Extracellular lipase from <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> JCM5962(T): Isolation,identification, and characterization

The study was done to isolate, identify, and characterize a good lipolytic strain from soil. Lipolytic strain isolation was done using tributyrin agar medium. The biochemical testing and 16S rRNA gene sequencing analysis was done for identification. The enzyme was purified using ammonium sulfate precipitation and column chromatography. Results have shown a novel high lipolytic strain of P. aeruginosa JCM5962(T), isolated from soil of sugarcane field. The 16S rRNA sequence analysis confirmed the strain as P. aeruginosa JCM5962(T); further, the sequence was submitted to Genbank (KX946966.1). The isolate produced an extracellular lipase which was purified as single band of 31 kDa. Maximum lipase activity was observed at 50 °C and pH 8.0. Activity was enhanced in the presence of cobalt and benzene solvent, whereas mercury, sodium dodecyl sulfate, and chloroform inhibited it. The enzyme’s marked stability and activity at high temperature, alkaline pH and organic solvents suggest that this can be effectively used in a variety of applications in industries and as biotechnological tools.     

Cadmium increases catechol 2,3-dioxygenase activity in Variovorax sp. 12S, a metal-tolerant and phenol-degrading strain

A Gram-negative bacterium, designated as strain 12S, was isolated from a heavy metal-polluted soil. According to the biochemical characteristics, FAME analysis, and 16S rRNA gene sequence analysis, the isolated strain was identified as Variovorax sp. 12S. In the presence of 0.1 mM cadmium, 12S was able to completely utilize up to 1.5 mM of phenol as the sole carbon and energy source in an MSM–TRIS medium. Degradation of phenol was accompanied by a slow bacterial growth rate and an extension of the lag phase. The cells grown on phenol showed catechol 2,3-dioxygenase (C23O) activity. The activity of C23O from 12S cultivated in medium with Cd²⁺ was almost 20 % higher than in the control. Since environmental contamination with aromatic compounds is often accompanied by the presence of heavy metals, Variovorax sp. 12S and its C23O appear to be very powerful and useful tools in the biotreatment of wastewaters and soil decontamination.     

Staurosporine from the endophytic Streptomyces sp. strain CNS-42 acts as a potential biocontrol agent and growth elicitor in cucumber

Chinese medicinal plants and their surrounding rhizospheric soil serve as promising sources of actinobacteria. A total of 180 actinobacteria strains were isolated from the rhizosphere soil, leaves, stems, and roots of nine selected plants and have been identified as potential biocontrol agents against Fusarium oxysporum f. sp. cucumerinum. An endophytic strain CNS-42 isolated from Alisma orientale showed the largest zone of inhibition demonstrating a potent effect against F. oxysporum f. sp. cucumerinum and a broad antimicrobial activity against bacteria, yeasts, and other pathogenic fungi. The in vivo biocontrol assays showed that the disease severity index was significantly reduced (P < 0.05), and plant shoot fresh weight and height increased greatly (P < 0.05) in plantlets treated with strain CNS-42 compared to the negative control. This isolate was identified as Streptomyces sp. based on cultural, physiological, morphological characteristics, and 16S rRNA gene analysis. Further bioassay-guided isolation and purification revealed that staurosporine was responsible for its antifungal and plant growth promoting activities and the latter property of staurosporine is reported for the first time. The in vivo assay was further performed and indicated that staurosporine showed good growth promoting effect on the plant shoot biomass of cucumber. This is the first critical evidence identifying CNS-42 as a biocontrol agent for the soil borne pathogen, F. oxysporum f. sp. cucumerinum.     

