Salt Mine In Yunnan Research Articles

Halomicrococcus hydrotolerans gen. nov., sp. nov., an extremely halophilic archaeon isolated from a subterranean salt deposit

A halophilic archaeon, strain H22T, was isolated from a subterranean salt deposit sampled at Yunnan salt mine, PR China. Colonies of strain H22T were light pink-pigmented. Cells were coccus, non-motile, Gram-stain-negative, and did not lyse in distilled water. The strain was aerobic and grew at 20-55 °C (optimum, 37 °C), in the presence of 10-30 % (w/v) NaCl (20 %) and at pH 6.5-9.0 (pH 7.0). Mg2+ was required for growth (optimum, 0.005 M). Major polar lipids were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and sulfated mannosyl-glucosyl-glycerol diether-1. Sequence similarity search based on the multiple 16S rRNA genes (rrnA, rrnB and rrnC) of strain H22T revealed that it was most closely related to species of the genera Haloarchaeobius, Haladaptatus, Halorussus and Halorubellus with relative low sequence similarities (91.9-93.7 %). The strain, however, shared highest rpoB' gene sequence identities with Halorussus rarus TBN4T (90.8 % rpoB' gene sequence similarity). Phylogenetic trees based on 16S rRNA and rpoB' gene sequences revealed a robust lineage of the strain H22T with members of related genera of the family Halobacteriaceae. The DNA G+C content of strain H22T was 62.9 mol%. Genome-based analysis of average nucleotide identity (ANI) and in silico DNA-DNA hybridization (DDH) between strains H22T and its closest relative were equal or lower than 77.7 and 22.4 %, respectively, which were far below the threshold for delineation of a new species. Based on ANI values, in silico DDH, and distinct morphological and physiological differences from the previously described taxa, we suggest that strain H22T represents a novel species of a new genus within the family Halobacteriaceae, for which the name Halomicrococcus hydrotolerans gen. nov., sp. nov. is proposed. The type strain is H22T (=CGMCC 1.16291T=NBRC 113231T).

Haloterrigena salifodinae sp. nov., an extremely halophilic archaeon isolated from a subterranean rock salt.

A novel extremely halophilic strain, designated ZY19T, was isolated from a rock salt sample from Yunnan salt mine, PR China. Strain ZY19T is neutrophilic, non-motile and requires at least 10% (w/v) NaCl for growth. Optimal growth is observed at 20-25% (w/v) NaCl, pH 7.5-8.0 and 42°C. Mg2+ is not required for growth. The cells do not lyse in distilled water. On the basis of 16S rRNA gene sequence analysis, strain ZY19T belongs to the genus Haloterrigena (Htg.) and is closely related to Haloterrigena salina XH-65T (98.5% sequence similarity) and Haloterrigena turkmenica DSM 5511T (97.9%). Phylogenetic and phylogenomic analysis showed that strain ZY19T clusters with the species Htg. salina and Htg. turkmenica forming an independent clade separated from other members of the genus. The value of genomic average nucleotide identity (ANI) between strains ZY19T and its close relative, Htg. salina XH-65T was 94.2%. DNA-DNA relatedness between strains ZY19T and Htg. salina XH-65T revealed by in silico DNA-DNA hybridization (DDH) was 56.3%. Both the ANI value and the degree of in silico DDH are below the accepted threshold for members of the same species. The major polar lipids were found to consist of phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, sulfated diglycosyl-diether-1 and mannose-2, 6-disulfate (1 → 2)-glucose glycerol diether. The genomic DNA G+C content was determined to be 64.5mol%. Based on the results of the phenotypic, chemotaxonomic, genetic similarity and inferred phylogeny, strain ZY19T is distinct from other validly named species and thus represents a novel species within the genus Haloterrigena, for which the name Haloterrigena salifodinae sp. nov. is proposed. The type strain is ZY19T (=CGMCC 1.16114T=NBRC 112981T).

