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2,3,5-Triphenyl tetrazolium chloride (TTC) staining is a method to distinguish the mitochondrial activity of cells based on the color: colorless TTC turns red when under reducing conditions. Although the assay reflects the mitochondrial activity of cells, which enzyme(s) in the electron transport system contribute to TTC reduction has been unclear. TTC staining assays using gene disruptants related to the electron transport system in Saccharomyces cerevisiae revealed those disruptants related to electron transport from each electron donor to ubiquinone (red colonies) and disruptants that were related to ubiquinol-cytochrome c oxidoreductase and cytochrome c oxidase (white colonies). In addition, when the enzyme activities of ubiquinol-cytochrome c oxidoreductase and cytochrome c oxidase were measured using TTC as the electron acceptor, only ubiquinol-cytochrome c oxidoreductase showed TTC reduction activity, and the activity was enhanced by potassium cyanide, an inhibitor of cytochrome c oxidase. These results indicated that ubiquinol-cytochrome c oxidoreductase is involved in TTC reduction in S. cerevisiae. The fermentation profiles of BY4741UΔcor1 and BY4741UΔcox4, which exhibited no TTC staining activity, were almost identical to that of the parental strain BY4741U. However, cell growth and ethanol and succinate production of the ura3-mutated strain BY4741, which also exhibited no TTC staining activity, was altered compared to those of BY4741U, indicating that the fermentation profile varies among strains that show no TTC staining activity. The relationship between uracil metabolism and TTC staining activity was also determined based on metabolome analysis.

Fourteen novel lipomycetaceous yeasts species were isolated from soil samples collected from the Hokkaido, Chiba and Okinawa prefectures of Japan. Phylogenetic analyses of the D1/D2 domains of the large subunit rRNAs and translation elongation factor 1 alpha genes (TEF1-α) revealed that five strains of two species from the soil in Furano-shi, Hokkaido were related to Dipodascopsis anomala and 29 strains representing 12 species from soils in Kamogawa-shi, Chiba and Iriomote Island, Okinawa were in the Myxozyma clade. The two species of Dipodascopsis form globose or ellipsoid ascospores in their sac-like ascus and pseudohyphae. Furthermore, these species produce ascospores in their pseudohyphae and do not produce an acicular ascus, which is common among the three species including D. anomala. Therefore, we propose transferring D. anomala to the genus Babjevia and amending Babjevia. Two novel species were described and included in the genus Babjevia: Babjevia hyphoforaminiformans sp. nov. (holotype NBRC 111233; MycoBank no. MB 829051) and Babjevia hyphasca sp. nov. (holotype NBRC 112965; MycoBank no. MB 829053). The 12 species in the Myxozyma clade produce neither ascospores nor pseudohyphae and have different characteristics in assimilating several carbon sources from each other. Thus, we propose that the novel species of Lipomyces be classified as forma asexualis (f.a.). From Kamogawa-shi, Chiba (19 strains representing five species): Lipomyces melibiosiraffinosiphilus f.a., sp. nov. (holotype NBRC 111411; MycoBank no. MB 829034), Lipomyces kiyosumicus f.a., sp. nov. (holotype NBRC 111424; MycoBank no. MB 829035), Lipomyces chibensis f.a., sp. nov. (holotype NBRC 111413; MycoBank no. MB 829036), Lipomyces kamogawensis f.a., sp. nov. (holotype NBRC 112967; MycoBank no. MB 829037), Lipomyces amatsuensis f.a., sp. nov. (holotype NBRC 111420; MycoBank no. MB 829041). From Iriomote island, Okinawa (10 strains representing seven species): Lipomyces taketomicus f.a., sp. nov. (holotype NBRC 112966; MycoBank no. MB 829042), Lipomyces yaeyamensis f.a., sp. nov. (holotype NBRC 110433; MycoBank no. MB 829050), Lipomyces iriomotensis f.a., sp. nov. (holotype NBRC 110436; MycoBank no. MB 829045), Lipomyces haiminakanus f.a., sp. nov. (holotype NBRC 110435; MycoBank no. MB 829046), Lipomyces komiensis f.a., sp. nov. (holotype NBRC 110440; MycoBank no. MB 829047), Lipomyces nakamensis f.a., sp. nov. (holotype NBRC 110434; MycoBank no. MB 829048), Lipomyces sakishimensis f.a., sp. nov. (holotype NBRC 110439; MycoBank no. MB 829049).