Azospirillum himalayense sp. nov., a nifH bacterium isolated from Himalayan valley soil,India

Gram-negative, free-living bacterial strain ptl-3^T was isolated from Himalayan valley soil, India. Polyphasic taxonomy was performed including morphological characterization, fatty acid analysis, biochemical tests, 16S rRNA and nifH gene sequence analyses. 16S rRNA gene sequence analysis showed that the strain ptl-3^T belonged to the genus Azospirillum and was closely related to A. brasilense (98.7 % similarity) and A. rugosum (97 % similarity). 16S rRNA gene sequence similarity (96–95 %) was shown with other members of the genus Azospirillum. Major fatty acid 18:1ω7c was also similar to the genus Azospirillum. DNA–DNA relatedness value between strain ptl-3^T and A. brasilense was found to be 47 %. Various biochemical tests showed that the strain ptl-3^T differed from its closely related species A. brasilense. On the basis of phenotypic, chemotaxonomic and molecular genetics evidence, a bacterium with the type strain ptl-3^T is proposed as a novel species of the genus Azospirillum. The name of bacterial strain ptl-3^T has been proposed as Azospirillum himalayense sp. nov. The type strain of ptl-3^T (CCUG 58760^T, KCTC 23189^T) has been submitted to two culture collection centres. The accession numbers for 16S rRNA and nifH gene are GQ 284588 and GQ 249665. respectively.     

Oceanobacillus halophilum sp. nov. Isolated from a Mangrove Forest Soil

A halophilic, aerobic bacterium, designated GD01^T, was isolated from a mangrove forest soil near the South China Sea. Cells of strain GD01^T were Gram staining positive, oxidase positive, and catalase positive. The strain was rod shaped and motile by means of peritrichous flagella and produced ellipsoidal endospores. The strain was able to grow with NaCl at concentrations of 0.5–12 % (optimum 3–5 %, w/v), at temperatures of 20–50 °C (optimum 30 °C), and at pH 6.0–8.5 (optimum pH 7.0). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain GD01^T formed a cluster with O. profundus DSM 18246^T (96.4 % 16S rRNA gene sequence similarity), O. caeni KCTC 13061^T (95.4 %), and O. oncorhynchi JCM 12661^T (94.5 %). The G+C content of strain GD01^T was 38.7 mol%. The major respiratory quinone was MK-7. The major cellular fatty acids (>5 %) were anteiso-C_15:0, iso-C_16:0 (13.7 %), anteiso-C_17:0 (12.6 %), iso-C_15:0 (9.9 %), iso-C_14:0 (9.5 %), and C_16:0 (5.0 %). The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, glycolipid, four unknown lipids, and four unknown phospholipids. Based on phenotypic characteristics, chemotaxonomic features, and phylogenetic analysis based on 16S rRNA gene sequences, the strain was identified to represent a distinct novel species in the genus Oceanobacillus, and the name proposed is Oceanobacillus halophilum sp. nov. with type train GD01^T (=CCTCC AB 2012863^T = KCTC 33101^T).     

Paenibacillus beijingensis sp. nov., a novel nitrogen-fixing species isolated from jujube garden soil

A novel Gram-positive, rod-shaped, motile, spore-forming, nitrogen-fixing bacterium, designated strain 7188^T, was isolated from jujube rhizosphere soil in Beijing, China. The strain grew at 4–40 °C and pH 6–12, with an optimum of 30 °C and pH 7.0, respectively. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain 7188^T is a member of the genus Paenibacillus. Levels of 16S rRNA gene sequence similarities between strain 7188^T and the type strains of all recognized members of the genus Paenibacillus were below 96 %. The major cellular fatty acids were anteiso-C_15:0, anteiso-C_17:0 and C_16:0. The predominant menaquinone was MK-7. The DNA G+C content of strain 7188^T was 60.3 mol%. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and unknown aminophospholipids. The diamino acid in the cell wall peptidoglycan is meso-diaminopimelic acid. On the basis of these results, strain 7188^T is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus beijingensis sp. nov. is proposed. The type strain is 7188^T (=ACCC 03082^T = DSM 24997^T).     

Vulcaniibacterium tengchongense gen. nov., sp. nov. isolated from a geothermally heated soil sample, and reclassification of Lysobacter thermophilus Wei et al. 2012 as Vulcaniibacterium thermophilum comb. nov.

