Methylovorus mays sp. nov.: A new species of aerobic, obligately methylotrophic bacteria associated with plants

A bacterial strain utilizing methanol as the sole source of carbon and energy was isolated from the maize phyllosphere. Cells are nonpigmented gram-negative motile rods that do not form spores or prosthecae and reproduce by binary fission. The strain does not require vitamins or supplementary growth factors. It is obligately aerobic and urease-, oxidase-, and catalase-positive. The optimum growth temperature is 35–40°C; the optimum pH is 7.0–7.5. The doubling time is 2 h. The bacterium implements the ribulose monophosphate pathway and possesses NAD⁺-dependent 6-phosphogluconate dehydrogenase and enzymes of the glutamate cycle. α-Ketoglutarate dehydrogenase and enzymes of the glyoxylate cycle (isocitrate lyase and malate synthase) are absent. Fatty acids are dominated by palmitic (C_16:0) and palmitoleic (C_16:1) acids. The major phospholipids are phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylcholine. Cardiolipin is present in minor amounts. The dominant ubiquinone is Q₈ The bacterial genome contains genes controlling the synthesis and secretion of cytokinins. The G+C content of DNA is 57.2 mol %, as determined from the DNA thermal denaturation temperature T_m. The bacterium shows low DNA homology (<10%) with restricted facultative methylotrophic bacteria of the genusMethylophilus (M. methylotrophus NCIMB 10515^T andM. leisingerii VKM B-2013¹) and with the obligate methylotrophic bacterium (Methylobacillus glycogenes ATCC 29475^T). DNA homology with the type representative of the genusMethylovorus, M. glucosetrophus VKM B-1745^T, is high (58%). The new isolate was classified as a new species,Methylovorus mays sp. nov.     

Halophilic and halotolerant aerobic methylobacteria from the technogenic Solikamsk biotopes

Seven strains of moderately halophilic and halotolerant aerobic methylobacteria from the technogenic Solikamsk biotopes (Perm krai, Russia) were isolated in pure cultures and characterized. The isolates were represented by gram-negative and gram-positive (strain 2395B) cells. All the cells were shown to multiply by binary fission without formation of spores or prosthecae. All isolates except strain 2395B were able to oxidize methanol by a classical methanol dehydrogenase. The ribulose monophosphate (RMP) (strain LS), serine (strains S12, S3, 2395A), or ribulose bisphosphate (strains SK15 and S3270) pathways of C₁-assimilation were used. In strain 2395B, the key enzymes of the RMP and serine metabolic pathways were determined. Using polyphasic taxonomy, three strains were identified as representatives of the known species: Arthrobacter protophormiae 2395B, Methylophaga thalassica LS, and Ancylobacter rudongensis S3270. Three more strains were identified as members of new species: Methylopila oligotropha sp. nov. (strain 2395AT; VKM B-2788^T = CCUG 63805^T), Ancylobacter defluvii sp. nov. (strain SK15^T; VKM B-2789^T = CCUG 63806^T), and Paracoccus communis sp. nov. (strain S3^T; VKM B-2787^T = CCUG 63804^T). According to the results of 16S rRNA gene sequencing, the obligately methylotrophic strain S12 had less than 94% similarity with the known genera of the Proteobacteria and was probably a representative of a novel genus.     

Methylobacillus arboreus sp. nov., and Methylobacillus gramineus sp. nov., novel non-pigmented obligately methylotrophic bacteria associated with plants

Two newly isolated obligate methanol-utilizing bacteria (strains Iva^T and Lap^T) with the ribulose monophosphate pathway of C₁ assimilation are described. The isolates are strictly aerobic, Gram negative, asporogenous, motile rods multiplying by binary fission, mesophilic and neutrophilic, synthesize indole-3-acetate. The prevailing cellular fatty acids are straight-chain saturated C_16:0 and unsaturated C_16:1 acids. The major ubiquinone is Q-8. The predominant phospholipids are phosphatidylethanolamine, phosphatidylglycerol and cardiolipin. Ammonia is assimilated by glutamate dehydrogenase. The DNA G+C contents of strains Iva^T and Lap^T are 54.0 and 50.5 mol% (T_m), respectively.Based on 16S rRNA gene sequence analysis and DNA–DNA relatedness (38–45%) with type strains of the genus Methylobacillus, the novel isolates are classified as the new species of this genus and named Methylobacillus arboreus Iva^T (VKM B-2590^T, CCUG 59684^T, DSM 23628^T) and Methylobacillus gramineus Lap^T (VKM B-2591^T, CCUG 59687^T, DSM 23629^T).The GenBank accession numbers for the 16S rRNA gene and mxaF gene sequences of the strains Iva^T and Lap^T are GU937479, GU937478 and HM030736, HM030735, respectively.     

