Phosphatidylglycerol Phosphate Research Articles

Halorubrum halodurans sp. nov., an extremely halophilic archaeon isolated from a hypersaline lake.

Two extremely halophilic archaea, strains Cb34T and C170, belonging to the genus Halorubrum, were isolated from the brine of the hypersaline lake Aran-Bidgol in Iran. Cells of the two strains were motile, pleomorphic rods, stained Gram-variable and produced red-pigmented colonies. Strains Cb34T and C170 required 25 % (w/v) salts, pH 7.0 and 37 °C for optimal growth under aerobic conditions; 0.3 M Mg2+ was required. Cells of both isolates were lysed in distilled water and hypotonic treatment with < 10 % NaCl provoked cell lysis. Phylogenetic analysis based on 16S rRNA gene sequence similarities showed that these two strains were closely related to Halorubrum cibi B31T (98.8 %) and other members of the genus Halorubrum. In addition, studies based on the rpoB' gene revealed that strains Cb34T and C170 are placed among the species of Halorubrum and are closely related to Halorubrum cibi B31T, with rpoB' gene sequence similarity less than or equal to 95.7 %. The polar lipid patterns of both strains consisted of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and sulfated mannosyl glucosyl diether. The DNA G+C content was 62.1-62.4 mol%. DNA-DNA hybridization studies confirmed that strains Cb34T and C170 constitute a distinct species. Data obtained in this study show that the two strains represent a novel species, for which the name Halorubrum halodurans sp. nov. is proposed. The type strain is Cb34T ( = CECT 8745T = IBRC-M 10233T).

Halobellus ramosii sp. nov., an extremely halophilic archaeon isolated from a saline-wetland wildfowl reserve.

An extremely halophilic archaeon, strain S2FP14T, was isolated from a brine sample from the inland hypersaline lake Fuente de Piedra, a saline-wetland wildfowl reserve located in the province of Málaga in southern Spain. Colonies were red-pigmented and the cells were Gram-staining-negative, motile and pleomorphic. S2FP14T was able to grow in media containing 12.5-30 % (w/v) total salts (optimum 20 %) at pH 7-8.5 (optimum 7.5) and at 25-50 °C (optimum 37 °C). The 16S rRNA gene sequence analysis indicated that this strain represented a member of the genus Halobellus. S2FP14T showed a similarity of 99.5 % to Halobellus inordinatus YC20T, 96.1 % to Halobellus litoreus GX31T, 95.9 % to Halobellus limi TBN53T, 95.5 % to Halobellus rarus YC21T, 95.2 % to Halobellus rufus CBA1103T, 94.6 % to Halobellus salinus CSW2.24.4T and 94.6 % to Halobellus clavatus TNN18T. The rpoB' gene sequence similarity of strain S2FP14T was 97.4 % to 87.6 % with members of genus Halobellus. The major phospholipids of strain S2FP14T were phosphatidylglycerol phosphate methyl ester and phosphatidylglycerosulfate, plus a very small amount of phosphatidylglycerol and an archaeal analogue of bisphosphatidylglycerol. With regard to glycolipid composition, the most abundant glycolipids were the sulfated diglycosyl diphytanilglyceroldiether and a glycosyl-cardiolipin. The G+C content of strain S2FP14T genomic DNA was 61.4 mol%. The DNA-DNA hybridization between strain S2FP14T and Halobellus inordinatus JCM 18361T was 51 %. Based on the phylogenetic, phenotypic and chemotaxonomic features, a novel species, Halobellus ramosii sp. nov. is proposed. The type strain is S2FP14T ( = CECT 8167T = DSM 26177T).

Lipidomics analysis of follicular fluid by ESI-MS reveals potential biomarkers for ovarian endometriosis.

