Halorubellus salinus gen. nov., sp. nov. and Halorubellus litoreus sp. nov., novel halophilic archaea isolated from a marine solar saltern

Two extremely halophilic archaeal strains GX3(T) and GX26(T) were isolated from the Gangxi marine solar saltern near the Weihai city of Shandong Province, China. Cells from the two strains were pleomorphic and stained Gram-negative, colonies were red-pigmented. Strains GX3(T) and GX26(T) were able to grow at 25-50 °C (optimum 37 °C), at 1.4-5.1M NaCl (optimum 3.1M), at pH 5.5-9.5 (optimum pH 7.0) and neither strain required Mg(2+) for growth. Cells lyse in distilled water and the minimal NaCl concentration to prevent cell-lysis was 8% (w/v). The major polar lipids of the two strains were PA (phosphatidic acid), PG (phosphatidylglycerol), PGP-Me (phosphatidylglycerol phosphate methyl ester) and three major glycolipids (GL1, GL2 & GL3) chromatographically identical to S-TGD-1 (sulfated galactosyl mannosy glucosyl diether), S-DGD-1 (sulfated mannosyl glucosyl diether), and DGD-1 (mannosyl glucosyl diether) respectively, an unidentified lipid (GL4) was also detected in strain GX26(T). Phylogenetic analysis based on 16S rRNA gene revealed that strain GX3(T) and strain GX26(T) formed a distinct clade with the closest relative, Haladaptatus paucihalophilus (89.9-92.4% and 90.4-92.7, respectively). The rpoB' gene similarities between strains GX3(T) and GX26(T), and between the two strains and the closest relative, Halorussus rarus TBN4(T) are 96.5%, 84.3% and 83.9%, respectively. The DNA G+C contents of strain GX3(T) and strain GX26(T) are 67.3 mol% and 67.2 mol%, respectively. The DNA-DNA hybridization value between strain GX3(T) and strain GX26(T) was 44%. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain GX3(T) and strain GX26(T) represent two novel species in a new genus within the family Halobacteriaceae, Halorubellus salinus gen. nov., sp. nov. (type strain GX3(T)=CGMCC 1.10384(T)=JCM 17115(T)) and Halorubellus litoreus sp. nov. (type strain GX26(T)=CGMCC 1.10386(T)=JCM 17117(T)).     

Halobacterium rubrum sp. nov., isolated from a marine solar saltern

Halophilic archaeal strain TGN-42-S1^T was isolated from the Tanggu marine solar saltern, China. Cells from strain TGN-42-S1^T were observed to be pleomorphic rods, stained Gram-negative, and formed red-pigmented colonies on solid media. Strain TGN-42-S1^T was found to be able to grow at 20–50 °C (optimum 35–37 °C), at 1.7–4.8 M NaCl (optimum 3.1 M), at 0–1.0 M MgCl₂ (optimum 0.1 M), and at pH 5.0–9.0 (optimum pH 7.0–7.5). The cells lysed in distilled water, and the minimal NaCl concentration to prevent cell-lysis was found to be 10 % (w/v). The major polar lipids of the strain were phosphatidic acid, phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, galactosyl mannosyl glucosyl diether (TGD-1), sulfated galactosyl mannosyl glucosyl diether (S-TGD-1), sulfated galactosyl mannosyl galactofuranosyl glucosyl diether (S-TeGD), and three unidentified glycolipids which were chromatographically identical to those of the Halobacterium species. The 16S rRNA gene and rpoB′ gene of strain TGN-42-S1^T were phylogenetically related to the corresponding genes of Halobacterium jilantaiense CGMCC 1.5337^T (98.8 and 93.5 % nucleotide identity, respectively), Halobacterium salinarum CGMCC 1.1958^T (98.4 and 91.9 %), and Halobacterium noricense JCM 15102^T (96.9 and 91.1 %). The DNA G + C content of strain TGN-42-S1^T was determined to be 69.2 mol %. Strain TGN-42-S1^T showed low DNA–DNA relatedness with Hbt. jilantaiense CGMCC 1.5337^T and Hbt. salinarum CGMCC 1.1958^T, the most closely related members of the genus Halobacterium. The phenotypic, chemotaxonomic, and phylogenetic properties suggested that strain TGN-42-S1^T (=CGMCC 1.12575^T =JCM 19908^T) represents a new species of Halobacterium, for which the name Halobacterium rubrum sp. nov. is proposed.     