清酒の呈味や香りに影響を与える化合物は主にアルコール発酵を担う酵母によって生成される。酵母は減数分裂の過程でゲノムの再編成が起こることから，胞子形成後の一倍体株には親株と異なる性質を示す株が現れることが知られている。そこで本研究では，酒質の多様化を目指し，清酒酵母であるきょうかい酵母K701を親株として一倍体株を取得し，清酒小仕込み試験により親株の醸造特性と異なる株と類似した株を取得した。特に，H14株は発酵能と酢酸イソブチル生成能が優れていた。今後，これらの一倍体株を用いた交配により，多様な醸造特性を発揮する2倍体株への育種が期待される。, Since sake yeasts produce not only ethanol but also compounds related to flavor and taste during sake brewing, the sake yeasts exhibiting various types of fermentation products are intensively isolated. One of the major breeding methods is mating of haploid strains because of the genome rearrangement which occurrs during meiosis and haploid strains showing different properties against the parental strains. In this study, we isolated a high isobutylacetate producing haploid stain H14 from diploid sake yeast Kyokai No. 701 as a parental strain. The strain H14 also showed high ethanol productivity during sake brewing. Using this haploid strain H14 showing high ethanol and isobutylacetate productivity as a parental strain leads to development of new diploid sake yeasts.

Fourteen novel lipomycetaceous yeasts species were isolated from soil samples collected from the Hokkaido, Chiba and Okinawa prefectures of Japan. Phylogenetic analyses of the D1/D2 domains of the large subunit rRNAs and translation elongation factor 1 alpha genes (

A bacterial strain, designated 5GHs33-3T, was isolated from greenhouse soil collected from Yongin region, Gyeonggi province, South Korea. The strain was an aerobic, Gram-stain-positive, flagellated, rod-shaped bacterium. Strain 5GHs33-3T grew at 4–37 °C (optimally at 28–30 °C), pH 6.0–10.0 (optimally at pH 7.0) and with 0–7 % (w/v) NaCl (optimally with 0 %). The 16S rRNA gene sequence analysis revealed that strain 5GHs33-3T had high sequence similarity with Actinotalea suaedae EGI 60002T (98.4 %), Actinotalea ferrariae CF5-4T (96.4 %) and Actinotalea fermentans DSM 3133T (96.2 %), and less than 95.5 % sequence similarity against all the other species with validly published names. The phylogenetic tree revealed that strain 5GHs33-3T formed a robust independent monophyletic line with Actinotalea suaedae EGI 60002T. The predominant fatty acids of strain 5GHs33-3T were anteiso-C15 : 0 and iso-C14 : 0. The only quinone was MK-8(H4). Polar lipids were diphosphatidylglycerol, phosphatidylinositol, an unknown phosphoglycolipid and unknown lipids. The peptidoglycan type was A4β, with ornithine as the diagnostic diamino acid and an interpeptide bridge comprising l-Glu. The DNA G+C content is 69.0 mol%. Based on phylogenetic evidence and the results of phenotypic, genotypic and chemotaxonomic analyses, strain 5GHs33-3T represents a novel species of a new genus of the family Cellulomonadaceae , for which the name Pseudactinotalea terrae gen. nov., sp. nov. is proposed. The type strain of the type species is 5GHs33-3T (=KACC 16542T=NBRC 111006T). We also propose the reclassification of Actinotalea suaedae as Pseudactinotalea suaedae comb. nov. (type strain EGI 60002T=JCM 19624T=KACC 17839T=KCTC 29256T).

Fluorescence in situ hybridization (FISH) has been employed to identify microorganisms at the single cell level under a microscope. Extensive efforts have been made to improve and extend the FISH technique; however, the development of a widely applicable protocol is a continuing challenge. The present study evaluated the effects of divalent cations in the hybridization solution on the FISH-based detection of various species of bacteria and archaea with rRNA-targeted probes. A flow cytometric analysis after FISH with a standard hybridization buffer detected positive signals from less than 30% of Escherichia coli IAM 1264 cells. However, the number of cells with positive signals increased to more than 90% after the addition of calcium chloride to the hybridization buffer. Mn2+ also had positive effects, whereas Mg2+ did not. The positive effects of Ca2+ were similarly observed for bacteria belonging to Enterobacteriaceae, including Enterobacter sakazakii IAM 12660T, E. aerogenes IAM 12348, Klebsiella planticola IAM 14202, and Salmonella enterica subsp. enterica serovar Typhimurium strain LT2. These results indicate that the supplementation of Ca2+ to the hybridization buffer for FISH contributes to the efficient detection of Enterobacteriaceae cells.

A novel spherical actinobacterium, designated RS-2-3T, was isolated from the rhizosphere of a mangrove growing on Rambut Island, Indonesia, and its taxonomic position was investigated using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that strain RS-2-3T was related to the members of the genus Kocuria. The highest 16S rRNA gene sequence similarity value was observed with Kocuria marina KMM 3905T (97.0 %). The peptidoglycan type of strain RS-2-3T was found to be A3α with an interpeptide bridge comprising l-Ala4-5. The predominant menaquinone was MK-7(H2) and the major fatty acids were anteiso-C15 : 0 and iso-C15 : 0. The DNA G+C content was 71.8 mol%. These characteristics were consistent with those of members of the genus Kocuria. Meanwhile, physiological and biochemical characteristics revealed that strain RS-2-3T differed from the species of the genus Kocuria with validly published names. Therefore, strain RS-2-3T represents a novel species of the genus Kocuria, for which the name Kocuria pelophila sp. nov. is proposed. The type strain is RS-2-3T (=NBRC 110990T=InaCC A704T).

