- Volume 61, Issue 5, 2011

In the context of studying the effects of transgenic fructan-producing potatoes on the community structure of phyllosphere bacteria, a group of strains closely related to the species Leifsonia ginsengi was isolated. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the new isolates and L. ginsengi DSM 19088T formed a lineage at the genus level and this finding was supported by chemotaxonomic characterization. The peptidoglycan type of the representative isolate, K134/01T, and L. ginsengi DSM 19088T was B2γ, with d- and l-diaminobutyric acid as the diagnostic diamino acid and glycine, alanine and threo-3-hydroxyglutamic acid. The almost-complete substitution of glutamic acid by threo-3-hydroxyglutamic acid supported the differentiation of the new strains from recognized species of the genus Leifsonia. Furthermore, the detection of substantial amounts of the fatty acid cyclohexyl-C17 : 0 in the new isolates and L. ginsengi DSM 19088T was a prominent chemotaxonomic feature for a clear demarcation of these strains from all genera of the family Microbacteriaceae that display the B2γ cell-wall type. Comparative phylogenetic and phenotypic analyses of the isolates and L. ginsengi DSM 19088T revealed the separate species status of the isolates. On the basis of these results, it is proposed that L. ginsengi should be classified as the type species of a novel genus, Herbiconiux gen. nov., with the name Herbiconiux ginsengi gen. nov., comb. nov. (type strain wged11T = CGMCC 4.3491T = JCM 13908T = DSM 19088T = NBRC 104580T). The phyllosphere isolates are assigned to a novel species, Herbiconiux solani sp. nov. (type strain K134/01T = DSM 19813T = LMG 24387T = NBRC 106740T).

A novel actinomycete strain, designated YIM 63587T, was isolated from surface-sterilized roots of Artemisia annua L. collected from Yunnan province, south-west China. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 63587T was affiliated to the genus Pseudonocardia. 16S rRNA gene sequence similarities between strain YIM 63587T and type strains of species of the genus Pseudonocardia were 96.6–93.8 %. The diagnostic cell-wall diamino acid in the peptidoglycan layer of strain YIM 63587T was meso-diaminopimelic acid and the whole-cell sugars were arabinose, galactose, mannose and ribose. The predominant menaquinone was MK-8(H4) (97.7 %). The phospholipids were diphosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, phosphatidylinositol mannosides, phosphatidylinositol and an unknown phospholipid. The major cellular fatty acids (>5 %) were iso-C16 : 0 (44.7 %), iso-C14 : 0 (10.3 %), iso-C16 : 1 H (9.8 %) and iso-C15 : 0 (7.7 %). The G+C content of the genomic DNA was 68.2 mol%. On the basis of phylogenetic, physiological and chemotaxonomic data, strain YIM 63587T represents a novel species of the genus Pseudonocardia, for which the name Pseudonocardia artemisiae sp. nov. is proposed. The type strain is YIM 63587T ( = DSM 45313T = CCTCC AA 208081T).

A Gram-staining-positive bacterium, designated AS-0823T, which formed spiral spore chains on the aerial mycelium, was isolated from the intestinal tract of Armadillidium vulgare, a small terrestrial crustacean commonly found on the ground around houses in Japan. 16S rRNA gene sequence analysis showed that the isolate belonged to the genus Streptomyces and was most closely related to Streptomyces longisporus ISP 5166T (98.6 % 16S rRNA gene sequence similarity), Streptomyces curacoi NBRC 12761T (98.4 %) and Streptomyces griseoruber NBRC 12873T (98.4 %). The affiliation of strain AS-0823T to the genus Streptomyces was supported by chemotaxonomic data: iso-C16 : 0, anteiso-C15 : 0, C16 : 0, iso-C15 : 0 and anteiso-C17 : 0 as the major cellular fatty acids, ll-diaminopimelic acid as the characteristic diamino acid in the peptidoglycan and the absence of mycolic acids. DNA–DNA hybridization and physiological and biochemical analysis supported the differentiation of strain AS-0823T from S. longisporus JCM 4395T. Therefore, strain AS-0823T represents a novel species, for which the name Streptomyces coacervatus sp. nov. is proposed. The type strain is AS-0823T ( = IFM 11055T  = DSM 41983T  = JCM 17138T).