Halobellus rarus sp. nov., a halophilic archaeon from an inland salt lake of China

Two halophilic archaeal strains, YC21^T and YC77, were isolated from an inland salt lake of China. Both have pleomorphic rod-shaped cells that lyse in distilled water, stain Gram-negative and form red-pigmented colonies. They are neutrophilic, require at least 2.1 M NaCl for growth under the optimum growth temperature of 37 °C. The major polar lipids of the two strains were phosphatidylglycerol (PG), phosphatidylglycerol phosphate methyl ester (PGP-Me), phosphatidylglycerol sulfate (PGS), two major glycolipids (GL1 and GL2) chromatographically identical to sulfated mannosyl glucosyl diether (S-DGD-1) and mannosyl glucosyl diether (DGD-1), respectively. Trace amounts of two unidentified lipids (GL0-1 and GL0-2) were also detected. The 16S rRNA gene sequences of the two strains are 99.9 % identical, show 94.0–98.9 % similarity to the closest relative members of Halobellus of the family Halobacteriaceae. The rpoB′ gene similarity between strains YC21^T and YC77 is 99.8 % and show 90.3–95.3 % similarity to the closest relative members of Halobellus. The DNA G+C content of strains YC21^T and YC77 were 66.1 and 66.2 mol%, respectively. The DNA–DNA hybridization value between strain YC20^T and strain YC77 was 89 %, and the two strains showed low DNA–DNA relatedness with Halobellus limi TBN53^T, the most related member of Halobellus. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strains YC21^T and YC77 represent a novel species of the genus Halobellus, for which the name Halobellus rarus sp. nov. is proposed. The type strain is YC21^T (=CGMCC 1.12121^T = JCM 18362^T).     

Halobellus litoreus sp. nov., a Halophilic Archaeon Isolated from a Chinese Marine Solar Saltern

Halophilic archaeal strain GX31^T was isolated from a marine solar saltern of China. The cells of the strain were rod-shaped and lysed in distilled water, stain Gram-negative and formed red-pigmented colonies. It was neutrophilic, and required at least 0.9 M NaCl and 0–1.0 M MgCl₂ for growth under the optimum growth temperature of 37 °C. The major polar lipids of the strain were phosphatidylglycerol (PG), PG phosphate methyl ester, PG sulphate, and two major glycolipids chromatographically identical to sulphated mannosyl glucosyl diether (S-DGD-1) and mannosyl glucosyl diether (DGD-1), respectively. Trace amounts of two unidentified lipids were also detected. On the basis of 16S rRNA gene sequence analysis, strain GX31^T was closely related to the members of Halobellus of the family Halobacteriaceae with similarities of 94.1–98.7 %. Strain GX31^T showed 89.8–95.4 % of the rpoB′ gene similarity to the members of Halobellus. The DNA G+C content of strain GX31^T was 66.8 mol%. Strain GX31^T showed low DNA–DNA relatedness with two most related members of the genus Halobellus. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain GX31^T represent a novel species of the genus Halobellus, for which the name Halobellus litoreus sp. nov. is proposed. The type strain is GX31^T (=CGMCC 1.10387^T = JCM 17118^T).     

Haloplanus litoreus sp. nov. and Haloplanus ruber sp. nov., from a marine solar saltern and an aquaculture farm,respectively

Two halophilic archaea, strains GX21^T and R35^T, were isolated from a marine solar saltern and an aquaculture farm in China, respectively. Cells of the two strains were observed to be pleomorphic, flat, to contain gas vesicles, stain Gram-negative and produce red-pigmented colonies. Strain GX21^T was found to be able to grow at 25–50 °C (optimum 37 °C), at 2.6–4.8 M NaCl (optimum 3.4 M NaCl), at 0.05–1.0 M MgCl₂ (optimum 0.1 M MgCl₂) and at pH 6.0–8.5 (optimum pH 6.5) while strain R35^T was found to be able to grow at 25–45 °C (optimum 37 °C), at 2.1–4.8 M NaCl (optimum 3.1 M NaCl), at 0–0.7 M MgCl₂ (optimum 0.03 M MgCl₂) and at pH 5.5–9.5 (optimum pH 6.5–7.0). The cells of both isolates were observed to lyse in distilled water. The minimum NaCl concentrations that prevented cell lysis were determined to be 15 % (w/v) for strain GX21^T and 12 % (w/v) for strain R35^T. The major polar lipids of the two strains were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, one major glycolipid and a minor lipid chromatographically identical to sulfated mannosyl glucosyl diether and mannosyl glucosyl diether, respectively. 16S rRNA gene sequence analysis revealed that strains GX21^T and R35^T show 97.1 % sequence similarity to each other and are closely related to Haloplanus aerogenes TBN37^T (96.8 and 95.8 %), Haloplanus vescus RO5-8^T (96.7 and 96.1 %), Haloplanus salinus YGH66^T (96.4 and 95.8 %) and Haloplanus natans JCM 14081^T (96.3 and 95.4 %). The rpoB′ gene similarity between strains GX21^T and R35^T is 90.5 % and show 88.5–90.8 % similarity to the Haloplanus species with validly published names. The DNA G+C content of strain GX21^T and R35^T were determined to be 65.8 and 66.0 mol%, respectively. The DNA–DNA hybridization values between strain GX21^T and strain R35^T, and the two strains with the Haloplanus species with validly published names, showed less than 50 % DNA–DNA relatedness. It was concluded that strain GX21^T (=CGMCC 1.10456^T = JCM 17092^T) and strain R35^T (=CGMCC 1.10594 ^T = JCM 17271^T) represent two new species of Haloplanus, for which the names Haloplanus litoreus sp. nov. and Haloplanus ruber sp. nov. are proposed.     

