Differential stability of bacterial photosynthetic apparatus of Rhodobacter alkalitolerans strain JA916T under alkaline and light environment.

In purple bacteria, photosynthesis is performed by densely packed pigment-protein complexes, including the light-harvesting complexes known as RC-LH1 and LH2, with carotenoids to assist in the functioning of photosynthesis. Most photosynthetic bacteria are exposed to various abiotic stresses such as light, temperature, alkalinity-acidity, and salinity. Rhodobacter (R.) alkalitolerans was discovered from the alkaline pond; here, we report the comparative study of the photosynthetic apparatus of R. alkalitolerans in various light intensities in relation to its high pH tolerance ability. With increased light intensity, the stability of photosystem complexes decreased in normal pH (npH pH 6.80 ± 0.05) conditions, whereas in high pH (hpH pH 8.60 ± 0.05), acclimation was observed to high light. The content of bacteriochlorophyll a, absorbance spectra, and circular dichroism data shows that the integrity of photosystem complexes is less affected in hpH compared with npH conditions. Large pore blue native polyacrylamide gel electrophoresis of photosystem protein complexes and sucrose density gradient of n-dodecyl ß-D-maltoside solubilized intracytoplasmic membranes show that LH2 is more affected in npH than in hpH, whereas RC-LH1 monomer or dimer has shown interplay between monomer and dimer in hpH, although the dimer and monomer both increased in npH. Increased content and expression level of ATPase protein complex and subunit-"c" of ATPase, fast relaxation kinetics of p515, and relatively higher membrane lipid content in hpH along with less photooxidative stress and subsequently lesser superoxide dismutase activity exemplify photoprotection in hpH. Furthermore, the increased expression levels of antiporter NhaD in hpH signify its role in the maintenance of homeostatic balance in hpH.

Sinomonas cellulolyticus sp. nov., isolated from Loktak lake.

A novel cellulolytic strain JC656T was isolated from the rhizosphere soil of Alisma plantago-aquatica of floating island (Phumdis) of Loktak lake, Manipur, India. The 16S rRNA gene sequence similarities between strain JC656T and other Sinomonas type strains ranged between 98.5 and 97.3%, wherein strain JC656T exhibited the highest sequence similarity (98.5%) to Sinomonas notoginsengisoli KCTC 29237T. Colonies were yellow-colored and grew aerobically. Cells were gram-positive, rod-shaped and non-motile. The optimal growth of the strain JC656T occured at 28 °C and pH 7. Strain JC656T contained MK-9 as the predominant isoprenoid quinone and anteiso-C15:0, iso-C16:0 and anteiso-C17:0 as the major fatty acids. Diphosphatidylglycerol, phosphatidylinositol, phosphatidylglycerol, phosphatidylmonomethylethanolamine and a glycolipid were the polar lipids. Strain JC656T contained lysine, alanine, glutamine, diaminopimelic acid (DAP) and two unidentified amino acids as characteristic cell wall amino acids. The genome size of strain JC656T was 3.9 Mb with a DNA G + C content of 69.9 mol %. For the affirmation of the strain's taxonomic status, a detailed phylogenomic study was done. Based on its phylogenetic position and morphological, physiological, and genomic features, strain JC656T represents a new species of the genus Sinomonas, for which we propose the name Sinomonas cellulolyticus sp. nov. The type strain JC656T = (KCTC 49339T = NBRC 114142T).

Draft Genome Sequence of Afifella sp. Strain JA880, Isolated from a Salt Pond.

We report the draft genome sequence of Afifella sp. strain JA880, which was isolated from a saltwater pond near Pata, Gujarat, India. The genome assembly contains 3,794,364 bp, with a GC content of 63.5%. The genome sequence provides insights into the metabolic potential of Afifella sp. strain JA880.

Neoroseomonas marina sp. nov., Isolated from a Beach Sand.

