- Volume 63, Issue Pt_10, 2013

A Gram-stain-positive, aerobic organism, isolated from a blood sample from a 51-year-old woman, was studied for its taxonomic position. Based on 16S rRNA gene sequence similarity comparisons, strain CCUG 53762T was grouped into the genus Alicyclobacillus , most closely related to the type strain of Alicyclobacillus pohliae (94.7 %). The 16S rRNA gene sequence similarity to other species of the genus Alicyclobacillus was ≤91 % and similarity to species of the genus Tumebacillus was 91.3–93 %. The occurrence of menaquinone MK-7 as the major respiratory quinone, meso-diaminopimelic acid as the diagnostic diamino acid of the cell wall and the fatty acid profile supported the allocation of the strain to the genus Alicyclobacillus . Major fatty acids were iso- and anteiso-branched fatty acids. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and three unknown phospholipids. The absence of the iso-branched fatty acids iso-C16 : 0 and iso-C17 : 0 allowed differentiation of strain CCUG 53762T from A. pohliae CIP 109385T. In addition, the results of physiological and biochemical tests also allowed phenotypic differentiation of strain CCUG 53762T from this most closely related species. The G+C content of the DNA was 47 mol%. Strain CCUG 53762T therefore represents a novel species of the genus Alicyclobacillus , for which we propose the name Alicyclobacillus consociatus sp. nov., with CCUG 53762T ( = CCM 8439T) as the type strain.

Two Gram-stain-negative, facultatively anaerobic and endospore-forming rod-shaped bacterial strains, THMBG22T and R24, were isolated from decomposing algal scum. Phylogenetic analysis of 16S rRNA gene sequences showed that the two strains were closely related to each other (99.7 % similarity) and that they were also closely related to Paenibacillus sacheonensis DSM 23054T (97–97.1 %) and Paenibacillus phyllosphaerae DSM 17399T (96.1–96.4 %). This affiliation was also supported by rpoB-based phylogenetic analyses. Growth was observed at 20–40 °C (optimum, 30–37 °C) and at pH 5.0–9.0 (optimum, pH 6.0–7.0). The cells contained MK-7 as the sole respiratory quinone and anteiso-C15 : 0 as the major cellular fatty acid. Their cellular polar lipids were composed of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and 12 unidentified polar lipids. The diamino acid of their cell-wall peptidoglycan was meso-diaminopimelic acid. The DNA–DNA hybridization value between THMBG22T and R24 was 84 %, and DNA–DNA relatedness to the most closely related species with a validly published name ( P. sacheonensis ) was 35–37 %. These results supported the assignment of the new isolates to the genus Paenibacillus and also distinguished them from the previously described species of the genus Paenibacillus . Hence, it is proposed that strains THMBG22T and R24 represent a novel species of the genus Paenibacillus , with the name Paenibacillus taihuensis sp. nov. The type strain is THMBG22T ( = CGMCC 1.10966T = NBRC 108766T).

A Gram-stain-positive, spore-forming, rod-shaped, motile, strictly aerobic bacterial strain, designated CAU 1051T, was isolated from a sand dune and its taxonomic position was investigated using a polyphasic approach. Strain CAU 1051T grew optimally at pH 5.0 and 30 °C. NaCl was not required for growth but up to 10.0 % (w/v) NaCl was tolerated. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CAU 1051T formed a distinct lineage within the genus Oceanobacillus and was most closely related to Oceanobacillus profundus CL-MP28T, Oceanobacillus caeni S-11T, and Oceanobacillus picturae LMG 19492T (96.8 %, 95.6 % and 95.3 % similarity, respectively). DNA–DNA reassociation analysis showed that strain CAU 1051T displayed 28.2±0.7 % relatedness to O. profundus KCTC 13625T. Strain CAU 1051T contained MK-7 as the only isoprenoid quinone and anteiso-C15 : 0 as the major fatty acid. The cell wall peptidoglycan of strain CAU 1051T contained meso-diaminopimelic acid. The polar lipids were composed of diphosphatidylglycerol, phosphatidylglycerol, six unidentified phospholipids, an unidentified glycolipid, and six unidentified polar lipids. The major whole-cell sugars were glucose and ribose. The DNA G+C content was 36.3 mol%. On the basis of phenotypic data and phylogenetic inference, strain CAU 1051T represents a novel species of the genus Oceanobacillus for which the name Oceanobacillus chungangensis sp. nov. is proposed. The type strain is CAU 1051T ( = KCTC 33035T = CCUG 63270T).