A novel bacterium, strain 7515T-26T, was isolated from an air sample collected in Taean region, Republic of Korea. Cells were aerobic, Gram-stain-positive, non-flagellated cocci, growing in the temperature, pH and NaCl ranges of 10-33 °C, pH 5.0-9.0 and 0-2 % (w/v). It shared high 16S rRNA gene sequence similarity with Friedmanniella lacustris EL-17AT (97.6 %), Friedmanniella lucida FA2T (96.9 %) and Friedmanniella luteola FA1T (96.9 %), showing high sequence similarities of 96.5-97.6 % with members of the genus Friedmanniella. Phylogenetic trees based on 16S rRNA gene sequences showed that strain 7515T-26T and members of the genus Friedmanniella formed a compact cluster separable from other genera. The isolate contained anteiso-C15 : 0, iso-C14 : 0 3-OH and iso-C15 : 0 as the major cellular fatty acids, and MK-9(H4) as the predominant isoprenoid quinone. Polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, two unknown phospholipids and one unknown lipid, and the DNA G+C content was 73.1 mol%. The peptidoglycan type was A3γ. It showed DNA-DNA hybridization values of less than 70 % with F. lacustris EL-17AT. On the basis of the evidence from this polyphasic study, a novel species, Friedmanniella aerolata sp. nov., is proposed. The type strain is 7515T-26T ( = KACC 17306T = DSM 27139T).

Ru.bro.bac.te'ri.a. N.L. masc. n. Rubrobacter type genus of the type order; suff. -ia ending to denote a class; N.L. pl. neut. n. Rubrobacteria the Rubrobacterales class. Actinobacteria / Rubrobacteria Keywords: Rubrobacter; Rubrobacteria; Rubrobacterales

Ther.mo.le.o.phi'li.a. N.L. neut. n. Thermoleophilum type genus of the type order; suff. -ia ending to denote a class; N.L. pl. neut. n. Thermoleophilia the Thermoleophilales class. Actinobacteria / Thermoleophilia Keywords: Thermoleophilia class. nov.; Thermoleophilales; Actinobacteria phyl. nov.

A bacterial strain, PSGM2-16T, was isolated from a pot of paddy soil grown with rice in Suwon region, Republic of Korea, and was characterized as having aerobic, Gram-stain-positive, short-rod-shaped cells with one polar flagellum. The 16S rRNA gene sequence of strain PSGM2-16T revealed the highest sequence similarities with Knoellia locipacati DMZ1T (97.4 %), Fodinibacter luteus YIM C003T (97.2 %) and Lapillicoccus jejuensis R-Ac013T (97.0 %), and the phylogenetic tree showed that strain PSGM2-16T formed a subgroup with Ornithinibacter aureus HB09001T and F. luteus YIM C003T within the family Intrasporangiaceae. The major fatty acids (>10 % of the total fatty acids) of strain PSGM2-16T were iso-C16 : 0, C17 : 1ω8c and iso-C14 : 0. The predominant menaquinone was MK-8(H4). The polar lipids present were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, three aminophospholipids and two phospholipids. The peptidoglycan was type A4γ with meso-diaminopimelic acid as the diagnostic diamino acid. DNA–DNA hybridization values between strain PSGM2-16T and closely related taxa were much less than 70 %. The genomic DNA G+C content of strain PSGM2-16T was 70.0 mol%. On the basis of the evidence presented, it is concluded that strain PSGM2-16T represents a novel species of a new genus in the family Intrasporangiaceae, for which the name Oryzobacter terrae gen. nov., sp. nov. is proposed. The type strain of the type species is PSGM2-16T ( = KACC 17299T = DSM 27137T = NBRC 109598T).

Genome-based analysis of type-I polyketide synthase and nonribosomal peptide synthetase gene clusters in a novel strain taxonomically close to the genus Salinispora

