- Volume 68, Issue 4, 2018

A cultivation-based study of the microbial diversity of cellular phone screens led to the isolation of a Gram-stain-positive, aerobic, rod-shaped and non-endospore-forming bacterium, designated S2T63T, exhibiting phenotypic and genotypic characteristics unique to the type strains of closely related species. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain is a member of Microbacterium , and most closely related to Microbacterium aurantiacum IFO 15234T and Microbacterium kitamiense Kitami C2T. The DNA–DNA relatedness values of the strain S2T63T to M. aurantiacum KACC 20510T, M. kitamiense KACC 20514Tand Microbacterium laevaniformans KACC 14463T were 65 % (±4), 29.5 % (±3) and 55.9 % (±4), respectively. The genomic DNA G+C content was 71.8 mol%. The major fatty acids were anteiso-C15 : 0, iso-C16 : 0, C16 : 0 and anteiso-C17 : 0. The main polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and two unidentified polar lipids. The peptidoglycan contained the amino acids glycine, lysine, alanine and glutamic acid, with substantial amounts of hydroxy glutamic acid detected, which is characteristic of peptidoglycan type B1α. The predominant menaquinones were MK-12 and MK-13. Rhamnose, fucose and galactose were the whole-cell sugars detected. The strain also showed biofilm production, estimated by using crystal violet assay. Based on the results of the phenotypic and genotypic characterizations, it was concluded that the new strain represents a novel species of the genus Microbacterium , for which the name Microbacteriumtelephonicum is proposed, with S2T63T (=MCC 2967T=KACC 18715T=LMG 29293T) as the type strain.

A novel coccus-shaped, Gram-stain-positive, non-motile and aerobic bacterium, designated strain NSG39T, was isolated from the intestine of a Korean rockfish, Sebastes schlegelii. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the newly isolated strain NSG39T was closely related to Tessaracoccus flavus RP1T (98.0 %). The isolate grew at 15–37 °C, pH 7–9 and 0–4 % (w/v) salinity, with optimal growth at 30 °C, pH 8 and 0 % (w/v) salinity. The cell wall of the organism contained ll-diaminopimelic acid as a diagnostic diamino acid, and ribose, mannose, glucose and galactose as diagnostic sugars. The polar lipid comprised diphosphatidylglycerol, phosphatidylglycerol, three glycolipids and four unidentified polar lipids. The major cellular fatty acid was anteiso-C15 : 0 (47.2 %). The major menaquinone was MK-9 (H4). The DNA G+C content of the isolate was 68.8 mol%. The genome-based orthologous average nucleotide identity value for strain NSG39T and T. flavus RP1T was 76.6 %. Based on the phylogenetic analysis and its biological characteristics, strain NSG39T is considered to represent a novel species of the genus Tessaracoccus , for which the name Tessaracoccus aquimaris is proposed. The type strain is NSG39T (=KACC 17540T=JCM 19289T).

A novel endophytic actinomycete, strain WPS1-2T, isolated from a root of Globba winitii C. H. Wright, was characterized taxonomically by using a polyphasic approach. Strain WPS1-2T exhibited identical characteristics to the members of the genus Micromonospora . Single spores were observed directly on substrate mycelia. The cell-wall peptidoglycan of the strain contained meso-diaminopimelic acid and 3-OH-meso-diaminopimelic acid. Whole-cell hydrolysates contained glucose, ribose, arabinose and xylose. The predominant menaquinones were MK-10(H8) and MK-10(H10). The major cellular fatty acids consisted of iso-C15 : 0, iso-C16 : 0 and anteiso-C15 : 0. According to the 16S rRNA gene sequence of the strain, WPS1-2T showed highest similarity to Micromonospora costi CS1-12T (99.02 %). Phylogenetic analysis of the gyrase subunit B (gyrB) gene indicated that the strain was related to M. costi CS1-12T. The DNA G+C content was 73.7 mol%. The strain could be distinguished from closely related type strains by using a combination of morphological, chemotaxonomic, physiological and biochemical data together with DNA–DNA relatedness values. Based on these observations, strain WPS1-2T is considered to represent a novel species of the genus Micromonospora , for which the name Micromonospora globbae sp. nov. is proposed. The type strain is WPS1-2T (=KCTC 39787T=NBRC 112325T=TISTR 2405T).