A thermotolerant Gram-staining negative and aerobic bacterium, designated strain YIM 77520^T, was isolated from a geothermally heated soil sample collected at Rehai National Park, Tengchong, Yunnan Province, South-West China. Cells of the strain were found to be rod-shaped and colonies were light beige and circular. The strain was found to grow in the presence of 0–2 % (w/v) total salts (optimum, 0 %), at pH 6.0–8.0 (optimum, pH 7.0) and 25–55 °C (optimum, 45 °C). The only quinone detected was Q-8 and the genomic DNA G+C content was determined to be 66.9 mol%. The major fatty acids (>10 %) were identified as iso-C_16:0 and iso-C_15:0. The phospholipids were found to consist of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, five unknown phospholipids and three aminophospholipids. Based on the 16S rRNA gene sequence analysis, strain YIM 77520^T was found to form a cluster with Lysobacter thermophilus YIM 77875^T and showed the highest 16S rRNA gene sequence similarity to L. thermophilus YIM 77875^T (96.0 %). These two strains formed a distinct lineage of the family ‘Xanthomonadaceae’. On the basis of the morphological and chemotaxonomic characteristics, as well as genotypic data, a new genus, Vulcaniibacterium gen. nov. is proposed with Vulcaniibacterium tengchongense sp. nov. as the type species. The type strain of V. tengchongense sp. nov. is strain YIM 77520^T (=DSM 25623^T = CCTCC AB 2011152^T). Furthermore we propose that L. thermophilus Wei et al. 2012 is reclassified in the new genus as Vulcaniibacterium thermophilum comb. nov. (type strain YIM 77875^T = CCTCC AB 2012064^T = KCTC 32020^T) based on polyphasic data.     

Halococcus sediminicola sp. nov., an extremely halophilic archaeon isolated from a marine sediment

A novel, red-pigmented and coccoid haloarchaeon, designated strain CBA1101^T, was isolated from a marine sediment. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain CBA1101^T is most closely related to the genus Halococcus in the family Halobacteriaceae. Strain CBA1101^T had a highest 16S rRNA gene sequence similarity of 98.4 % with Halococcus dombrowskii DSM 14522^T, followed by 93.7–98.3 % with sequences of other type strains in the genus Halococcus. The RNA polymerase subunit B′ gene sequence similarity of strain CBA1101^T with that of Halococcus qingdaonensis JCM 13587^T is 89.5 % and lower with those of other members of the genus Halococcus. Strain CBA1101^T was observed to grow at 25–40 °C, pH 6.0–9.0 and in the presence of 15–30 % (w/v) NaCl, with optimal growth at 35–40 °C, pH 7.0 and with 20 % NaCl. The cells of strain CBA1101^T are Gram-negative and did not lyse in distilled water. The major polar lipids were identified as phosphatidylglyerol, phosphatidylglycerol phosphate methyl ester, sulfated diglycosyl diether, unidentified phospholipids and unidentified glycolipids. The genomic DNA G+C content was determined 66.0 mol%. The DNA–DNA hybridization experiment showed that there was less than 40 % relatedness between strain CBA1101^T and the reference species in the genus Halococcus. Based on this polyphasic taxonomic analysis, strain CBA1101^T is considered to represent a new species in the genus Halococcus, for which the name Halococcus sediminicola sp. nov. is proposed. The type strain is CBA1101^T (=JCM 18965^T = CECT 8275^T).     

Pontibacter soli sp. nov., isolated from the soil of a Populus rhizosphere in Xinjiang, China

A Gram-stain negative, rod-shaped, non-motile and pink-pigmented bacterial strain, designated strain HYL7-26^T, was isolated from a soil in the Desert Park of Huyang forest located in Xinjiang, China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain HYL7-26^T belongs to the genus Pontibacter in the family Cytophagaceae. The 16S rRNA gene sequence similarity between strain HYL7-26^T and type strains of Pontibacter species ranged from 93.2 to 96.0 %. Strain HYL7-26^T was found to contain iso-C_15:0 (15.9 %), iso-C_17:0 3-OH (9.5 %) and summed feature 4 (comprising anteiso-C_17:1 B and/or iso-C_17:1 I, 21.0 %, as defined by the MIDI system) as the major cellular fatty acids. The major respiratory quinone was identified as MK-7 and the DNA G+C content was determined to be 43.8 mol%. sym-Homospermidine was the major polyamine observed in the cells. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain HYL7-26^T is considered to represent a novel species of the genus Pontibacter, for which the name Pontibacter soli sp. nov. is proposed. The type strain is HYL7-26^T (=CCTCC AB 206240^T = NRRL B-59490^T).     