A novel thermotolerant bacterium, designated SgZ-8T, was isolated from a compost sample. Cells were non-motile, endospore-forming, Gram-staining positive, oxidase-negative and catalase-positive. The isolate was able to grow at 20–65 °C (optimum 50 °C) and pH 6.0–9.0 (optimum 6.5–7.0), and tolerate up to 9.0 % NaCl (w/v) under aerobic conditions. Anaerobic growth occurred with anthraquinone-2,6-disulphonate (AQDS), fumarate and NO3 - as electron acceptors. Phylogenetic analysis based on the16S rRNA and gyrB genes grouped strain SgZ-8T into the genus Bacillus , with the highest similarity to Bacillus badius JCM 12228T (96.2 % for 16S rRNA gene sequence and 83.5 % for gyrB gene sequence) among all recognized species in the genus Bacillus . The G+C content of the genomic DNA was 49.3 mol%. The major isoprenoid quinone was menaquinone 7 (MK-7) and the polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified phospholipid. The major cellular fatty acid was iso-C16 : 0. On the basis of its phenotypic and phylogenetic properties, chemotaxonomic analysis and the results of physiological and biochemical tests, strain SgZ-8T ( = CCTCC AB 2012108T = KACC 16706T) was designated the type strain of a novel species of the genus Bacillus , for which the name Bacillus thermotolerans sp. nov. is proposed.

A novel anaerobic bacterium that could ferment amino acids and organic acids was isolated from an anaerobic, propionate-oxidizing enrichment culture originating from soil of a rice field in Japan. Cells of the isolate, designated strain 4F6ET, were Gram-staining-negative, oxidase- and catalase-negative, non-spore-forming, vibrio-shaped, motile rods (0.8×2.0–2.5 µm) with two or three lateral flagella. Growth occurred at 20–42 °C (optimum at 37–40 °C), at pH 6.4–8.4 (optimum at pH 7.3) and at 0–1.5 % (w/v) NaCl (optimum at 0–0.5 %). Good growth occurred on glycine, serine, cysteine, pyruvate and citrate, whereas poor growth was observed on threonine, glutamine, l-malate, α-ketoglutarate, peptone and Casamino acids. In co-culture with the hydrogen-utilizing methanogen Methanobacterium formicicum JCM 10132T, strain 4F6ET oxidized alanine, valine, leucine, isoleucine, methionine, aspartate, glutamate, histidine, asparagine and fumarate. Yeast extract was required for growth. The G+C content of genomic DNA was 61.9 mol%. A phylogenetic analysis based on comparison of the 16S rRNA gene sequence showed that the type strains of Fretibacterium fastidiosum , Aminobacterium colombiense and Aminobacterium mobile , members of the family Synergistaceae , were the closest relatives of strain 4F6ET, with low sequence similarities (89.3, 89.5 and 86.2 %, respectively). Strain 4F6ET contained iso-C13 : 0 (24.43 %), iso-C15 : 0 (16.47 %) and C19 : 1ω11c/C19 : 1ω9c (16.32 %) as the major fatty acids, which differed from those of F. fastidiosum , Aminobacterium colombiense and Aminobacterium mobile . On the basis of phenotypic, chemotaxonomic and phylogenetic differences between strain 4F6ET and the type strains of F. fastidiosum and Aminobacterium species, we propose that strain 4F6ET represents a novel genus and species, Aminivibrio pyruvatiphilus gen. nov., sp. nov. The type strain of Aminivibrio pyruvatiphilus is strain 4F6ET ( = JCM 18417T = DSM 25964T).