Two strains of species of the genus Marmoricola , designated KIS18-7T and JOS5-1T, were isolated from soil samples in Korea. The 16S rRNA gene sequence of strain KIS18-7T showed highest similarities with Marmoricola scoriae Sco-D01T (97.8 %), Marmoricola aequoreus SST-45T (97.6 %) and Marmoricola aurantiacus BC 361T (97.3 %), and strain JOS5-1T had highest sequence similarities with M. aequoreus SST-45T (97.5 %) and Marmoricola bigeumensis MSL-05T (97.3 %). The sequence similarity between KIS18-7T and JOS5-1T was 98.1 %. Phylogenetic analysis showed that these strains grouped with species of the genus Marmoricola . The major fatty acids of strain KIS18-7T were iso-C16 : 0 and C17 : 1ω8c, and C17 : 1ω8c, C18 : 0 10-methyl (TBSA), C18 : 1ω9c, C17 : 0 10-methyl and C16 : 0 2-OH for strain JOS5-1T. Strain KIS18-7T contained the polar lipids, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylcholine, while strain JOS5-1T contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, one unknown aminolipid and two unknown phospholipids. The peptidoglycan of both strains contained ll-diaminopimelic acid as the diagnostic diamino acid and a single glycine residue as the interpeptide bridge (type A3γ). The major menaquinone of both strains was MK-8(H4). The G+C contents of the DNA of strains KIS18-7T and JOS5-1T were 68.0 mol% and 62.9 mol%, respectively. These data demonstrate that strains KIS18-7T and JOS5-1T are representatives of two novel species of the genus Marmoricola , for which the names Marmoricola solisilvae sp. nov. (type strain KIS18-7T = KACC 17307T = DSM 27140T = NBRC 109601T) and Marmoricola terrae sp. nov. (type strain JOS5-1T = KACC 17308T = DSM 27141T = NBRC 109602T) are proposed.

A novel Gram-stain-positive actinobacterium, designated PS-14-7T, was isolated from the rhizosphere of a mangrove on Pramuka Island, Indonesia, and its taxonomic position was investigated using a polyphasic approach. The peptidoglycan type of strain PS-14-7T was A4α and lysine was the diagnostic diamino acid of the peptidoglycan. The predominant menaquinone was MK-8(H4) and the major fatty acids were anteiso-C15 : 0, C16 : 0 and iso-C16 : 0. The DNA G+C content was 72.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain PS-14-7T was closely related to Serinibacter salmoneus Kis4-28T (99.6 %). However, DNA–DNA hybridization and phenotypic characteristics revealed that strain PS-14-7T differed from Serinibacter salmoneus . Therefore, strain PS-14-7T represents a novel species of the genus Serinibacter , for which the name Serinibacter tropicus sp. nov. is proposed. The type strain is PS-14-7T ( = NBRC 110108T = InaCC A 515T). An emended description of the genus Serinibacter is also proposed.

Two novel Gram-stain positive actinobacteria, designated PS-14-16(T) and RS-7-1, were isolated from the rhizosphere of a mangrove and sea sediment, respectively, and their taxonomic positions were investigated by a polyphasic approach. Both strains were observed to form vegetative hyphae in the early phase of growth but the hyphae eventually fragment into short rods to coccoid cells. The peptidoglycan type of both strains was found to be A4α. Their predominant menaquinone was identified as MK-9(H4) and the major fatty acid as anteiso-C(15:0). The DNA G+C content was determined to be 68.4-68.5 mol%. 16S rRNA gene sequencing revealed that strains PS-14-16(T) and RS-7-1 were related to members of the family Cellulomonadaceae. Their nearest phylogenetic neighbour was found to be Sediminihabitans luteus, which is currently the only species of the genus Sediminihabitans, with a similarity of 97.94%. However, strains PS-14-16(T) and RS-7-1 were distinguishable from the members of the genus Sediminihabitans and the other genera within the family Cellulomonadaceae in terms of chemotaxonomic characteristics and phylogenetic relationship. The results of DNA-DNA hybridization experiments indicated that strains PS-14-16(T) and RS-7-1 belong to the same species. Strains PS-14-16(T) and RS-7-1 are concluded to represent a novel genus and species of the family Cellulomonadaceae, for which the name Tropicihabitans flavus gen. nov., sp. nov. is proposed. The type strain of T. flavus is PS-14-16(T) (=NBRC 110109(T) = IanCC A 516(T)). [corrected].

Type strains of 72 validated Nocardia species were phylogenetically analyzed based on the multilocus sequence analysis (MLSA) concatenated atpD-groL1-groL2-recA-rpoA-secY-sodA-ychF. Furthermore, their similarity based on digital DNA-DNA hybridization (dDDH) was calculated. Nocardia soli, Nocardia cummidelens and Nocardia salmonicida, Nocardia nova and Nocardia elegans, Nocardia exalbida and Nocardia gamkensis, and Nocardia coubleae and Nocardia ignorata formed coherent clades, respectively. Moreover, each set showed over 70% relatedness by dDDH and shared common phenotypic characteristics. Therefore, we propose a reclassification of Nocardia soli and Nocardia cummidelens as a later heterotypic synonym of Nocardia salmonicida, Nocardia elegans as a later heterotypic synonym of Nocardia nova, Nocardia gamkensis as a later heterotypic synonym of Nocardia exalbida, and Nocardia coubleae as a later heterotypic synonym of Nocardia ignorata.