Halalkalicoccus subterraneus sp. nov., an extremely halophilic archaeon isolated from a subterranean halite deposit.

An extremely halophilic archaeon, designated strain GSM28T, was isolated from a subterranean halite deposit in a Yunnan salt mine, China. Cells of the strain were observed to be cocci, non-motile and Gram-variable, and to require at least 15% (w/v) NaCl for growth (optimum 20%). Growth was found to occur in the ranges of 20-45°C (optimum 42°C) and pH 6.0-8.5 (optimum 7.5). Cells did not lyse in distilled water. Phylogenetic analysis based on 16S rRNA gene sequences indicated that this strain belongs to the genus Halalkalicoccus and shows 99.1% similarities with its close phylogenetic relative Halalkalicoccus paucihalophilus DSM 24557T. Genomic ANI analysis showed that the DNA-DNA relatedness between strain GSM28T and the closely related species Hac. paucihalophilus DSM 24557T and Halalkalicoccus jeotgali B3T was 83.7% and 83.1%, respectively. The major polar lipids were determined to be phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, sulfated mannosyl-glucosyl-glycerol diether-1 and two unidentified glycolipids. The DNA G + C content was determined to be 61.8mol%. On the basis of physiological, biochemical tests and phylogenetic differentiations, strain GSM28T is concluded to represent a novel species in the genus Halalkalicoccus, for which the name Halalkalicoccus subterraneus sp. nov. is proposed. The type strain is GSM28T (= CGMCC 1.16344T = NBRC 113432T).

Halorubrum glutamatedens sp. nov., a Halophilic Archaeon Isolated from a Rock Salt.

An extremely halophilic archaeon, strain ZY8T, was isolated from a rock salt of Yunnan salt mine. It was able to grow at 12-30% (w/v) NaCl (optimum, 15-20%), pH 7.0-9.0 (optimum, pH 8.5), and 20-45°C (optimum, 42°C). Sequence similarity search of its 16S rRNA gene showed that strain ZY8T belonged to the genus Halorubrum, and it is closely related to species of H. aethiopicum SAH-A6T (98.6%), H. aquaticum EN-2T (98.6%), and H. halodurans Cb34T (98.5%), respectively. Strain ZY8T contained phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and phosphatidylglycerol sulfate as its major phospholipids, and a sulfated diglycosyl diether as its major glycolipid. The DNA G+C content was 66.7mol%. DNA-DNA relatedness between strains ZY8T and closely related species were far below 70%. Based on the phenotypic and phylogenetic analyses, it is proposed that strain ZY8T represents a novel species of the genus Halorubrum, for which the name Halorubrum glutamatedens sp. nov. is proposed. The type strain is ZY8T (=CGMCC 1.16026T=NBRC 112866T).

Halococcus salsus sp. nov., a novel halophilic archaeon isolated from rock salt.

Two pink-pigmented halophilic archaea, designated strains ZJ1T and J81, were isolated from rock salt of Yunnan Salt Mine, China, and commercial salt imported from Bolivia, respectively. Cells were non-motile, coccoid, approximately 0.8-1.6 µm in diameter, stained Gram-negative and often occurred in pairs. Colonies were wet, opaque and smooth-edged. Strain ZJ1T grew optimally with 20 % (w/v) NaCl, at pH 7.5 and at 38-40 °C, which was the same as for strain J81. 16S rRNA gene sequence similarity between strains ZJ1T and J81 was 99.7 %. Sequence similarity searches based on the 16S rRNA gene and cell morphology suggested that strains ZJ1T and J81 belong to the genus Halococcus in the family Halococcaceae. The major polar lipids of the type strain, ZJ1T, were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and sulfated diglycosyl-diether-1. The profile of polar lipids, cell shape, motility and lack of lysis of cells in distilled water show that strains ZJ1T and J81 were similar to other members of the genus Halococcus. Strain ZJ1T shared the highest 16S rRNA gene and rpoB' gene sequence similarities of 99.0 and 95.3 % with Halococcus hamelinensis 100A6T, respectively, followed by less than 94.6 % with sequences of other species in the genus Halococcus. DNA-DNA relatedness between strains ZJ1T and J81 was 90.1±0.7 %, while 27±0.5 % was found between strain ZJ1T and H. hamelinensis JCM 12892T (=100A6T), and 29.0±0.5 % between strains J81 and H. hamelinensis JCM 12892T. The DNA G+C content of strain ZJ1T was 66.5 mol% (Tm). The stable phylogenetic position, differential physiological and biochemical properties and extensive sequence divergence suggest that strains ZJ1T and J81 represent a novel species, for which the name Halococcus salsus sp. nov. is proposed. The type strain is ZJ1T (=CGMCC 1.16025T=NBRC 112867T).