Three Gram-stain-positive, obligately anaerobic, non-motile, non-spore-forming, spindle-shaped bacterial strains (HT03-11T, KO-38 and TT-111), isolated from human faeces were characterized by phenotypic and molecular taxonomic methods. Comparative 16S rRNA gene sequencing showed that the strains were highly related to each other genetically (displaying >99 % sequence similarity) and represented a previously unknown subline within the Blautia coccoides rRNA group of organisms (cluster XIVa). The closest phylogenetic neighbours of strain HT03-11T were Clostridium bolteae WAL 16351T (93.7 % 16S rRNA gene sequence similarity) and Clostridium saccharolyticum WM1T (93.7 % similarity). All isolates produced lactic acid, formic acid, acetic acid and succinic acid as fermentation end products from glucose. Their chemotaxonomic properties included lysine as the cell wall diamino acid and C16 : 0, C18 : 1ω7c DMA and C16 : 0 DMA as the major fatty acids. The G+C contents of the genomic DNA were 46.9–47.2 mol% (HPLC). Several phenotypic and chemotaxonomic characteristics could be readily used to differentiate the isolates from phylogenetically related clostridia. Therefore, strains HT03-11T, KO-38 and TT-111 represent a novel species in a new genus of the family Lachnospiraceae , for which the name Fusicatenibacter saccharivorans gen. nov., sp. nov. is proposed. The type strain of the type species is HT03-11T ( = YIT 12554T = JCM 18507T = DSM 26062T).

Lactobacillus delbrueckii is divided into five subspecies based on phenotypic and genotypic differences. A novel isolate, designated ZN7a-9T, was isolated from malted sorghum wort used for making an alcoholic beverage (dolo) in Burkina Faso. The results of 16S rRNA gene sequencing, DNA–DNA hybridization and peptidoglycan cell-wall structure type analyses indicated that it belongs to the species L. delbrueckii . The genome sequence of isolate ZN7a-9T was determined by Illumina-based sequencing. Multilocus sequence typing (MLST) and split-decomposition analyses were performed on seven concatenated housekeeping genes obtained from the genome sequence of strain ZN7a-9T together with 41 additional L. delbrueckii strains. The results of the MLST and split-decomposition analyses could not establish the exact subspecies of L. delbrueckii represented by strain ZN7a-9T as it clustered with L. delbrueckii strains unassigned to any of the recognized subspecies of L. delbrueckii . Strain ZN7a-9T additionally differed from the recognized type strains of the subspecies of L. delbrueckii with respect to its carbohydrate fermentation profile. In conclusion, the cumulative results indicate that strain ZN7a-9T represents a novel subspecies of L. delbrueckii closely related to Lactobacillus delbrueckii subsp. lactis and Lactobacillus delbrueckii subsp. delbrueckii for which the name Lactobacillus delbrueckii subsp. jakobsenii subsp. nov. is proposed. The type strain is ZN7a-9T = DSM 26046T = LMG 27067T.

Two bacterial strains, designated IWT246T and IWT248, were isolated from orchardgrass (Dactylis glomerata L.) silage from Iwate prefecture, Japan, and examined for a taxonomic study. Both organisms were rod-shaped, Gram-stain-positive, catalase-negative, facultatively anaerobic and homofermentative. The cell wall did not contain meso-diaminopimelic acid and the major fatty acids were C18 : 1ω9c and C19 cyclo 9,10/:1. Comparative analyses of 16S rRNA, pheS and rpoA gene sequences revealed that these strains were novel and belonged to the genus Lactobacillus . Based on 16S rRNA gene sequence similarity, the isolates were most closely related to the type strains of the following members of the genus Lactobacillus : Lactobacillus coryniformis subsp. coryniformis (96.7 % similarity), L. coryniformis subsp. torquens (96.6 %), L. bifermentans (95.5 %) and L. rennini (94.1 %). However, the 16S rRNA gene sequences of both IWT246T and IWT248 were 99.7 % similar to that of ‘ Lactobacillus backi’ JCM 18665; this name has not been validly published. Genotypic, phenotypic and chemotaxonomic analyses confirmed that these novel strains occupy a unique taxonomic position. DNA–DNA hybridization experiments demonstrated genotypic separation of the novel isolates from related Lactobacillus species. The name Lactobacillus iwatensis sp. nov. is proposed for the novel isolates, with strain IWT246T ( = JCM 18838T = DSM 26942T) as the type strain. Our results also suggest that ‘L. backi’ does represent a novel Lactobacillus species. The cells did not contain meso-diaminopimelic acid in their cell-wall peptidoglycan and the major fatty acids were C16 : 0, C19 cyclo 9,10/:1 and summed feature 10 (one or more of C18 : 1ω11c, C18 : 1ω9t, C18 : 1ω6t and unknown ECL 17.834). We therefore propose the corrected name Lactobacillus backii sp. nov., with the type strain JCM 18665T ( = LMG 23555T = DSM 18080T = L1062T).