Strain CN3T, a Coriaria nepalensis isolate, appears to form hyphae and sporangia typical of members fo the genus Frankia . However, it failed to form vesicles, to reduce acetylene and to induce nodules on its original host plant. A polyphasic approach was used here to determine the taxonomic status of strain CN3T. The 16S rRNA gene sequence of strain CN3T showed the highest sequence identity with Frankia asymbiotica type strain M16386T (99.4 %). Digital DNA–DNA hybridization between strains CN3T and M16386T was 25.7 %, which is clearly below the accepted cut-off point of 70 %. The G+C content of DNA was 71.8 mol%. Whole-cell hydrolysates of strain CN3T were rich in meso-diaminopimelic acid. Cell-wall sugars were composed of galactose, glucose, mannose, rhamnose and traces of ribose. The polar lipid profile contained phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, phosphoglycolipids, phospholipid, six uncharacterized glycolipids and two uncharacterized lipids. The predominant menaquinone (>25 %) was MK-9(H6). Major fatty acids (>15 %) of strain CN3T consisted of iso-C16 : 0, C17 : 1ω8c and C15 : 0. Based on 16S rRNA gene phylogeny, genome sequence analysis and phenotypic results, strain CN3T (=DSM 105290T=CECT 9314T) is proposed to represent the type strain of a novel species, Frankia saprophytica sp. nov.

A novel actinobacterial strain, designated N237T, was isolated from sediment soil of wetlands at Meonmulkkak, Dongbaek-Dongsan, the lava forest, Gotjawal, Jeju, Republic of Korea. Cells of strain N237T were Gram-stain-positive, non-motile rods and formed pale yellow colonies on ten-fold diluted Reasoner’s 2A agar. Strain N237T contained iso-C16 : 0 and C17 : 1 ω8c as the major fatty acids, MK-9(H4) as the predominant isoprenoid quinone and meso-DAP as the diamino acid in the peptidoglycan. It contained diphosphatidylglycerol, phosphatidylinositol polymannosides, an unidentified phospholipid, an unidentified aminophospholipid, an unidentified aminolipid, two unidentified glycophospholipids, three unidentified glycolipids and two unidentified lipids as polar lipids. The DNA G+C content was 68.1 mol%. Strain N237T formed a separate lineage in the genus Jatrophihabitans , as demonstrated by phylogenetic analysis based on 16S rRNA sequencing. It was most closely related to Jatrophihabitans soli KIS75-12T (95.6 % sequence similarity). The combined phenotypic, chemotaxonomic and phylogenetic characteristics supported the conclusion that strain N237T represents a novel species in the genus Jatrophihabitans , for which the name Jatrophihabitans telluris sp. nov. is proposed. The type strain is N237T (=KCTC 39922T=NRRL B-65477T).

Strain SYSU D8004T was isolated from a sample collected from an arid area in Saudi Arabia. The isolate was Gram-stain-positive, non-motile, aerobic and non-spore-forming. It could grow at 4–45 °C, at pH 6.0–10.0 and in the presence of up to 17 % (w/v) NaCl. Pairwise comparison of the 16S rRNA gene sequences indicated that strain SYSU D8004T shared highest sequence similarity with Georgenia halophila YIM 93316T (96.5 %). Menaquinone MK-8(H4) was detected as the respiratory quinone. The polar lipid profile of strain SYSU D8004T consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, two phosphatidylinositol mannosides, two unidentified phospholipids and an unidentified glycolipid. Strain SYSU D8004T contained anteiso-C15 : 0, iso-C15 : 0 and C14 : 0 as the predominant fatty acids (>10 %). Galactose, glucose and rhamnose were detected as whole-cell sugars. Based on analyses of the phenotypic, genotypic and phylogenetic characteristics, it was determined that strain SYSU D8004T could be differentiated from other closely related members of the genus Georgenia . Strain SYSU D8004T is therefore considered to represent a novel species of the genus Georgenia , for which the name Georgenia deserti sp. nov. is proposed. The type strain is SYSU D8004T (=CGMCC 1.15793T=KCTC 39987T).