A pink-pigmented bacterium (strain JC162T = KCTC 32190T) was isolated from a beach sand sample. Cells were Gram-stain-negative, coccoid, non-motile, and strictly aerobic. EzBioCloud BLAST search of 16S rRNA gene sequence showed that strain KCTC 32190T had the highest sequence identity to the members of the genus Neoroseomonas and was closely related to N. oryzicola YC6724T (99.8%), N. sediminicola FW-3T (98.5%), N. soli 5N26T (98.2%), and other members of the genus Neoroseomonas (< 97.9%) in the family Acetobacteriaceae within the class of Alphaproteobacteria. Chemo-organoheterotrophy was the only growth mode and growth was possible on a wide range of organic substrates. Strain KCTC 32190T was positive for catalase and oxidase. Fatty acid composition of strain KCTC 32190T includes (in decreasing %) C18:1ω7c, cyclo-C19:0ω8c, C18:02-OH, C16:0, C18:03-OH, C16:1ω7c/C16:1ω6c, C16:02-OH and C16:1ω5c. Polar lipids comprised of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, an unidentified amino lipid, and three unidentified lipids. The genomic DNA G+C content of the strain KCTC 32190T was 70.9 mol%. Strain KCTC 32190T has a low ANI value of < 92.7% and genome reassociation (based on digital DNA-DNA hybridization) value of < 48.8% with the nearest type strains. The genome relatedness is supported by other polyphasic taxonomic data to propose strain KCTC 32190T as a new species in the genus Neoroseomonas with the name Neoroseomonas marina sp. nov. The type strain is strain JC162T (KCTC 32190T = CGMCC1.12364T).

Tryptophan, a non-canonical melanin precursor: New L-tryptophan based melanin production by Rubrivivax benzoatilyticus JA2.

Melanins are chemically diverse ubiquitous pigments found across the life forms synthesized via different biochemical pathways mainly from L-tyrosine or acetyl CoA. Though few reports suggest the possibility of tryptophan-based melanin synthesis, however, such tryptophan-based melanin and its biosynthesis remained a biochemical riddle. Here we report tryptophan-based melanin production by bacterium, Rubrivivax benzoatilyticus JA2. Aerobic cultures of strain JA2 produced brown pigment when grown on L-tryptophan-containing media. Purified pigment showed typical physico-chemical properties of melanin. Further, extensive spectroscopic studies revealed that pigment is an amorphous, indole-type polymer with stable free radical centers. Further, hydrolysis of the brown pigment revealed the presence of indole moiety, confirming the indolic nature of the pigment. Demonstration of in vitro and in vivo pigment synthesis directly from L-tryptophan or hydroxytryptophan confirms tryptophan-based melanin synthesis in strain JA2. Interestingly, canonical melanin biosynthetic inhibitors did not affect the pigment synthesis indicating possible non-canonical tryptophan-based melanin biosynthesis in strain JA2. Further, the exometabolite profiling and precursor feeding studies suggests that L-tryptophan converted to hydroxytryptophan/hydroxyindoles and their subsequent polymerization lead to the formation of melanin. The current study sheds light on biosynthetic diversity of melanins and L-tryptophan can be a potential precursor for melanin synthesis in life forms.

New insights into aniline toxicity: Aniline exposure triggers envelope stress and extracellular polymeric substance formation in Rubrivivax benzoatilyticus JA2.

Aniline is a major environmental pollutant of serious concern due to its toxicity. Although microbial metabolism of aniline is well-studied, its toxic effects and physiological responses of microorganisms to aniline are largely unexplored. Rubrivivax benzoatilyticus JA2, an aniline non-degrading bacterium, tolerates high concentrations of aniline and produces extracellular polymeric substance(EPS). Surprisingly, strain JA2 forms EPS only when exposed to aniline and other toxic compounds like organic solvents and heavy metals indicating that EPS formation is coupled to cell toxicity. Further, extensive reanalysis of the previous proteomic data of aniline exposed cells revealed up-regulation of envelope stress response(ESR) proteins such as periplasmic protein folding, envelope integrity, transmembrane complex, and cell-wall remodelling proteins. In silico analysis and molecular modeling of three highly up-regulated proteins revealed that these proteins were homologous to CpxARP proteins of ESR signalling pathway. Furthermore, EPS formation to known ESR activators(Triton-X-100, EDTA) suggests that envelope stress possibly regulating the EPS production. The present study suggests that aniline triggers envelope stress; to counter this strain JA2 activates ESR pathway and EPS production. Our study revealed the hitherto unknown toxic effects of aniline as an acute envelope stressor thus toxicity of aniline may be more profound to life-forms than previously thought.

