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1 College of Life Sciences, Zhejiang University, Hangzhou 310058, People's Republic of China
2 Altun Mountain National Nature Reserve Administrative, Kuerle 841000, People's Republic of China
Correspondence
Min Wu
wumin{at}zju.edu.cn
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ABSTRACT |
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Results of one-dimensional TLC of polar lipids of Hrr. saccharovorum CGMCC 1.2147T and strain AJ201T are available as supplementary material in IJSEM Online.
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; Kamekura & Dyall-Smith, 1995) are widely distributed in hypersaline habitats (Grant & Larsen, 2001). At the time of writing, there are 16 species with validly published names in the genus Halorubrum, including three alkaliphilic species, Halorubrum vacuolatum (Mwatha & Grant, 1993; Kamekura et al., 1997), Hrr. tibetense (Fan et al., 2004) and Hrr. alkaliphilum (Feng et al., 2005), and 13 neutrophilic species, Halorubrum saccharovorum (the type species; Tomlinson & Hochstein, 1976), Hrr. sodomense (Oren, 1983), Hrr. lacusprofundi (Franzmann et al., 1988), Hrr. coriense (Nuttall & Dyall-Smith, 1993), Hrr. trapanicum (McGenity & Grant, 1995; Trüper, 2003), Hrr. distributum (Zvyagintseva & Tarasov, 1987; Oren & Ventosa, 1996), Hrr. tebenquichense (Lizama et al., 2002), Hrr. terrestre (Ventosa et al., 2004), Hrr. xinjiangense (Feng et al., 2004), Hrr. ezzemoulense (Kharroub et al., 2006), Hrr. orientale (Castillo et al., 2006), Hrr. lipolyticum (Cui et al., 2006) and Hrr. aidingense (Cui et al., 2006). Here, we report the taxonomic characterization of a neutrophilic strain belonging to the genus Halorubrum.
Strain AJ201T was isolated from mud of Ayakekum salt lake (37° 37' N 89° 29' E; 3884 m altitude) located in the Altun Mountain, in the northern part of the Qinghai–Tibet Plateau. The lake environment was described previously in detail (Xu et al., 2005a). The sample was incubated with S-G medium (Sehgal & Gibbons, 1960) for approximately 1 week and the liquid was plated by using a tenfold dilution series method. After 1–2 weeks of incubation at 37 °C, a representative red colony was picked and maintained on S-G medium. The strain was purified by repeated restreaking; purity was confirmed by the uniformity of cell morphology. The optimal conditions for growth were determined in S-G medium modified with 0.85–5.1 M NaCl or 0–1.0 M Mg2+.
Phenotypic tests and lipid analysis were performed according to Xu et al. (2005b) as mentioned previously in the minimal standards for the description of new taxa in the order Halobacteriales (Oren et al., 1997). Detailed results of cell morphology, physiological tests and antibiotic sensitivity are given in the species description. The major polar lipids of strain AJ201T were determined to be C20C20 derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and sulfated mannosyl-glucosyl-glycerol diether (Supplementary Fig. S1 in IJSEM Online).
The 16S rRNA gene was amplified and analysed as described previously (Xu et al., 2005b). The almost-complete 16S rRNA gene sequence of strain AJ201T (1470 nt; nucleotide positions 6–1540 in the Escherichia coli numbering) showed 92.9–97.5 % similarity to those of the type strains of Halorubrum species. A phylogenetic tree was constructed by the neighbour-joining method with the MEGA3 program package (Kumar et al., 2004), after multiple alignment of the data by CLUSTAL_X (Thompson et al., 1997). The neighbour-joining tree (Fig. 1) showed that strain AJ201T clustered with the neutrophilic Halorubrum species. The DNA G+C content of strain AJ201T determined by thermal denaturation (Tm) (Marmur & Doty, 1962) was 65.7±0.5 mol%. DNA–DNA hybridizations were carried out in 2x SSC at 80 °C by following the procedure of De Ley et al. (1970), as modified by Huß et al. (1983), using a Beckman Coulter DU800 spectrophotometer. DNA–DNA hybridization experiments (Table 1) showed hybridization values between strain AJ201T and closely related species of the genus Halorubrum ranging between 14 and 46 %, thus indicating that this strain represents a novel species of the genus Halorubrum.