The aim of the present study was to analyze the lipid profile of follicular fluid from patients with endometriosis and endometrioma who underwent in vitro fertilization treatment (IVF). The control group (n = 10) was composed of women with tubal factor or minimal male factor infertility who had positive pregnancy outcomes after IVF. The endometriosis group consisted of women with endometriosis diagnosed by videolaparoscopy (n = 10), and from the same patients, the endometriomas fluids were collected, which composed the endometrioma group (n = 10). From the follicular fluid and endometriomas, lipids were extracted by the Bligh and Dyer method, and the samples were analyzed by tandem mass spectrometry. We observed phosphatidylglycerol phosphate, phosphatidylcholine, phosphatidylserine, and phosphatidylnositol bisphosphate in the control group. In the endometriosis group, sphingolipids and phosphatidylcholines were more abundant, while in the endometrioma group, sphingolipids and phosphatidylcholines with different m/z from the endometriosis group were found in high abundance. This analysis demonstrated that there is a differential representation of these lipids according to their respective groups. In addition, the lipids found are involved in important mechanisms related to endometriosis progress in the ovary. Thus, the metabolomic approach for the study of lipids may be helpful in potential biomarker discovery.

Halorubrum rutilum sp. nov. isolated from a marine solar saltern.

A halophilic archaeal strain, YJ-18-S1(T), was isolated from Yangjiang marine solar saltern, Guangxi Province, China. Cells were pleomorphic, stained Gram-negative and formed red-pigmented colonies on agar plates. Strain YJ-18-S1(T) was able to grow at 20-55 °C (optimum 37 °C), at 0.9-4.8 M NaCl (optimum 2.6 M NaCl), at 0.005-1.0 M MgCl2 (optimum 0.3 MgCl2) and at pH 5.5-8.5 (optimum pH 7.0). The cells were lysed in distilled water, and the minimal NaCl concentration to prevent cell lysis was found to be 5 % (w/v). The major polar lipids of the strain were phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and sulfated mannosyl glucosyl diether. The 16S rRNA gene and rpoB' gene of strain YJ-18-S1(T) were phylogenetically related to the corresponding genes of Halorubrum members (94.3-98.0 and 86.7-96.1 % similarities, respectively). The DNA G+C content of strain YJ-18-S1(T) was 66.2 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain YJ-18-S1(T) (=CGMCC 1.12554(T) = JCM 30030(T)) represents a new species of Halorubrum, for which the name Halorubrum rutilum sp. nov. is proposed.

Halorussus amylolyticus sp. nov., isolated from an inland salt lake.

A halophilic archaeal strain, YC93T, was isolated from Yuncheng salt lake in Shanxi Province, China. Cells were pleomorphic rods, stained Gram-negative and formed light-red-pigmented colonies on agar plates. Strain YC93T was able to grow at 25–50 °C (optimum 37 °C), with 1.4–4.8 M NaCl (optimum 2.0 M), with 0–1.0 M MgCl2 (optimum 0.05 M) and at pH 6.0–9.5 (optimum pH 7.0). Cells lysed in distilled water and the minimal NaCl concentration to prevent cell lysis was 8 % (w/v). 16S rRNA gene sequence analysis revealed that strain YC93T had two dissimilar 16S rRNA genes both of which were phylogenetically related to those of the two recognized members of the genus Halorussus (93.0–95.3 % similarity). The rpoB′ gene of strain YC93T was phylogenetically related to the corresponding gene of Halorussus rarus TBN4T (91.3 % similarity) and Halorussus ruber YC25T (90.5 %). The major polar lipids were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and five glycolipids chromatographically identical to those of Halorussus rarus CGMCC 1.10122T. The DNA G+C content of strain YC93T was 64.6 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain YC93T represents a novel species of the genus Halorussus, for which the name Halorussus amylolyticus sp. nov. is proposed. The type strain is YC93T ( = CGMCC 1.12126T = JCM 18367T).

Halorientalis brevis sp. nov., Isolated from an Inland Salt Lake of China.