Precursor-feeding and altered-growth conditions reveal novel blue pigment production by Rubrivivax benzoatilyticus JA2.

OBJECTIVE: To explore the secondary metabolite biosynthetic potential of Rubrivivax benzoatilyticus JA2 using a new metabolite mining strategy. RESULTS: Combination of precursor-feeding and altered growth conditions were used to mine new biomolecules. Strain JA2 utilised L-phenylalanine as sole source of nitrogen and showed pigments production only under phenylalanine-amended aerobic cultures. Stable isotope based precursor feeding studies indicated the blue pigment consists of 4-phenyl rings derived from L-phenylalanine. The purified blue pigment displayed characteristic visible-absorption and pH-dependent color variations. Precursor-feeding under altered growth conditions activated the plausible novel aromatic pigment production in strain JA2. CONCLUSION: Our approach unraveled the previously unknown pigment synthesis in strain JA2 and demonstrated the potential of mining strategy in discovering the hidden secondary metabolite repertoire in microorganisms.

Phylogenetic diversity of sulfate-reducing bacteria of sediments of Chilika Lake, India, determined through analysis of the dissimilatory sulfite reductase (dsr AB) gene.

In this study, the sulfate-reducing bacteria, (SRB) were identified and reported for the first time through analysis of functional gene dsrAB, from the DNA of sediment samples collected from 10 sites of the Chilika lake. The finding illustrates Forty six Operational Taxonomic Units (OTUs), identified from the DGGE which were obtained from the 10 sediment samples. Of these, 34 OTUs exhibited around 78-96% sequence similarity and 12 OTUs showed 97 to 100% sequence similarity to the dsrAB gene of reported type strains of SRB. The sequence information obtained revealed the presence and distribution of diverse types of SRB which include phylotypes related to Desulfovibrio, Desulfonatronovibrio, Desulfomicrobium, Desulfobotulous and Desulfobacca. Upon comparison of dsrAB gene sequences of SRB obtained through this study with those collected from the GenBank, and through the dendrogram constructed, it was observed that except 13 OTUs that clustered closely with the reported type strains, all other 36 OTUs clustered distantly and had no representative member of SRB. This indicated the presence of phylogenetically diverse groups of SRB inhabiting the lake Chilika.

iTRAQ-based quantitative proteomics reveals insights into metabolic and molecular responses of glucose-grown cells of Rubrivivax benzoatilyticus JA2.

Anoxygenic photosynthetic bacteria thrive under diverse habitats utilising an extended range of inorganic/organic compounds under different growth modes. Although they display incredible metabolic flexibility, their responses and adaptations to changing carbon regimes is largely unexplored. In the present study, we employed iTRAQ-based global proteomic profiling and physiological studies to uncover the adaptive strategies of a phototrophic bacterium, Rubrivivax benzoatilyticus JA2 to glucose. Strain JA2 displayed altered growth rates, reduced cell size and progressive loss of pigmentation when grown on glucose compared to malate under photoheterotrophic condition. A ten-fold increase in the saturated to unsaturated fatty acid ratio of glucose-grown cells indicates a possible membrane adaptation. Proteomic profiling revealed extensive metabolic remodelling in the glucose-grown cells wherein signal-transduction, selective-transcription, DNA-repair, transport and protein quality control processes were up-regulated to cope with the changing milieu. Proteins involved in DNA replication, translation, electron-transport, photosynthetic machinery were down-regulated possibly to conserve the energy. Glycolysis/gluconeogenesis, TCA cycle and pigment biosynthesis were also down-regulated. The cell has activated alternative energy metabolic pathways viz., fatty acid ß-oxidation, glyoxylate, acetate-switch and Entner-Doudoroff pathways. Overall, the present study deciphered the molecular/metabolic events associated with glucose-grown cells of strain JA2 and also unraveled how a carbon source modulates the metabolic phenotypes. SIGNIFICANCE: Anoxygenic photosynthetic bacteria (APB) exhibit incredible metabolic flexibility leading to diverse phenotypes. They thrive under diverse habitat using an array of inorganic/organic compounds as carbon sources, yet their metabolic adaptation to varying carbon regime is mostly unexplored. Present study uncovered the proteomic insights of the cellular responses of strain JA2 to changing carbon sources viz. malate and glucose under photoheterotrophic conditions. Our study suggests that carbon source can also determine the metabolic fate of the cells and reshape the energy dynamics of APB. Here, for the first time study highlighted the plausible carbon source (glucose) mediated regulation of photosynthesis in APB. The study sheds light on the plausible cellular events and adaptive metabolic strategies employed by strain JA2 in presence of non-preferred carbon source. It also revealed new insights into the metabolic plasticity of APB to the changing milieu.