A Gram-stain-negative, aerobic, catalase- and oxidase-positive, non-flagellated, rod-shaped bacterium, designated strain P-50-3T, was isolated from seawater of the Pacific. The strain grew at 10–40 °C (optimum at 30 °C) and with 0–12 % (w/v, optimum 2 %) NaCl. It reduced nitrate to nitrite but did not hydrolyse gelatin, starch or Tween 80. Analysis of 16S rRNA gene sequences showed that strain P-50-3T clustered tightly with the genus Albimonas and shared the highest 16S rRNA gene sequence similarity (94.3 %) with the type strain of Albimonas donghaensis . The major respiratory quinone was Q-10 and the major cellular fatty acids were C18 : 1ω7c, C18 : 0, 11-methyl C18 : 1ω7c and C16 : 0. Polar lipids included phosphatidylglycerol (PG), phosphatidylcholine (PC), two unidentified aminolipids and an unidentified lipid. The genomic DNA G+C content of strain P-50-3T was 69.0 mol%. On the basis of the data obtained in this polyphasic study, strain P-50-3T represents a novel species within the genus Albimonas , for which the name Albimonas pacifica sp. nov. is proposed. The type strain of Albimonas pacifica is P-50-3T ( = KACC 16527T = CGMCC 1.11030T). An emended description of the genus Albimonas Lim et al. 2008 is also proposed.

Four strains (08HL01032T, 09HG994, 10HP82-6 and 10HL1960) were isolated from water of air-conditioning systems of various cooling towers in Guangzhou city, China. Cells were Gram-stain-negative coccobacilli without flagella, catalase-positive and oxidase-negative, showing no reduction of nitrate, no hydrolysis of urea and no production of H2S. Growth was characteristically enhanced in the presence of l-cysteine, which was consistent with the properties of members of the genus Francisella . The quinone system was composed of ubiquinone Q-8 with minor amounts of Q-9. The polar lipid profile consisted of the predominant lipids phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, two unidentified phospholipids (PL2, PL3), an unidentified aminophospholipid and an unidentified glycolipid (GL2). The polyamine pattern consisted of the major compounds spermidine, cadaverine and spermine. The major cellular fatty acids were C10 : 0, C14 : 0, C16 : 0, C18 : 1ω9c and C18 : 1 3-OH. A draft whole-genome sequence of the proposed type strain 08HL01032T was generated. Comparative sequence analysis of the complete 16S and 23S rRNA genes confirmed affiliation to the genus Francisella , with 95 % sequence identity to the closest relatives in the database, the type strains of Francisella philomiragia and Francisella noatunensis subsp. orientalis . Full-length deduced amino acid sequences of various housekeeping genes, recA, gyrB, groEL, dnaK, rpoA, rpoB, rpoD, rpoH, fopA and sdhA, exhibited similarities of 67–92 % to strains of other species of the genus Francisella . Strains 08HL01032T, 09HG994, 10HP82-6 and 10HL1960 exhibited highly similar pan-genome PCR profiles. Both the phenotypic and molecular data support the conclusion that the four strains belong to the genus Francisella but exhibit considerable divergence from all recognized Francisella species. Therefore, we propose the name Francisella guangzhouensis sp. nov., with the type strain 08HL01032T ( = CCUG 60119T = NCTC 13503T).

The gut of the Western honeybee, Apis mellifera, is colonized by a characteristic set of bacteria. Two distinct gammaproteobacteria are consistent members of this unique microbial community, and one has recently been described in a new genus and species with the name Gilliamella apicola . Here, we present the isolation and characterization of PEB0191T, a strain belonging to the second gammaproteobacterial species present in the honeybee gut microbiota, formerly referred to as ‘Gammaproteobacterium-2’. Cells of strain PEB0191T were mesophilic and had a mean length of around 2 µm, and optimal growth was achieved under anaerobic conditions. Growth was not obtained under aerobic conditions and was reduced in a microaerophilic environment. Based on 16S rRNA gene sequence analysis, strain PEB0191T belongs to the family Orbaceae , and its closest relatives, with around 95 % sequence similarity, are species of the genera Orbus and Gilliamella . Phylogenetic analyses suggest that PEB0191T is more closely related to the genus Orbus than to the genus Gilliamella . In accordance with its evolutionary relationship, further similarities between strain PEB0191T and other members of the family Orbaceae were revealed based on the respiratory quinone type (ubiquinone 8), the fatty acid profile and the DNA G+C content. Interestingly, like strains of the genus Gilliamella , PEB0191T exhibited a high level of resistance to oxytetracycline. The similar levels of sequence divergence from the genera Gilliamella and Orbus and its uncertain phylogenetic position within the family Orbaceae indicate that strain PEB0191T represents a novel species of a new genus, with the proposed name Frischella perrara gen. nov., sp. nov. The type strain of Frischella perrara is PEB0191T ( = NCIMB 14821T = ATCC BAA-2450T).