Halophilic archaeal strain YC89(T) was isolated from Yuncheng salt lake in Shanxi, China. Cells from strain YC89(T) were short rods, lysed in distilled water, stained Gram-negative and formed red-pigmented colonies on agar plate. Strain YC89(T) was able to grow at 25-50°C (optimum 37°C), at 1.4-4.8 M NaCl (optimum 2.6-3.1 M), at 0-1.0 M MgCl2 (optimum 0.3 M) and at pH 6.0-9.5 (optimum pH 7.5). The major polar lipids are phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, sulfated mannosyl glucosyl diether and two unknown glycolipids. 16S rRNA gene analysis revealed that strain YC89(T) was phylogenetically related to Halorientalis persicus D108(T) (95.6% nucleotide identity) and H. regularis TNN28(T) (95.3% nucleotide identity). The rpoB' gene similarities between strain YC89(T) and H. persicus IBRC-M 10043(T) and H. regularis TNN28(T) were 88.1 and 88.0%, respectively. The DNA G+C content of strain YC89(T) was determined to be 61.3 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain YC89(T) (=CGMCC 1.12125(T) = JCM 18366(T)) represents a new species of Halorientalis, for which the name H. brevis sp. nov. is proposed.

Haloarchaeobius amylolyticus sp. nov., isolated from a marine solar saltern.

Halophilic archaeal strain XD48(T) was isolated from a Chinese marine solar saltern. Cells were pleomorphic, stained Gram-negative and formed red-pigmented colonies on solid media. Strain XD48(T) was found to be able to grow at 25-50 °C (optimum 37 °C), at 0.9-4.8 M NaCl (optimum 3.1 M NaCl), at 0-1.0 M MgCl2 (optimum 0.3 MgCl2) and at pH 5.0-9.5 (optimum pH 6.5). The cells lysed in distilled water, and the minimal NaCl concentration to prevent cell lysis was found to be 5% (w/v). The major polar lipids of the strain were phosphatidic acid (PA), phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me), phosphatidylglycerol sulfate (PGS), sulfated galactosyl mannosyl glucosyl diether (S-TGD-1), sulfated mannosyl glucosyl diether (S-DGD-1) and six unknown glycolipids. The 16S rRNA gene and rpoB' gene of strain XD48(T) were phylogenetically related to the corresponding genes of Haloarchaeobius members (92.4-93.9 and 89.6-90.5% similarities, respectively). The DNA G + C content of strain XD48(T) was determined to be 65.3 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain XD48(T) (=CGMCC 1.12230(T) = JCM 18642(T)) represents a new species of Haloarchaeobius, for which the name Haloarchaeobius amylolyticus sp. nov. is proposed.

Halosimplex litoreum sp. nov., isolated from a marine solar saltern.

A halophilic archaeal strain, YGH94(T), was isolated from the Yinggehai marine solar saltern near the Shanya city of Hainan Province, China. Cells of the strain were observed to be short rods, stain Gram-negative and to form red-pigmented colonies on solid media. Strain YGH94(T) was found to grow at 25-50 °C (optimum 40 °C), at 0.9-4.8 M NaCl (optimum 3.1 M), at 0-1.0 M MgCl2 (optimum 0.05 M) and at pH 5.0-9.0 (optimum pH 7.5). The cells were found to lyse in distilled water and the minimal NaCl concentration to prevent cell-lysis was determined to be 5 % (w/v). The major polar lipids were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and four major glycolipids (disulfated mannosyl glucosyl diether, sulfated mannosyl glucosyl diether and two unidentified glycolipids chromatographically identical to glycolipids in Halosimplex carlsbadense JCM 11222(T)). Strain YGH94(T) was found to possess two heterogeneous 16S rRNA genes (rrnA and rrnB) and both are related to those of Hsx. carlsbadense JCM 11222(T) (92.7-98.6 % similarities), Halosimplex pelagicum R2(T) (94.6-99.2 % similarities) and Halosimplex rubrum R27(T) (92.9-98.8 % similarities). The rpoB' gene similarity between strain YGH94(T) and Hsx. carlsbadense JCM 11222(T), Hsx. pelagicum R2(T) and Hsx. rubrum R27(T) are 95.4, 94.9 and 95.1 %, respectively. The DNA G+C content of strain YGH94(T) was determined to be 64.0 mol%. Strain YGH94(T) showed low DNA-DNA relatedness (35-39 %) with the current three members of the genus Halosimplex. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain YGH94(T) (=CGMCC 1.12235(T) = JCM 18647(T)) represents a new species of the genus Halosimplex, for which the name Halosimplex litoreum sp. nov. is proposed.

Halovenus salina sp. nov., an extremely halophilic archaeon isolated from a saltern.

