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Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, PR China
Correspondence
Shuang-Jiang Liu
shuangjiang{at}hotmail.com
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ABSTRACT |
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The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain BD-1T is AY510707.
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) are widely distributed in hypersaline environments such as salt and/or soda lakes and solar salterns (Ochsenreiter et al., 2002, as cited therein). Being fast-growing species and active in the conversion of sugars and amino acids, Halorubrum species are considered to play important roles in the carbon and nitrogen cycles of hypersaline environments. Currently, the genus Halorubrum contains nine species with validly published names: Halorubrum sodomense (Oren, 1983), Halorubrum lacusprofundi (Franzmann et al., 1988), Halorubrum saccharovorum (Tomlinson & Hochstein, 1976), Halorubrum trapanicum, Halorubrum coriense (Kamekura & Dyall-Smith, 1995), Halorubrum distributum, Halorubrum vacuolatum (Kamekura et al., 1997; Grant & Larsen, 1989), Halorubrum tebenquichense (Lizama et al., 2002) and Halorubrum tibetense (Fan et al., 2004). Strain BD-1T was isolated from sludge samples of Xiao-Er-Kule saline lake in Xinjiang, China, by enrichment and subsequent plating of the enriched cultures on agar medium containing the following ingredients (l–1): Casamino acids, 1 g; yeast extract, 1 g; trisodium citrate, 3 g; NaCl, 200 g; KCl, 2 g; MgSO4.7H2O, 5 g; MnSO4.7H2O, 0·2 mg; FeSO4.7H2O, 0·05 g; glucose, 10 g (pH adjusted to 7·0 with 1 M NaOH solution). This strain grew over a temperature range of 10–54 °C (optimum 40 °C) as determined by using a temperature gradient incubator and over a pH range of 6·0–10·0 (optimum 7·0–7·5) as determined with various pH buffers. Routine cultivation was conducted at 40 °C and pH 7·5. The requirements for NaCl and magnesium for growth were determined in media with 0·9–5·2 M NaCl or 0–0·5 M MgSO4.
Phenotypic tests were performed according to the proposed minimal standards for the description of new taxa in the order Halobacteriales (Oren et al., 1997). Cell motility and morphology were examined by phase-contrast and transmission electron microscopy of exponentially growing liquid cultures. Gram staining was carried out as described by Dussault (1955). Colony morphology was observed on optimal growth agar medium after incubation at 40 °C for 4 days. Anaerobic growth was tested in the presence of 5 g potassium nitrate, L-arginine or DMSO l–1 in filled, stoppered tubes. Tests for catalase and oxidase activities and hydrolysis of starch and Tween 80 were performed as described previously (Gonzalez et al., 1978). Nitrate reduction, H2S formation, indole formation and the utilization of sugars, alcohols, amino acids and organic acids were examined as described by Oren et al. (1997). Phospholipids and glycolipids were separated on silica gel plates (10x10 cm) by TLC and were analysed according to Kamekura et al. (1997) and Xin et al. (2001).
DNA base composition was determined by thermal denaturation (Tm). The 16S rRNA gene sequence was amplified with archaeon-specific primers under conditions described previously (Zhang et al., 2003). 16S rRNA gene sequence alignments were performed by using CLUSTAL W version 1.3b (Van de Peer & De Wachter, 1994). A phylogenetic tree (Fig. 1) was constructed by the neighbour-joining method with Kimura's two-parameter calculation model in TREECON W version 1.3b. DNA–DNA hybridizations of strain BD-1T to the type strains of Hrr. trapanicum and Hrr. distributum were performed by thermal denaturation and the renaturation method of De Ley et al. (1970) as modified by Huß et al. (1983).
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). Cluster 1 contains Hrr. saccharovorum and Hrr. lacusprofundi (with similarity of 98·3 %). Cluster 2 contains Hrr. sodomense, Hrr. trapanicum, Hrr. coriense and Hrr. distributum (with similarity >97·7 %). Cluster 3 is phylogenetically distant from the above two clusters and contains Hrr. vacuolatum. The recent discovery of Hrr. tebenquichense (Lizama et al., 2002), Hrr. tibetense (Fan et al., 2004) and strain BD-1T (Hrr. xinjiangense sp. nov.) significantly reduced the bootstrap support (49 %) of clusters 1 and 3 (Fig. 1
), while cluster 2 is still supported by 100 % bootstrap. Strain BD-1T clearly belongs to cluster 2, and the closest relative was Hrr. trapanicum (98·4 %). The DNA–DNA relatedness of strain BD-1T to Hrr. trapanicum and Hrr. distributum was 47 and 24 %, respectively.
Description of Halorubrum xinjiangense sp. nov.
Halorubrum xinjiangense (xin.ji.ang.en'se. N.L. neut. adj. xinjiangense pertaining to Xinjiang, where the strain was isolated).
Cells are short rods (0·8–1·2x1·8–2·6 µm) and Gram-negative. Colonies on agar plates containing 25 % (w/v) total salts are red, elevated and round. Chemo-organotrophic and aerobic. Growth occurs at 2·0–5·2 M NaCl, pH 6–10 and 10–54 °C, with optimal growth at 3·1–3·4 M NaCl, pH 7·0–7·5 and 40 °C. Magnesium is not required for growth. Catalase and oxidase are positive. Anaerobic growth with nitrate, arginine and DMSO does not occur and nitrate reduction to nitrite or dinitrogen is not observed. H2S is produced from cysteine, but indole formation is negative. Tween 80 and starch are not hydrolysed. Amino acids are not required for growth. Fructose, glucose, sucrose and maltose are utilized with production of acids. No growth or acid production is observed on mannose, lactose, raffinose, rhamnose, galactose, D-ribose, glycerol, mannitol or sorbitol. Organic acids are not utilized. Cells contain C20C20 derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and sulfated mannosyl-glucosyl glycerol diether. The G+C content of DNA is 68·0 % (Tm).