Haloplanus salinus sp. nov., an extremely halophilic archaeon from a Chinese marine solar saltern

Halophilic archaeal strain YGH66^T was isolated from the Yinggehai marine solar saltern near the Sanya city of Hainan Province, China. Cells were pleomorphic, flat, stained Gram-negative, and produced pink-pigmented colonies. Strain YGH66^T was able to grow at 20–50 °C (optimum 37 °C), at 0.9–4.8 M NaCl (optimum 3.1 M NaCl), at 0.005–1.0 M MgCl₂ (optimum 0.05 M MgCl₂), and at pH 6.0–8.0 (optimum pH 7.0). The cells of strain YGH66^T were lysed in distilled water, and the minimum NaCl concentration that prevented cell lysis was 5 % (w/v). The major polar lipids of the strain were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, one major glycolipid (GL1) chromatographically identical to sulfated mannosyl glucosyl diether and a minor unidentified lipid (GL2), respectively. On the basis of 16S rRNA gene sequence analysis, strain YGH66^T was closely related to Haloplanus natans JCM 14081^T, Haloplanus aerogenes TBN37^T, and Haloplanus vescus RO5-8^T with the similarities of 98.0, 97.6, and 96.9 %, respectively. The rpoB′ gene similarity between strain YGH66^T and the current three members of Haloplanus were 90.4–92.8 %. The DNA G+C content of strain YGH66^T was 67.2 mol %. The DNA–DNA hybridization values between strain YGH66^T and three members of Haloplanus, H. natans JCM 14081^T, H. aerogenes TBN37^T, H. vescus RO5-8^T were 50, 46 and 39 %, respectively. It was concluded that strain YGH66^T represents a novel species of the genus Haloplanus, for which the name Haloplanus salinus sp. nov. is proposed. The type strain is YGH66^T (=CGMCC 1.12127^T = JCM 18368^T).     

Halolamina rubra sp. nov., a haloarchaeon isolated from non-purified solar salt

Two Gram-stain negative, rod-shaped and motile extreme halophiles, designated CBA1107^T and CBA1108, were isolated from non-purified solar salt. Based on the phylogenetic analysis, strains CBA1107^T and CBA1108 were shown to belong to the genus Halolamina, with similarities for the 16S rRNA gene sequences between strains CBA1107^T and Halolamina pelagica TBN21^T , Halolamina salina WSY15-H3^T and Halolamina salifodinae WSY15-H1^T of 98.3, 97.6 and 97.3 %, respectively; the similarities for the rpoB′ gene sequences between the same strains were 96.0, 95.3 and 94.6 %, respectively. The colonies of both strains were observed to be red pigmented on growth medium. Strain CBA1107^T was observed to grow at 20–50 °C, in the presence of 15–30 % NaCl, at pH 6.0–9.0, and with 0.005–0.5 M Mg²⁺. The cells of both strains lysed in distilled water. The DNA–DNA hybridization experiments showed that strain CBA1107^T shared 97 % relatedness with CBA1108 and <50 % relatedness with H. pelagica JCM 16809^T, H. salina JCM 18549^T and H. salifodinae JCM 18548^T. The genomic DNA G+C content of strain CBA1107^T was determined to be 65.1 mol%. The major polar lipids of the two strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and glycolipids including sulfated mannosyl glucosyl diether and mannosyl glucosyl diether. Based on the polyphasic taxonomic analyses, the strains are considered to represent a new taxon for which the name Halolamina rubra sp. nov. is proposed, with the type strain CBA1107^T (=CECT 8421^T =JCM 19436^T).     