Rhodobacter alkalitolerans sp. nov., isolated from an alkaline brown pond.

An alkali-tolerant, Gram-stain-negative, motile, rod-to-oval-shaped, yellowish brown-colored, phototrophic bacterium, designated as strain JA916T, was isolated from an alkaline brown pond in Gujarat, India. The DNA G + C content of the strain JA916T was 65.1 mol%. Strain JA916T grew well at pH 10. Respiratory quinone was Q-10 and major fatty acid was C18:1ω7c/C18:1ω6c, with significant quantities of C15:02OH observed. Strain JA916T shared the highest 16S rRNA gene sequence similarity with the type strains of Rhodobacter johrii (98.4%), followed by Rhodobacter megalophilus (98.3%), Rhodobacter sphaeroides (98.3%), Rhodobacter azotoformans (97.9%) and other members of the genus Rhodobacter (< 97%). 16S rRNA gene-based phylogenetic tree shows that strain JA916T formed a distinct sub-clade with Rhodobacter johrii, Rhodobacter megalophilus, Rhodobacter sphaeroides and Rhodobacter azotoformans. Further, rpoB-based phylogenetic analysis showed lower similarity with closely related species (≤ 93.0%) of the genus Rhodobacter, which suggests that JA916T is a novel species of the genus Rhodobacter. DNA-DNA hybridization values between strain JA916T and related type strains were less than 40%. Phenotypic, chemotaxonomical and phylogenetic differences showed that strain JA916T was distinct from other species of the genus Rhodobacter, suggesting strain JA916T represents a new species of the genus for which the name Rhodobacter alkalitolerans sp. nov. is proposed. Type strain is JA916T (= KCTC 15473T = LMG 28749T).

Insights into the carbonic anhydrases and autotrophic carbon dioxide fixation pathways of high CO2 tolerant Rhodovulum viride JA756.

Biofixation of CO2 is being extensively investigated to solve the global warming problem. Purple non-sulfur bacteria are fast growers that consume CO2 and produce beneficial biomass. Better the growth at higher CO2 levels, more efficient are the strains for biofixation. Nine among fifty strains that were analyzed at elevated CO2 levels responded with better growth. Considering its enhanced growth at high CO2 and metabolic versatility, Rhodovulum viride strain JA756 was chosen to make further studies. Strain JA756 tolerates up to 50% (v/v) CO2 with its optimum between 20-40% (v/v), yielding a biomass of 3.4 g. L-1. The pattern of specific enzyme activity of carbonic anhydrase corresponded well with that of its growth. To gain insights into the genomic composition and genes related to carbonic anhydrases and CO2 fixation, draft genome sequencing of JA756 was carried out which revealed the presence of two non-homologous genes encoding for ß and γ carbonic anhydrases, both of which are assumed to be implicated in maintaining intracellular inorganic carbon concentration at equilibrium. Most of the genes involved in the Calvin pathway, reductive tricarboxylic acid pathway, 3-hydroxypropionate bicycle and C4 pathways were found in the draft genome. While the experimental determinations of active roles of two of these pathways are still underway, the expression of key genes of Calvin and C4 pathway suggest their functional role in the organism. Owing to its metabolic versatility, JA756 can be advantageous for biological CO2 assimilation facilities located by the coastline, inland and also at wide ranges of CO2 concentrations.

Stable Isotope-Assisted Metabolic Profiling Reveals Growth Mode Dependent Differential Metabolism and Multiple Catabolic Pathways of l-Phenylalanine in Rubrivivax benzoatilyticus JA2.