Lysobacter thermophilus sp. nov., isolated from a geothermal soil sample in Tengchong, south-west China

A Gram-negative and aerobic bacterium, designated YIM 77875^T, was isolated from a geothermal soil sample collected at Rehai National Park, Tengchong, Yunnan Province, south-west China. Bacterial growth occurred from 37 to 65 °C (optimum 50 °C), pH 6.0–8.0 (optimum pH 7.0) and 0–1 % NaCl (w/v). Cells were rod-shaped and colonies were convex, circular, smooth, yellow and non-transparent. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain YIM 77875^T belongs to the genus Lysobacter. The 16S rRNA gene sequence similarity values between strain YIM 77875^T and other species of the genus Lysobacter were all below 94.7 %. The polar lipids of strain YIM 77875^T were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and five unknown phospholipids. The predominant respiratory quinone was Q-8 and the G+C content was 68.8 mol%. Major fatty acids were iso-C_16:0, iso-C_15:0 and iso-C_11:0. On the basis of the morphological and chemotaxonomic characteristics, as well as genotypic data, strain YIM 77875^T represents a novel species, Lysobacter thermophilus sp. nov., in the genus Lysobacter. The type strain is YIM 77875^T (CCTCC AB 2012064^T = KCTC 32020^T).     

Paenibacillus brassicae sp. nov., isolated from cabbage rhizosphere in Beijing, China

A novel Gram-positive, rod-shaped, motile, spore-forming, nitrogen-fixing bacterium, designated strain 112^T, was isolated from cabbage rhizosphere in Beijing, China. The strain was found to grow at 10–40 °C and pH 4–11, with an optimum of 30 °C and pH 7.0, respectively. Phylogenetic analysis based on a fragment of the full-length 16S rRNA gene sequence revealed that strain 112^T is a member of the genus Paenibacillus. High levels of 16S rRNA gene similarities were found between strain 112^T, Paenibacillus sabinae DSM 17841^T (97.82 %) and Paenibacillus forsythiae DSM 17842^T (97.22 %). However, the DNA–DNA hybridization values between strain 112^T and the type strains of these two species were 10.36 and 6.28 %, respectively. The predominant menaquinone was found to be menaquinone 7 (MK-7). The major fatty acids were determined to be anteiso-C_15:0 and C_16:0. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and unknown aminophospholipids. The cell wall peptidoglycan was found to contain meso-diaminopimelic acid. The DNA G+C content was determined to be 55.4 mol%. On the basis of its phenotypic characteristics, 16S rRNA gene sequence analysis and the value of DNA–DNA hybridization, strain 112^T is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus brassicae sp. nov. is proposed. The type strain is 112^T (= ACCC 01125^T = DSM 24983^T).     

Pseudomonas silesiensis sp. nov. strain A3T isolated from a biological pesticide sewage treatment plant and analysis of the complete genome sequence

Microorganisms classified in to the Pseudomonas genus are a ubiquitous bacteria inhabiting variety of environmental niches and have been isolated from soil, sediment, water and different parts of higher organisms (plants and animals). Members of this genus are known for their metabolic versatility and are able to utilize different chemical compounds as a source of carbon, nitrogen or phosphorus, which makes them an interesting microorganism for bioremediation or bio-transformation. Moreover, Pseudomonas sp. has been described as a microorganism that can easily adapt to new environmental conditions due to its resistance to the presence of high concentrations of heavy metals or chemical pollution. Here we present the isolation and analysis of Pseudomonas silesiensis sp. nov. strain A3^T isolated from peaty soil used in a biological wastewater treatment plant exploited by a pesticide packaging company. Phylogenetic MLSA analysis of 4 housekeeping genes (16S rRNA, gyrB, rpoD and rpoB), complete genome sequence comparison (ANIb, Tetranucleotide identity, digital DDH), FAME analysis, and other biochemical tests indicate the A3^T strain (type strain PCM 2856^T = DSM 103370^T) differs significantly from the closest relative species and therefore represents a new species within the Pseudomonas genus. Moreover, bioinformatic analysis of the complete sequenced genome showed that it consists of 6,823,539 bp with a 59.58 mol% G + C content and does not contain any additional plasmids. Genome annotation predicted the presence of 6066 genes, of which 5875 are coding proteins and 96 are RNA genes.     