<Emphasis Type="Italic">Methylobacillus caricis</Emphasis> sp. nov., an obligate methylotroph isolated from roots of sedge (<Emphasis Type="Italic">Carex</Emphasis> sp.)

A new obligately methylotrophic bacterium, strain OV^T, was isolated from roots of sedge (Carex sp.). The isolate was represented by aerobic gram-negative motile, non-spore-forming rods, which divided by binary fission. Optimal growth occurred at 22?29°C and pH 7.5?8.5 in the presence of 0.5?2% NaCl; growth was inhibited by 3.5% NaCl. Strain OV^T utilized methanol as the only carbon and energy source. The organism used the KDPG variant of the ribulose monophosphate pathway (RuMP) of С₁ metabolism. Ammonium was assimilated by reductive amination of α-ketoglutarate. The major cellular fatty acids were C_16:0 (45.5%), C_16:1ω7c (40.7%), and C_17cyc (8.0%). The major ubiquinone was Q₈. The dominant phospholipids were phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol. The DNA G + C content of strain OV^T was 51.4 mol % (T_m). While the 16S rRNA gene sequence of strain OV^T exhibited high similarity to those of Methylobacillus species: M. gramineus Lap^T (99.6%) and M. glycogenes TK 0113^T (98.7%), the DNA-DNA hybridization level between strain OV^T and M. gramineus Lap^T was only 52%. Based on the data obtained, strain OV^T was assigned to the new species Methylobacillus caricis sp. nov. (=VKM B-3158^T = JCM 32031^T).     

Stenotrophomonas chelatiphaga sp. nov., a new aerobic EDTA-degrading bacterium

A new ethylenediaminetetraacetic acid (EDTA)-utilizing gammaproteobacterial strain LPM-5^T was isolated from municipal sewage sludge. Aerobic, gram-negative, motile rods multiply by binary fission. Neutrophilic and mesophilic, these are unable to grow in the presence of 3% NaCl (w/v), and unable to reduce nitrate to nitrite, and are oxidase and catalase positive, but lipase negative. The major cellular fatty acids are C_i15:0, C_a15:0 and C_16:1w7c. The dominant phospholipids are phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol (cardiolipin). The DNA G+C content is 68.3 mol% (T_m). The 16S rRNA gene sequence analysis showed a high similarity of strain LPM-5^T to the species members of genus Stenotrophomonas: S. maltophilia LMG 958^T (98.6%), S. rhizophila CCUG 47042^T (98.3%), S. koreensis TR6-01^T (97.6%) and S. acidaminiphila CIP 106456^T (97.0%). Based on these results and modest DNA–DNA hybridization levels with S. maltophilia VKM B-591^T (=LMG 958^T) (51%) and S. rhizophila CCUG 47042^T (52%), the isolate was classified as a novel species, Stenotrophomonas chelatiphaga sp. nov. (type strain LPM-5^T=VKM B-2486=DSM-21508=CCUG 57178).     