The number of tacrolimus (FK506)-producing isolates has increased remarkably because of the clinical importance of tacrolimus as an immunosuppressant. However, the taxonomy of several of these isolates has not been studied. The taxonomic status of strain ATCC 55098T, a tacrolimus-producing strain, was determined in this study. The genotypic, phenotypic and chemotaxonomic properties were consistent with the inclusion of strain ATCC 55098T in the genus Streptomyces. The 16S rRNA gene sequence of strain ATCC 55098T was determined and used to generate phylogenetic trees with corresponding sequences of the most closely related type strains (≥98 % 16S rRNA gene sequence similarity) of species of the genus Streptomyces. Strain ATCC 55098T formed a distinct phylogenetic branch adjacent to a cluster comprising Streptomyces fulvissimus NBRC 13482T and Streptomyces flavofungini NBRC 13371T. However, morphological and physiological tests and DNA–DNA relatedness distinguished strain ATCC 55098T from its closest phylogenetic neighbours. On the basis of these results, strain ATCC 55098T represents a novel species of the genus Streptomyces, for which the name Streptomyces tacrolimicus sp. nov. is proposed. The type strain is ATCC 55098T ( = CECT 7664T).

A Gram-type-positive, strictly aerobic actinobacterium, designated strain MON 2.2T, was isolated from the surface of a sandstone monument. Cells with a coccoid shape, arranged in pairs or clusters, were non-motile and did not produce spores. The 10 closest 16S rRNA gene sequence matches (~95 % similarity) found in the public databases were uncultured actinobacteria, while the closest cultured members indicated a phylogenetic relationship with members of the family Propionibacteriaceae (92–95 % similarity). Subsequent phylogenetic analysis placed the new isolate within the radiation of the genera Friedmanniella and Microlunatus, but forming an independent branch. Chemotaxonomic markers were consistent with the classification of strain MON 2.2T in the family Propionibacteriaceae, amongst the genera containing ll-diaminopimelic acid in their peptidoglycan. Characteristic fatty acids iso-C15 : 0 and anteiso-C15 : 0 also supported its affiliation to this taxon; however, polar lipid and menaquinone compositions clearly differentiated strain MON 2.2T from other genera in the family. On the basis of these results and additional physiological data obtained in the present study, it is proposed that strain MON 2.2T be classified in a novel species in a new genus, for which the name Auraticoccus monumenti gen. nov., sp. nov. is proposed. The type strain of Auraticoccus monumenti is MON 2.2T ( = CECT 7672T  = DSM 23257T  = LMG 25551T).

A novel actinomycete, strain X4T, was isolated from a phosphate mine in Youssoufia, 100 km north of Marrakesh, Morocco. The taxonomic status of this strain was evaluated by a polyphasic approach. Strain X4T had white aerial mycelium with Rectiflexibiles spore chains bearing smooth-surfaced spores and did not produce diffusible pigments. Chemotaxonomic analysis showed that the cell wall of strain X4T contained ll-diaminopimelic acid and glycine. Phylogenetic analysis based on the almost complete 16S rRNA gene sequence indicated that strain X4T belongs to the Group I streptomycetes, branching off next to Streptomyces ramulosus NRRL B-2714T and Streptomyces kasugaensis M338-M1T. DNA–DNA relatedness and phenotypic data enabled strain X4T to be distinguished from the phylogenetically most closely related type strains. It is therefore proposed that strain X4T represents a novel species of the genus Streptomyces, for which the name Streptomyces youssoufiensis sp. nov. is proposed; the type strain is X4T ( = CCMM B709T  = DSM 41920T).

A Gram-positive bacterium, strain M-6-3T, was isolated from salt-fermented seafood in Korea. The organism grew in 0–10 % (w/v) NaCl and at 25–37 °C, with optimal growth occurring in 5 % NaCl and at 28–30 °C. The peptidoglycan type was variation A4γ with meso-diaminopimelic acid as the diagnostic cell-wall diamino acid. The polar lipid profile of strain M-6-3T consisted of diphosphatidylglycerol, phosphatidylglycerol, an unidentified phospholipid and an unknown glycolipid. Strain M-6-3T contained MK-7 as the major component of the quinone system and anteiso-C15 : 0 (62.1 %) as the predominant fatty acid. Based on 16S rRNA gene sequence similarity studies, strain M-6-3T was most closely related to Brachybacterium rhamnosum LMG 19848T (98.5 %). The G+C content of the genomic DNA was 71.5 mol% and the mean DNA–DNA hybridization value with reference strains was 14.32±2.0 %. Based on phenotypic, genotypic and phylogenetic analyses, it is proposed that strain M-6-3T represents a novel species for which the name Brachybacterium squillarum sp. nov. is proposed; the type strain is M-6-3T ( = KACC 14221T  = JCM 16464T).

