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1 State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, 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
shuangjiang{at}hotmail.com
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ABSTRACT |
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2)-glucose glycerol diether (S2-DGD). The isolates were different in that strain ABH32T did not require magnesium and had strong ability to assimilate sugars for growth, whereas strain AX-7T required magnesium and did not assimilate glucose, sucrose or maltose for growth. Both strain ABH32T and strain AX-7T were most closely related to members of the genera Haloterrigena and Natrinema. On the basis of phenotypic and genotypic characteristics, especially their cellular polar lipids, the two isolates were assigned to the genus Haloterrigena. Moreover, the DNA relatedness values for strains ABH32T and AX-7T with respect to members of the genus Haloterrigena were found to be well below 70 %. Thus, strains ABH32T and AX-7T represent two novel species within the genus Haloterrigena, for which the names Haloterrigena longa sp. nov. (type strain ABH32T=CGMCC 1.5334T=JCM 13562T) and Haloterrigena limicola sp. nov. (type strain AX-7T=CGMCC 1.5333T=JCM 13563T) are proposed.
2)-glucose glycerol dietherThe GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of strains ABH32T and AX-7T are DQ367242 and DQ367241, respectively.
Thin-layer chromatograms showing the polar lipids of strains ABH32T and AX-7T are available as supplementary figures in IJSEM Online.
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as accommodating a single species, Haloterrigena turkmenica. Haloterrigena thermotolerans (Montalvo-Rodríguez et al., 2000) and Haloterrigena saccharevitans (Xu et al., 2005) were described subsequently. In phylogenetic trees based on 16S rRNA gene sequences, species of the genera Haloterrigena and Natrinema sometimes cluster together (Montalvo-Rodríguez et al., 2000; Xin et al., 2000; Tindall, 2003) depending on the selection of sequences used. However, there are differences in the polar lipid composition among species of these two genera. All type strains of the species of Haloterrigena possess mannose-2,6-disulfate (12)-glucose glycerol diether (S2-DGD), but lack phosphatidylglycerol sulfate (or contain levels below the limit of detection) (Ventosa et al., 1999; Montalvo-Rodríguez et al., 2000). The opposite is true for the species of Natrinema (McGenity et al., 1998; Xin et al., 2000), and two strains of Htg. turkmenica, JCM 9743 and strain GSL-11, have been shown to possess glycolipids other than S2-DGD (Ventosa et al., 1999). Thus, on the basis of a combination of phylogenetic, morphological and chemotaxonomic characteristics, the currently described species (represented by their type strains) of these genera can be distinguished from each other. During our surveys on the biodiversity of the halophilic archaea of Aibi salt lake in the Xin-Jiang region of China (Cui et al., 2006), we obtained two strains that showed the same polar lipid pattern as that of members of the genus Haloterrigena. In this paper, we describe the characterization and the taxonomy of the two strains, ABH32T and strain AX-7T. The results indicated that these two stains represent two novel species of the genus Haloterrigena.
Strain ABH32T and strain AX-7T were isolated from sediment of Aibi salt lake. The isolation procedure was the same as that described previously (Xin et al., 2000; Feng et al., 2004, 2005; Xu et al., 2005). The characterization of strains ABH32T and AX-7T was performed according to the minimal standards proposed for the description of novel taxa of the order Halobacteriales (Oren et al., 1997). Morphological observation, growth characteristic tests, miscellaneous biochemical tests, nutrition tests, sensitivity to antimicrobial agents, lipid analysis and nucleic acid characterization were performed as described or cited by Xu et al. (2005).
The cells of strains ABH32T and AX-7T are rods, Gram-negative and able to grow in a wide range of salinities (1.7–5.1 M NaCl). Colonies on agar medium are red-pigmented. Strain ABH32T is different from strain AX-7T in that the former does not require magnesium, is unable to reduce nitrate and is able to assimilate sugars (glucose, sucrose and maltose). Detailed results of morphological analyses, assessments of growth characteristics, nutritional experiments, antibiotic-sensitivity determinations and miscellaneous biochemical tests conducted with strains ABH32T and AX-7T are given in the species descriptions. The physiological and biochemical differences among strain ABH32T, strain AX-7T and other members of the genus Haloterrigena are listed in Table 1.
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) of strain ABH32T and strain AX-7T were 63.2 and 61.9 mol%, respectively.
The 16S rRNA gene sequence similarity between strain ABH32T and strain AX-7T was 94.0 %. Phylogenetic analysis based on 16S rRNA gene similarities showed that strains AX-7T and ABH32T are closely related to Htg. thermotolerans PRST (97.0 and 95.4 % similarity, respectively), Htg. saccharevitans AB14T (96.2 and 94.7 %), Htg. turkmenica VKM B-1734T (96.0 and 93.9 %), Natrinema pallidum NCIMB 777T (96.4 and 95.1 %), Natrinema versiforme XF10T (96.2 and 95.3 %), Natrinema pellirubrum NCIMB 786T (96.0 and 94.7 %) and Natrinema altunense AJ2T (96.0 and 94.6 %). A phylogenetic tree constructed on the basis of the 16S rRNA gene showed that neither strain ABH32T nor strain AX-7T clustered with Natrinema; they formed a loose group with other members of the genus Haloterrigena (Fig. 1). As strains ABH32T and AX-7T contain bis-sulfated diglycosyl diether, a polar lipid characteristic of the genus Haloterrigena, we concluded that these strains should be classified within the genus Haloterrigena.