Methylovorus mays--novel species of aerobic, obligatory methylotrophic bacteria associated with plants

A bacterial strain utilizing methanol as the sole source of carbon and energy was isolated from the maize phyllosphere. Cells are nonpigmented gram-negative motile rods that do not form spores or prosthecae and reproduce by binary fission. The strain does not require vitamins or supplementary growth factors. It is obligately aerobic and urease-, oxidase-, and catalase-positive. The optimum growth temperature is 35-40 degrees C; the optimum pH is 7.0-7.5. The doubling time is 2 h. The bacterium implements the ribulose monophosphate pathway and possesses NAD(+)-dependent 6-phosphogluconate dehydrogenase and enzymes of the glutamate cycle. alpha-Ketoglutarate dehydrogenase and enzymes of the glyoxylate cycle (isocitrate lyase and malate synthase) are absent. Fatty acids are dominated by palmitic (C16:0) and palmitoleic (C16:1) acids. The major phospholipids are phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylcholine. Cardiolipin is present in minor amounts. The dominant ubiquinone is Q8. The bacterial genome contains genes controlling the synthesis and secretion of cytokinins. The G + C content of DNA is 57.2 mol %, as determined from the DNA thermal denaturation temperature (Tm). The bacterium shows low DNA homology (< 10%) with restricted facultative methylotrophic bacteria of the genus Methylophilus (M. methylotrophus NCIMB 10515T and M. leisingerii VKM B-2013T) and with the obligate methylotrophic bacterium (Methylobacillus glycogenes ATCC 29475T). DNA homology with the type representative of the genus Methylovorus, M. glucosetrophus VKM B-1745T, is high (58%). The new isolate was classified as a new species, Methylovorus mays sp. now.     

Analysis of the Anaerobic Microbial Community Capable of Degrading p-Toluene Sulfonate

Three strains of Clostridium sp., 14 (VKM B-2201), 42 (VKM B-2202), and 21 (VKM B-2279), two methanogens, Methanobacterium formicicum MH (VKM B-2198) and Methanosarcina mazei MM (VKM B-2199), and one sulfate-reducing bacterium, Desulfovibrio sp. SR1 (VKM B-2200), were isolated in pure cultures from an anaerobic microbial community capable of degrading p-toluene sulfonate. Strain 14 was able to degrade p-toluene sulfonate in the presence of yeast extract and bactotryptone and, like strain 42, to utilize p-toluene sulfonate as the sole sulfur source with the production of toluene. p-Toluene sulfonate stimulated the growth of Ms. mazei MM on acetate. The sulfate-reducing strain Desulfovibrio sp. SR1 utilized p-toluene sulfonate as an electron acceptor. The putative scheme of p-toluene sulfonate degradation by the anaerobic microbial community is discussed.     

Fontivita pretiosa gen. nov., sp. nov., a thermophilic planctomycete of the order Tepidisphaerales from a hot spring of Baikal lake region

A novel facultatively anaerobic moderately thermophilic bacterium, strain B-254^T, was isolated from a terrestrial hot spring near the town of Goryachinsk in the Baikal lake region (Russian Federation). Motile spherical cells of the strain were present as single cocci, in pairs, or aggregates. The cells had a Gram negative cell wall and reproduced by binary fission. The isolate grew at 30–57 °C (opt. 50–54 °C) and at pH 5.1–8.4 (opt. 6.6–7.1). Strain B-254^T was a chemoorganoheterotroph, growing on mono-, di- and polysaccharides (xylan, starch, galactan, galactomannan, xyloglucan, arabinan, curdlan, beta-glucan, locust bean gum, xanthan gum). Sodium chloride or yeast extract were not required for growth. Major cellular fatty acids were iso-C_16:0, anteiso-C_17:0, and C_20:0; major polar lipid was phosphatidylethanolamine. The complete genome of strain B-254^T was 5.54 Mb; its GC content was 64 %. According to the results of 16S rRNA gene sequence-based phylogenetic analysis and the conserved proteins sequences-based phylogenomic analysis strain B-254^T was on a separate lineage within the order Tepidisphaerales (Phycisphaerae, Planctomycetes). Based on phylogenetic and phylogenomic analyses of Phycisphaerae, whole genome comparisons of Tepidisphaerales as well as distinctive phenotypic features of the strain, it was assigned to a novel genus and species for which the name Fontivita pretiosa gen. nov. sp. nov. is proposed. Strain B-254^T = KCTC 82380^T = VKM B-3507^T.     