An extremely halophilic archaeon was isolated from a water sample of Isla Bacuta saltern in Huelva, Spain. Strain ASP54(T) is a novel red-pigmented, motile, rod-shaped, Gram-stain-negative and strictly aerobic haloarchaeon. Strain ASP54(T) grew in media containing 15-30% (w/v) salts and optimally with 25% (w/v) salts. It grew between pH 5.0 and 9.0 (optimally at pH 7.5) and at 20-40 °C (optimally at 37 °C). Phylogenetic analysis based on multi-locus sequence analysis (MLSA) and the comparison of 16S rRNA gene sequences revealed that strain ASP54(T) is most closely related to the genus Halovenus. The closest relatives were Halovenus aranensis EB27(T) (92.1% 16S rRNA gene sequence similarity), Halorientalis regularis TNN28(T) (92.1%), and Halorientalis persicus D108(T) (92.0%). The polar lipid pattern of strain ASP54(T) consisted of biphosphatidylglycerol, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, sulfated mannosyl glucosyl diether and a minor-phospholipid. The predominant respiratory quinone was menaquinone-8 (MK-8) (83%), and a minor amount of MK-8(VIII-H2) (17%) was also detected. The G+C content of the genomic DNA of this strain was 63.1 mol%. Based on the phenotypic, chemotaxonomic and phylogenetic data presented in this study, strain ASP54(T) represents a novel species of the genus Halovenus, for which the name Halovenus salina sp. nov. is proposed. The type strain is ASP54(T) ( = CEC(T) 8749(T) = IBRC-M 10946(T) = JCM 30072(T)).

Halovarius luteus gen. nov., sp. nov., an extremely halophilic archaeon from a salt lake.

An extremely halophilic archaeon, strain DA50T, was isolated from a brine sample of Urmia lake, a hypersaline environment in north-west Iran. Strain DA50T was orange-pigmented, motile, pleomorphic and required at least 2.5 M NaCl but not MgCl2 for growth. Optimal growth was achieved at 4.0 M NaCl and 0.3 M MgCl2. The optimum pH and temperature for growth were pH 7.0 and 45 °C, while it was able to grow over a pH range of 6.5-8.0 and a temperature range of 25-50 °C. Analysis of 16S rRNA gene sequences revealed that strain DA50T is a member of the family Halobacteriaceae, showing a low level of similarity with other members of this family. Highest similarities, 94.4, 94.0 and 93.9 %, were obtained with the 16S rRNA gene sequences of the type strains of Natrialba aegyptia, Halobiforma lacisalsi and Halovivax asiaticus, respectively. Polar lipid analyses revealed that strain DA50T contains phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester. Four unidentified glycolipids and two minor phospholipids were also observed. The only quinone present was MK-8(II-H2). The G+C content of its DNA was 62.3 mol%. On the basis of the data obtained, the new isolate could not be classified in any recognized genus. Strain DA50T is thus considered to represent a novel species of a new genus within the family Halobacteriaceae, order Halobacteriales, for which the name Halovarius luteus gen. nov., sp. nov. is proposed. The type strain of Halovarius luteus is DA50T ( = IBRC-M 10912T = CECT 8510T).

Halorubrum gandharaense sp. nov., an alkaliphilic haloarchaeon from commercial rock salt.

A Gram-stain-negative, non-motile, pleomorphic rod-shaped, orange-red-pigmented, facultatively aerobic and haloalkaliphilic archaeon, strain MK13-1T, was isolated from commercial rock salt imported from Pakistan. The NaCl, pH and temperature ranges for growth of strain MK13-1T were 3.0-5.2 M NaCl, pH 8.0-11.0 and 15-50 °C, respectively. Optimal growth occurred at 3.2-3.4 M NaCl, pH 9.0-9.5 and 45 °C. Addition of Mg2+ was not required for growth. The major polar lipids of the isolate were C20C20 and C20C25 archaeol derivatives of phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester. Glycolipids were not detected. The DNA G+C content was 64.1 mol%. The 16S rRNA gene sequence of strain MK13-1T was most closely related to those of the species of the genus Halorubrum, Halorubrum luteum CECT 7303T (95.9% similarity), Halorubrum alkaliphilum JCM 12358T (95.3%), Halorubrum kocurii JCM 14978T (95.3%) and Halorubrum lipolyticum JCM 13559T (95.3%). The rpoB' gene sequence of strain MK13-1T had < 90% sequence similarity to those of other members of the genus Halorubrum. Based on the phylogenetic analysis and phenotypic characterization, strain MK13-1T may represent a novel species of the genus Halorubrum, for which the name Halorubrum gandharaense sp. nov. is proposed, with the type strain MK13-1T ( = JCM 17823T = CECT 7963T).