Halorubrum salinum sp. nov., isolated from a marine solar saltern

The halophilic archaeal strain GX71^T was isolated from the Gangxi marine solar saltern near the Weihai city of Shandong Province, China. Cells of the strain were pleomorphic and lysed in distilled water, stained Gram-negative and formed red-pigmented colonies. Strain GX71^T was able to grow at 25–45 °C (optimum 30 °C), in the presence of 1.7–4.8 M NaCl (optimum 2.6 M NaCl), with 0.005–0.7 M MgCl₂ (optimum 0.05 M MgCl₂) and at pH 5.5–9.5 (optimum pH 7.0–7.5). Cells lysed in distilled water and the minimal NaCl concentration to prevent cell lysis was 10 % (w/v). The major polar lipids of the strain were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, one major glycolipid chromatographically identical to sulfated mannosyl glucosyl diether (S-DGD-3) and an unidentified lipid was also detected. The 16S rRNA gene sequence of strain GX71^T showed 94.0–97.0 % similarity to members of the genus Halorubrum of the family Halobacteriaceae. The rpoB′ gene sequence of strain GX71^T was 87.3–93.4 % similarity to current members of the genus Halorubrum. The DNA G+C content of GX71^T was 67.1 mol%. Strain GX71^T showed low DNA–DNA relatedness with Halorubrum lipolyticum CGMCC 1.5332^T, Halorubrum saccharovorum CGMCC 1.2147^T, Halorubrum kocurii CGMCC 1.7018^T and Halorubrum arcis CGMCC 1.5343^T, the most closely related members of the genus Halorubrum. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain GX71^T represents a novel species of the genus Halorubrum, for which the name Halorubrum salinum sp. nov. is proposed. The type strain is GX71^T (= CGMCC 1.10458^T = JCM 17093^T).     

<Emphasis Type="Italic">Salinarchaeum laminariae</Emphasis> gen. nov., sp. nov.: a new member of the family <Emphasis Type="Italic">Halobacteriaceae</Emphasis> isolated from salted brown alga <Emphasis Type="Italic">Laminaria</Emphasis>

Halophilic archaeal strains R26^T and R22 were isolated from the brown alga Laminaria produced at Dalian, Liaoning Province, China. Cells from the two strains were pleomorphic rods and Gram negative, and colonies were red pigmented. Strains R26^T and R22 were able to grow at 20–50°C (optimum 37°C) in 1.4–5.1 M NaCl (optimum 3.1–4.3 M) at pH 5.5–9.5 (optimum pH 8.0–8.5) and neither strain required Mg²⁺ for growth. Cells lyse in distilled water and the minimum NaCl concentration required to prevent cell lysis was 8% (w/v) for strain R26^T and 12% (w/v) for strain R22. The major polar lipids of the two strains were phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester and minor phosphatidylglycerol sulfate; glycolipids were not detected. Phylogenetic analyses based on 16S rRNA genes and rpoB′ genes revealed that strains R26^T and R22 formed a distinct clade with the closest relative, Natronoarchaeum mannanilyticum. The DNA G+C content of strains R26^T and R22 was 65.8 and 66.4 mol%, respectively. The DNA–DNA hybridization value between strains R26^T and R22 was 89%. The phenotypic, chemotaxonomic and phylogenetic properties suggest that the strains R26^T and R22 represent a novel species in a new genus within the family Halobacteriaceae, for which the name Salinarchaeum laminariae gen. nov., sp. nov. is proposed. The type strain is R26^T (type strain R26^T = CGMCC 1.10590^T = JCM 17267^T, reference strain R22 = CGMCC 1.10589).     

Salinarubrum litoreum gen. nov., sp. nov.: a new member of the family Halobacteriaceae isolated from Chinese marine solar salterns

Three halophilic archaeal strains, XD46^T, YJ-63-S1 and ZS-1-H, were isolated from three Chinese marine solar salterns. All were observed to have pleomorphic cells that lysed in distilled water, stained Gram-negative and formed red-pigmented colonies. They were found to grow optimally at 37 °C, at pH 7.0 and in the presence of 2.6 M NaCl and 0.05 M Mg²⁺. The major polar lipids were identified as those typical for members of the Halobacteriaceae but also included major glycolipids chromatographically identical to sulfated mannosyl glucosyl diether (S-DGD-1), mannosyl glucosyl diether (DGD-1) and two unidentified ones. The 16S rRNA gene sequences of the three strains were 99.8–100 % identical, showing most similarity to sequences of members of the family Halobacteriaceae, and clustering together as a distinct clade in phylogenetic tree reconstructions. The rpoB′ gene similarities between the three strains were 98.7–100 % and lower to the sequences of other halobacteria. Their DNA G+C contents were determined to be 65.1–65.5 mol%. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strains XD46^T (=CGMCC 1.12237^T = JCM 18649^T), YJ-63-S1 (=CGMCC 1.12574) and ZS-1-H (=CGMCC 1.12544) represent a novel species in a new genus within the family Halobacteriaceae, for which the name Salinarubrum litoreum gen. nov., sp. nov. is proposed.     