Halorubrum depositum sp. nov., a Novel Halophilic Archaeon Isolated from a Salt Deposit.

A non-motile, pleomorphic rod-shaped or oval, red-pigmented (nearly scarlet), extremely halophilic archaeon, strain Y78T, was isolated from a salt deposit of Yunnan salt mine, China. Analysis of the 16S rRNA gene sequence showed that it was phylogenetically related to species of the genus Halorubrum, with a close relationship to Halorubrum rutilum YJ-18-S1T (98.6%), Halorubrum yunnanense Q85T (98.3%), and Halorubrum lipolyticum 9-3T (98.1%). The temperature, NaCl, and pH ranges for growth were 25-50°C, 12-30% (w/v), and 6.5-9.0, respectively. Mg2+ was required for growth. The polar lipids of strain Y78T were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, and a sulfated diglycosyl diether. The DNA G+C content was 66.6mol%. DNA-DNA hybridization values between strain Y78T and two closely related species of the genus Halorubrum were far below 70%. Based on the data presented in this study, strain Y78T represents a novel species for which the name Halorubrum depositum sp. nov. is proposed; the type strain is Y78T (= CGMCC 1.15456T = JCM 31272T).

Marivibrio halodurans gen. nov., sp. nov., a marine bacterium in the family Rhodospirillaceae isolated from underground rock salt.

Gram-negative, spiral or curved rod-shaped cells of a bacterial strain, designated ZC80T, were isolated from a rock salt sample collected at Yunnan salt mine, China. Analysis of the strain's 16S rRNA gene sequence revealed a clear affiliation of this novel strain within the family Rhodospirillaceae. Strain ZC80T formed a robust cluster with Pelagibius litoralis CL-UU02T at a 16S rRNA gene sequence similarity level of 88.1 %. Strain ZC80T shared no more than 91.0 % 16S rRNA gene sequence similarity with the type strains of other species in the family Rhodospirillaceae. Strain ZC80T was able to grow in the presence of 2-15 % (w/v) NaCl, and grew at 10-50 °C and pH 6.0-10.0. The major fatty acids were C19 : 0 cyclo ω8c (41.3 %). The major isoprenoid quinone was ubiquinone 10 (Q-10). The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol and an unidentified aminolipid. The DNA G+C content of strain ZC80T was 60.8 mol%. On the basis of phylogenetic analyses and chemotaxonomic and physiological data, strain ZC80T is considered to represent a novel species of a new genus in the family Rhodospirillaceae, for which the name Marivibrio halodurans gen. nov., sp. nov. is proposed. The type strain of Marivibrio halodurans is ZC80T (=CGMCC 1.15697T=NBRC 112461T).

