Novel 16S rRNA gene sequences retrieved from highly saline brine sediments of Kebrit Deep, Red Sea

Abstract In this study, we report on first 16S rRNA gene sequences from highly saline brine sediments taken at a depth of 1,515 m in the Kebrit Deep, northern Red Sea. Microbial DNA extracted directly from the sediments was subjected to PCR amplification with primers specific for bacterial and archaeal 16S rRNA gene sequences. The PCR products were cloned, and a total of 11 (6 bacterial and 5 archaeal) clone types were determined by restriction endonuclease digestion. Phylogenetic analysis revealed that most of the cloned sequences were unique, showing no close association with sequences of cultivated organisms or sequences derived from environmental samples. The bacterial clone sequences form a novel phylogenetic lineage (KB1 group) that branches between the Aquificales and the Thermotogales. The archaeal clone sequences group within the Euryarchaeota. Some of the sequences cluster with the group II and group III uncultivated archaea sequence clones, while two clone groups form separate branches. Our results suggest that hitherto unknown archaea and bacteria may thrive in highly saline brines of the Red Sea under extreme environmental conditions.

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

[2]  N. Pace,et al.  Microbial ecology and evolution: a ribosomal RNA approach. , 1986, Annual review of microbiology.

[3]  M. Schoell,et al.  New Deeps with Brines and Metalliferous Sediments in the Red Sea , 1972 .

[4]  C. Woese,et al.  Methanopyrus kandleri: an archaeal methanogen unrelated to all other known methanogens. , 1991, Systematic and applied microbiology.

[5]  C R Woese,et al.  Archaeal phylogeny: reexamination of the phylogenetic position of Archaeoglobus fulgidus in light of certain composition-induced artifacts. , 1991, Systematic and applied microbiology.

[6]  K. Schleifer,et al.  Detection and in situ identification of representatives of a widely distributed new bacterial phylum. , 1997, FEMS microbiology letters.

[7]  R. Zierenberg,et al.  Isotopic constraints on the origin of the Atlantis II, Suakin and Valdivia brines, Red Sea , 1986 .

[8]  E. Bonatti Punctiform initiation of seafloor spreading in the Red Sea during transition from a continental to an oceanic rift , 1985, Nature.

[9]  E. Delong Archaea in coastal marine environments. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[10]  H. Noller,et al.  Gene organization and primary structure of a ribosomal RNA operon from Escherichia coli. , 1981, Journal of molecular biology.

[11]  J. Fuhrman,et al.  Widespread Archaea and novel Bacteria from the deep sea as shown by 16S rRNA gene sequences , 1997 .

[12]  A. Hiraishi,et al.  Phylogenetic Evidence for the Existence of Novel Thermophilic Bacteria in Hot Spring Sulfur-Turf Microbial Mats in Japan , 1998, Applied and Environmental Microbiology.

[13]  R. Huber,et al.  The Order Thermotogales , 1992 .

[14]  P. Stoffers,et al.  Hydrographic structure of brine-filled deeps in the Red Sea—new results from the Shaban, Kebrit, Atlantis II, and Discovery Deep , 1998 .

[15]  D. B. Nedwell,et al.  Phylogenetic diversity of Archaea in sediment samples from a coastal salt marsh , 1997, Applied and environmental microbiology.

[16]  W. Liesack,et al.  Occurrence of novel groups of the domain Bacteria as revealed by analysis of genetic material isolated from an Australian terrestrial environment , 1992, Journal of bacteriology.

[17]  R. Huber,et al.  Thermocrinis ruber gen. nov., sp. nov., a Pink-Filament-Forming Hyperthermophilic Bacterium Isolated from Yellowstone National Park , 1998, Applied and Environmental Microbiology.

[18]  E. Delong,et al.  High abundance of Archaea in Antarctic marine picoplankton , 1994, Nature.

[19]  N. Pace,et al.  The Analysis of Natural Microbial Populations by Ribosomal RNA Sequences , 1986 .

[20]  N. Pace,et al.  Remarkable archaeal diversity detected in a Yellowstone National Park hot spring environment. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[21]  G. Pautot d'ouverture océanique réalisée à l'aide du Seabeam , 1983 .

[22]  R. Huber,et al.  Aquifex pyrophilus gen. nov. sp. nov., Represents a Novel Group of Marine Hyperthermophilic Hydrogen-Oxidizing Bacteria , 1992 .

[23]  R. Huber,et al.  Isolation of a hyperthermophilic archaeum predicted by in situ RNA analysis , 1995, Nature.

[24]  E. Delong,et al.  Vertical distribution and phylogenetic characterization of marine planktonic Archaea in the Santa Barbara Channel , 1997, Applied and environmental microbiology.

[25]  G. Olsen,et al.  A phylogenetic analysis of Aquifex pyrophilus. , 1992, Systematic and applied microbiology.

[26]  P. Styles,et al.  Two Stage Red Sea Floor Spreading , 1974, Nature.

[27]  M Weizenegger,et al.  Bacterial phylogeny based on comparative sequence analysis (review) , 1998, Electrophoresis.

[28]  K. Stetter,et al.  Reclassification of the crenarchael orders and families in accordance with 16S rRNA sequence data. , 1997, International journal of systematic bacteriology.

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

[30]  James R. Cole,et al.  A new version of the RDP (Ribosomal Database Project) , 1999, Nucleic Acids Res..