We have developed butanol-producing consolidated bioprocessing from cellulosic substrates through coculture of cellulolytic clostridia and butanol-producing Clostridium saccharoperbutylacetonicum strain N1-4. However, the butanol fermentation by strain N1-4 (which has an optimal growth temperature of 30°C) is sensitive to the higher cultivation temperature of 37°C; the nature of this deleterious effect remains unclear. Comparison of the intracellular metabolites of strain N1-4 cultivated at 30°C and 37°C revealed decreased levels of multiple primary metabolites (notably including nucleic acids and cofactors) during growth at the higher temperature. Supplementation of the culture medium with 250 mg/liter adenine enhanced both cell growth (with the optical density at 600 nm increasing from 4.3 to 10.2) and butanol production (increasing from 3.9 g/liter to 9.6 g/liter) at 37°C, compared to those obtained without adenine supplementation, such that the supplemented 37°C culture exhibited growth and butanol production approaching those observed at 30°C in the absence of adenine supplementation. These improved properties were based on the maintenance of cell viability. We further showed that adenine supplementation enhanced cell viability during growth at 37°C by maintaining ATP levels and inhibiting spore formation. This work represents the first demonstration (to our knowledge) of the importance of adenine-related metabolism for clostridial butanol production, suggesting a new means of enhancing target pathways based on metabolite levels. IMPORTANCE Metabolomic analysis revealed decreased levels of multiple primary metabolites during growth at 37°C, compared to 30°C, in C. saccharoperbutylacetonicum strain N1-4. We found that adenine supplementation restored the cell growth and butanol production of strain N1-4 at 37°C. The effects of adenine supplementation reflected the maintenance of cell viability originating from the maintenance of ATP levels and the inhibition of spore formation. Thus, our metabolomic analysis identified the depleted metabolites that were required to maintain cell viability. Our strategy, which is expected to be applicable to a wide range of organisms, permits the identification of the limiting metabolic pathway, which can serve as a new target for molecular breeding. The other novel finding of this work is that adenine supplementation inhibits clostridial spore formation. The mechanism linking spore formation and metabolomic status in butanol-producing clostridia is expected to be the focus of further research.

Culture collections have played important roles in the preservation and distribution of microbial cultures to end users and contributed significantly toward microbial taxonomy as acting depositories of type strains that are used to identify new relatives of these strains. Quality management requirement of maintaining cultures is increasingly becoming important with the progress of molecular biology and genomics accompanying databases in life sciences and biotechnology. At the same time, culture collections are expected to work for the infrastructure to ensure compliance with national and international laws and regulations for handling and transferring microbial resources. Microbiologists must pay particular attention to the national policies in their respective countries concerning the Convention on Biological Diversity for access into microbial resources. Biological resource centers (BRCs) are the advanced stage of culture collections and expected to provide infrastructural support for the activities of microbiologists in relation to social accountability as well as the scientific activities.

A novel Gram-stain-positive, short rod-shaped, non-flagellated and mesophilic strain, KIS12-7T, isolated from a soil sample collected from Daecheong-Island in Ongjin County, Republic of Korea, was studied using a polyphasic approach. Phylogenetic trees based on the 16S rRNA gene sequence revealed that the novel strain was a member of the genus Gryllotalpicola , showing more than 97.0 % sequence similarity with Gryllotalpicola daejeonensis RU-04T (98.0 %), Gryllotalpicola koreensis RU-16T (97.7 %) and Gryllotalpicola kribbensis PU-02T (97.3 %). However, DNA–DNA relatedness values demonstrated that strain KIS12-7T could be clearly distinguished from closely related species of the genus Gryllotalpicola . The cell-wall peptidoglycan of strain KIS12-7T was of the type B2 and the acyl type was acetyl. The predominant menaquinones were MK-11 and MK-10. Polar lipids were diphosphatidylglycerol, phosphatidylglycerol, one unknown phosphoglycolipid, one unknown glycolipid, one unknown phospholipid and one unknown lipid. The G+C content of the genomic DNA was 72.1 mol%. On the basis of the evidence presented, strain KIS12-7T is a representative of a novel species of the genus Gryllotalpicola , and the name Gryllotalpicola soli sp. nov. is proposed; the type strain is KIS12-7T ( = DSM 27182T = KACC 17302T = NBRC 109659T).