Halorubrum rubrum sp. nov., an extremely halophilic archaeon from a Chinese salt lake

Two halophilic archaeal strains, YC87^T and YCA11, were isolated from Yuncheng salt lake in Shanxi, China. Cells of the two strains were observed to be pleomorphic rod-shaped, stained Gram-negative and produced red-pigmented colonies. Strain YC87^T was able to grow at 20–50 °C (optimum 37 °C), at 1.4–4.8 M NaCl (optimum 2.1 M NaCl), at 0.05–1.0 M MgCl₂ (optimum 0.3 M MgCl₂) and at pH 6.0–9.0 (optimum pH 7.0) while strain YCA11 was able to grow at 20–50 °C (optimum 37 °C), at 2.1–4.8 M NaCl (optimum 3.1 M NaCl), at 0.01–0.7 M MgCl₂ (optimum 0.1 M MgCl₂) and at pH 6.0–9.0 (optimum pH 7.5). The cells of both isolates were observed to lyse in distilled water. The minimum NaCl concentrations that prevented cell lysis were determined to be 8 % (w/v) for strain YC87^T and 12 % (w/v) for strain YCA11. The major polar lipids of the two strains were identified as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and one major glycolipid chromatographically identical to sulfated mannosyl glucosyl diether; another major glycolipid and trace amounts of several unidentified lipids were also detected. The 16S rRNA gene sequences of the two strains were 99.8 % identical, showing 93.2–98.2 % similarity to members of the genus Halorubrum of the family Halobacteriaceae. The rpoB′ gene similarity between strains YC87^T and YCA11 was 99.3 % and showed 87.5–95.2 % similarity to the closest relative members of the genus Halorubrum. The DNA G+C content of strains YC87^T and YCA11 were determined to be 64.9 and 64.5 mol%, respectively. The DNA–DNA hybridization value between strain YC20^T and strain YC77 was 87 % and the two strains showed low DNA–DNA relatedness with Halorubrum cibi JCM 15757^T and Halorubrum aquaticum CGMCC 1.6377^T, the most related members of the genus Halorubrum. The phenotypic, chemotaxonomic and phylogenetic properties suggest that strains YC87^T and YCA11 represent a novel species of the genus Halorubrum, for which the name Halorubrum rubrum sp. nov. is proposed. The type strain is YC87^T (=CGMCC 1.12124^T = JCM 18365^T).     

Halobellus rufus sp. nov., an extremely halophilic archaeon isolated from non-purified solar salt

A halophilic archaeon, designed strain CBA1103^T, was isolated from non-purified solar salt. The cells of strain CBA1103^T were observed to be Gram-stain negative and pleomorphic, and the colonies appear red. Strain CBA1103^T was observed to grow between 20 and 55 °C (optimum 37 °C), and in NaCl concentrations of 10–30 % (w/v) (optimum 15 %) with 0–0.5 M MgSO₄·7H₂O (optimum 0.1 M) and at pH 6.0–9.0 (optimum pH 7.0). Additionally, the cells lyse in distilled water. The major polar lipids of strain CBA1103^T are phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate and two glycolipids chromatographically identical to sulfated mannosyl glucosyl diether and manosyl glucosyl diether. Strain CBA1103^T is shown to belong to the Halobellus genus and exhibits similarity to related taxa; the 16S rRNA gene sequence similarity between strain CBA1103^T and Halobellus rarus 18362^T, Hbs. limi 16811^T, Hbs. litoreus JCM 17118^T, Hbs. inordinatus YC20^T, Hbs. clavatus TNN18^T and Hbs. salinus CSW2.24.4^T is 97.3, 96.5, 96.5, 94.5, 94.5 and 93.7 %, respectively. The RNA polymerase subunit B gene sequence of strain CBA1103^T shows 93.7 % similarity with the sequence of Hbs. litoreus JCM 17118^T; the similarity is lower with sequences from the type strains of other species of Halobellus. The genomic DNA G+C content of strain CBA1103^T was determined to be 67.0 mol% a value which is in the range of the genomic DNA G+C content of members of the genus Halobellus (61.5–69.2 mol%). These results suggest that strain CBA1103^T should be considered to represent a new taxon for which the name Halobellus rufus sp. nov. is proposed, with the type strain CBA1103^T (=CECT 8423^T =JCM 19434^T).     

