal of bacteriology.

[15]  F. Ayala Molecular systematics , 2004, Journal of Molecular Evolution.

[16]  N. Pace,et al.  Novel Division Level Bacterial Diversity in a Yellowstone Hot Spring , 1998, Journal of bacteriology.

[17]  K. Horikoshi,et al.  Bacterial diversity in deep-sea sediments from different depths , 1999, Biodiversity & Conservation.

[18]  James R. Cole,et al.  The Ribosomal Database Project (RDP-II): previewing a new autoaligner that allows regular updates and the new prokaryotic taxonomy , 2003, Nucleic Acids Res..

[19]  S. Ekendahl,et al.  16S rRNA gene diversity of attached and unattached bacteria in boreholes along the access tunnel to the Äspö hard rock laboratory, Sweden , 1996 .

[20]  S. Poe Sensitivity of phylogeny estimation to taxonomic sampling. , 1998, Systematic biology.

[21]  K. Schleifer,et al.  ARB: a software environment for sequence data. , 2004, Nucleic acids research.

[22]  Philip Hugenholtz,et al.  Microbial Diversity in a Hydrocarbon- and Chlorinated-Solvent-Contaminated Aquifer Undergoing Intrinsic Bioremediation , 1998, Applied and Environmental Microbiology.