Two thermophilic, strictly anaerobic, Gram-negative bacteria, designated strains AZM34c06T and AZM44c09T, were isolated from terrestrial hot springs in Japan. The optimum growth conditions for strain AZM34c06T were 60 °C, pH 7.4 and 0 % additional NaCl, and those for strain AZM44c09T were 70 °C, pH 7.4 and 0 % additional NaCl. Complete genome sequencing was performed for both strains, revealing genome sizes of 2.19 Mbp (AZM34c06T) and 2.01 Mbp (AZM44c09T). Phylogenetic analyses based on 16S rRNA gene sequences and the concatenated predicted amino acid sequences of 33 ribosomal proteins showed that both strains belonged to the genus Thermotoga . The closest relatives of strains AZM34c06T and AZM44c09T were the type strains of Thermotoga lettingae (96.0 % similarity based on the 16S rRNA gene and 84.1 % similarity based on ribosomal proteins) and Thermotoga hypogea (98.6 and 92.7 % similarity), respectively. Using blast, the average nucleotide identity was 70.4–70.5 % when comparing strain AZM34c06T and T. lettingae TMOT and 76.6 % when comparing strain AZM44c09T and T. hypogea NBRC 106472T. Both values are far below the 95 % threshold value for species delineation. In view of these data, we propose the inclusion of the two isolates in the genus Thermotoga within two novel species, Thermotoga profunda sp. nov. (type strain AZM34c06T = NBRC 106115T = DSM 23275T) and Thermotoga caldifontis sp. nov. (type strain AZM44c09T = NBRC 106116T = DSM 23272T).

Two bacterial strains, KIS66-7(T) and 5GH26-15(T), were isolated from soil samples collected in the South Korean cities of Tongyong and Gongju, respectively. Both strains were aerobic, Gram-stain-positive, mesophilic, flagellated, and rodshaped. A phylogenetic analysis revealed that both strains belonged to the family Microbacteriaceae of the phylum Actinobacteria. The 16S rRNA gene sequence of strain KIS66-7(T) had the highest similarities with those of Labedella gwakjiensis KSW2-17(T) (97.3%), Cryobacterium psychrophilum DSM 4854T (97.2%), Leifsonia lichenia 2Sb(T) (97.2%), Leifsonia naganoensis JCM 10592(T) (97.0%), and Cryobacterium mesophilum MSL-15(T) (97.0%). Strain 5GH26-15(T) showed the highest sequence similarities with Leifsonia psychrotolerans LI1T (97.4%) and Schumannella luteola KHIAT (97.1%). The 16S rRNA gene sequence from KIS66-7(T) exhibited 96.4% similarity with that from 5GH26-15(T). Strain KIS66-7(T) contained a B2γ type peptidoglycan structure with D-DAB as the diamino acid; MK-13, MK-12, and MK-14 as the respiratory quinones; ai-C15:0, ai-C17:0, and i-C16:0 as the major cellular fatty acids; and diphosphatidylglycerol, phatidylglycerol, and glycolipids as the predominant polar lipids. Strain 5GH26-15T had a B2β type peptidoglycan structure with D-DAB as the diamino acid; MK-14 and MK-13 as the respiratory quinones; ai-C15:0, i-C16:0, and ai-C{vn17:0} as the major cellular fatty acids; and diphosphatidylglycerol, phatidylglycerol, and glycolipids as the predominant polar lipids. Both strains had low DNA-DNA hybridization values (

A novel Gram-stain-positive actinobacterium, designated HR08-44T, was isolated from a sea-sediment sample collected from the foreshore of Rishiri Island, Japan, and its taxonomic position was investigated by a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain HR08-44T was closely related to the members of the genus Janibacter , with pairwise sequence similarities of 97.3–98.8 %. Strain HR08-44T had peptidoglycan type A1γ, with meso-diaminopimelic acid as the diagnostic diamino acid. The predominant menaquinone was MK-8(H4) and the major fatty acids were iso-C16 : 0, C17 : 1ω8c, C17 : 0 and C18 : 1ω9c. These data supported the affiliation of the novel strain to the genus Janibacter . Meanwhile, the results of DNA–DNA hybridization and physiological and biochemical tests indicated that strain HR08-44T can be distinguished from recognized species of the genus Janibacter . Therefore, strain HR08-44T represents a novel species of the genus Janibacter , for which the name Janibacter cremeus sp. nov. is proposed; the type strain is HR08-44T ( = NBRC 107693T = DSM 26154T).

A novel actinomycete strain, 7402J-48T, was isolated from an air sample collected from Jeju island, Republic of Korea. Cells were Gram-positive, aerobic, flagellated, short rods. Strain 7402J-48T showed highest 16S rRNA gene sequence similarity (98.6 %) with Angustibacter luteus TT07R-79T, and had relatively low sequence similarities (below 95.1 %) with other members of the family Kineosporiaceae . The cell wall of strain 7402J-48T contained alanine, glutamic acid and 2,6-diaminopimelic acid, suggesting A1γ-type peptidoglycan. The menaquinones were MK-9(H4) and MK-8(H4). The acyl type of the cell-wall muramic acid was acetyl. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside and an unknown lipid were present. The cellular fatty acid profile comprised a large amount of anteiso-C15 : 0, moderate amounts of C16 : 0, summed feature 3 (including C16 : 1ω7c and/or C16 : 1ω6c), C17 : 0, iso-C15 : 0, C16 : 0 N alcohol and C17 : 1ω8c, and small amounts of other fatty acids. The DNA G+C content of strain 7402J-48T was 73 mol%. Based on its phenotypic and genotypic characteristics, strain 7402J-48T ( = KACC 15527T = NBRC 108730T) is proposed as the type strain of a novel species, Angustibacter aerolatus sp. nov. An emended description of the genus Angustibacter is provided.