Zhihengliuella salsuginis sp. nov., a moderately halophilic actinobacterium from a subterranean brine

A novel moderately halophilic, alkaliphilic, non-motile, non-sporulating, catalase-positive, oxidase-negative, aerobic, coccus-shaped, Gram-positive bacterium, designated strain JSM 071043^T, was isolated from a subterranean brine sample collected from a salt mine in Hunan Province, China. Growth occurred with 0.5–20% (w/v) NaCl (optimum 5–10%) at pH 6.5–10.5 (optimum pH 8.5) and at 10–40°C (optimum 25–30°C). Good growth also occurred in the presence of 0.5–20% (w/v) KCl (optimum 5–8%) or 0.5–25% (w/v) MgCl₂·6H₂O (optimum 5–10%). The peptidoglycan type was A4α (l-Lys–l-Ala–l-Glu) and major cell-wall sugars were tyvelose and mannose. The major cellular fatty acids were anteiso-C_15:0, iso-C_16:0 and anteiso-C_17:0. Strain JSM 071043^T contained MK-9 and MK-8 as the predominant menaquinones and diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol as the major polar lipids. The DNA G + C content was 67.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain JSM 071043^T was a member of the suborder Micrococcineae, and was most closely related to Zhihengliuella halotolerans YIM 70185^T (sequence similarity 98.9%) and Zhihengliuella alba YIM 90734^T (98.2%), and the three strains formed a distinct branch in the phylogenetic tree. The combination of phylogenetic analysis, DNA–DNA relatedness values, phenotypic characteristics and chemotaxonomic data supports the proposal that strain JSM 071043^T represents a novel species of the genus Zhihengliuella, for which the name Z. salsuginis sp. nov. is proposed. The type strain is JSM 071043^T (= DSM 21149^T = KCTC 19466^T).     

<Emphasis Type="Italic">Yaniella soli</Emphasis> sp. nov., a new actinobacterium isolated from non-saline forest soil in China

A novel Gram-stain-positive, slightly halophilic, facultatively alkaliphilic, non-motile, non-sporulating, catalase-positive, oxidase-negative, aerobic bacterium, designated strain JSM 070026^T, was isolated from non-saline forest soil in China. Growth occurred with 0–20% (w/v) NaCl (optimum, 2–4%) and at pH 6.0–10.5 (optimum, pH 8.0) and 5–40°C (optimum, 30°C). Good growth also occurred in the presence of 0–28% (w/v) KCl (optimum, 2–5%) or 0–25% (w/v) MgCl₂·6H₂O (optimum, 1–4%). The peptidoglycan type was A4α (l-Lys–Gly–l-Glu). Cell-wall sugars contained mannose and xylose. The major cellular fatty acids were anteiso-C15:0 and iso-C15:0. Strain JSM 070026^T contained menaquinone 8 as the major respiratory quinone and diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol as the major polar lipids. The DNA G + C content of strain JSM 070026^T was 56.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain JSM 070026^T was a member of the suborder Micrococcineae and most closely related to Yaniella flava YIM 70178^T (sequence similarity 99.4%) and Yaniella halotolerans YIM 70085^T (97.9%). The three strains formed a distinct branch in the phylogenetic tree. The combination of phylogenetic analysis, DNA–DNA relatedness values, phenotypic characteristics and chemotaxonomic data supports the proposal that strain JSM 070026^T represents a novel species of the genus Yaniella, for which the name Yaniella soli sp. nov. is proposed. The type strain is JSM 070026^T (=DSM 22211^T = KCTC 13527^T).     

Halobellus ruber sp. nov., a deep red-pigmented extremely halophilic archaeon isolated from a Korean solar saltern