Methylobacillus rhizosphaerae sp. nov., a novel plant-associated methylotrophic bacterium isolated from rhizosphere of red pepper

A novel plant-associated obligate methylotrophic bacterium, designated strain Ca-68^T, was isolated from the rhizosphere soil of field-grown red pepper from India. The isolates are strictly aerobic, Gram negative, motile rods multiplying by binary fission and formaldehyde is assimilated via the ribulose monophosphate pathway. A comparative 16S rRNA gene sequence-based phylogenetic analysis placed the strain in a clade with the species Methylobacillus flagellatus, Methylobacillus glycogens and Methylobacillus pratensis, with which it showed pairwise similarity of 97.8, 97.4 and 96.2 %, respectively. The major fatty acids are C_16:0, C_10:0 3OH and C_16:1 ω7c. The G+C content of the genomic DNA is 59.7 mol%. The major ubiquinone is Q-8. Dominant phospholipids are phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. Based on 16S rRNA gene sequence analysis and DNA–DNA relatedness (14–19 %) with type strains of the genus Methylobacillus, the novel isolate was classified as a new species of this genus and named Methylobacillus rhizosphaerae Ca-68^T (=KCTC 22383^T = NCIMB 14472^T).     

Characterization of an organic solvent-tolerant lipase from Haloarcula sp. G41 and its application for biodiesel production

A haloarchaeal strain G41 showing lipolytic activity was isolated from the saline soil of Yuncheng Salt Lake, China. Biochemical and physiological characterizations along with 16S rRNA gene sequence analysis placed the isolate in the genus Haloarcula. Lipase production was strongly influenced by the salinity of growth medium with maximum in the presence of 20 % NaCl or 15 % Na₂SO₄. The lipase was purified to homogeneity with a molecular mass of 45 kDa. Substrate specificity test revealed that it preferred long-chain p-nitrophenyl esters. The lipase was highly active and stable over broad ranges of temperature (30–80 °C), pH (6.0–11.0), and NaCl concentration (10–25 %), with an optimum at 70 °C, pH 8.0, and 15 % NaCl, showing thermostable, alkali-stable, and halostable properties. Enzyme inhibition studies indicated that the lipase was a metalloenzyme, with serine and cysteine residues essential for enzyme function. Moreover, it displayed high stability and activation in the presence of hydrophobic organic solvents with log P _ow?≥?2.73. The free and immobilized lipases from strain G41 were applied for biodiesel production, and 80.5 and 89.2 % of yields were achieved, respectively. This study demonstrated the feasibility of using lipases from halophilic archaea for biodiesel production.     

Salinicola peritrichatus sp. nov., isolated from deep-sea sediment

A Gram stain-negative, aerobic and rod-shaped bacterium, strain DY22^T, was isolated from a deep-sea sediment collected from the east Pacific Ocean. The isolate was found to grow in the presence of 0–20.0 % (w/v) NaCl and at pH 4.5–8.5; optimum growth was observed with 0.5–2.0 % (w/v) NaCl and at pH 5.0–7.0. Chemotaxonomic analysis showed the presence of ubiquinone-9 as predominant respiratory quinone and C_16:0, C_19:0 ω8c cyclo and C_12:0 3-OH as major cellular fatty acids. The genomic DNA G+C content was determined to be 59.6 mol%. Comparative 16S rRNA gene sequence analysis revealed that the novel isolate belongs to the genus Salinicola. Strain DY22^T exhibited the closest phylogenetic affinity to the type strain of Salinicola salarius with 97.2 % sequence similarity and less than 97 % sequence similarity with respect to other Salinicola species with validly published names. The DNA–DNA reassociation values between strain DY22^T and S. salarius DSM 18044^T was 52 ± 4 %. On the basis of phenotypic, chemotaxonomic and genotypic data, strain DY22^T represents a novel species of the genus Salinicola, for which the name Salinicola peritrichatus sp. nov. (type strain DY22^T = CGMCC 1.12381^T = JCM 18795^T) is proposed.     