Strain 2CBEGH3T, which is an obligately anaerobic, non-pigmented, non-spore-forming, non-motile, Gram-stain-positive coccobacillus, was isolated from a faecal sample of a healthy Japanese man. The 16S rRNA gene sequence analysis showed that strain 2CBEGH3T represented a member of the family Atopobiaceae and formed a monophyletic cluster with Olsenella uli DSM 7084T (93.6 % sequence similarity), Olsenella umbonata strain lac31T (93.0 %), Olsenella profusa JCM 14553T (92.7 %) and Olsenella scatoligenes strain SK9K4T (92.7 %) as closest neighbours and Atopobium species. The hsp60 gene sequence analysis supported the phylogenetic relationships based on the 16S rRNA gene sequences, with sequence similarity values of 82.1–84.7 % to the four species described above. A unique three-base (one amino acid residue) insertion was found in the alignment regions of the hsp60 gene sequence of strain 2CBEGH3T. The major end products from d-glucose were d- and l-lactic acids produced at the ratio of 75 : 25, while four species of the genus Olsenella produced d- and l-lactic acids at ratios of 94–98 : 2–6. The isolate formed characteristic crater-like colonies on Eggerth–Gagnon agar plates. The major cellular fatty acids were C18 : 1ω9c, C18 : 1ω9c dimethyl acetal (DMA) and C16 : 0 DMA. The G+C content of the genomic DNA was 68.4 mol%. On the basis of these data, strain 2CBEGH3T represents a novel species in a novel genus of the family Atopobiaceae, for which the name Parolsenella catena gen. nov., sp. nov. is proposed. The type strain of P. catena is 2CBEGH3T (=JCM 31932T=DSM 105194T).

A psychrophilic, Gram-stain-positive, rod-shaped bacterium, designated strain Hh31T, was isolated from Xinjiang No. 1 Glacier in China. Strain Hh31T was catalase-positive, oxidase-negative and able to grow at between 0–18 °C. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Hh31T belonged to the genus Cryobacterium and was most closely related to the type strains of Cryobacterium levicorallinum , Cryobacterium luteum and Cryobacterium flavum . DNA–DNA hybridization, calculation of average nucleotide identity and digital DNA–DNA hybridization revealed that strain Hh31T was distinct from its closest phylogenetic neighbours. The major cellular fatty acids of strain Hh31T were anteiso-C15 : 0, anteiso-C15 : 1, iso-C15:0, iso-C16 : 0 and anteiso-C17 : 0. The predominant menaquinones of strain Hh31T were MK-9 and MK-10. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, one unidentified phospholipid, one unidentified glycolipid and another unidentified lipid. Physiological tests such as carbon source utilization, showed phenotypic differentiation of strain Hh31T from the closest related phylogenetic neighbours. Based on a polyphasic approach, a novel species, Cryobacterium aureum sp. nov., is proposed, with Hh31T (=NBRC 107882T=CGMCC 1.11213T) as the type strain.

A novel, non-motile, coccoid, Gram-stain-positive actinobacterium, designated BMG 862T, was isolated from a marble sample collected from the Bulla Regia monument, Northern Tunisia. Its taxonomic position was determined using a polyphasic approach. Results from chemotaxonomic analyses showed MK-9(H4), MK-8(H4) and MK-9(H2) as the predominant menaquinones. The major polar lipids comprised diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, glycophosphatidylinositol, hydroxy-phosphatidylethanolamine and three unidentified phospholipids. The fatty acids consisted of significant amounts (≥10 %) of iso-C16 : 0, C17 : 1ω8c, iso-C15 : 0 and C16 : 1ω7c. Phylogenetic analysis on the basis of 16S rRNA gene sequence comparisons showed that strain BMG 862T belongs to the genus Blastococcus , being most closely related to Blastococcus saxobsidens (=DSM 44509T) (99.5 %) and Blastococcus capsensis (=DSM 46835T=CECT 8876T) (99.3 %). The genomic DNA G+C content of the organism was 74.7 mol%. Results of DNA–DNA hybridization and physiological tests allowed differentiation of strain BMG 862T from related species. The strain was also characterized by its ability to hydrolyse xanthine. On the basis of phenotypic and molecular characteristics, strain BMG 862T (=DSM 46842T=CECT 8884T) represents the type strain of a novel species of the genus Blastococcus , for which the name Blastococcus xanthinilyticus sp. nov. is proposed.