Anoxygenic phototrophic bacteria are metabolically versatile and survive under different growth modes using diverse organic compounds, yet their metabolic diversity is largely unexplored. In the present study, we employed stable-isotope-assisted metabolic profiling to unravel the l-phenylalanine catabolism in Rubrivivax benzoatilyticus JA2 under varying growth modes. Strain JA2 grows under anaerobic and aerobic conditions by utilizing l-phenylalanine as a nitrogen source. Furthermore, ring-labeled 13C6-phenylalanine feeding followed by liquid chromatography-mass spectrometry exometabolite profiling revealed 60 labeled metabolic features (M + 6, M + 12, and M + 18) derived solely from l-phenylalanine, of which 11 were identified, 7 putatively identified, and 42 unidentified under anaerobic and aerobic conditions. However, labeled metabolites were significantly higher in aerobic compared to anaerobic conditions. Furthermore, detected metabolites and enzyme activities indicated multiple l-phenylalanine catabolic routes mainly Ehrlich, homogentisate-dependent melanin, benzenoid, and unidentified pathways operating under anaerobic and aerobic conditions in strain JA2. Interestingly, the study indicated l-phenylalanine-dependent and independent benzenoid biosynthesis in strain JA2 and a differential flux of l-phenylalanine to Ehrlich and benzenoid pathways under anaerobic and aerobic conditions. Additionally, unidentified labeled metabolites strongly suggest the presence of unknown phenylalanine catabolic routes in strain JA2. Overall, the study uncovered the l-phenylalanine catabolic diversity in strain JA2 and demonstrated the potential of stable isotope-assisted metabolomics in unraveling the hidden metabolic repertoire.

Desertibacillus haloalkaliphilus gen. nov., sp. nov., isolated from a saline desert.

Two Gram-stain-positive, rod-shaped and endospore-forming bacteria that represent a single species, designated strains KJ1-10-99T and KJ1-10-93, were isolated from a saline desert of Little Rann of Kutch, Gujarat, India. Analysis of 16S rRNA gene sequences revealed that the isolates belonged to the family Bacillaceae and were closely related to each other with 16S rRNA gene sequence similarity of 99.9 %. However, these two isolates formed a novel phylogenetic branch within this family. Both strains were aerobic, catalase and oxidase positive, and could grow optimally at 37 °C and pH 9. Further, strains KJ1-10-99T and KJ1-10-93 grew optimally at a NaCl concentration of 7.5 and 15 % (w/v), respectively. Both strains shared highest sequence similarity with Fermentibacillus polygoni IEB3T (96.90 %) followed by Bacillus nanhaiisediminis NH3T (96.3 %) and Bacillus alkalinitrilicus ANL-iso4T (96.3 %). The major cellular fatty acids were anteiso-C15 : 0, anteiso-C17:0, C16 : 0, and iso-C15 : 0. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol in both strains. The predominant isoprenoid quinone was MK-7 in both the strains. The peptidoglycan contained meso-diaminopimelic acid (meso-DAP) as the diagnostic diamino acid. The DNA G+C content of strains KJ1-10-99T and KJ1-10-93 were 48.7 and 48.9 mol% respectively. Both strains could be distinguished from closest phylogenetic neighbours based on a number of phenotypic properties. On the basis of polyphasic taxonomic analysis and phylogenetic data, we conclude that the strains KJ1-10-99T (=LMG 29918T=KCTC 33878T) and KJ1-10-93 (=LMG 29919=KCTC 33877) represent a novel species of a new genus in the family Bacillaceae, order Bacillales, for which the name Desertibacillus haloalkaliphilus gen. nov., sp. nov. is proposed.

Bacillus catenulatus sp. nov., an alkalitolerant bacterium isolated from a soda lake.