A novel endophytic actinomycete, designated strain 202201T, was isolated from an Acanthus illicifolius root collected from the mangrove reserve zone in Hainan Province, China. Phylogenetic analysis based on 16S rRNA gene sequences suggested that strain 202201T fell within the family Micromonosporaceae. The strain formed an extensively branched substrate mycelium, which carried uneven warty-surfaced spores. Cell walls of strain 202201T contained meso-diaminopimelic acid and xylose, mannose, arabinose, ribose and glucose were detected as whole-cell sugars. The acyl type of the cell-wall polysaccharides was glycolyl. The major menaquinones were MK-9(H4), MK-9(H6), MK-9(H8) and MK-10(H4). The polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol mannoside and phosphatidylserine. The major cellular fatty acids were 10-methyl-C17 : 0, iso-C15 : 0, iso-C16 : 0 and C17 : 1ω8c. The DNA G+C content was 72.3 mol%. On the basis of the morphological and chemotaxonomic characteristics, phylogenetic analysis and characteristic patterns of 16S rRNA gene signature nucleotides, strain 202201T ( = CGMCC 4.5597T  = DSM 45430T) represents a novel species of a new genus within the family Micromonosporaceae, for which the name Jishengella endophytica gen. nov., sp. nov. is proposed.

A Gram-positive, aerobic actinomycete, designated strain NEAU 119T, was isolated from the rhizosphere of a fig tree and was characterized using a polyphasic approach. The isolate formed branching, non-fragmenting vegetative hyphae and produced black pigment on yeast extract/malt extract (ISP medium 2). The G+C content of the DNA was 76.6 mol%. The organism had chemotaxonomic characteristics typical of the genus Actinoalloteichus and was closely related to the type strains of Actinoalloteichus cyanogriseus, Actinoalloteichus spitiensis and Actinoalloteichus hymeniacidonis, currently the only three recognized species of the genus Actinoalloteichus, sharing 16S rRNA gene similarities of 96.4, 96.6 and 98.1 %, respectively. However, the results of DNA–DNA hybridization studies demonstrated that the novel strain showed only 46.8 % relatedness with the type strain of A. hymeniacidonis. In addition, a set of phenotypic characteristics also readily distinguished strain NEAU 119T from the type strains of recognized species of the genus Actinoalloteichus. According to the above data, it is proposed that strain NEAU 119T represents a novel species, Actinoalloteichus nanshanensis sp. nov. The type strain of Actinoalloteichus nanshanensis is NEAU 119T ( = CGMCC 4.5714T = NBRC 106685T).

A novel actinomycete, strain P0402T, was isolated from peat swamp forest soil collected in Thailand. Its taxonomic position was determined by using a polyphasic taxonomic approach. The chemotaxonomic characteristics of this strain matched those of the genus Micromonospora, i.e. the presence of meso-diaminopimelic acid and N-glycolyl muramic acid in the peptidoglycan, whole-cell sugar pattern D, phospholipid type II, and cellular fatty acid type 3b. Phylogenetic analysis based on 16S rRNA gene sequences revealed a close relationship between strain P0402T and Micromonospora coxensis JCM 13248T (99.0 % similarity), Micromonospora eburnea JCM 12345T (99.0 %), Micromonospora marina JCM 12870T (98.9 %), Micromonospora halophytica JCM 3125T (98.7 %), Micromonospora chalcea JCM 3031T (98.7 %), Micromonospora purpureochromogenes JCM 3156T (98.6 %) and Micromonospora aurantiaca JCM 10878T (98.5 %). It could be clearly distinguished from these type strains based on low levels of DNA–DNA relatedness and phenotypic differences. On the basis of the data presented, strain P0402T is suggested to represent a novel species of the genus Micromonospora, for which the name Micromonospora humi sp. nov. is proposed. The type strain is P0402T ( = JCM 15292T  = PCU 315T  = TISTR 1883T).

An actinomycete strain, HM7T, was isolated from sandy soil from the banks of the Arvand River, Khoramshahr, Iran. In a study conducted using a polyphasic taxonomic approach, it was found that this strain represented a novel species in the genus Nocardiopsis. Strain HM7T had long, branched substrate mycelia and the aerial mycelium completely fragmented to long chains of rod-shaped spores. The cell wall of strain HM7T contained meso-diaminopimelic acid, galactose and ribose. The predominant phospholipids were phosphatidylglycerol, phosphatidylethanolamine diphosphatidylglycerol, and phosphatidylcholine. The main menaquinones were MK-10(H2), MK-10(H4), MK-10(H0) and MK-9(H2). Strain HM7T showed the highest degree of 16S rRNA gene sequence similarity with Nocardiopsis sinuspersici UTMC 00102T (99.8 %) and Nocardiopsis quinghaiensis YIM 28A4T (99.3 %), but differed significantly from these type strains based on DNA–DNA hybridization data (37.6 and 38.9 % relatedness, respectively). Physiological, biochemical and phenotypic characteristics allowed strain HM7T to be differentiated from related species. On the basis of results from phenotypic and phylogenetic analyses, strain HM7T is considered to represent a novel species of the genus Nocardiopsis, for which the name Nocardiopsis arvandica sp. nov. is proposed; the type strain of this species is HM7T ( = UTMC 00103T = DSM 45278T = CCUG 58831T).