An aerobic actinobacterium, strain AN130378T, was isolated from a soil sample collected in Korea and subjected to taxonomic investigation using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequence data showed that strain AN130378T is a member of the genus Stackebrandtia , with sequence similarities of 97.3 % to Stackebrandtia albiflava YIM 45751T and 97.1 % to Stackebrandtia endophytica YIM 64602T. The whole-cell hydrolysates contained meso-diaminopimelic acid as the diagnostic diamino acid, and galactose, glucose and xylose. The major menaquinones were MK-11(H4), MK-10(H4) and MK-11(H6), while the major fatty acids were identified as iso-C15 : 0, anteiso-C17 : 0, anteiso-C15 : 0, iso-C17 : 0 and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c). The polar lipids were identified as diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmethylethanolamine, an unidentified aminophospholipid, four unidentified glycolipids, three unidentified phospholipids and two unidentified polar lipids. The G+C content of the genomic DNA was determined to be 67.7 mol%. All chemotaxonomic and genotypic data indicated that the strain belongs to the genus Stackebrandtia . On the basis of morphological, chemotaxonomic data and phylogenetic analysis, strain AN130378T is considered to represent a novel species within the genus Stackebrandtia , for which the name Stackebrandtia soli sp. nov. is proposed. The type strain is AN130378T (=DSM 103573T=KCTC 39809T).

A novel endophytic actinobacterium, designated strain EGI 6500139T, was isolated from the surface-sterilized roots of Anabasis aphylla L., collected from Xinjiang, northwest PR China, and subjected to polyphasic taxonomic characterization. Strain EGI 6500139T formed sparse aerial mycelium with rod-like spores. Whole-cell hydrolysates of the isolate contained meso-diaminopimelic acid as the cell-wall diamino acid, glucose as major sugar, and mannose, galactose, xylose and ribose as minor sugars. The polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside, one unidentified glycolipid, one unidentified phospholipid and four unidentified polar lipids. The major fatty acids identified were anteiso-C17 : 0, anteiso-C15 : 0, iso-C15 : 0 and iso-C16 : 0. The predominant menaquinones detected were MK-11 and MK-11(H2). The G+C content of the genomic DNA of strain EGI 6500139T was 70.4 mol%. Strain EGI 6500139T showed the highest 16S rRNA gene sequence similarity to Glycomyces lacisalsi XHU 5089T (96.3 %). Phylogenetic analysis showed that strain EGI 6500139T fell within the clade of the genus Glycomyces , and formed a clade with G. lacisalsi XHU 5089T and G. albus CCTCC AA 2013004T. Based on phenotypic, chemotaxonomic and phylogenetic data, strain EGI 6500139T represents a novel species of the genus Glycomyces , for which the name Glycomyces anabasis sp. nov. (type strain EGI 6500139T=JCM 30088T=KCTC 29495T) is proposed.