Halorubrum persicum sp. nov., an extremely halophilic archaeon isolated from sediment of a hypersaline lake.

An extremely halophilic archaeon belonging to the genus Halorubrum, strain C49T, was isolated from sediment of the hypersaline lake Aran-Bidgol in Iran. Phylogenetic analysis based on 16S rRNA gene sequence similarities showed that strain C49T was closely related to Halorubrum saccharovorum JCM 8865T (99.5 %) and other species of the genus Halorubrum. Studies based on multilocus sequence analysis revealed that strain C49T is placed among the species of Halorubrum; the strain constituted a defined branch in comparison with the type strains of species of Halorubrum, while the 16S rRNA gene sequence divergence could not define the status of the newly isolated strain. For optimum growth, strain C49T required 20 % (w/v) salts at pH 7.0 and 37 °C under aerobic conditions. Mg2+ was not required. The cells were pleomorphic rods, motile and stained Gram-variable. Colonies of the strain were pink. Hypotonic treatment with <12 % NaCl provoked cell lysis. The polar lipid pattern of strain C49T consisted of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester derived from both C20C20 and C20C25 archaeol, phosphatidylglycerol sulfate and sulfated mannosyl glucosyl diether. The DNA G+C content was 64.2 mol%. DNA-DNA hybridization studies and average nucleotide identity confirmed that strain C49T constitutes a distinct genospecies. Data obtained in this study show that strain C49T represents a novel species, for which the name Halorubrum persicum sp. nov. is proposed. The type strain is C49T ( = IBRC-M 10232T = JCM 30541T).

Halomicroarcula salina sp. nov., isolated from a marine solar saltern.

Halophilic archaeal strain YGHS18(T) was isolated from the Yinggehai marine solar saltern near Shanya city of Hainan Province, China. Cells from the strain were observed to be pleomorphic rods, stained Gram-negative, and formed red-pigmented colonies on solid media. Strain YGHS18(T) was found to be able to grow at 20-50 °C (optimum 37 °C), with 0.9-4.8 M NaCl (optimum 2.1 M) and at pH 5.5-9.0 (optimum pH 7.0). The cells lysed in distilled water and the minimum NaCl concentration to prevent cell lysis was found to be 0.9 M. The major polar lipids of the strain were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, glucosyl mannosyl glucosyl diether and a diglycosyl diether (DGD-2). Strain YGHS18(T) possessed two heterogeneous 16S rRNA genes (rrnA and rrnB) and both were related to those of members of the genera Haloarcula (93.1-96.9% sequence similarity) and Halomicroarcula (92.7-96.1% similarity). The rrnA gene (orthologous gene) of strain YGHS18(T) clustered phylogenetically with members of the genus Halomicroarcula while the rrnB gene formed a paraphyly with members of the genera Halomicroarcula and Haloarcula . The rpoB' gene of strain YGHS18(T) was related phylogenetically to species of the genera Halomicroarcula (91.6-92.7% sequence similarity) and Haloarcula (91.5-92.4% similarity). EF-2 gene analysis revealed that strain YGHS18(T) was related phylogenetically to species of the genus Halomicroarcula (92.2-92.9% sequence similarity) rather than to those of the genus Haloarcula (90.9-91.7% similarity). The DNA G+C content of strain YGHS18(T) was determined to be 64.5 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strain YGHS18(T) ( =CGMCC 1.12128(T) =JCM 18369(T)) represents a novel species of the genus Halomicroarcula , for which the name Halomicroarcula salina sp. nov. is proposed.

Halocalculus aciditolerans gen. nov., sp. nov., an acid-tolerant haloarchaeon isolated from commercial salt.