Halorubrum salsamenti sp. nov., a Novel Halophilic Archaeon Isolated from a Brine of Salt Mine

A non-motile, spherical or oval extremely halophilic archaeon, strain Y69T, was isolated from a brine of the Yunnan salt mine, China. Colonies on JCM 168 agar plate were round (1-2mm in diameter), moist, and orange-pigmented. Phylogenetic analysis of the almost-complete 16S rRNA gene sequence showed that the isolate belonged to the species of the genus Halorubrum, with a close relationship to Halorubrum aidingense 31-hongT (98.5%), Halorubrum lacusprofundi ATCC 49239T (98.2%), and Halorubrum kocurii BG-1T (98.0%). The major polar lipids of strain Y69T were phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and a sulfated diglycosyl diether. Strain Y69T grew in 15-30% (w/v) NaCl. The temperature and pH ranges for growth were 25-50°C and 6.5-9.0, respectively. Optimal growth occurred at 20% (w/v) NaCl, 42°C, and pH 8.0. Mg2+ was required for growth. The DNA G+C content was determined to be 65.1mol% by the thermal denaturation method. DNA-DNA hybridization values between strain Y69T and the closely related species were lower than 70%. Based on the data presented in this study, strain Y69T represents a novel species for which the name Halorubrum salsamenti sp. nov. is proposed. The type of the strain is Y69T (=CGMCC 1.15455T=JCM 31270T).

Complete genome sequence of Halomonas ventosae virulent halovirus QHHSV-1.

A virulent halovirus QHHSV-1 which lyses Halomonas ventosae QH52-2 originating from the Qiaohou salt mine in Yunnan, Southwest China was characterized. The complete genome of QHHSV-1 is composed of a circular double-stranded DNA of 37,270 base pairs in length, with 66.8% G+C content and 69 putative open reading frames (ORFs), which were classified into five functional groups, including morphogenesis, replication/regulation, packaging, lysis and lysogeny. A putative Cro repressor gene and an integrase gene were found in the genome, showing that QHHSV-1 may utilize a lambda-like repression system under unfavorable conditions. QHHSV-1 is the first report of the whole genome sequence of the virulent Halomonas phage belonging to the family Siphoviridae.

Halorubrum trueperi sp. nov., a halophilic archaeon isolated from a salt mine.

A novel, extremely halophilic archaeon, strain Y73T, was isolated from a salt mine in Yunnan, China. Colonies formed on solid medium were circular (2-3 mm in diameter), smooth, orange, glistening and convex (~1 mm in elevation). Cells were Gram-stain-negative, non-motile and pleomorphic. Mg2+ was required for growth (optimum at 0.05 M). Optimal growth was observed at 20 % (w/v) NaCl, 42-45 °C and pH 7.5-8.5 under aerobic conditions. 16S rRNA gene sequence comparison showed that strain Y73T was closely related to Halorubrum halophilum B8T (similarity: 98.1 %), Halorubrum lipolyticum 9-3T (97.9 %) and Halorubrum saccharovorum JCM 8865T (97.6 %). Levels of rpoB' gene sequence similarity between strain Y73T and H. halophilum B8T, H. lipolyticum 9-3T and H. saccharovorum JCM 8865T were 93.6, 93.8 and 94.7 %, respectively. DNA-DNA relatedness between strain Y73T and H. halophilum B8T was 38.7±0.5 %, while that between strain Y73T and H. saccharovorum JCM 8865T was 31.0±0.9 % . The DNA G+C content of strain Y73T was 61.9 mol%. The major polar lipids of strain Y73T were phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and phosphatidylglycerol sulfate as phospholipids, and sulfated diglycosyl diether-1 as glycolipid. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain Y73T represents a novel species, for which the name Halorubrum trueperi sp. nov is proposed. The type strain is Y73T (=CGMCC 1.15503T=JCM 31271T).

Halobaculum roseum sp. nov., isolated from underground salt deposits.