Two novel (18CT and 6C) Gram-stain-positive, rod shaped, motile and endospore-forming bacterial strains were isolated from Lonar soda lake, India. Based on 16S rRNA gene sequence analysis, strains 18CT and 6C were identified as belonging to the class Firmibacteria, and were most closely related to Bacillus cohnii KCTC 3572T (99.3 and 99.9%, respectively), Bacillus zhanjiangensis KCTC 13713T (97.4 and 98.0%, respectively), Bacillus halmapalus LMG 17950T (97.0 and 97.6%, respectively) and other members in the genus Bacillus (<97.0%). However, the DNA-DNA relatedness between 18CT and 6C and B. cohnii KCTC 3572T (49.6 ± 0.9 and 51.6 ± 0.7, respectively), B. zhanjiangensis KCTC 13713T (42.9 ± 0.8 and 47.1 ± 0.3, respectively) and B. halmapalus LMG 17950T (39.9 ± 0.8 and 40.8 ± 0.3, respectively) indicated that the novel strains were distantly related to these strains. Further, the high 16S rRNA gene sequence similarity (100%) and DNA-DNA relatedness (90 ± 5%) suggested that strains 18CT and 6C were members of a genomospecies. The strains grew optimally at a pH of 7.5 with 2-3% (w/v) NaCl and temperature of 37 °C. Strains 18CT and 6C were catalase and oxidase negative. The cell wall of strain 18CT contained meso-diaminopimelic acid as the diagnostic diamino acid, which was in contrast with its nearest neighbour B. cohnii KCTC 3572T, which contained ornithine and aspartic acid. Polar lipids include diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), an unknown phospholipid (PL) and three unknown lipids (L1-3). The predominant isoprenoid quinone was MK-7. iso-C15:0 (32.5%) was the predominant fatty acid and significant proportions of anteiso-C15:0 (19.5%), C16:0 (11.5%), iso-C17:0 (9.5%) and anteiso-C17:0 (6.3%) were also detected. The DNA G + C content of strains 18CT and 6C were 39.3 and 39.2 mol%, respectively. The results of molecular, biochemical and chemotaxonomic tests showed a clear differentiation of strains 18CT and 6C from all other members of the genus Bacillus, for which the name Bacillus catenulatus sp. nov. is proposed. The type strain is 18CT (=KCTC 33781T = CGMCC 1.15475T).

Nesterenkonia cremea sp. nov., a bacterium isolated from a soda lake.

A Gram-stain-positive, aerobic, non-motile, rod-shaped, non-endospore-forming bacterial strain, 10CT, was isolated from Lonar soda lake in India. Based on the 16S rRNA gene sequence analysis, this strain was identified as belonging to the genus Nesterenkonia and was most closely related to the type strains of Nesterenkonia lacusekhoensis (99.1 %, sequence similarity), Nesterenkonia aethiopica (96.9 %), Nesterenkonia flava (96.9 %) and related of the genus Nesterenkonia (<96.6 %, sequence similarity). However, the DNA-DNA relatedness of strain 10CT with N. lacusekhoensis KCTC 19283T was only 34.6±0.9. The DNA G+C content of strain 10CT was 68.6 mol%. Strain 10CT was an aerobic microbe with optimal growth at 37 °C, pH 7.5-8.0 and 5-6 % (w/v) NaCl. The cell-wall peptidoglycan of strain 10CT was of the type A4α (l-Lys-l-Glu). The major polar lipids present were phosphatidylglycerol, diphosphatidylglycerol and phosphatidylcholine. The major isoprenoid quinones were MK-7, MK-8 and MK-9. Major fatty acids of strain 10CT were anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The results of phylogenetic, chemotaxonomic and biochemical tests allowed a clear differentiation of strain 10CT, which represents a novel member of the genus Nesterenkonia for which the name Nesterenkonia cremea sp. nov. is proposed. The type strain is 10CT (=LMG 29100T=KCTC 39636T=CGMCC 1.15388T).

Bacillus alcaliphilum sp. nov., a bacterium isolated from a soda lake.