A novel actinomycete, designated strain NW8-21T, was isolated from mountain soil in Mae-Wong National Park, Nakornsawan province, Thailand, and was taxonomically characterized by using a polyphasic approach. Based on 16S rRNA gene sequence analysis, the strain was revealed to have the closest similarity to Pseudonocardia endophytica YIM 56035T with the highest 16S rRNA gene sequence similarity value of 98.7 %, followed by Pseudonocardia nantongensis KLBMP 1282T (98.0 %). The chemotaxonomic properties, i.e. arabinose and galactose as the diagnostic reducing sugar in cells, MK-8 (H4) as a major menaquinone, iso-C16 : 0 as the main cellular fatty acid component and phosphatidylethanolamine, phosphatidylmethylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylinositol, phosphatidylcholine as the characteristic phospholipids, confirmed a taxonomic affiliation of the strain that was consistent with those of the genus Pseudonocardia . Several phenotypic differences and the DNA–DNA hybridization results (less than 40 % relatedness value) indicated that strain NW8-21T shoud be considered to represent a novel species of the genus Pseudonocardia , for which the name Pseudonocardia soli is proposed. The type strain is strain NW8-21T (=BCC 58125T=NBRC 109519T).

A Gram-stain-positive, flagellated, catalase- and cytochrome c oxidase-positive bacterial strain, designated S20-100T, was isolated from alpine forest soil. Growth occurred at a temperature range of 0–30 °C, at pH 6–9 and in the presence of 0–1 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain S20-100T was related to the genus Marmoricola and had the highest 16S rRNA gene sequence similarity to Marmoricola ginsengisoli Gsoil 097T (98.4 %) and Marmoricola solisilvae KIS18-7T (98.3 %). The cell-wall peptidoglycan of strain S20-100T contained ll-diaminopimelic acid (ll-Dpm) as the diagnostic diamino acid and was of the type A3γ ll-Dpm – Gly. The strain contained MK-8(H4) as the predominant isoprenoid quinone and diphosphatidylglycerol, phosphatidylglycerol, four unidentified phospholipids and three unidentified lipids in lower amounts. The major cellular fatty acids (>10 %) were iso-C16 : 0, C17 : 1ω6c and C18 : 1ω9c. The genomic DNA G+C content was 66.2 mol%. Combined data of phylogenetic, phenotypic and chemotaxonomic analyses demonstrated that strain S20-100T represents a novel species of the genus Marmoricola , for which the name Marmoricola silvestris sp. nov. is proposed. The type strain is S20-100T (=DSM 104694T=LMG 30008T).

A Gram-stain-positive, aerobic, non-motile, non-spore-forming and short-rod-shaped actinobacterium, designated strain 1T4Z-3T, was isolated from a piece of surface-sterilized branch of Aegiceras corniculatum collected from the Cotai Ecological Zones in Macao, China. Comparative 16S rRNA gene sequence analysis showed that strain 1T4Z-3T was clearly affiliated to the genus Amnibacterium and exhibited 97.9 % gene sequence similarity to Amnibacterium kyonggiense JCM 16463T, 97.3 % gene sequence similarity to Amnibacterium soli JCM 19015T and less than 96.4 % gene sequence similarities to other genera of the family Microbacteriaceae. Strain 1T4Z-3T had L-2,4-diaminobutyric acid as the diagnostic cell-wall diamino acid. The major fatty acids (>10 % of total fatty acids) were iso-C16 : 0 (46.6 %) and anteiso-C15 : 0 (27.3 %). The predominant menaquinones of strain 1T4Z-3T were MK-11 (81.4 %) and MK-12 (14.1 %). The polar lipids comprised diphosphatidylglycerol, phosphatidylglycerol, six unidentified glycolipids, four unidentified phospholipids and two unidentified lipids. The DNA G+C content of strain 1T4Z-3T was 71.4 mol%. Based on the phylogenetic, phenotypic and chemotaxonomic features, strain 1T4Z-3T is considered to represent a novel species of the genus Amnibacterium , for which the name Amnibacterium endophyticum sp. nov. is proposed. The type strain of Amnibacterium endophyticum is 1T4Z-3T (=KCTC 39983T=CGMCC 1.16066T).