Two extremely halophilic archaea, strains D90T and D93, were isolated from underground salt deposits of Yunnan salt mine, China. Both strains were pleomorphic or short rods, non-motile, Gram-negative and required 1.7 M NaCl for growth, with optimum at 3.4 M. Mg2+ was not required for growth. Multiple copies of the 16S rRNA gene were obtained for both strains. Sequence similarity analysis of 16S rRNA genes revealed that strains D90T and D93 were closely related to Halobaculum magnesiiphilum MGY-184T and Halobaculum gomorrense DSM 9297T with the sequence similarity between 96.2-98.1 %. The sequence similarity of the rpoB' gene between strain D90T and Halobaculum gomorrenseJCM 9908T was 94.1 %. The DNA G+C contents of strains D90T and D93 were 65.9 and 67.6 mol%, respectively. The major polar lipids of both strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and glycolipid. The DNA-DNA relatedness value between strains D90T and D93 was 90.1±0.5 %, while that between strain D90T and Halobaculum gomorrenseJCM 9908T was 30.0±0.7 %. The comparison of physiological and biochemical characteristics, including the requirements of NaCl, Mg2+, pH, etc., differentiated strains D90T and D93 from Halobaculum magnesiiphilum MGY-184T and Halobaculum gomorrenseJCM 9908T. Therefore, strains D90T and D93 represent a novel species of the genus Halobaculum, for which the name Halobaculum roseum sp. nov. is proposed. The type strain is D90T (=CGMCC 1.15501T=JCM 31273T).

Halorubrum yunnanense sp. nov., isolated from a subterranean salt mine.

Two halophilic archaeal strains, Q85(T) and Q86, were isolated from a subterranean salt mine in Yunnan, China. Cells were rod-shaped, Gram-stain-negative and motile. Colonies were red, smooth, convex and round (1.0-2.0 mm in diameter). The orthologous 16S rRNA and rpoB9 gene sequences of these two strains were almost identical (99.5 and 99.7 % similarities). Their closest relatives were Halorubrum kocurii BG-1(T) (98.0-98.1 % 16S rRNA gene sequence similarity), Halorubrum aidingense 31-hong(T) (97.6–97.7 %) and Halorubrum lipolyticum 9-3T (97.5-97.6 %). The level of DNA-DNA relatedness between strains Q85(T) and Q86 was 90 %, while that between Q85(T) and other related Halorubrum strains was less than 30 % (29 % for H. kocurii BG-1(T), 25 % for H. aidingense 31-hong(T) and 22 % for H. lipolyticum 9-3(T)). Optimal growth of the two novel strains was observed with 20 % (w/v) NaCl and at 42–45 8C under aerobic conditions, with a slight difference in optimum Mg2+ concentration (0.7 M for Q85(T),0.5 M for Q86) and a notable difference in optimum pH (pH 7.5 for Q85(T), pH 6.6 for Q86). Anaerobic growth occurred with nitrate, but not with L-arginine or DMSO. The major polar lipids of the two strains were identical, including phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and sulfated diglycosyl diether, which are the major lipids of the genus Halorubrum. The G+C contents of strains Q85(T) and Q86 were 66.3 and 66.8 %, respectively. Based on the phenotypic, chemotaxonomic and phylogenetic properties of strains Q85(T) and Q86, a novel species, Halorubrum yunnanense sp. nov., is proposed. The type strain is Q85(T) (=5CGMCC 1.15057(T)=JCM 30665(T)).

A novel Halomonas ventosae-specific virulent halovirus isolated from the Qiaohou salt mine in Yunnan, Southwest China.

Although Halomonas phages belonging to the families Myoviridae and Siphoviridae have been reported, no virulent Halomonas siphoviruses are known. In this study, a virulent bacteriophage, QHHSV-1, of the family Siphoviridae that specifically infects H. ventosae QH52-2 was isolated from the Qiaohou salt mine. Restriction analysis indicated that QHHSV-1 is a dsDNA virus with a genome size of 33.5-39.5kb. Transmission electron microscopy showed that QHHSV-1 is a typical representative of the Siphoviridae, with an icosahedral head (47nm in diameter) and a non-contractile tail (75nm in length). We also assessed the adsorption rate of QHHSV-1 for the host bacterium and found significant inhibition after the addition of 10mM CaCl2. Based on a one-step growth curve, we determined a latent period of 30min and a burst size of 73 PFU/infected cell. At the optimal pH of 8.0, 25.9 and 15.2% of the phages survived after a 60-min incubation at 50 and 60°C, respectively. Phage replication was possible at a wide range of salt concentrations, from 2.0 to 20% (w/v), with an optimum concentration of 5%. The survival of QHHSV-1 at different salt concentrations decreased with time and 25% survival after 25days at 30% salt concentration.