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as well as 16S rRNA gene sequence analysis and DNA–DNA hybridization data justify the creation of a novel species, for which we propose the name Halorubrum arcis sp. nov.
Description of Halorubrum arcis sp. nov.
Halorubrum arcis (ar'cis. L. gen. n. arcis of a height, summit or peak, referring to the isolation of the type strain from a saline lake on the Qinghai–Tibet Plateau).
Gram-negative and motile. Young cultures show rod-like cells (0.5–1x2–4 µm). Colonies on complex agar medium are 1–2 mm in diameter, smooth, circular, elevated and red. Halophilic. Cells lyse immediately in distilled water. NaCl concentration required for growth is 2.2–5.2 M, with an optimum at 3.4–3.9 M. Magnesium is not required for growth, and the optimum Mg2+ concentration is 20–200 mM. The pH and temperature for growth are pH 6.0–8.5 (optimum pH 7.5) and 25–55 °C (optimum 42 °C). Chemo-organotrophic and aerobic. Oxidase- and catalase-positive. Anaerobic growth does not occur with arginine or DMSO. Nitrate is reduced to nitrite. H2S is produced from thiosulfate. Starch is hydrolysed. Tween 80 and casein are not hydrolysed. Gelatin is not liquefied. Casamino acids are required for growth. The following substrates are utilized for growth: glucose, maltose, glycerol and starch. No growth is observed on arabinose, fructose, galactose, lactose, mannitol, mannose, rhamnose, ribose, sorbitol, sucrose, xylose, alanine, arginine, aspartate, glycine, glutamate, lysine, ornithine, acetate, citrate, fumarate, lactate, malate, propionate, pyruvate or succinate. Acid is produced from glucose, maltose and glycerol. Sensitive to novobiocin, bacitracin, anisomycin and rifampicin, but not to ampicillin, chloramphenicol, erythromycin, neomycin or penicillin. The major polar lipids are C20C20 derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and sulfated mannosyl-glucosyl-glycerol diether. The DNA G+C content of the type strain is 65.7±0.5 mol% (Tm).
The type strain is strain AJ201T (=CGMCC 1.5343T=JCM 13916T), isolated from a salt lake on the Qinghai–Tibet Plateau.
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ACKNOWLEDGEMENTS |
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REFERENCES |
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TOPABSTRACTMAIN TEXTREFERENCES |
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Cui, H.-L., Tohty, D., Zhou, P.-J. & Liu, S.-J. (2006). Halorubrum lipolyticum sp. nov. and Halorubrum aidingense sp. nov., isolated from two salt lakes in Xin-Jiang, China. Int J Syst Evol Microbiol 56, 1631–1634.
De Ley, J., Cattoir, H. & Reynaerts, A. (1970). The quantitative measurement of DNA hybridization from renaturation rates. Eur J Biochem 12, 133–142.[Medline]
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.
Feng, J., Zhou, P.-J. & Liu, S.-J. (2004). Halorubrum xinjiangense sp. nov., a novel halophile isolated from saline lakes in China. Int J Syst Evol Microbiol 54, 1789–1791.
Feng, J., Zhou, P., Zhou, Y.-G., Liu, S.-J. & Warren-Rhodes, K. (2005). Halorubrum alkaliphilum sp. nov., a novel haloalkaliphile isolated from a soda lake in Xin-Jiang, China. Int J Syst Evol Microbiol 55, 149–152.
Franzmann, P. D., Stackebrandt, E., Sanderson, K., Volkman, J. 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.
Grant, W. D. & Larsen, H. (2001). Genus VII. Halorubrum. In Bergey's Manual of Systematic Bacteriology, 2nd edn, vol. 1, pp. 320–324. Edited by D. R. Boone, R. W. Castenholz & G. M. Garrity. New York: Springer.
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Kharroub, K., Quesada, T., Ferrer, R., Fuentes, S., Aguilera, M., Boulahrouf, A., Ramos-Cormenzana, A. & Monteoliva-Sánchez, M. (2006). Halorubrum ezzemoulense sp. nov., a halophilic archaeon isolated from Ezzemoul sabkha, Algeria. Int J Syst Evol Microbiol 56, 1583–1588.
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60 % are shown. Bar, 0.05 substitutions per nucleotide position.