Micromonospora halotolerans sp. nov., isolated from the rhizosphere of a Pisum sativum plant

A filamentous actinomycete strain designated CR18^T was isolated on humic acid agar from the rhizosphere of a Pisum sativum plant collected in Spain. This isolate was observed to grow optimally at 28 °C, pH 7.0 and in the presence of 5 % NaCl. Phylogenetic analyses based on the 16S rRNA gene sequence indicated a close relationship with the type strains of Micromonospora chersina and Micromonospora endolithica. A further analysis based on a concatenated DNA sequence stretch of 4,523 bp that included partial sequences of the atpD, gyrB, recA, rpoB and 16S rRNA genes clearly differentiated the new strain from recognized Micromonospora species compared. DNA–DNA hybridization studies further supported the taxonomic position of strain CR18^T as a novel genomic species. Chemotaxonomic analyses which included whole cell sugars, polar lipids, fatty acid profiles and menaquinone composition confirmed the affiliation of the new strain to the genus Micromonospora and also highlighted differences at the species level. These studies were finally complemented with an array of physiological tests to help differentiate between the new strain and its phylogenetic neighbours. Consequently, strain CR18^T (= CECT 7890^T = DSM 45598^T) is proposed as the type strain of a novel species, Micromonospora halotolerans sp. nov.     

Halomonas socia sp. nov., isolated from high salt culture of Dunaliella salina

A moderately halophilic bacteria designed strain NY-011^T was isolated from the high salt culture of Dunaliella salina in Chengdu of Sichuan Province, China. The isolate was Gram-negative, nonmotile, rod-shaped and 12.5–21.6 μm in length. Colonies on solid media are circular, wet, smooth and cream. The strain grew optimally at 37 °C, pH 7.0 and in the presence of 8 % NaCl. Acid was produced from glycerol, d-arabinose, glucose, trehalose, inositol, mannose, mannitol, sucrose, maltose and sorbitol. Catalase is produced but not oxidase. The major fatty acids are C18: 1ω7c (37.59 %), C19: 0 cyclo ω8c (18.29 %), C16: 0 (16.05 %) and C6: 0 (12.43 %). The predominant respiratory lipoquinone found in strain NY-011^T is ubiquinone with nine isoprene units (Q-9). The genomic DNA G + C content of strain NY-011^T was 62.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain NY-011^T belonged to the genus Halomonas. The highest levels of 16S rRNA gene sequence similarity were found between the strain NY-011^T and H. pantelleriensis (sequence similarity 98.43 %). However, the levels of DNA–DNA relatedness between them were only 23.1 %. In addition, the strain NY-011^T had a phenotypic profile that readily distinguished it from H. pantelleriensis. The strain NY-011^T therefore represents a new species of the genus Halomonas, for which the name Halomonas socia sp. nov. is proposed, with NY-011^T (=CCTCC AB 2011033^T = KCTC 23671^T) as the type strain.     

Halorubrum halophilum sp. nov., an extremely halophilic archaeon isolated from a salt-fermented seafood