A novel halophilic archaeon, strain MBLA0160^T, was isolated from a solar saltern in Sorae, Republic of Korea. The cells are deep-red pigmented, Gram-negative, rod shaped, motile, and lysed in distilled water. The strain MBLA0160^T grew at 25–45 °C (optimum 37 °C), in 15–30% (w/v) NaCl (optimum 20%) and 0.1–1.0 M MgCl₂ (optimum 0.3–0.5 M) at pH 5.0–9.0 (optimum 7.0). Phylogenetic analysis based on the 16S rRNA sequence showed that this strain was related to two species within the genus Halobellus (Hbs.), with 98.4% and 95.8% similarity to Hbs. salinus CSW2.24.4 ^T and Hbs. clavatus TNN18^T, respectively. The major polar lipids of the strain MBLA160^T were phosphatidylglycerol, phosphatidylglycerol sulfate, and phosphatidylglycerol phosphate methyl ester. The genome size, G?+?C content, and N50 value of MBLA0160^T were 3.49 Mb, 66.5 mol%, and 620,127 bp, respectively. According to predicted functional proteins of strain MBLA0160^T, the highest category was amino acid transport and metabolism. Genome rapid annotation showed that amino acid and derivatives was the most subsystem feature counts. Pan-genomic analysis showed that strain MBLA0160^T had 97 annotated unique KEGG, which were mainly included metabolism and environmental information processing. Ortholog average nucleotide identities (OrthoANI) and in silico DNA-DNA hybridization (isDDH) values between the strain MBLA0160^T and other strains of the genus Halobellus were under 84,4% and 28.1%, respectively. The genome of strain MBLA0160^T also contain the biosynthetic gene cluster for C₅₀ carotenoid as secondary metabolite. Based on the phylogenetic, phenotypic, chemotaxonomic properties, and comparative genomic analyses, strain MBLA0160^T is considered to represent a novel species of the genus Halobellus, for which the name Halobellus ruber sp. nov. is proposed. The type strain is MBLA0160^T (=?KCTC 4291 ^T?=?JCM 34172 ^T).

     

Alkalilactibacillus ikkensis, gen. nov., sp. nov., a novel enzyme-producing bacterium from a cold and alkaline environment in Greenland

Three novel Gram-positive, endospore-forming bacteria were isolated from a cold and alkaline environment. Phylogenetic analysis showed that the strains were almost identical, and that they were related to Natronobacillus azotifigens 24KS-1^T (95.8% identity), Paraliobacillus quinghaiensis YIM-C158^T (95.1%), Paraliobacillus ryukyuensis O15-7^T (94.5%), and Halolactibacillus miurensis M23-1^T (93.9%). The isolates produced amylase, α-galactosidase, β-galactosidase, and β-glucuronidase, and showed optimal growth at pH 10, at 20°C, and at 2–8% (w/v) NaCl. Major fatty acids were C_14:0 (10.6–11.6%), anteiso-C_15:0 (25.7–32.7%), C_16:1 ω11c (12.2–16.0%), and C_16:0 (14.0–20.4%). The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol, and meso-diaminopimelic acid was found in the cell-wall peptidoglycan. The G+C content was 38.4%. DNA–DNA hybridization between strain GCM68^T and H. miurensis M23-1^T was 32.4%, while hybridization to N. azotifigens 24KS-1^T, Amphibacillus tropicus Z-7792^T, and Paraliobacillus ryukyuensis O15-7^T was below 30%. The phylogenetic analysis and G+C content place strain GCM68^T in relation to species belonging to Bacillus rRNA group 1, but phylogenetic and physiologic data combined with chemotaxonomic analyses support our proposal for a new genus, Alkalilactibacillus, gen. nov., with the novel species Alkalilactibacillus ikkensis, sp. nov. (type strain is GCM68^T = DSM 19937 = LMG 24405).     

Thermovorax subterraneus, gen. nov., sp. nov., a thermophilic hydrogen-producing bacterium isolated from geothermally active underground mine

A thermophilic, rod-shaped, motile, Gram-positive, spore-forming bacterium strain 70B^T was isolated from a geothermally active underground mine in Japan. The temperature and pH range for growth was 50–81°C (optimum 71°C) and 6.2–9.8 (optimum pH 7–7.5), respectively. Growth occurred in the presence 0–2% NaCl (optimum 1% NaCl). Strain 70B^T could utilize glucose, fructose, mannose, mannitol, pyruvate, cellobiose and tryptone as substrates. Thiosulfate was used as electron acceptor. Major whole-cell fatty acids were iso-C_15:0, C_16:0 DMA (dimethyl acetal), C_16:0 and anteiso-C_15:0. The G+C mol% of the DNA was 44.2%. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that the closest relatives of strain 70B^T were Thermosediminibacter oceani DSM 16646^T (94% similarity) and Thermosediminibacter litoriperuensis DSM 16647 (93% similarity). The phenotypic, chemotaxonomic and phylogenetic properties suggest that strain 70B^T represents a novel species in a new genus, for which the name Thermovorax subterraneus gen. nov., sp. nov. is proposed. The type strain of Thermovorax subterraneus is 70B^T (=DSM 21563 = JCM 15541).     

