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1 State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, ZhongGuanCun, Haidian, Beijing 100080, People's Republic of China
2 Graduate University of Chinese Academy of Sciences, Beijing 100049, People's Republic of China
3 College of Life Sciences, Xin-Jiang Normal University, Urumqi 830053, People's Republic of China
Correspondence
Shuang-Jiang Liu
liusj{at}sun.im.ac.cn
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ABSTRACT |
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A thin-layer chromatogram showing the glycolipids/phospholipids of strains 9-3T and 31-hongT and members of the genus Halorubrum is available as a supplementary figure in IJSEM Online.
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), Hrr. saccharovorum (Tomlinson & Hochstein, 1976), Hrr. coriense (Kamekura & Dyall-Smith, 1995; McGenity & Grant, 1995), Hrr. distributum (Zvyagintseva & Tarasov, 1987; Oren & Ventosa, 1996), Hrr. sodomense (Oren, 1983; Oren & Ventosa, 1996), Hrr. tebenquichense (Lizama et al., 2002), Hrr. terrestre (Ventosa et al., 2004), Hrr. trapanicum (McGenity & Grant, 1995), Hrr. xinjiangense (Feng et al., 2004), Hrr. alkaliphilum (Feng et al., 2005), Hrr. tibetense (Fan et al., 2004) and Hrr. vacuolatum (Mwatha & Grant, 1993; Kamekura et al., 1997). A phylogenetic tree based on the 16S rRNA gene sequences of these species indicates that the 12 species belonged to three distinct clusters (Fig. 1). Cluster 1 contains neutrophilic species, including Hrr. coriense, Hrr. distributum, Hrr. sodomense, Hrr. tebenquichense, Hrr. terrestre, Hrr. trapanicum and Hrr. xinjiangense. Cluster 2 consists of two other neutrophilic species, Hrr. lacusprofundi and Hrr. saccharovorum. Cluster 3 contains three alkaliphilic species, Hrr. alkaliphilum, Hrr. tibetense and Hrr. vacuolatum. During our surveys on the halophilic archaeal diversity of two salt lakes in the Xin-Jiang region of China (Cui et al., 2006), two strains that clustered tightly with members of cluster 2 were obtained. In this paper, we present the taxonomy of the two strains, strains 9-3T and 31-hongT.
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; Feng et al., 2004). Characterization of strains 9-3T and 31-hongT was performed according to the proposed minimal standards for the description of novel taxa in the order Halobacteriales (Oren et al., 1997). The morphology and growth characteristics were assessed, miscellaneous biochemical and nutrition tests were performed, sensitivity to antimicrobial agents was determined, lipids were analysed and nucleic acid characterization and DNA–DNA hybridization (thermal denaturation and renaturation method) were performed as described and cited by Feng et al. (2004, 2005).
The cells of strains 9-3T and 31-hongT are motile, Gram-negative rods. Both strains are able to grow in a wide range of salinities (1.7–4.3 M salt; optimal growth at 2.6 M). The cells lyse in distilled water. Colonies on agar medium are red-pigmented. Details of the morphology, growth characteristics, nutrition, antibiotic sensitivity and biochemistry of strains 9-3T and 31-hongT are given in the species descriptions. Differences between strains 9-3T and 31-hongT and other members of the genus Halorubrum are shown in Table 1.
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Polar lipid analysis indicated that both strains contained C20–C20 derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and phosphatidylglycerol sulfate. Glycolipids of strain 9-3T migrated with the same RF values as those of Hrr. saccharovorum and Hrr. lacusprofundi, but not those of strain 31-hongT (see Supplementary Fig. S1 available in IJSEM Online). The DNA G+C contents of strains 9-3T and 31-hongT are 65.9 and 64.2 mol% (Tm), respectively. The DNA–DNA relatedness values for strain 9-3T with respect to Hrr. saccharovorum JCM 8865T and Hrr. lacusprofundi JCM 8891T were 51.6 and 25.1 %, and those for strain 31-hongT with respect to Hrr. saccharovorum JCM 8865T and Hrr. lacusprofundi JCM 8891T were 29.4 and 44.9 %, respectively. The DNA–DNA relatedness between strains 9-3T and 31-hongT was 54 %.
On the basis of these results, it is concluded that strains 9-3T and 31-hongT represent two novel species of the genus Halorubrum, for which the names Halorubrum lipolyticum sp. nov. and Halorubrum aidingense sp. nov. are proposed.
Description of Halorubrum lipolyticum sp. nov.
Halorubrum lipolyticum (li.po.ly'ti.cum. Gr. n. lipos fat; N.L. neut. adj. lyticum from Gr. adj. lutikos able to loosen, able to dissolve; N.L. neut. adj. lipolyticum fat-dissolving, referring to the property of being able to hydrolyse lipids).