Three halophilic archaeal strains, MH2-243-1(T), MH2-93-1 and MH2-91-1 were isolated from commercial salt samples from Japan, Australia, and Bolivia. Strain MH2-243-1(T) was able to grow in the presence of 12-30% (w/v) NaCl (optimum, 18% NaCl), at pH 4.5-7.0 (optimum, pH 6.0) and at 20-60 °C (optimum, 40 °C). Strains MH2-91-1 and MH2-93-1 grew in slightly different ranges. The orthologous 16S rRNA gene sequences of the three strains were almost identical (99.8-99.9% similarities), and the closest relative was Salarchaeum japonicum JCM 16327(T) with 94.2-94.3% 16S rRNA gene sequence similarities, followed by strains of members of the closely related genera Halobacterium and Halarchaeum . The RNA polymerase subunit B' gene (rpoB') sequence also showed the highest similarity (86.6%) to that of Salarchaeum japonicum JCM 16327(T). The DNA G+C contents of strains MH2-243-1(T), MH2-93-1 and MH2-91-1 were 68.5, 68.8 and 68.3 mol%, respectively. DNA-DNA relatedness values amongst the three strains were 97-99%. The polar lipids of the three strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, and at least seven unidentified glycolipids. The polar lipid composition differed from those of Salarchaeum japonicum and species of the genera Halobacterium and Halarchaeum . Based on the phenotypic and phylogenetic analyses, it is proposed that the isolates represent a novel species of a new genus, for which the name Halocalculus aciditolerans gen. nov., sp. nov. is proposed. The type strain of the type species is MH2-243-1(T) ( = JCM 19596(T) =KCTC 4149(T)) isolated from solar salt produced in Japan. MH2-93-1 ( = JCM 19595) and MH2-91-1 ( = JCM 19594) are additional strains of the type species.

Halococcus agarilyticus sp. nov., an agar-degrading haloarchaeon isolated from commercial salt.

Two agar-degrading halophilic archaeal strains, 62 E(T) and 197 A, were isolated from commercial salt samples. Cells were non-motile cocci, approximately 1.2-2.0 µm in diameter and stained Gram-negative. Colonies were pink-pigmented. Strain 62 E(T) was able to grow with 24-30% (w/v) NaCl (optimum, 27%), at pH 6.5-8.5 (optimum, pH 7.5) and at 22-47 °C (optimum, 42 °C). The 16S rRNA gene sequences of strains 62 E(T) and 197 A were identical, and the level of DNA-DNA relatedness between them was 90 and 90% (reciprocally). The closest relative was Halococcus saccharolyticus JCM 8878(T) with 99.7% similarity in 16S rRNA orthologous gene sequences, followed by Halococcus salifodinae JCM 9578(T) (99.6%), while similarities with other species of the genus Halococcus were equal to or lower than 95.1%. The rpoB' gene tree strongly supported that the two strains were members of the genus Halococcus . Mean DNA-DNA relatedness between strain 62 E(T) and H. saccharolyticus JCM 8878(T) and H. salifodinae JCM 9578(T) was 46 and 44%, respectively. The major polar lipids were archaeol derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, derived from both C20C20 and C20C25 archaeol, and sulfated diglycosyl archaeol-1. Several unidentified glycolipids were present. Based on the phenotypic and phylogenetic analyses, the isolates are considered to represent a novel species of the genus Halococcus , for which the name Halococcus agarilyticus sp. nov. is proposed. The type strain is 62 E(T) ( = JCM 19592(T) =KCTC 4143(T)).

Haloarchaeobius salinus sp. nov., isolated from an inland salt lake, and emended description of the genus Haloarchaeobius