An alginate lyase-producing bacterium, designated AlyHP32T, was isolated from the gut of sea urchin (Hemicentrotus pulcherrimus) obtained from the South Sea, Republic of Korea. Cells of strain AlyHP32T were Gram-reaction-negative and motile with a single polar flagellum. The strain grew with 1–6 % (w/v) NaCl (optimum 2–4 %) and at 4–30 °C (optimum 15–25 °C). Phylogenetic analysis based on sequences of the 16S rRNA gene and five housekeeping genes (atpA, pyrH, recA, rpoA and rpoD) revealed that strain AlyHP32T belonged to the genus Vibrio and formed a compact clade with the Vibrio splendidus group. However, DNA–DNA hybridization and fingerprints using the repetitive primers BOX and REP indicated that strain AlyHP32T was distinct from closely related species of the genus Vibrio . The major fatty acids were summed feature 3 (C16:1ω7c and/or C16:1ω6c) and C16:0. The DNA G+C content was 44.1 mol%. The predominant quinone was ubiquinone Q-8. Based on genotypic, phenotypic and DNA–DNA hybridization analysis, strain AlyHP32T represents a novel species of the genus Vibrio ; the name Vibrio hemicentroti sp. nov. (type strain AlyHP32T = KCTC 32085T = DSM 26178T) is proposed for this novel taxon.

A novel Gram-stain-negative, aerobic, rod-shaped strain designated PG04T was isolated from the rhizosphere of watermelon plants cultivated in Beijing, China. A polyphasic taxonomic study was performed on the new isolate. On the basis of 16S rRNA gene sequence similarity studies, isolate PG04T belonged clearly to the genus Hansschlegelia and was most closely related to Hansschlegelia zhihuaiae (97.3 % similarity to the type strain). The predominant respiratory quinone was ubiquinone 10 (Q-10) and the polar lipid profile was composed of the major lipids diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidylcholine. The major fatty acids were C18 : 1ω7c (41.3 %), C19 : 0 cyclo ω8c (30.6 %) and C16 : 0 (19.1 %). The G+C content of the DNA was about 64.4 mol%. DNA–DNA hybridization experiments showed 34.4 % relatedness between strain PG04T and H. zhihuaiae DSM 18984T. The results of physiological and biochemical tests and differences in fatty acid profiles allowed clear phenotypic differentiation of strain PG04T from the most closely related species in the genus, H. zhihuaiae . Strain PG04T therefore represents a novel species within the genus Hansschlegelia , for which the name Hansschlegelia beijingensis sp. nov. is proposed, with the type strain PG04T ( = DSM 25481T = ACCC 05759T).

A novel nitrogen-fixing strain, designated DQS-4T, was isolated from oil-contaminated soil in Taiwan and was characterized using a polyphasic taxonomic approach. Cells of strain DQS-4T stained Gram-negative, contained poly-β-hydroxybutyrate granules and were motile rods, surrounded by a thin capsule. Cells displayed a strictly aerobic type of metabolism and fixed nitrogen microaerobically. Growth occurred at 10–45 °C (optimum, 35–40 °C), at pH 7.0–8.0 (optimum, pH 7.0) and with 0–2 % NaCl (optimum, 0.5–1 %). Phylogenetic analyses based on 16S rRNA gene sequences showed that strain DQS-4T belonged to the genus Azoarcus , and its closest neighbours were Azoarcus indigens VB32T and Azoarcus communis SWub3T, with sequence similarities of 97.4 and 96.4 %, respectively. The major cellular fatty acids of strain DQS-4T were summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c), C16 : 0 and C18 : 1ω7c. The major cellular hydroxy fatty acid was C10 : 0 3-OH. The DNA G+C content was 64.5 mol%. The polar lipid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and several uncharacterized aminophospholipids and phospholipids. The mean level of DNA–DNA relatedness between strain DQS-4T and A. indigens LMG 9092T was 27.4 %. On the basis of the genotypic and phenotypic data, strain DQS-4T represents a novel species in the genus Azoarcus , for which the name Azoarcus olearius sp. nov. is proposed. The type strain is DQS-4T ( = BCRC 80407T = KCTC 23918T = LMG 26893T).