The original description of Arthrobacter nasiphocae M597/99/10T demonstrated that it is distantly related to the type species of the genus Arthrobacter, Arthrobacter globiformis, and that this phylogenetic relationship is reflected by the distinct peptidoglycan type [Lys-Ala2-Gly2-3-Ala(Gly)] and the features of the quinone system, which is composed of menaquinones MK-9(H2) and MK-8(H2). Here, we report a re-evaluation of the taxonomic status of A. nasiphocae. Phylogenetically, it was observed to be only distantly related to the genus Arthrobacter and to the type species of related genera. Re-analysis confirmed the quinone system menaquinones MK-9(H2) and MK-8(H2) in A. nasiphocae. Analysis of cell polar lipids showed a profile consisting of the predominant lipids diphosphatidylglycerol, phosphatidylglycerol, dimannosylglyceride, an unidentified phospholipid and an unidentified aminophosphoglycolipid, and several minor unidentified lipids. This profile clearly is different from that of Arthrobacter species. The cell fatty acid profile also showed characteristics that distinguished A. nasiphocae from Arthrobacter species. The phylogenetic distance of A. nasiphocae from any type species of genera within the family Micrococcaceae and the distinct chemotaxonomic traits warrant the reclassification of A. nasiphocae within a novel genus, for which we propose the name Falsarthrobacter nasiphocae gen. nov., comb. nov. The type strain is M597/99/10T (=CCUG 42953T=CIP 107054T=DSM 13988T=JCM 11677T).

A novel hyperthermophilic archaeon of strain HS-3T, belonging to the family Sulfolobaceae , was isolated from an acidic terrestrial hot spring in Hakone Ohwaku-dani, Japan. Based on 16S rRNA gene sequence analysis, the closest phylogenetic relatives of strain HS-3T were, first, Sulfolobus solfataricus (96.4 %) and, second, Sulfolobus shibatae (96.2 %), indicating that the strain belongs to the genus Sulfolobus . However, the sequence similarity to the type species of the genus Sulfolobus ( Sulfolobus acidocaldarius ) was remarkably low (91.8 %). In order to determine whether strain HS-3T belongs to the genus Sulfolobus , its morphological, biochemical and physiological characteristics were examined in parallel with those of S. solfataricus and S. shibatae . Although there were some differences in chemolithotrophic growth between strain HS-3T, S. solfataricus and S. shibatae , their temperature, pH and facultatively anaerobic characteristics of growth, and their utilization of various sugars were almost identical. In contrast, the utilization of various sugars by S. acidocaldarius was quite different from that of HS-3T, S. solfataricus and S. shibatae . Phylogenetic evidence based on the 16S and the 23S rRNA gene sequences also clearly distinguished the monophyletic clade composed of strain HS-3T, S. solfataricus , and S. shibatae from S. acidocaldarius . Based on these results, we propose a new genus and species, Saccharolobus caldissimus gen. nov., sp. nov., for strain HS-3T, as well as two reclassifications, Saccharolobus solfataricus comb. nov. and Saccharolobus shibatae comb. nov. The type strain of Saccharolobus caldissimus is HS-3T (=JCM 32116T and InaCC Ar80T). The type species of the genus is Saccharolobus solfataricus.

A psychrotolerant, methylotrophic methanogen, strain YSF-03T, was isolated from the saline meromictic Lake Shira in Siberia. Cells of strain YSF-03T were non-motile, irregular cocci and 0.8–1.2 µm in diameter. The methanogenic substrates utilized by strain YSF-03T were methanol and trimethylamine. The temperature range of growth for strain YSF-03T was from 0 to 37 °C. The optimum growth conditions were 30–37 °C, pH 7.0–7.4 and 0.17 M NaCl. The G+C content of the genome of strain YSF-03T was 41.3 mol%. Phylogenetic analysis revealed that strain YSF-03T was most closely related to Methanolobus profundi MobMT (98.15 % similarity in 16S rRNA gene sequence). Genome relatedness between strain YSF-03T and MobMT was computed using the Genome-to-Genome Distance Calculator and average nucleotide identity, which gave values of 23.5 and 79.3 %, respectively. Based on the morphological, phenotypic, phylogenetic and genomic relatedness data presented here, it is evident that strain YSF-03T represents a novel species of the genus Methanolobus , for which the name Methanolobus psychrotolerans sp. nov. is proposed. The type strain is YSF-03T (=BCRC AR10049T=DSM 104044T=NBRC 112514T).