The halophilic archaeal strain, YC82(T), was isolated from Yuncheng salt lake in Shanxi, PR China. Cells from strain YC82(T) were Gram-stain negative, pleomorphic rods, which lysed in distilled water and formed light-red colonies on solid media. Strain YC82(T) grew at 25-50 °C (optimum 37 °C), in 1.4-4.8 M NaCl (optimum 2.0 M), in 0-1.0 M MgCl2 (optimum 0.05 M) and at pH 6.0-9.5 (optimum pH 7.5). The major polar lipids of strain YC82(T) were phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and three glycolipids, which were chromatographically identical to those of Haloarchaeobius iranensis IBRC-M 10013(T) and Haloarchaeobius litoreus CGMCC 1.10390(T). 16S rRNA gene analysis revealed that strain YC82(T) had two dissimilar 16S rRNA genes and that it was phylogenetically related to Hab. iranensis IBRC-M 10013(T) (94.3-99.0 % nucleotide identity) and Hab. litoreus CGMCC 1.10390(T) (94.1-98.8 % nucleotide identity). The rpoB' gene similarities between strain YC82(T) and Hab. iranensis IBRC-M 10013(T) and Hab. litoreus CGMCC 1.10390(T) were 96.5 % and 95.7 %, respectively. The DNA G+C content of strain YC82(T) was 63.7 mol%. Strain YC82(T) showed low DNA-DNA relatedness with Hab. iranensis IBRC-M 10013(T) and Hab. litoreus CGMCC 1.10390(T). The phenotypic, chemotaxonomic and phylogenetic properties of strain YC82(T) ( = CGMCC 1.12232(T) = JCM 18644(T)) suggest that it represents a novel species of the genus Haloarchaeobius, for which the name Haloarchaeobius salinus sp. nov. is proposed. An emended description of the genus Haloarchaeobius is also presented.

Halostagnicola bangensis sp. nov., an alkaliphilic haloarchaeon from a soda lake.

An extremely haloalkaphilic archaeon, strain T26(T), belonging to the genus Halostagnicola, was isolated from sediment of the soda lake Bange in the region of Tibet, China. Phylogenetic analysis based on 16S rRNA gene sequence similarities showed that strain T26(T) was closely related to Halostagnicola alkaliphila 167-74(T) (98.4 %), Halostagnicola larsenii XH-48(T) (97.5 %) and Halostagnicola kamekurae 194-10(T) (96.8 %). Strain T26(T) grew optimally in media containing 25 % (w/v) salts, at pH 9.0 and 37 °C in aerobic conditions. Mg(2+) was not required for growth. The cells were motile, pleomorphic and Gram-stain-variable. Colonies of this strain were pink pigmented. Hypotonic treatment caused cell lysis. The polar lipids of the isolate consisted of C20C20 and C20C25 derivatives of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and minor phospholipids components. Glycolipids were not detected, in contrast to the two neutrophilic species of this genus. The genomic DNA G+C content of strain T26(T) was 60.1 mol% and DNA-DNA hybridization showed a relatedness of 19 and 17 % with Halostagnicola alkaliphila CECT 7631(T) and Halostagnicola larsenii CECT 7116(T), respectively. The comparison of 16S rRNA gene sequences, detailed phenotypic characterization, polar lipid profile and DNA-DNA hybridization studies revealed that strain T26(T) belongs to the genus Halostagnicola, and represents a novel species for which the name Halostagnicola bangensis sp. nov. is proposed. The type strain is T26(T) ( = CECT 8219(T) = IBRC-M 10759(T) = JCM 18750(T)).

Natribaculum breve gen. nov., sp. nov. and Natribaculum longum sp. nov., halophilic archaea isolated from saline soil.

Two halophilic archaeal strains, TRM20010(T) and TRM20345(T), were isolated from saline soil of the Lop Nur region in Xinjiang, north-west China. Cells from the two strains were pleomorphic rods, stained Gram-negative and produced red-pigmented colonies. Strains TRM20010(T) and TRM20345(T) were able to grow at 30-62 °C (optimum 37 °C), 0.9-5.1 M NaCl (optimum 2.6 and 3.4 M, respectively) and pH 6.0-10.0 (optimum pH 7.0-7.5) and neither strain required Mg(2+) for growth. The major polar lipids of the two strains were phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me), two glycolipids chromatographically identical to galactosyl mannosyl glucosyl diether (TGD-1) and disulfated mannosyl glucosyl diether (S2-DGD). Phylogenetic analysis based on 16S rRNA and rpoB' genes revealed that strains TRM20010(T) and TRM20345(T) clustered together and formed a distinct clade separated from the related genera Halovivax, Haloterrigena, Halostagnicola, Natronolimnobius and Natrinema. The DNA G+C contents of strains TRM20010(T) and TRM20345(T) were 63.9 and 63.8 mol%, respectively. The DNA-DNA hybridization value between strain TRM20010(T) and strain TRM20345(T) was 42.8 %. The phenotypic, chemotaxonomic and phylogenetic properties suggested that strains TRM20010(T) and TRM20345(T) represent two novel species in a new genus within the family Halobacteriaceae, for which the names Natribaculum breve gen. nov., sp. nov. (type strain TRM20010(T) = CCTCC AB2013112(T) = NRRL B-59996(T)) and Natribaculum longum sp. nov. (type strain TRM20345(T) = CCTCC AB2013113(T) = NRRL B-59997(T)) are proposed.