An aerobic, Gram-stain-negative, spiral or rod-shaped, non-spore-forming, diazotrophic bacterium (strain CC-LY743T) was isolated from a fermentative tank in Taiwan. Strain CC-LY743T was able to grow at 20–37 °C and pH 6.0–8.0 and tolerated up to 3.0 % (w/v) NaCl. It was positive for nitrogen fixation, with activity of 10.6 nmol ethylene h−1. 16S rRNA gene sequence analysis of strain CC-LY743T showed highest similarity to Azospirillum picis DSM 19922T (96.1 %), Azospirillum oryzae JCM 21588T (96.0 %) and Azospirillum rugosum DSM 19657T (96.0 %) and lower similarity (<96.0 %) to all other Azospirillum species. Highest nifH gene sequence similarities were obtained with Azospirillum brasilense BCRC 12270T (92.0 %), Azospirillum formosense BCRC 80273T (92.3 %) and A. rugosum DSM 19657T (91.8 %). It was positive in the rapid identification by a genus-specific primer set. The predominant quinone system was ubiquinone 10 (Q-10) and the DNA G+C content was 69.6±0.1 mol%. The major fatty acids found in strain CC-LY743T were n-C16 : 0, C19 : 0 cyclo ω8c, C14 : 0 3-OH/C16 : 1 iso I, C16 : 1ω7c/C16 : 1ω6c and C18 : 1ω7c/C18 : 1ω6c. Based on its phylogenetic, phenotypic and chemotaxonomic features, strain CC-LY743T is considered to represent a novel species within the genus Azospirillum for which the name Azospirillum fermentarium sp. nov. is proposed. The type strain is CC-LY743T ( = BCRC 80505T = JCM 18688T = LMG 27264T).

A budding prosthecate bacterial strain, designated NL23T, was isolated from a methanol-fed denitrification system treating seawater at the Montreal Biodome, Canada. Phylogenetic analysis based on 16S rRNA (rRNA) gene sequences showed that the strain was affiliated with the genus Hyphomicrobium of the Alphaproteobacteria and was most closely related to Hyphomicrobium zavarzinii with 99.4 % sequence similarity. Despite this high level of 16S rRNA gene sequence similarity, DNA–DNA hybridization assays showed that strain NL23T was only distantly related to H. zavarzinii ZV-622T (12 %). Strain NL23T grew aerobically, but also had the capacity to grow under denitrifying conditions in the presence of nitrate without nitrite accumulation. Growth occurred at pH 7.0–9.5, with 0–1 % NaCl and at temperatures of 15–35 °C. Major fatty acids were C18 : 1ω7c or ω6c (84.6 %) and C18 : 0 (8.5 %), and major quinones were Q8 (5 %) and Q9 (95 %). The complete genome of the strain was sequenced and showed a DNA G+C content of 63.8 mol%. Genome analysis predicted open reading frames (ORF) encoding the key enzymes of the serine pathway as well as enzymes involved in methylotrophy. Also, ORF encoding a periplasmic nitrate reductase (Nap), a nitrite reductase (Nir), a nitric oxide reductase (Nor) and a nitrous oxide reductase (Nos) were identified. Our results support that strain NL23T represents a novel species within the genus Hyphomicrobium , for which the name Hyphomicrobium nitrativorans sp. nov. is proposed. The type strain is NL23T ( = ATCC BAA-2476T = LMG 27277T).