<Emphasis Type="Italic">Singulispaera mucilagenosa</Emphasis> sp. nov., a novel acid-tolerant representative of the order <Emphasis Type="Italic">Planctomycetales</Emphasis>

Two novel strains of budding bacteria, Z-0071^T and Z-0072, were isolated from dystrophic humified waters formed by xylotrophic fungi in the course of spruce wood degradation. The cells of both strains are coccoid (0.95–1.80 μm), nonmotile, single or arranged in pairs. The cells have a complex system of intracellular membranes and are covered with fimbriae and surrounded by a mucous capsule up to 0.3 μm thick. Both strains are aerobic organoheterotrophic, mesophilic, and acid-tolerant microorganisms that are able to grow under microaerobic conditions. They utilize N-acetyl-glucosamine, carbohydrates, and lactate as growth substrates. The strains grow in a pH range of 4.0–7.5 with an optimum at 6.0–6.5. The temperature range for growth is 4–30°C, with an optimum at 25–28°C. Strains Z-0071^T and Z-0072, inhabitants of dystrophic low-mineral waters, are NaCl-sensitive: the NaCl content in the media above 0.5 g/l inhibited growth. The main fatty acids of strains Z-0071^T and Z-0072 are C_16:0, C_18:1ω9c, and C_{18:2ω9c, 12c}. The DNA G + C base content is 51.2–51.7 mol %. The sequences of the 16S rRNA gene fragments (1310 bp) of strains Z-0071^T and Z-0072 were found to be identical. The obtained sequences showed a 94.3% similarity with the sequences of the type strain of the most closely related species Singulisphaera acidiphila MOB10≅T. The phenotypic and phylogenetic properties of strains Z-0071^T and Z-0072 support classification of these strains within the genus Singulisphaera as a new species Singulisphaera mucilagenosa sp. nov., with the type strain Z-0071^T (VKM B-2626).     

Cell wall teichoic acids of Bacillus licheniformis VKM B-511T, Bacillus pumilus VKM B-508T, and other strains previously assigned to Bacillus pumilus

The teichoic acids (TAs) of type strains, viz. Bacillus licheniformis VKM B-511^T and Bacillus pumilus VKM B-508^T, as well as phylogenetically close bacteria VKM B-424, VKM B-1554, and VKM B-711 previously assigned to Bacillus pumilus on the basis of morphological, physiological, and biochemical properties, were investigated. Three polymers were found in the cell wall of each of the 5 strains under study. Strains VKM B-508^T, VKM B-424, and VKM B-1554 contained polymers of the same core: unsubstituted 1,3-poly(glycerol phosphate) (TA I) and 1,3-poly(glycerol phosphate) with O-D-Ala and N-acetyl-??-D-glucosamine substituents (TA II and TA III??, respectively). The cell walls of two remaining strains contained TA I, TA II, and a poly(glycosylpolyol phosphate) with the following structure of repeating units: -6)-??-D-GlcpNAc(1??1)-snGro-(3-P-(TA III?) in ??Bacillus pumilus?? VKM B-711 (100% 16S rRNA gene similarity with the type strain of Bacillus safensis) and -6)-??-D-Galp-(1??2)-snGro-(3-P-(TA III?) in Bacillus licheniformis VKM B-511^T. The simultaneous presence of three different TAs in the cell walls was confirmed by the NMR spectroscopic DOSY methods. The structure of the polymers and localization of O-D-Ala residues were investigated by the chemical and NMR spectroscopic methods.     

Methyloligella halotolerans gen. nov., sp. nov. and Methyloligella solikamskensis sp. nov., two non-pigmented halotolerant obligately methylotrophic bacteria isolated from the Ural saline environments