Halomonas qiaohouensis sp. nov., isolated from salt mine soil in southwest China.

Two Gram-stain negative, moderately halophilic, aerobic, motile bacteria, designated strains YIM QH88(T) and YIM QH103, were isolated from the Qiaohou salt mine in Yunnan, southwest China. Cells of the strains were observed to be rod-shaped and produce creamy-coloured colonies. Growth of the two strains was observed at 10-45°C (optimum 25-37°C), at pH 6.0-10.0 (optimum 7.0-8.0), and in the presence of 0.5-20% (w/v) NaCl (optimum 2-6%). The two strains were found to contain summed feature 8 (C18:1 ω7c/ω6c), C19:0 cyclo ω8c and C16:0 as the major cellular fatty acids. The polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and unknown phospholipid. The G+C content of the genomic DNA of strains YIM QH88(T) and YIM QH103 were determined to be 64.6 and 64.2mol%, respectively, and the predominant respiratory quinone detected was ubiquinone 9. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strains YIM QH88(T) and YIM QH103 formed a distinct lineage within the genus Halomonas and were most closely related to Halomonas pantelleriensis DSM 9661(T) with 97.3 and 97.5% of 16S rRNA sequence similarity respectively. The DNA-DNA hybridization relatedness value for strains YIM QH88(T) and YIM QH103 was 95.2±0.8%. The levels of DNA-DNA relatedness between each of these two strains and the type strains of phylogenetically closely related Halomonas species were clearly below 70%. On the basis of their phylogenetic analysis, DNA-DNA hybridization relatedness, phenotypic and chemotaxonomic characteristics, strains YIM QH88(T) and YIM QH103 should be classified as a novel species of the genus Halomonas, for which the name Halomonas qiaohouensis sp. nov. is proposed. The type strain is YIM QH88(T) (=DSM 26770(T) =CCTCC AB 2012965(T)).

Gracilimonas mengyeensis sp. nov., a moderately halophilic bacterium isolated from a salt mine in Yunnan, south-western China

A facultatively anaerobic, Gram-staining-negative, pale red-pigmented, non-motile, rod-shaped, moderately halophilic bacterium, designated strain YIM J14(T), was isolated from a sediment sample from a salt mine in Yunnan, south-western China. Growth occurred at NaCl concentrations of between 2 % and 15 % (w/v) and optimally with 5-9 % NaCl. The optimum temperature and pH for growth of the strain were 28 °C and pH 7.5. The major cellular fatty acids were iso-C15 : 0, anteiso-C15 : 0 and iso-C17 : 1ω9c/10-methyl-C16 : 0. The polar lipid profile was composed predominantly of diphosphatidylglycerol, phosphatidylcholine and one unknown phospholipid. Minor amounts of other lipids were also detectable. The genomic DNA G+C content was 47.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparisons indicated that strain YIM J14(T) was related to Gracilimonas tropica in the phylum Bacteroidetes. The level of 16S rRNA gene sequence similarity between strain YIM J14(T) and Gracilimonas tropica CL-CB462(T) was 96.9 %. A DNA-DNA hybridization experiment between strain YIM J14(T) and Gracilimonas tropica indicated levels of relatedness of 28 %. Chemotaxonomic data supported the placement of strain YIM J14(T) in the genus Gracilimonas. DNA-DNA hybridization and biochemical and physiological characterization allowed strain YIM J14(T) to be differentiated from Gracilimonas tropica. It is therefore considered to represent a novel species of the genus Gracilimonas, for which the name Gracilimonas mengyeensis sp. nov. is proposed. The type strain YIM J14(T) ( = ACCC 10717(T) = DSM 21985(T)).