Two novel (14BT and 7B) Gram-stain-positive, rod-shaped, motile and endospore-forming bacterial strains were isolated from Lonar soda lake, India. Based on 16S rRNA gene sequence analysis, the strains 14BT and 7B were identified as belonging to the class Firmibacteria and were most closely related to Bacillus halodurans LMG 7121T (99.7 and 99.8%, respectively), Bacillus okuhidensis LMG 22468T (99.1 and 99.2%, respectively) and other members in the genus Bacillus (<97.0%). However, the DNA-DNA relatedness studies indicated that the strains 14BT and 7B were distantly related to B. halodurans LMG 7121T (49.1 ± 0.6 and 45.7 ± 0.6, respectively) and B. okuhidensis LMG 22468T (40.9 ± 0.9 and 42.1 ± 0.5, respectively). The high 16S rRNA gene sequence similarity (99.9%) and DNA-DNA relatedness (88 ± 9) indicated that strains 14BT and 7B were members of a single species. The strains grew optimally at a pH of 9.0-9.5 with 2-5% (w/v) NaCl and temperature of 37 °C. Strains 14BT and 7B were catalase positive and oxydase negative. The cell wall of strain 14BT contained meso-diaminopimelic acid as the diagnostic diamino acid. Polar lipids include diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), an unknown aminophospholipid (APL1) and three unknown lipids (L1-3). The predominant isoprenoid quinone is MK-7. anteiso-C15:0 (30.8%) was the predominant fatty acid, and significant proportions of iso-C15:0 (24.9%), iso-C16:0 (17.9%) and anteiso-C17:0 (12.3%) were also detected in strains 14BT and 7B. The DNA G+C content of strains 14BT and 7B was 41.6 and 41.3 mol%, respectively. The results of molecular, physiological and biochemical tests allowed a clear differentiation of strains 14BT and 7B from all other members of the genus Bacillus, for which the name Bacillus alcaliphilum sp. nov. is proposed. The type strain is 14BT (=KCTC 33777T = CGMCC 1.15474T).

Tersicoccus solisilvae sp., nov., a bacterium isolated from forest soil.

A Gram-stain-positive, aerobic, non-motile, coccoid bacterial strain, 36AT, was isolated from Munnar, in India. Based on the results of 16S rRNA gene sequence analysis, this strain was identified as representing a member of the genus Tersicoccus and was most closely related to the type strain of Tersicoccus phoenicis (98.9 %, sequence similarity), the only other member of the genus, and to members of the related genus Arthrobacter (<96.1 %, sequence similarity). However, the DNA-DNA relatedness of strain 36AT with T. phoenicis DSM 30849T was only 31.9±0.8. The DNA G+C content of strain 36AT was 70.9 mol%. Strain 36AT was an aerobic microbe with optimal growth at 37 °C, pH 6.0-8.0 and NaCl 0.5-3 % (w/v). Cell-wall peptidoglycan of strain 36AT was of type A11.20 (Lys-Ser-Ala2). Polar lipids present were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol, two phospholipids, a glycolipid and four unknown lipids. The major isoprenoid quinones were MK-9 (H2) and MK-8 (H2). Major fatty acids of strain 36AT were anteiso-C15 : 0, anteiso-C17 : 0, iso-C16 : 0 and iso-C15 : 0. The results of phylogenetic, chemotaxonomic and biochemical tests allowed a clear differentiation of strain 36AT, which represents a novel member of the genus Tersicoccus, for which the name Tersicoccus solisilvae sp. nov., is proposed. The type strain is 36AT (=KCTC 33776T=CGMCC 1.15480T).

Pontibacillus salipaludis sp. nov., a slightly halophilic bacterium isolated from a salt pan.

Two novel Gram-stain-positive, facultatively anerobic, rod-shaped, motile, endospore-forming bacterial strains, 9DMT and 6DM, were isolated from Tuticorn, India. The high 16S rRNA gene sequence similarity (99.9 %) and DNA-DNA relatedness (88±2 %) indicated that strains 9DMT and 6DM were members of a single species. Based on 16S rRNA gene sequence analysis these strains were identified as belonging to the genus Pontibacillus and were related most closely to the type strains of Pontibacillus chungwhensis (99.7 % sequence similarity) and Pontibacillus marinus (97.9 %). The DNA G+C content of strains 9DMT and 6DM was 41.6 and 41.4 mol%, respectively. However, the level of DNA-DNA relatedness of these strains with P. chungwhensis KCTC 3890T and P. marinus KCTC 3917T was only 40.9 + 0.8 and 39.8 + 0.9 %, respectively. Strains 9DMT and 6DM were facultative anaerobes with optimal growth at 37 °C, at pH 7.0-8.0 and with 2-3 % (w/v) NaCl. The cell-wall peptidoglycan of strain 9DMT contained meso-diaminopimelic acid. Polar lipids included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, a phospholipid and three unknown lipids. The isoprenoid quinone was MK-7. Major fatty acids of strain 9DMT were anteiso-C15 : 0, iso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. The results of phylogenetic, chemotaxonomic and biochemical tests allowed the clear differentiation of strains 9DMT and 6DM, which are considered to represent a novel member of the genus Pontibacillus, for which the name Pontibacillus salipaludis sp. nov. is proposed. The type strain is 9DMT (= KCTC 33742T=LMG 29102T= CGMCC 1.15353T).