Two newly isolated halotolerant obligately methylotrophic bacteria (strains C2^T and SK12^T) with the serine pathway of C₁ assimilation are described. The isolates are strictly aerobic, Gram negative, asporogenous, non-motile rods, forming rosettes, multiplying by binary fission. Mesophilic and neutrophilic, accumulate intracellularly compatible solute ectoine and poly-β-hydroxybutyrate. The novel strains are able to grow at 0 up to 16% NaCl (w/v), optimally at 3–5% NaCl. The major cellular fatty acids are C_18:1ω7c and C_19:0cyc and the prevailing quinone is Q-10. The predominant phospholipids are phosphatidylcholine, phosphatidylglycerol, phosphatidyldimethylethanolamine and phosphatidylethanolamine. Assimilate NH₄⁺ by glutamate dehydrogenase and via the glutamate cycle (glutamine synthetase and glutamate synthase). The DNA G + C contents of strains C2^T and SK12^T are 60.9 and 60.5 mol% (Tm), respectively. 16S rRNA gene sequence similarity between the two new isolates are 99% but below 94% with other members of the Alphaproteobacteria thus indicating that they can be assigned to a novel genus Methyloligella. Rather low level of DNA–DNA relatedness (53%) between the strains C2^T and SK12^T indicates that they represent two separate species of the new genus, for which the names Methyloligella halotolerans gen. nov., sp. nov. and Methyloligella solikamskensis sp. nov. are proposed. The type strain of Methyloligella halotolerans is C2^T (=VKM B-2706^T = CCUG 61687^T = DSM 25045^T) and the type strain of Methyloligella solikamskensis is SK12^T (=VKM B-2707^T = CCUG 61697^T = DSM 25212^T).     

Ancylobacter dichloromethanicus sp. nov. – a new aerobic facultatively methylotrophic bacterium utilizing dichloromethane

A novel aerobic facultative methylotroph was isolated from contaminated soil. The organism (strain DM16) is a Gram-negative asporogenous non-motile curved rod multiplying by binary fission. Cells are neutrophilic and mesophilic. This strain utilized dichloromethane, methanol, formate and formaldehyde along with a variety of polycarbon compounds. Strain DM16 employs the ribulosebisphosphate pathway for C₁ assimilation. The DNA G+C content is 64.5 mol%. The major ubiquinone is Q-10. The dominant cellular fatty acids are 18:1ω7c (58.6%), cyclo-19:0ω8c (34.8%) and 16:0 (3.2%). Sequencing of the 16S rRNA gene and DNA–DNA hybridization experiments clearly indicated that this methylotroph should be classified as a new species within genus Ancylobacter – Ancylobacter dichloromethanicus sp. nov. with the type strain DM16^T (DSM 21507^T=VKM B-2484^T).     

<Emphasis Type="Italic">Sphingomonas jejuensis</Emphasis> sp. nov., isolated from marine sponge <Emphasis Type="Italic">Hymeniacidon flavia</Emphasis>

A Gram-negative, non-motile, rod shaped, and orange-pigmented chemoheterotrophic bacterium, strain MS-31^T was isolated from the marine sponge Hymeniacidon flavia, collected from near Jeju Island, Korea. The Strain MS-31^T was subjected to a polyphasic taxonomic study. The phylogenetic analysis based on the 16S rRNA gene sequences revealed that the novel isolate could be affiliated within the genus Sphingomonas. The strain MS-31^T showed 95.6% of 16S rRNA gene sequence similarity with the most closely related species Sphingomonas koreensis JSS26^T. The DNA G+C content of the strain MS-31^T was 69.4 mol%. The major isoprenoid quinone was ubiqunone 10 and predominant cellular fatty acids were summed feature 7 (comprising C_18:1 ω7c, C_18:1 Ω9t and/or C_18:1 ωl2t, 39.7%), C_16:0 (16.3%), C_14:0 2OH (15.9%) and summed feature 3 (comprising C_16:1 ω7c and/or C_15:0 iso 2OH, 11.7%). The polar lipids were sphingoglycolipid, phosphatidyletha-nolamine, phosphatidylglycerol, diphosphatidylglycerol and unidentified glycolipid. Based on the evidence from the polyphasic taxonomic study, the strain should be classified as a new species of the genus Sphingomonas. As a result, the name Sphingomonas jejuensis sp. nov. (type strain MS-31^T =KCTC 23321^T =NBRC 107775^T) is proposed.     