The class Thermoplasmata harbors huge uncultured archaeal lineages at the order level, so-called Groups E2 and E3. A novel archaeon Kjm51a affiliated with Group E2 was enriched from anaerobic sludge in the present study. Clone library analysis of the archaeal 16S rRNA and mcrA genes confirmed a unique archaeal population in the enrichment culture. The 16S rRNA gene-based phylogeny revealed that the enriched archaeon Kjm51a formed a distinct cluster within Group E2 in the class Thermoplasmata together with Methanomassiliicoccus luminyensis B10(T) and environmental clone sequences derived from anaerobic digesters, bovine rumen, and landfill leachate. Archaeon Kjm51a showed 87.7% 16S rRNA gene sequence identity to the closest cultured species, M. luminyensis B10(T), indicating that archaeon Kjm51a might be phylogenetically novel at least at the genus level. In fluorescence in situ hybridization analysis, archaeon Kjm51a was observed as coccoid cells completely corresponding to the archaeal cells detected, although bacterial rod cells still coexisted. The growth of archaeon Kjm51a was dependent on the presence of methanol and yeast extract, and hydrogen and methane were produced in the enrichment culture. The addition of 2-bromo ethanesulfonate to the enrichment culture completely inhibited methane production and increased hydrogen concentration, which suggested that archaeon Kjm51a is a methanol-reducing hydrogenotrophic methanogen. Taken together, we propose the provisional taxonomic assignment, named Candidatus Methanogranum caenicola, for the enriched archaeon Kjm51a belonging to Group E2. We also propose to place the methanogenic lineage of the class Thermoplasmata in a novel order, Methanomassiliicoccales ord. nov.

The aim of this study was to reclarify the taxonomic relationship among Streptomyces hygroscopicus subsp. hygroscopicus, Streptomyces endus and Streptomyces sporocinereus. Whole genome shotgun sequencing was performed for the type strains of these three taxa. Average nucleotide identity and digital DNA-DNA hybridization values among the three taxa were greater than the thresholds for bacterial species delineation, indicating that they belong to the same genomospecies. In addition, the phenotypic data previously reported also support the synonymy. Therefore, S. endus and S. sporocinereus should be reclassified as later heterotypic synonyms of S. hygroscopicus subsp. hygroscopicus.

A novel and extremely halophilic archaeon, designated strain 2a_47_2T, was isolated from a solar saltern sample collected in Indonesia. Cells of the strain were Gram-stain-negative, non-motile and pleomorphic and formed orange-red pigmented colonies. Strain 2a_47_2T grew at 20-48 °C (optimum 38-41 °C), pH 6.0-8.5 (optimum pH 7.5), >1.7 M NaCl (optimum 2.6 M) and

The electron transport system in Pseudomonas hydrogenothermophila strain TH-1, a thermophilic, aerobic hydrogen-oxidizing bacterium, was studied using a membrane fraction, an ubiquinone-depleted membrane fraction (Q(-) MF), and an ubiqunone-reconstituted membrane fraction (Q(+) MF). From spectrophotometric and TLC analyses, cytochromes b and c, ubiquinone-8 (Q8), and cytochrome o were detected as respiratory components in the membrane fraction of P. hydrogenothermophila. Spectrophotometric analyses also showed that reduction of all the components in the membrane fraction depends on a membrane-bound hydrogenase reaction. In Q(-) MF, reduction of cytochromes b and c depending on membrane-bound hydrogenase reaction could not be observed. However, the reduction was restored in Q(+) MF. From these results, a possible electron transport system in P. hydrogenothermophila strain TH-1 is proposed.

Here, we report the draft genome sequence of strain NBRC 16556, deposited as Streptomyces hygroscopicus subsp. hygroscopicus into the NBRC culture collection. An average nucleotide identity analysis confirmed that the taxonomic identification is correct. The genome sequence will serve as a valuable reference for genome mining to search new secondary metabolites.