A Gram-reaction-negative, aerobic, motile by one polar flagellum, yellow-pigmented, rod-shaped bacterium, designated strain THG-B117T, was isolated from soil of a cornus fruit field of Hoengseong province in South Korea and its taxonomic position was investigated by a polyphasic study. Strain THG-B117T grew well at 25–30 °C and at pH 6.0–8.0 in the absence of NaCl on nutrient agar. On the basis of 16S rRNA gene sequence similarity, strain THG-B117T was shown to belong to the family Xanthomonadaceae and be related to Dyella japonica XD53T (98.7 % similarity), Dyella terrae JS14-6T (98.0 %), Dyella koreensis BB4T (96.9 %), Dyella soli JS12-10T (96.9 %) and Dyella thiooxydans ATSB10T (96.7 %). DNA–DNA hybridization experiments showed that DNA relatedness between strain THG-B117T and its phylogenetically closest neighbours was below 45.1 %. The G+C content of the genomic DNA of strain THG-B117T was 64.8 mol%. The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, unidentified aminolipids, unidentified aminophospholipids and unidentified phospholipids. Phenotypic and chemotaxonomic data (major ubiquinone was Q-8, and major fatty acids were iso-C15 : 0, iso-C16 : 0, iso-C17 : 0 and iso-C17 : 1ω9c) supported the affiliation of strain THG-B117T with the genus Dyella . The results of physiological and biochemical tests suggested that strain THG-B117T was different genotypically and phenotypically from recognized species of the genus Dyella , and represents a novel species of this genus. The name Dyella kyungheensis sp. nov. is proposed, with the type strain THG-B117T ( = KACC 16981T = JCM 18747T).

A Gram-stain-negative, rod-shaped bacterium was isolated and designated strain L6-8T during a study of endophytic bacterial communities in lesser duckweed (Lemna aequinoctialis). Cells of strain L6-8T were motile with peritrichous flagella. The analysis of the nearly complete 16S rRNA gene sequence indicated that strain L6-8T was phylogenetically related to species of the genus Rhizobium . Its closest relatives were Rhizobium borbori DN316T (97.6 %), Rhizobium oryzae Alt 505T (97.3 %) and Rhizobium pseudoryzae J3-A127T (97.0 %). The sequence similarity analysis of housekeeping genes recA, glnII, atpD and gyrB showed low levels of sequence similarity (<91.5 %) between strain L6-8T and other species of the genus Rhizobium with validly published names. The pH range for growth was 4.0–9.0 (optimum 6.0–7.0), and the temperature range for growth was 20–45 °C (optimum 30 °C). Strain L6-8T tolerated NaCl up to 2 % (w/v) (optimum 1 % NaCl). The predominant components of cellular fatty acids were C19 : 0 cyclo ω8c (31.32 %), summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c; 25.39 %) and C16 : 0 (12.03 %). The DNA G+C content of strain L6-8T was 60.4 mol% (T m). nodC and nifH were not amplified in strain L6-8T. DNA–DNA relatedness between strain L6-8T and R. borbori DN316T, R. oryzae Alt505T and R. pseudoryzae J3-A127T was between 11.2 and 18.3 %. Based on the sequence similarity analyses, phenotypic, biochemical and physiological characteristics and DNA–DNA hybridization, strain L6-8T could be readily distinguished from its closest relatives and represents a novel species of the genus Rhizobium , for which the name Rhizobium paknamense sp. nov. is proposed. The type strain is L6-8T ( = NBRC 109338T = BCC 55142T).

An obligately aerobic, chemoheterotrophic, mesophilic prosthecate bacterium, designated strain CGM1-3ENT, was isolated from the enrichment cultures of forest soil from Cheonggyesan Mountain, Republic of Korea. Cells were Gram-reaction-negative, motile rods (1.3–2.4 µm long by 0.30–0.75 µm wide) with single flagella. The strain grew at 10–37 °C (optimum 25–30 °C) and at pH 4.5–9.5 (optimum 5.0–7.0). The major cellular fatty acids were C16 : 0, C18 : 1ω7c 11-methyl, C12 : 1 3-OH and summed feature 8 (comprising C18 : 1ω7c/C18 : 1ω6c). The genomic DNA G+C content of strain CGM1-3ENT was 63.7 mol%. The closest phylogenetic neighbour to strain CGM1-3ENT was identified as Asticcacaulis biprosthecium DSM 4723T (97.2 % 16S rRNA gene sequence similarity) and the DNA–DNA hybridization value between strain CGM1-3ENT and A. biprosthecium DSM 4723T was less than 24.5 %. Strain CGM1-3ENT used d-glucose, d-fructose, sucrose, maltose, trehalose, d-mannose, d-mannitol, d-sorbitol, d-galactose, cellobiose, lactose, raffinose, fumarate, pyruvate, dl-alanine and glycerol as carbon sources. Based on data from the present polyphasic study, the forest soil isolate CGM1-3ENT is considered to represent a novel species of the genus Asticcacaulis , for which the name Asticcacaulis solisilvae sp. nov. is proposed. The type strain is CGM1-3ENT ( = AIM0088T = KCTC 32102T = JCM 18544T).