Pelagirhabdus alkalitolerans gen. nov., sp. nov., an alkali-tolerant and thermotolerant bacterium isolated from beach sediment, and reclassification of Amphibacillus fermentum as Pelagirhabdus fermentum comb. nov.

A novel bacterial strain, designated S5T, was isolated from Pingaleshwar beach, in India. Cells were Gram-stain-positive, rod-shaped, non-motile and non-endospore-forming. Based on 16S rRNA gene sequence analysis, the strain was identified as belonging to the class Firmibacteria and was related most closely to Amphibacillus fermentum DSM 13869T (97.6 % sequence similarity). However, it shared only 93.1 % 16S rRNA gene sequence similarity with Amphibacillus xylanus NBRC 15112T, the type species of the genus, indicating that strain S5T might not be a member of the genus Amphibacillus. The DNA-DNA relatedness between strain S5T and Amphibacillus fermentum DSM 13869T was 39 %. The cell-wall peptidoglycan contained meso-diaminopimelic acid. Polar lipids included diphosphatidylglycerol, phosphatidylglycerol and two phospholipids. Isoprenoid quinones were absent from strain S5T. Fatty acid analysis revealed that anteiso-C15 : 0, C16 : 0 and iso-C15 : 0 were the predominant fatty acids present. The results of phylogenetic, chemotaxonomic and biochemical tests allowed the clear differentiation of strain S5T, which is considered to represent a novel species of a new genus in the family Bacillaceae, for which the name Pelagirhabdus alkalitolerans gen. nov., sp. nov. is proposed. The type strain of Pelagirhabdus alkalitolerans is S5T ( = KCTC 33632T = CGMCC 1.15177T). Based on the present study, it is also suggested to transfer Amphibacillus fermentum to this new genus, as Pelagirhabdus fermentum comb. nov. The type strain of Pelagirhabdus fermentum is Z-7984T = (DSM 13869T = UNIQEM 210T).

Cellulosimicrobium aquatile sp. nov., isolated from Panagal reservoir, Nalgonda, India.

A Gram-stain positive, non-spore forming, non-motile, yellow-pigmented, rod shaped and coccoid aerobic strain designated 3bp(T) was isolated from Panagal reservoir, at Nalgonda, Telangana, India. Based on 16S rRNA gene sequence analysis it was identified that strain 3bp(T) belongs to the class Actinobacteria and is closely related to Cellulosimicrobium funkei VTT E-072700(T), Cellulosimicrobium cellulans LMG16221(T) and Cellulosimicrobium terreum KCTC 19206(T). The DNA-DNA relatedness data demonstrated that strain 3bp(T) is distinguishable from the above three species of the genus. Further, the chemotaxonomic properties of strain 3bp(T) were consistent with those of the genus Cellulosimicrobium: MK-9(H4) was the predominant menaquinone, anteiso-C15 : 0, iso-C15:0,  anteiso-C17 : 0 and iso- C16:0 were the predominant cellular fatty acids and the cell-wall peptidoglycan contains L-lysine. The DNA G+C content of strain 3bp(T) was 73.8 mol%. The results of the polyphasic analysis allowed a clear differentiation of strain 3bp(T) from all other members of the genus Cellulosimicrobium. Strain 3bp(T) is thus considered to represent a novel member of the genus Cellulosimicrobium, for which the name Cellulosimicrobium aquatile sp. nov. is proposed. The type strain is 3bp(T) (=KCTC 39527(T) = MCC 2761(T) = LMG 28646(T)).