Halorussus ruber sp. nov., isolated from an inland salt lake of China.

Halophilic archaeal strain YC25(T) was isolated from Yuncheng salt lake in Shanxi, China. Cells of strain YC25(T) were observed to be pleomorphic rods, stained Gram-negative, and formed red-pigmented colonies on solid media. Strain YC25(T) was found to be able to grow at 25-50°C (optimum 37°C), at 1.4-4.8M NaCl (optimum 1.7M), at 0-1.0M MgCl2 (optimum 0.01M), and at pH 5.5-9.0 (optimum pH 6.5). The cells lysed in distilled water, and the minimal NaCl concentration to prevent cell lysis was found to be 8% (w/v). The major polar lipids of the strain were phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me), phosphatidylglycerol sulfate (PGS), sulfated galactosyl mannosyl glucosyl diether (S-TGD-1), sulfated mannosyl glucosyl diether (S-DGD-1), galactosyl mannosyl glucosyl diether (TGD-1), mannosyl glucosyl diether (DGD-1), and an unknown diglycosyl diether (DGD-2) chromatographically identical to those of Halorussus rarus CGMCC 1.10122(T). The 16S rRNA gene and rpoB' gene of strain YC25(T) were phylogenetically related to the corresponding genes of Halorussus rarus CGMCC 1.10122(T) (94.3-95.4 and 91.5% nucleotide identity, respectively). The DNA G+C content of strain YC25(T) was determined to be 63.3mol%. The phenotypic, chemotaxonomic, and phylogenetic properties suggested that strain YC25(T) (=CGMCC 1.12122(T)=JCM 18363(T)) represents a new species of Halorussus, for which the name Halorussus ruber sp. nov. is proposed.

Halorubrum laminariae sp. nov., isolated from the brine of salted brown alga Laminaria.

Two halophilic archaeal strains, R60(T) and R61, were isolated from the brine of salted brown alga Laminaria. Cells of the two strains were observed to be rod-shaped, stain Gram-negative and to lyse in distilled water. Strain R60(T) was found to contain gas vacuoles and to produce pink-pigmented colonies, while strain R61 lacked gas vacuoles and produces red-pigmented colonies. Both strains were found to be able to grow at 20-50°C (optimum 30°C), at 1.7-4.8M NaCl (optimum 2.6-3.1M NaCl), at 0-1.0M MgCl2 (optimum 0.005-0.1M MgCl2) and at pH 6.0-9.5 (optimum pH 7.0). The major polar lipids were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and one major glycolipid chromatographically identical to a sulfated mannosyl glucosyl diether produced by Halorubrum members of the Halobacteriaceae. The 16S rRNA gene sequences of the two strains were 99.9% identical, showing 94.6-98.0% similarity to those of members of the genus Halorubrum. The EF-2 gene similarity between strains R60(T) and R60 was 100 % and showed 84.6-94.5 % similarity to those of members of the genus Halorubrum. The DNA G+C contents of the two strains were determined to be 63.0mol%. The DNA-DNA hybridization value between strain R60(T) and strain R61 was 92% and the two strains showed low DNA-DNA relatedness with the most related members of Halorubrum. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain R60(T) (=CGMCC 1.12689(T)=JCM 30040(T)) and strain R61 (=CGMCC 1.12696) represent a novel species of the genus Halorubrum, for which the name Halorubrum laminariae sp. nov. is proposed.