Virgibacillus zhanjiangensis sp. nov., a marine bacterium isolated from sea water

A Gram-positive, endospore-forming, catalase- and oxidase-positive, motile, rod-shaped, aerobic bacterium, designated strain JSM 079157^T, was isolated from surface seawater off the coastline of Naozhou Island in South China Sea. The organism was able to grow with 1–15% (w/v) total salts (optimum, 4–7%), and at pH 6.0–10.0 (optimum, pH 7.5) and 10–45°C (optimum, 30°C). meso-Diaminopimelic acid was present in the cell-wall peptidoglycan. The predominant menaquinone was MK-7, and the polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The major cellular fatty acids were anteiso-C_15:0 (45.1%) and anteiso-C_17:0 (16.2%), and the DNA G + C content was 39.5 mol%. A phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain JSM 079157^T should be assigned to the genus Virgibacillus, being related most closely to the type strains of Virgibacillus litoralis (97.4% sequence similarity), Virgibacillus necropolis (97.3%) and Virgibacillus carmonensis (97.1%). These four strains formed a distinct subcluster in the phylogenetic tree. The levels of DNA–DNA relatedness between the new isolate and the type strains of V. litoralis, V. necropolis and V. carmonensis were 30.4, 19.3 and 12.6%, respectively. The results of the phylogenetic analysis, combined with DNA–DNA relatedness data, phenotypic characteristics and chemotaxonomic information, support the suggestion that strain JSM 079157^T represents a new species of the genus Virgibacillus, for which the name Virgibacillus zhanjiangensis sp. nov. is proposed. The type strain is JSM 079157^T (=DSM 21084^T = KCTC 13227^T).     

Cohnella plantaginis sp. nov., a novel nitrogen-fixing species isolated from plantain rhizosphere soil

A novel bacterium capable of fixing nitrogen was isolated from plantain rhizosphere soil in China. The isolate, designated YN-83^T, is Gram-positive, aerobic, motile and rod-shaped (0.4–0.6 μm × 1.9–2.6 μm). Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain YN-83^T was a member of the genus Cohnella. High similarity of 16S rRNA gene sequence was found between YN-83^T and Cohnella ginsengisoli DSM18997^T (97.99%), whereas the similarity was below 96.0% between YN-83^T and the other Cohnella species. DNA–DNA relatedness between strain YN-83^T and C. ginsengisoli DSM18997^T was 27.4 ± 6.2%. The DNA G+C content of strain YN-83^T was 59.3 mol%. The predominant isoprenoid quinone was MK-7 and the major fatty acids were anteiso-C_15:0 (44.3%), iso-C_15:0 (11.3%), iso-C_16:0 (18.6%) and C_16:0 (7.7%). The polar lipids of strain YN-83^T consist of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, lyso- phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides. On the basis of phenotypic and chemotaxonomic properties, 16S rRNA gene sequence, G+C content and DNA–DNA hybridization, strain YN-83^T represents a novel species of the genus Cohnella, for which the name Cohnella plantaginis sp. nov. (type strain YN-83^T = DSM 25424^T = CGMCC 1.12047^T) is proposed.     

Aliidiomarina haloalkalitolerans sp. nov., a marine bacterium isolated from coastal surface seawater

A novel Gram-negative, rod shaped, motile, non-sporing strictly aerobic bacterium, designated strain AK5^T, was isolated from a sea water sample collected near Visakhapatnam coast, Bay of Bengal, India. Colonies on marine agar were circular, 3–4 mm in diameter, creamish and rose with entire margin. Growth occurred at 10–40°C, 0.5–12% (w/v) NaCl and pH of 7–11. Strain AK5^T was oxidase and catalase positive. The fatty acids were dominated by iso-branched saturated and unsaturated fatty acids with a high abundance of iso-C_15:0, iso-C_17:0 and summed feature 9 (as defined by MIDI). Q8 was found to be the major respiratory quinone and diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and four unidentified phospholipids as polar lipids. The DNA G+C content of strain AK5^T was 54.7 ± 0.2 mol%. Phylogenetic analysis revealed that strain AK5^T was a member of the genus Aliidiomarina and closely related to Aliidiomarina taiwanensis with a phylogenetic distance of 5.3% (94.7% 16S rRNA gene sequence similarity) and clustered with the same species. Results from the polyphasic taxonomy study support the conclusion that strain AK5^T represents a novel Aliidiomarina species, for which the name Aliidiomarina haloalkalitolerans sp. nov. is proposed. The type strain of A. haloalkalitolerans is AK5^T (= MTCC 11064^T = JCM 17359^T).