Vitamin B<Subscript>12</Subscript>-independent strains of <Emphasis Type="Italic">Methylophaga marina</Emphasis> isolated from Red Sea algae

Two strains (KM3 and KM5) of halophilic methylobacteria isolated from Red Sea algae do not require vitamin B₁₂ for growth and can use methanol, methylamine, dimethylamine, trimethylamine, dimethyl sulfide, and fructose as sources of carbon and energy. The cells of these strains are gram-negative motile monotrichous (strain KM3) or peritrichous (strain KM5) rods. The strains are strictly aerobic and require Na⁺ ions but not growth factors. They are oxidase-and catalase-positive and reduce nitrates to nitrites. Both strains can grow in a temperature range of 4 to 37°C (with optimal growth at 29–34°C), at pH between 5.5 and 8.5 (with optimal growth at pH 7.5–8.0), and in a range of salt concentrations between 0.5 and 15% NaCl (with optimal growth at 5–9% NaCl). The phospholipids of these strains are dominated by phosphatidylethanolamine and phosphatidylglycerol and also include phosphatidylcholine, phosphatidylserine, and cardiolipin. The dominant fatty acids are C_16:1ω7c and C_16:0. The major ubiquinone is Q₈. The cells accumulate ectoin, glutamate, and sucrose as intracellular osmoprotectants. The strains implement the 2-keto-3-deoxy-6-phosphogluconate-dependent variant of the ribulose monophosphate pathway. The G+C content of the DNA is 44.4–44.7 mol%. Analysis of the 16S rRNA genes showed that both strains belong to Gammaproteobacteria and have a high degree of homology (99.4%) to Methylophaga marina ATCC 35842^T. Based on the data of polyphasic taxonomy, isolates KM3 and KM5 are identified as new strains M. marina KM3 (VKM B-2386) and M. marina KM5 (VKM B-2387). The ability of these strains to produce auxins (indole-3-acetic acid) suggests their metabolic association with marine algae.     

<Emphasis Type="Italic">Henriciella marina</Emphasis> gen. nov., sp. nov., a novel member of the family <Emphasis Type="Italic">Hyphomonadaceae</Emphasis> isolated from the East Sea

A bacterial strain, designated Iso4^T, was isolated from the East Sea of Korea and was subjected to a poly-phasic taxonomy study including phenotypic and chemotaxonomic characteristics as well as 16S rRNA gene sequence analysis. Cells of the strain were Gram-negative, motile, non-budding, non-stalked, and strictly aerobic. Strain Iso4^T grew optimally at 20°C in the presence of 1∼2% (w/v) NaCl and at pH 6.9∼7.6. The major respiratory quinone was Q-10 and the major cellular fatty acids were C_18:1 ω7c (53.5%), C_17:1 ω5c (11.7%), C_17:1 ω6c (8.1%), C_16:0 (7.8%), C_17:0 (4.8%), C_15:0 (2.9%), and C_16:1 ω5c (2.2%). The DNA G+C content of strain Iso4^T was 56.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Iso4^T formed a monophyletic clade in the family Hyphomonadaceae, supported by high bootstrap value and was most closely related to the genus Hyphomonas (92∼94%), a member of marine bacteria in the family. The phenotypic, genotypic, and chemotaxonomic evidences also suggest strain Iso4^T represents a novel genus and species in the family Hyphomonadaceae, for which the name Henriciella gen. nov., sp. nov. is proposed. The type strain is Iso4^T (=KCTC 12513^T =DSM 19595^T =JCM 15116^T).     

Halonatronomonas betaini gen. nov., sp. nov., a haloalkaliphilic isolate from soda lake capable of betaine degradation and proposal of Halarsenatibacteraceae fam. nov. and Halothermotrichaceae fam. nov. within the order Halanaerobiales