A novel, Gram-stain-positive bacterial strain, Mer 29717T, was isolated from the branchia (gills) of a Japanese codling, Physiculus japonicus, collected from bottom waters of Suruga Bay in Shizuoka, Japan. Phylogenetic analysis based on 16S rRNA gene sequences indicated that this strain represents a distinct lineage within the family Dermacoccaceae and was related most closely to members of the genera Demetria and Yimella. It shared highest 16S rRNA gene sequence similarity (95.1 %) with Yimella lutea YIM 45900T. Strain Mer 29717T contained MK-8(H2) and MK-8(H4) as menaquinones, and iso-C16 : 0, C16 : 0, C17 : 1 cis-9, C17 : 0, C18 : 1 cis-9 and C19 : 1 cis-10 were the major cellular fatty acids. The cell-wall peptidoglycan of strain Mer 29717T was composed of l-Lys, d-Ser, l-Ser, Gly, d-Glu and d-Ala. Polar lipids were phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol and one unidentified phospholipid. Mycolic acids were not detected. The G+C content of the DNA of strain Mer 29717T was 68 mol%. On the basis of differential chemotaxonomic, physiological and biochemical data, strain Mer 29717T is considered to represent a novel species of a new genus, for which the name Branchiibius hedensis gen. nov., sp. nov. is proposed. The type strain of Branchiibius hedensis is Mer 29717T ( = NBRC 106121T  = DSM 22951T).

Two halophilic archaea, strains EN-2T and SH-4, were isolated from the saline lakes Erliannor and Shangmatala, respectively, in Inner Mongolia, China. Cells were strictly aerobic, motile rods. Colonies were red. Strains EN-2T and SH-4 were able to grow at 25–50 °C (optimum 35–40 °C), with 2.5–5.0 M NaCl (optimum 3.4 M NaCl) and at pH 6.0–9.0 (optimum pH 7.5). MgCl2 was not required for growth. Cells lysed in distilled water and the lowest NaCl concentration that prevented cell lysis was 12 % (w/v). On the basis of 16S rRNA gene sequence analysis, strains EN-2T and SH-4 were closely related to Halorubrum cibi B31T (97.9 and 98.0 % similarity, respectively), Hrr. tibetense 8W8T (97.3 and 97.7 %), Hrr. alkaliphilum DZ-1T (96.8 and 97.1 %), Hrr. luteum CGSA15T (96.8 and 97.0 %) and Hrr. lipolyticum 9-3T (96.8 and 97.0 %). DNA–DNA hybridization showed that strains EN-2T and SH-4 did not belong to the same species as any of these strains (≤45 % DNA–DNA relatedness) but that they are members of the same species (>70 % DNA–DNA relatedness). Polar lipid analysis revealed that strains EN-2T and SH-4 contained phosphatidylglycerol, phosphatidylglycerol phosphate methyl ester, sulfated diglycosyl diethers and several unidentified glycolipids. The DNA G+C content of both isolates was 62.1 mol%. It was concluded that strains EN-2T and SH-4 represent a novel species of the genus Halorubrum, for which the name Halorubrum aquaticum sp. nov. is proposed. The type strain is EN-2T ( = CECT 7174T  = CGMCC 1.6377T  = JCM 14031T).

A Gram-negative, pleomorphic, aerobic, haloalkaliphilic archaeon, strain 167-74T, was isolated from commercial rock salt imported into Japan from China. Phylogenetic analysis based on 16S rRNA gene sequence similarities showed that strain 167-74T is closely related to Halostagnicola larsenii XH-48T (98.3 %) and Halostagnicola kamekurae 194-10T (97.2 %). The major polar lipids of the isolate were C20C20 and C20C25 derivatives of phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester. A glycolipid was not detected, in contrast to the two existing, neutrophilic species of the genus Halostagnicola. The DNA G+C content of strain 167-74T was 60.7 mol%. and it gave DNA–DNA reassociation values of 19.5 and 18.8 %, respectively, with Hst. larsenii JCM 13463T and Hst. kamekurae 194-10T. Therefore, strain 167-74T represents a novel species, for which the name Halostagnicola alkaliphila sp. nov. is proposed, with the type strain 167-74T ( = JCM 16592T  = CECT 7631T).