A novel, red-pigmented, pleomorphic and short rod-shaped haloarchaeon, designated B8^T, was isolated from a salt-fermented seafood. Strain B8^T was found to be able to grow at 20–45 °C, in the presence of 15–30 % (w/v) NaCl and at pH 7.0–9.0. The optimum requirements were found to be a temperature range of 35–40 °C, pH 8.0 and the presence of 25 % NaCl. The cells of strain B8^T were observed to be Gram-staining negative and lysed in distilled water. Anaerobic growth did not occur in the presence of nitrate, l-arginine, dimethyl sulfoxide or trimethylamine N-oxide. The catalase and oxidase activities were found to be positive and nitrate was reduced in aerobic conditions. Tween 20, 40 and 80 were found to be hydrolyzed, whereas casein, gelatin and starch were not hydrolyzed. Indole or H₂S was not formed and urease activity was not detected. A phylogenetic analysis based on the 16S rRNA gene sequences indicated that strain B8^T is most closely related to members of the genus Halorubrum in the family Halobacteriaceae. Strain B8^T was found to have three 16S rRNA genes, rrnA, rrnB and rrnC; similarities between the 16S rRNA gene sequences are 99.0–99.8 %. Strain B8^T shared 99.0 % 16S rRNA gene sequence similarity with Halorubrum (Hrr.) lipolyticum JCM 13559^T and Hrr. saccharovorum DSM 1137^T, 98.8 % with Hrr. kocurii JCM 14978^T, 98.3 % with Hrr. lacusprofundi DSM 5036^T, 98.0 % with Hrr. arcis JCM 13916^T, 97.7 % with Hrr. aidingense JCM 13560^T and 97.0 % with Hrr. aquaticum JCM 14031^T, as well as 93.7–96.5 % with other type strains in the genus Halorubrum. The RNA polymerase subunit B′ gene sequence similarity of strain B8^T with Hrr. kocurii JCM 14978^T is 97.2 % and lower with other members of the genus Halorubrum. DNA–DNA hybridization experiments showed that strain B8^T shared equal or lower than 50 % relatedness with reference species in the genus Halorubrum. The genomic DNA G+C content of strain B8^T was determined to be 64.6 mol%. The major isoprenoid quinone of strain B8^T was identified as menaquinone-8 and the major polar lipids as phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, phosphatidylglycerol sulfate, sulfated mannosyl glucosyl diether and an unidentified phospholipid. Based on this polyphasic taxonomic study, strain B8^T is considered to represent a new species in the genus Halorubrum, for which the name Hrr. halophilum sp. nov. is proposed. The type strain is B8^T (=JCM 18963^T = CECT 8278^T).     

Roseomonas musae sp. nov., a new bacterium isolated from a banana phyllosphere

A Gram-negative, coccobacilli, non-spore forming and non-motile bacterium, designated PN1^T, was isolated from a banana leaf collected in Mattra island, Thailand. This isolate was observed to grow optimally at 30 °C and pH 7.0, and to grow with 0–3 % NaCl. Comparative 16S rRNA gene sequence analysis showed that strain PN1^T is closely related to members of the genus Roseomonas, exhibiting the highest 16S rRNA gene sequence similarity to Roseomonas aestuarii JC17^T (96.5 %). The DNA G + C content of strain PN1^T was determined to be 69.7 mol %. Based on physiological and biochemical tests, and genotypic differences between strain PN1^T and the validly named species of the genus Roseomonas, it is proposed that the strain be classified as a new species of Roseomonas for which the name Roseomonas musae sp. nov. is proposed. The type strain is PN1^T (= BCC 44863^T = NBRC 107870^T).     

Actinopolyspora righensis sp. nov., a novel halophilic actinomycete isolated from Saharan soil in Algeria

A novel halophilic actinomycete strain, H23^T, was isolated from a Saharan soil sample collected in Djamâa (Oued Righ region), El-Oued province, South Algeria. Strain H23^T was identified as a member of the genus Actinopolyspora by a polyphasic approach. Phylogenetic analysis showed that strain H23^T had 16S rRNA gene sequence similarities ranging from 97.8 % (Actinopolyspora xinjiangensis TRM 40136^T) to 94.8 % (Actinopolyspora mortivallis DSM 44261^T). The strain grew optimally at pH 6.0–7.0, 28–32 °C and in the presence of 15–25 % (w/v) NaCl. The substrate mycelium was well developed and fragmented with age. The aerial mycelium produced long, straight or flexuous spore chains with non-motile, smooth-surfaced and rod-shaped spores. Strain H23^T had MK-10 (H₄) and MK-9 (H₄) as the predominant menaquinones. The whole micro-organism hydrolysates mainly consisted of meso-diaminopimelic acid, galactose and arabinose. The diagnostic phospholipid detected was phosphatidylcholine. The major cellular fatty acids were anteiso-C_17:0 (37.4 %), iso-C_17:0 (14.8 %), iso-C_15:0 (14.2 %), and iso-C_16:0 (13.9 %). The genotypic and phenotypic data show that the strain represents a novel species of the genus Actinopolyspora, for which the name Actinopolyspora righensis sp. nov. is proposed, with the type strain H23^T (=DSM 45501^T = CCUG 63368^T = MTCC 11562^T).