A novel Gram stain positive actinobacterium, designated RS-7-4(T), was isolated from a sea sediment sample collected in Indonesia, and its taxonomic position was investigated using a polyphasic approach. Strain RS-7-4(T) was observed to form vegetative hyphae in the early phase of growth, but the hyphae eventually fragmented into short rods to coccoid cells. Growth occurred at 15-37 °C, pH 6.0-11.0 and in the presence of 0-7 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that strain RS-7-4(T) was closely related to the members of the genus Cellulosimicrobium, with a similarity range of 98.08-99.10 %. The peptidoglycan type of strain RS-7-4(T) was found to be A4α L-Lys-L-Thr-D-Asp. The predominant menaquinone was MK-9(H4), and the major fatty acids were anteiso-C15:0, iso-C15:0 and anteiso-C17:0. The DNA G+C content was 75.6 mol%. These chemotaxonomic features corresponded to those of the genus Cellulosimicrobium. Meanwhile, the results of DNA-DNA hybridization, and physiological and biochemical tests revealed that strain RS-7-4(T) was different from the recognized species of the genus Cellulosimicrobium. Therefore, strain RS-7-4(T) represents a novel species of the genus Cellulosimicrobium, for which the name Cellulosimicrobium marinum sp. nov. is proposed. The type strain is RS-7-4(T) (=NBRC 110994(T) =InaCC A726(T)).

Actinoplanes missouriensis Couch 1963 is a well-characterized member of the genus Actinoplanes, which is of morphological interest because its members typically produce sporangia containing motile spores. The sporangiospores are motile by means of flagella and exhibit chemotactic properties. It is of further interest that members of Actinoplanes are prolific sources of novel antibiotics, enzymes, and other bioactive compounds. Here, we describe the features of A. missouriensis 431(T), together with the complete genome sequence and annotation. The 8,773,466 bp genome contains 8,125 protein-coding and 79 RNA genes.

A novel isolate, designated 6408J-67(T), was isolated from an air sample collected from Jeju Island, Republic of Korea. Its phenotypic, genotypic, and chemotaxonomic properties were compared with those of members of the family Microbacteriaceae. The Gram-positive, aerobic, motile rod formed light yellow, smooth, circular and convex colonies. Optimal growth occurred at 30°C and pH 7.0. 16S rRNA gene sequence data showed that the isolate was a novel member of the family Microbacteriaceae, with the highest sequence similarity (97.4%) to Labedella gwakjiensis KSW2-17(T) and less (97%) sequence similarity with other taxa. The major cellular fatty acids (10% of the total) were anteiso-C(15:0), iso-C(14:0), and iso-C(16:0). The strain also contained MK-13, MK-12, and MK-14 as the major menaquinones, as well as diphosphatidylglycerol, phosphatidylglycerol, and two unknown glycolipids. Its peptidoglycan structure was B1β with 2,4-diaminobutyric acid as a diamino acid. Mycolic acids were absent. The DNA G+C content was 68.3 mol%. Based on these phenotypic and genotypic findings, strain 6408J-67(T) represents a novel species of a new genus within the family Microbacteriaceae, for which the name Diaminobutyricimonas aerilata gen. nov., sp. nov. is proposed. The type strain is 6408J-67(T) (=KACC 15518(T) =NBRC 108726(T)).

Oscillibacter valericigenes is a mesophilic, strictly anaerobic bacterium belonging to the clostridial cluster IV. Strain Sjm18-20(T) (=NBRC 101213(T) =DSM 18026(T)) is the type strain of the species and represents the genus Oscillibacter Iino et al. 2007. It was isolated from the alimentary canal of a Japanese corbicula clam (Corbicula japonica) collected on a seacoast in Shimane Prefecture in Japan. Phylogenetically, strain Sjm18-20(T) is closest to uncultured bacteria in digestive tracts, including the enriched cells thought to represent Oscillospira guilliermondii Chatton and Perard 1913. The isolated phylogenetic position and some distinct characteristics prompted us to determine the complete genome sequence. The 4,410,036 bp chromosome and the 60,586 bp plasmid were predicted to encode a total of 4,723 protein-coding genes.

A non-motile and non-endospore-forming rod, strain NBRC 16403T, was isolated from the phyllosphere of a sedge (Carex sp.). 16S rRNA gene sequence analysis indicated that strain NBRC 16403T was closely related to Herbiconiux solani DSM 19813T (98.6 % 16S rRNA gene sequence similarity), Herbiconiux ginsengi wged11T (97.8 %) and Herbiconiux moechotypicola RB-62T (97.8 %). The peptidoglycan (B2γ type) contained d- and l-2,4-diaminobutyric acids, d-alanine, glycine and threo-3-hydroxyglutamic acid, which replaced glutamic acid almost completely. The predominant menaquinones were MK-10 and MK-11. The polar lipid pattern comprised diphosphatidylglycerol, phosphatidylglycerol, three glycolipids and minor amounts of other polar lipids. The major fatty acids were anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0; no cyclohexyl-C17 : 0 was detected. The DNA G+C content was 71.0 mol%. The results of phylogenetic and DNA–DNA relatedness studies, along with phenotypic differences between strain NBRC 16403T and recognized members of the genus Herbiconiux, indicated that the isolate should be assigned to a novel species of the genus Herbiconiux, for which the name Herbiconiux flava sp. nov. is proposed. The type strain is NBRC 16403T ( = VKM Ac-2058T).