Fodinibius salinus gen. nov., sp. nov., a moderately halophilic bacterium isolated from a salt mine

A novel, moderately halophilic, rod-shaped bacterium, designated strain YIM D17(T), was isolated from a sample of sediment from a salt mine in Yunnan, south-western China. The taxonomy of strain YIM D17(T) was investigated using a polyphasic approach. Strain YIM D17(T) was Gram-stain-negative, strictly aerobic and non-motile and formed pink colonies on marine agar. Optimal growth occurred at 37 °C, pH 7.5-8.0 and in the presence of 10-15 % (w/v) NaCl. The major menaquinone was MK-7. The polar lipid profile was composed predominantly of diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, one phospholipid, one glycolipid and one aminolipid. Minor amounts of other lipids were also detectable. The predominant cellular fatty acids were iso-C(17 : 1)ω9c/10-methyl-C(16 : 0) (24.0 %), iso-C(15 : 0) (23.6 %) and C(16 : 1)ω7c/C(16 : 1)ω6c (13.8 %). The DNA G+C content was 43.0 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that the isolate formed a distinct clade with the genera Gracilimonas and Balneola (both in the phylum Bacteroidetes) and was related to the species Gracilimonas tropica, Balneola vulgaris and Balneola alkaliphila, with sequence similarities of 85.6 %, 83.0 % and 82.8 % to the respective type strains. On the basis of its phenotypic, chemotaxonomic and phylogenetic features, strain YIM D17(T) represents a novel species of a new genus, for which the name Fodinibius salinus gen. nov., sp. nov. is proposed. The type strain is YIM D17(T) ( = ACCC 10716(T) = DSM 21935(T)).

Salinarimonas rosea gen. nov., sp. nov., a new member of the α-2 subgroup of the Proteobacteria

A Gram-negative, rod-shaped, facultatively anaerobic, halotolerant bacterial strain, designated YIM YD3(T), was isolated from a salt mine in Yunnan, south-west China. The taxonomy of strain YIM YD3(T) was investigated by a polyphasic approach. Strain YIM YD3(T) was motile, formed pink colonies and was positive for catalase and oxidase activities. Q-10 was the predominant respiratory ubiquinone. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylcholine and two unknown phospholipids. The major fatty acids (>10 % of total fatty acids) were C(18 : 1)omega7c, C(18 : 1)omega9c, C(16 : 0) and C(19 : 0) cyclo omega8c. The DNA G+C content was 71.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that the isolate formed a distinct line within a clade containing the genera Balneimonas, Bosea, Chelatococcus and Microvirga in the order Rhizobiales, with highest levels of 16S RNA gene sequence similarity to the type strain of Balneimonas flocculans (93.5 %). On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain YIM YD3(T) represents a novel species in a new genus, for which the name Salinarimonas rosea gen. nov., sp. nov. is proposed, with strain YIM YD3(T) (=KCTC 22346(T)=CCTCC AA208038(T)) as the type strain.

Fodinicurvata sediminis gen. nov., sp. nov. and Fodinicurvata fenggangensis sp. nov., poly- -hydroxybutyrate-producing bacteria in the family Rhodospirillaceae

Two Gram-negatively stained, facultatively anaerobic, non-motile, vibrioid and rod-shaped, chemoheterotrophic bacterial strains, designated YIM D82(T) and YIM D812(T), were isolated from a salt mine in Yunnan, south-west China. DNA-DNA hybridization, genomic DNA G+C content and phylogenetic analyses based on 16S rRNA gene sequences divided the two isolates into two distinct genospecies that were also clearly differentiated by fatty acid profiles, carbon source utilization patterns, antibiotic susceptibility and biochemical characteristics. The two isolates grew in the presence of 1.5-20 % NaCl, and optimally at 28 degrees C and pH 7.5. The genomic DNA G+C contents of strains YIM D82(T) and YIM D812(T) were 61.5 and 62.3 mol%, respectively. Phylogenetic analyses based on 16S rRNA gene sequences indicated that strains YIM D82(T) and YIM D812(T) were members of the family Rhodospirillaceae and showed 90.5-90.6 % and 90.1-90.2 % similarities with their closest relatives, Rhodovibrio sodomensis and Rhodovibrio salinarum, respectively. Differential phenotypic and genotypic characteristics of the two isolates from recognized genera showed that the two strains should be classified as representing a new genus and two novel species for which the names Fodinicurvata sediminis gen. nov., sp. nov. (type strain YIM D82(T)=DSM 21159(T)=KCTC 22351(T)) and Fodinicurvata fenggangensis sp. nov. (type strain YIM D812(T)=CCTCC AA 208037(T)=DSM 21160(T)) are proposed.