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as modified by Huß et al. (1983). The DNA–DNA relatedness values for strain ABH32T and strain AX-7T with respect to Htg. turkmenica JCM 8891T, Htg. thermotolerans CGMCC 1.3709T and Htg. saccharevitans CGMCC 1.3730T were 43.1, 38.2, 40.3 % and 42.5, 45.9 and 43.0 %, respectively. The DNA–DNA relatedness between strain ABH32T and strain AX-7T was 32.3 %. On the basis of the phenotypic and genotypic data, it is concluded that strain ABH32T and strain AX-7T represent two novel species of the genus Haloterrigena, for which the names Haloterrigena longa sp. nov. and Haloterrigena limicola sp. nov., respectively, are proposed.
Description of Haloterrigena longa sp. nov.
Haloterrigena longa (lon'ga. L. fem. adj. longa long, referring to the production of long rods in liquid medium).
Cells are non-motile, rod-shaped (0.5–0.6x2.8–11 µm) and Gram-negative. Colonies on agar plates containing 3.1 M NaCl are red, elevated and round. Chemo-organotrophic and aerobic. Growth occurs at NaCl concentrations of 1.7–5.1 M, at Mg2+ concentrations of 0–1.0 M, at pH values in the range 6.5–9.0 and at temperatures of 30–56 °C. The optimal NaCl and Mg2+ concentrations for growth are 3.1 M and 0.1 M, respectively. Optimal pH and temperature values are 7.0–7.5 and 41–45 °C, respectively. Catalase- and oxidase-positive. Anaerobic growth with nitrate, arginine or DMSO does not occur. Nitrate reduction to nitrite is not observed. H2S is produced from Na2S2O3. Positive for indole formation. Tweens 20, 40 and 60 are hydrolysed but Tween 80 is not. Negative for caseinase, amylase and gelatinase. The following substrates are utilized as carbon sources: glucose, sucrose, maltose, pyruvate and acetate. Fructose, sorbose, xylose, galactose, mannose, D-ribose, lactose, mannitol and sorbitol are not utilized as carbon sources. Acid is produced in unbuffered medium from glucose and sucrose. Sensitive to rifampicin (5 µg per disc) and novobiocin (30 µg per disc). Resistant to the following antibiotics (µg per disc, unless otherwise indicated): erythromycin (15), neomycin (30), chloramphenicol (30), ampicillin (10), norfloxacin (10), ciprofloxacin (5), streptomycin (10), kanamycin (30), tetracycline (30), vancomycin (30), bacitracin (0.04 IU per disc) and penicillin G (10 IU per disc). The major polar lipids are the C20C20 and C20C25 derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and S2-DGD. The DNA G+C content of the type strain is 63.2 mol% (Tm).
The type strain, strain ABH32T (=CGMCC 1.5334T=JCM 13562T), was isolated from Aibi salt lake in Xin-Jiang, China.
Description of Haloterrigena limicola sp. nov.
Haloterrigena limicola (li.mi'co.la. L. n. limus mud; L. suff. -cola from L. n. incola dweller; N.L. masc. or fem. n. limicola mud-dweller).
Cells are motile, rod-shaped (0.6–0.8x1.8–3.6 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 1.7–5.1 M, at Mg2+ concentrations of 0.005–0.7 M, at pH values in the range 6.5–9.0 and at temperatures of 30–61 °C. Optimal NaCl concentration, Mg2+ concentration, pH and temperature for growth are 3.1 M, 0.05 M, pH 7.0 and 45–50 °C, respectively. Catalase- and oxidase-positive. Anaerobic growth with nitrate, arginine or DMSO does not occur. Nitrate reduction to nitrite is observed. H2S is produced from Na2S2O3. Negative for indole formation. Tween 20 is hydrolysed, but Tweens 40, 60 and 80 are not. Negative for caseinase, amylase and gelatinase. Glucose, sucrose, maltose, fructose, sorbose, xylose, galactose, mannose, D-ribose, lactose, mannitol and sorbitol are not utilized as carbon sources. Pyruvate and acetate are utilized for growth. Sensitive to rifampicin (5 µg per disc) and novobiocin (30 µg per disc). Resistant to the following antibiotics (µg per disc, unless otherwise indicated): erythromycin (15), neomycin (30), chloramphenicol (30), ampicillin (10), norfloxacin (10), ciprofloxacin (5), streptomycin (10), kanamycin (30), tetracycline (30), vancomycin (30), bacitracin (0.04 IU per disc) and penicillin G (10 IU per disc). The major polar lipids are the C20C20 and C20C25 derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and S2-DGD. The DNA G+C content of the type strain is 61.9 mol% (Tm).
The type strain, strain AX-7T (=CGMCC 1.5333T=JCM 13563T), was isolated from Aibi salt lake in Xin-Jiang, China.
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ACKNOWLEDGEMENTS |
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