A search for the organisms responsible for anaerobic betaine degradation in soda lakes resulted in isolation of a novel bacterial strain, designated Z-7014^T. The cells were Gram-stain-negative, non-endospore-forming rods. Growth occurred at 8–52 °C (optimum 40–45 °C), pH 7.1–10.1 (optimum pH 8.1–8.8) and 1.0–3.5 M Na⁺ (optimum 1.8 M), i.e. it can be regarded as a haloalkaliphile. The strain utilized a limited range of substrates, mostly peptonaceous but not amino acids, and was able to degrade betaine. Growth on betaine occurred only in the presence of peptonaceous substances which could not be replaced by vitamins. The G + C content of the genomic DNA of strain Z-7014^T was 36.1 mol%. The major cellular fatty acids (>5% of the total) were C_16:0 DMA, C_{18: 0} DMA, C_16:1ω8, C_16:0, C_18:1 DMA, C_16:1 DMA, C_18:1ω9, and C_18:0. Phylogenetic analysis of the 16S rRNA gene sequence revealed that strain Z-7014^T formed a distinct evolutionary lineage in the order Halanaerobiales with the highest similarity to Halarsenitibacter silvermanii SLAS-1^T (83.6%), Halothermothrix orenii H168^T (85.6%), and Halocella cellulosilytica DSM 7362^T (85.6%). AAI and POCP values between strain Z-7014^T and type strains of the order Halanaerobiales were 51.7–57.8%, and 33.8–58.3%, respectively. Based on polyphasic results including phylogenomic data, the novel strain could be distinguished from other genera, which suggests that strain Z-7014^T represents a novel species of a new genus, for which the name Halonatronomonas betaini gen. nov., sp. nov. is proposed. The type strain is Z-7014^T (=KCTC 25237^T = VKM B-3506^T). On the basis of phylogenomic data, it is also proposed to evolve two novel families Halarsenitibacteraceae fam. nov. and Halothermotrichaceae fam. nov. within the current order Halanaerobiales.     

Perlabentimonas gracilis gen. nov., sp. nov., a gliding aerotolerant anaerobe of the order Bacteroidales,isolated from a terrestrial mud volcano

A novel anaerobic bacterium (strain M08_MB^T) was isolated from a terrestrial mud volcano (Taman Peninsula, Russia). Gram-stain-negative cells were straight and slender rods with gliding motility, occasionally forming long filaments. The isolate was mesophilic, slightly halo- and alkaliphilic chemoorganoheterotroph, growing on carbohydrates (starch, dextrin, pectin, glucose, fructose, mannose, maltose, trehalose, lactose, sucrose) and proteinaceous compounds (peptone, tryptone, gelatin, casein and albumin). Strain M08_MB^T tolerated 3% oxygen in the gas phase while catalase negative. The dominant cellular fatty acids of strain M08_MB^T were C_15:0, C_15:1 and C_13:0 acids. 16S rRNA gene sequence analysis revealed that strain M08_MB^T belongs to the order Bacteroidales and only distantly related to other cultivated members of this order (85.12–90.01% 16S rRNA gene similarity). The genome of strain M08_MB^T had a size of 4.37 Mb with a DNA G + C content of 43.5 mol% (WGS). The genes involved in gliding motility, proteolysis, central carbon metabolism, and oxygen tolerance were listed in genome annotation. Based on the phenotypic and genotypic characteristics, strain M08_MB^T represents a novel species of a novel genus within family Tenuifilaceae, with proposed name Perlabentimonas gracilis gen. nov., sp. nov. The type strain is M08_ MB^T (=DSM 110720 ^T = VKM B-3471 ^T). This is the first representative of Bacteroidales isolated in pure culture from a mud volcano.     

<Emphasis Type="Italic">Thalassospira permensis</Emphasis> sp. nov., a new terrestrial halotolerant bacterium isolated from a naphthalene-utilizing microbial consortium

A halotolerant bacterium, strain SMB34^T, was isolated from a naphthalene-utilizing bacterial consortium obtained from primitive technogeneous soil (Verkhnekamsk salt deposit, Perm region, Russia) by enrichment procedure. The strain itself was unable to degrade naphthalene and grew at NaCl concentrations up to 11% (w/v). The 16S rRNA-based phylogenetic analysis showed that the strain belongs to the genus Thalassospira. The DNA-DNA hybridization values between SMB34^T and the type strains of phylogeneti-cally closest species (T. xiamenensis, T. profundimaris and T. tepidiphila) did not exceed 50%. The novel strain could be distinguished from the above species by the cell motility, MALDI/TOF mass spectra of whole cells and a range of physiological and biochemical characteristics. SMB34^T also considerably differs from the recently described species T. xianhensis, with the most striking differences in the DNA G + C content (53. ± 1.0 vs. 61.2 ± 1.0 mol %) and predominant ubiquinones (Q-10 vs. Q-9). The data obtained suggest strain SMB34^T (=VKM B-2527^T = NBRC 106175^T), designated as the type strain, represents a novel species, named Thalassospira permensis sp. nov.