Two Gram-stain-negative, non-motile, non-spore-forming, rod-shaped bacterial strains, designated 3B-2T and 10AOT, were isolated from a sand sample collected from the west coast of the Korean peninsula by using low-nutrient media, and their taxonomic positions were investigated in a polyphasic study. The strains did not grow on marine agar. They grew optimally at 30 °C and pH 6.5–7.5. Strains 3B-2T and 10AOT shared 97.5 % 16S rRNA gene sequence similarity and mean level of DNA–DNA relatedness of 12 %. In phylogenetic trees based on 16S rRNA gene sequences, strains 3B-2T and 10AOT, together with several uncultured bacterial clones, formed independent lineages within the evolutionary radiation encompassed by the phylum Bacteroidetes. Strains 3B-2T and 10AOT contained MK-7 as the predominant menaquinone and iso-C15 : 0 and C16 : 1ω5c as the major fatty acids. The DNA G+C contents of strains 3B-2T and 10AOT were 42.8 and 44.6 mol%, respectively. Strains 3B-2T and 10AOT exhibited very low levels of 16S rRNA gene sequence similarity (<85.0 %) to the type strains of recognized bacterial species. These data were sufficient to support the proposal that the novel strains should be differentiated from previously known genera of the phylum Bacteroidetes. On the basis of the data presented, we suggest that strains 3B-2T and 10AOT represent two distinct novel species of a new genus, for which the names Ohtaekwangia koreensis gen. nov., sp. nov. (the type species; type strain 3B-2T  = KCTC 23018T  = CCUG 58939T) and Ohtaekwangia kribbensis sp. nov. (type strain 10AOT  = KCTC 23019T  = CCUG 58938T) are proposed.

Strain 15-4T, a Gram-stain-negative, rod-shaped, non-motile bacterial strain that produced flexirubin-type pigments, was isolated from Tibet Province, China, and characterized by using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolate belonged to the family Chitinophagaceae, phylum ‘Bacteroidetes’, and was related to members of the genus Niabella, with sequence similarities ranging from 94.1 to 96.4 %. Strain 15-4T contained MK-7 as the predominant menaquinone and its DNA G+C content was 46.9 mol%. The major fatty acids of strain 15-4T were iso-C15 : 0 (41.3 %), iso-C15 : 1 G (14.9 %), iso-C17 : 0 3-OH (13.2 %) and summed feature 3 (comprising C16 : 1ω7c and/or iso-C15 : 0 2-OH, 16.0 %). These chemotaxonomic data supported the affiliation of strain 15-4T to the genus Niabella. However, a number of physiological and biochemical features enabled the isolate to be differentiated phenotypically from recognized speciesof the genus Niabella. On the basis of the evidence presented, it is proposed that strain 15-4T represents a novel species, Niabella tibetensis sp. nov.; the type strain is 15-4T ( = CCTCC AB 209167T = NRRL B-59394T). On the basis of these data, an emended description of the genus Niabella is also proposed.

A novel bacterial strain designated B3AT, isolated from shrimp paste, was investigated by a polyphasic taxonomic approach. Cells stained Gram-positive and were aerobic, non-pigmented, sporulating and rod-shaped with a polar flagellum. 16S rRNA gene sequence analysis indicated that strain B3AT belonged to the class Bacilli and was a member of the family Bacillaceae. Strain B3AT shared low levels of 16S rRNA gene sequence similarity (<94.0 %) with members of other genera in the family Bacillaceae and was most closely related to Halalkalibacillus halophilus BH2T (93.8 % sequence similarity). The isolate was able to grow at 20–45 °C, with 0.5–15 % NaCl and at pH 6–9. Menaquinone with seven isoprene units (MK-7) was the major respiratory quinone and 16 : 0 iso, 15 : 0 anteiso and 14 : 0 iso were the major fatty acids. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The characteristic diamino acid of the peptidoglycan was meso-diaminopimelic acid. The DNA G+C content was 45.3 mol%. On the basis of 16S rRNA gene sequence analysis in combination with chemotaxonomic and physiological data, strain B3AT represents a novel genus and species in the family Bacillaceae for which the name Allobacillus halotolerans gen. nov., sp. nov. is proposed. The type strain of Allobacillus halotolerans is B3AT ( = BCRC 17939T  = LMG 24826T).