Fodinibacter luteus gen. nov., sp. nov., an actinobacterium isolated from a salt mine

A Gram-positive-staining, aerobic, catalase- and oxidase-positive, irregular short rod-shaped actinobacterium, designated strain YIM C003T, was isolated from a salt mine in Yunnan, PR China. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM C003T was most closely related to strains of the genera Knoellia (94.2-96.0% similarity), Oryzihumus (95.6%), Terrabacter (94.9-95.4%), Janibacter (94.9-95.4%), Kribbia (95.0%), Lapillicoccus (95.0%) and Phycicoccus (94.2-95.0%) of the family Intrasporangiaceae and that it formed an independent monophyletic lineage with three strains of Oryzihumus leptocrescens. The DNA G+C content of strain YIM C003T was 72.0 mol%. The diagnostic cell-wall diamino acid was meso-diaminopimelic acid. The predominant menaquinone was MK-8(H4). Mycolic acids were not detected. The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and two unknown phospholipids. The major cellular fatty acids were C18:1omega9c and C16:0. These chemotaxonomic properties, together with data from phylogenetic analysis, enabled the novel isolate to be differentiated from all other members of the family. A novel species in a new genus, Fodinibacter luteus gen. nov., sp. nov., is proposed, with strain YIM C003T (=DSM 21208T=CCTCC AA 208036T) as the type strain of Fodinibacter luteus.

Sediminimonas qiaohouensis gen. nov., sp. nov., a member of the Roseobacter clade in the order Rhodobacterales

Two aerobic bacterial strains, YIM B024(T) and YIM B025, were isolated from a salt mine in Yunnan, south-west China. Both strains showed almost the same physiological properties. Cells were Gram-negative, non-motile, non-spore-forming rods. The novel strains grew at 15-37 degrees C, pH 6.5-9.0 and 0.25-20 % (w/v) NaCl; optimum growth was observed at 28-30 degrees C, pH 7.0-8.5 and 1.5-10 % NaCl. Oxidase, catalase and nitrate-reducing activities were detected. The two strains were closely related to each other with a 16S rRNA gene sequence similarity of 100 %. DNA-DNA hybridization experiments revealed high relatedness values (90+/-0.4 %) between strains YIM B024(T) and YIM B025, which suggested that these two new strains constituted a single species. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the two isolates formed a loose cluster with members of the genus Roseivivax in the Roseobacter clade, but were clearly separated from this genus. The levels of 16S rRNA gene sequence similarity between the two isolates and members of the genus Roseivivax ranged from 92.4 to 93.9 %. The major polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine and four unknown phospholipids. The major cellular fatty acids were C(18 : 1)omega7c, C(16 : 0), C(18 : 1)omega9c, 11-methyl C(18 : 1)omega7c and C(19 : 0) cyclo omega8c. The sole respiratory quinone was Q-10 and the genomic DNA G+C content was 63.0-64.1 mol%. The distinct phylogenetic position and a combination of phenotypic and chemotaxonomic characteristics supported the proposal of the new isolates as representing a novel species in a new genus, for which the name Sediminimonas qiaohouensis gen. nov., sp. nov. is proposed. The type strain of the type species is YIM B024(T) (=KCTC 22349(T)=CCTCC AA 208033(T)).