The type strain is BD-1T (=AS 1.3527T=JCM 12388T), isolated from Xiao-Er-Kule Lake in Xinjiang, China.
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ACKNOWLEDGEMENTS |
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REFERENCES |
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TOPABSTRACTMAIN TEXTREFERENCES |
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Dussault, H. P. (1955). An improved technique for staining halophilic bacteria. J Bacteriol 70, 484–485.
Fan, H., Xue, Y., Ma, Y., Ventosa, A. & Grant, W. D. (2004). Halorubrum tibetense sp. nov., a novel haloalkaliphilic archaeon from Lake Zabuye in Tibet, China. Int J Syst Evol Microbiol 54, 1213–1216.
Franzmann, P. D., Stackebrandt, E., Sanderson, K., Valkman, K., Cameron, D. E., Stevenson, P. L., McMeekin, T. A. & Burton, H. R. (1988). Halobacterium lacusprofundi sp. nov., a halophilic bacterium isolated from Deep Lake, Antarctica. Syst Appl Microbiol 11, 20–27.
Gonzalez, C., Gutierrez, C. & Ramirez, C. (1978). Halobacterium vallismortis sp. nov. An amylolytic and carbohydrate-metabolizing, extremely halophilic bacterium. Can J Microbiol 24, 710–715.[Medline]
Grant, W. D. & Larsen, H. (1989). Group III. Extremely halophilic archaebacteria. Order Halobacteriales ord. nov. In Bergey's Manual of Systematic Bacteriology, vol. 3, pp. 2216–2219. Edited by J. T. Staley, M. P. Bryant, N. Pfennig & J. G. Holt. Baltimore: Williams & Wilkins.
Huß, V. A. R., Festl, H. & Schleifer, K. H. (1983). Studies on the spectrometric determination of DNA hybridization from renaturation rates. Syst Appl Microbiol 4, 184–192.
Kamekura, M. & Dyall-Smith, M. L. (1995). Taxonomy of the family Halobacteriaceae and the description of two new genera Halorubrobacterium and Natrialba. J Gen Appl Microbiol 41, 333–350.
Kamekura, M., Dyall-Smith, M. L., Upasani, V., Ventosa, A. & Kates, M. (1997). Diversity of alkaliphilic halobacteria: proposals for transfer of Natronobacterium vacuolatum, Natronobacterium magadii, and Natronobacterium pharaonis to Halorubrum, Natrialba, and Natronomonas gen. nov., respectively, as Halorubrum vacuolatum comb. nov., Natrialba magadii comb. nov., and Natronomonas pharaonis comb. nov., respectively. Int J Syst Bacteriol 47, 853–857.[CrossRef][Medline]
Lizama, C., Monteoliva-Sánchez, M., Suárez-García, A., Rosselló-Mora, R., Aguilera, M., Campos, V. & Ramos-Cormenzana, A. (2002). Halorubrum tebenquichense sp. nov, a novel halophilic archaeon isolated from the Atacama Saltern, Chile. Int J Syst Evol Microbiol 52, 149–155.[Abstract]
McGenity, T. J. & Grant, W. D. (1995). Transfer of Halobacterium saccharovorum, Halobacterium sodomense, Halobacterium trapanicum NRC 34021 and Halobacterium lacusprofundi to the genus Halorubrum gen. nov., as Halorubrum saccharovorum comb. nov., Halorubrum sodomense comb. nov., Halorubrum trapanicum comb. nov., and Halorubrum lacusprofundi comb. nov. Syst Appl Microbiol 18, 237–243.
Ochsenreiter, T., Pfeifer, F. & Schleper, C. (2002). Diversity of archaea in hypersaline environments characterized by molecular-phylogenetic and cultivation studies. Extremophiles 6, 267–274.[CrossRef][Medline]
Oren, A. (1983). Halobacterium sodomense sp. nov., a Dead Sea halobacterium with an extremely high magnesium requirement. Int J Syst Bacteriol 33, 381–386.[CrossRef]
Oren, A., Ventosa, A. & Grant, W. D. (1997). Proposed minimal standards for description of new taxa in the order Halobacteriales. Int J Syst Bacteriol 47, 233–238.[CrossRef]
Tomlinson, G. A. & Hochstein, L. I. (1976). Halobacterium saccharovorum sp. nov., a carbohydrate-metabolizing, extremely halophilic bacterium. Can J Microbiol 22, 587–591.[Medline]
Van de Peer, Y. & De Wachter, R. (1994). TREECON for Windows: a software package for the construction and drawing of evolutionary trees for Microsoft Windows environment. Comput Appl Biosci 10, 569–570.
Xin, H., Itoh, T., Zhou, P., Suzuki, K. & Nakase, T. (2001). Natronobacterium nitratireducens sp. nov., a haloalkaliphilic archaeon isolated from a soda lake in China. Int J Syst Evol Microbiol 51, 1825–1829.[Abstract]
Zhang, D., Yang, H., Zhang, W., Huang, Z. & Liu, S.-J. (2003). Rhodocista pekingensis sp. nov., a cyst-forming phototrophic bacterium from a municipal wastewater treatment plant. Int J Syst Evol Microbiol 53, 1111–1114.
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