Cells are motile, rod-shaped (0.9–1.2x1.8–5.5 µm) and Gram-negative. Colonies on agar plates containing 2.6 M NaCl are red, elevated and round. Chemo-organotrophic and aerobic. Growth occurs at NaCl concentrations of 1.7–4.8 M, at pH 7.0–9.0 and temperatures of 25–58 °C. Optimal NaCl concentration, pH and temperature for growth are 2.6 M, pH 7.5 and 45–48 °C, respectively. Catalase- and oxidase-positive. Anaerobic growth with nitrate, arginine or DMSO does not occur. Nitrate reduction to nitrite is observed. H2S is not produced from Na2S2O3. Negative for indole formation. Tweens 20, 40, 60 and 80 are hydrolysed. Caseinase-negative. Amylase- and gelatinase-positive. Glucose, fructose, galactose, mannose, sucrose and maltose are utilized and acids are produced. No growth or acid production is observed on sorbose, xylose, lactose, D-ribose, mannitol or sorbitol. Sensitive to rifampicin, bacitracin and novobiocin. Resistant to erythromycin, neomycin, chloramphenicol, ampicillin, penicillin G, norfloxacin, ciprofloxacin, streptomycin, kanamycin, tetracycline and vancomycin. Cells contain C20–C20 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 65.9 mol% (Tm).
The type strain, 9-3T (=CGMCC 1.5332T=JCM 13559T), was isolated from Aibi salt lake in Xin-Jiang, China.
Description of Halorubrum aidingense sp. nov.
Halorubrum aidingense (ai.ding.en'se. N.L. neut. adj. aidingense from Aiding salt lake in China, where the type strain was isolated).
Cells are motile, rod-shaped (0.9–1.0x3.6–6.4 µm) and Gram-negative. Colonies on agar plates containing 2.6 M NaCl are red, elevated and round. Chemo-organotrophic and aerobic. Growth occurs at NaCl concentrations of 1.7–4.3 M, at pH 7.0–9.0 and at temperatures of 25–52 °C. Optimal NaCl concentration, pH and temperature for growth are 2.6 M, pH 7.5 and 40–42 °C, respectively. Catalase- and oxidase-positive. Anaerobic growth with nitrate, arginine or DMSO does not occur. Nitrate reduction to nitrite is observed. H2S is not produced from Na2S2O3. Positive for indole formation. Tweens 20, 40, 60 and 80 are hydrolysed. Casein and starch are not hydrolysed. Gelatin is not liquefied. Glucose, fructose, galactose, mannose, sucrose, maltose and mannitol are utilized and acids are produced. No growth or acid production is observed on sorbose, xylose, lactose, D-ribose or sorbitol. Sensitive to rifampicin, bacitracin and novobiocin. Resistant to erythromycin, neomycin, chloramphenicol, ampicillin, penicillin G, norfloxacin, ciprofloxacin, streptomycin, kanamycin, tetracycline and vancomycin. Cells contain C20–C20 derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate and phosphatidylglycerol phosphate sulfate. Sulfated mannosyl-glucosyl glycerol diether was not detected. The DNA G+C content of the type strain is 64.2 mol% (Tm).
The type strain, 31-hongT (=CGMCC 1.2670T=JCM 13560T), was isolated from Aiding salt lake in Xin-Jiang, China.
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ACKNOWLEDGEMENTS |
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REFERENCES |
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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. & 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., Liu, S.-J. & Warren-Rhodes, K. (2005). Halorubrum alkaliphilum sp. nov., a novel haloalkaliphile isolated from a soda lake in Xinjiang, 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.
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.[CrossRef]
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.
Lizama, C., Monteoliva-Sanchez, M., Suarez-Garcia, A., Rossello-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.
McGenity, T. J. & Grant, W. D. (2001). Genus VII. Halorubrum McGenity and Grant 1996, 362VP. 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.
Mwatha, W. E. & Grant, W. D. (1993). Natronobacterium vacuolata sp. nov., a haloalkaliphilic archaeon isolated from Lake Magadi, Kenya. Int J Syst Bacteriol 43, 401–404.
Oren, A. (1983). Halobacterium sodomense sp. nov., a Dead Sea halobacterium with an extremely high magnesium requirement. Int J Syst Bacteriol 33, 381–386.
Oren, A. & Ventosa, A. (1996). A proposal for the transfer of Halorubrobacterium distributum and Halorubrobacterium coriense to the genus Halorubrum as Halorubrum distributum comb. nov. and Halorubrum coriense comb. nov., respectively. Int J Syst Bacteriol 46, 1180.
Oren, A., Ventosa, A. & Grant, W. D. (1997). Proposal of minimal standards for the description of new taxa in the order Halobacteriales. Int J Syst Bacteriol 47, 233–238.
Tomlinson, G. A. & Hochstein, L. I. (1976). Halobacterium saccharovorum sp. nov., a carbohydrate-metabolizing, extremely halophilic bacterium. Can J Microbiol 22, 587–591.[Medline]
Ventosa, A., Gutiérrez, M. C., Kamekura, M., Zvyagintseva, I. S. & Oren, A. (2004). Taxonomic study of Halorubrum distributum and proposal of Halorubrum terrestre sp. nov. Int J Syst Evol Microbiol 54, 389–392.
Xin, H., Itoh, T., Zhou, P., Suzuki, K., Kamekura, M. & Nakase, T. (2000). Natrinema versiforme sp. nov., an extremely halophilic archaeon from Aibi salt lake, Xinjiang, China. Int J Syst Evol Microbiol 50, 1297–1303.[Abstract]
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