- Volume 67, Issue 8, 2017

A Gram-stain-negative, motile, rod-shaped bacterium designated OCN003T was cultivated from mucus taken from a diseased colony of the coral Montipora capitata in Kāne‘ohe Bay, O‘ahu, Hawai‘i. Colonies of OCN003T were pale yellow, 1–3 mm in diameter, convex, smooth and entire. The strain was heterotrophic, strictly aerobic and strictly halophilic. Cells of OCN003T produced buds on peritrichous prosthecae. Growth occurred within the pH range of 5.5 to 10, and the temperature range of 14 to 39 °C. Major fatty acids were 16 : 1ω7c, 16 : 0, 18 : 1ω7c, 17 : 1ω8c, 12 : 0 3-OH and 17 : 0. Phylogenetic analysis of 1399 nucleotides of the 16S rRNA gene nucleotide sequence and a multi-locus sequence analysis of three genes placed OCN003T in the genus Pseudoalteromonas and indicated that the nearest relatives described are Pseudoalteromonas spongiae , P. luteoviolacea , P. ruthenica and P. phenolica (97–99 % sequence identity). The DNA G+C content of the strain’s genome was 40.0 mol%. Based on in silico DNA–DNA hybridization and phenotypic differences from related type strains, we propose that OCN003T represents the type strain of a novel species in the genus Pseudoalteromonas , proposed as Pseudoalteromonas piratica sp. nov. OCN003T (=CCOS1042T=CIP 111189T). An emended description of the genus Pseudoalteromonas is presented.

Brine samples from Kulunda Steppe soda lakes (Altai, Russia) were inoculated into a hypersaline alkaline mineral medium with β-keratin (chicken feather) as a substrate. The micro-organisms dominating the enrichment culture were isolated by limiting serial dilution on the same medium with casein as a substrate. The cells of strain BSker1T were motile, curved rods. The strain was an obligately aerobic heterotroph utilizing proteins and peptides as growth substrates. The isolate was an obligate alkaliphile with a pH range for growth from pH 8.5 to 10.25 (optimum at pH 9.5), and it was extremely salt tolerant, growing with between 1 and 4.5 M total Na+ (optimally at 2–2.5 M). BSker1T had a unique composition of polar lipid fatty acids, dominated by two C17 species. The membrane polar lipids included multiple unidentified phospholipids and two aminolipids. According to phylogenetic analysis of the 16S rRNA gene sequence, the isolate forms a novel branch within the family Ectothiorhodospiraceae (class Gammaproteobacteria ) with the highest sequence similarity to the members of this family being 91 %. On the basis of distinct phenotypic and genotypic properties, strain BSker1T (=JCM 31341T=UNIQEM U1008T) is proposed to be classified as a representative of a novel genus and species, Natronospira proteinivora gen. nov., sp. nov.

A moderately halophilic bacterium designated strain M6-53T was isolated from water of a pond from a marine saltern located in Huelva, south-west Spain. Cells of the strain were Gram-stain-negative, strictly aerobic, motile, slightly curved rods, able to grow in media containing 5–25 % (w/v) NaCl (optimal growth at 10 %, w/v), at temperatures from 20 to 40 °C (optimally at 37 °C) and at pH 6.5–9 (optimally at pH 7.0). Phylogenetic analysis based on 16S rRNA gene sequences placed the new isolate within the genus Marinobacter , with the type strains of the most closely related species being Marinobacter persicus IBRC-M 10445T (98.5 % similarity), Marinobacter oulmenensis Set74T (97.2 %) and Marinobacter hydrocarbonoclasticus ATCC 49840T (97.1 %). The major fatty acids present in strain M6-53T were C18 : 1ω9c (29.5 %), C16 : 0 (26.7 %), C12 : 0 3-OH (15.1 %), C18 : 0 (10.2 %) and C16 : ω9c (9.6 %). The G+C content of the genomic DNA for this strain was determined to be 56.4 mol%. The DNA–DNA hybridization values between strain M6-53T and M. persicus CECT 7991T, M. oulmenensis CECT 7499T and M. hydrocarbonoclasticus DSM 50418 were 8, 41 and 38 %, respectively. These values are lower than the accepted 70 % threshold and showed that the new isolate represented a different species within the genus Marinobacter . Phylogenetic analysis based on the 16S rRNA gene sequence and the phenotypic, genotypic and chemotaxonomic features of this new isolate support the placement of strain M6-53T as a representative of a novel species of the genus Marinobacter , for which we propose the name Marinobacter aquaticus sp. nov., with strain M6-53T (=CECT 9228T=LMG 30006T) as the type strain.

A Gram-negative, aerobic, polar-flagellated and rod-shaped, sometimes slightly curved bacterium, designated MA5T, was isolated from the gut of an abalone of the species Haliotis gigantea collected in Japan. Phylogenetic analyses based on 16S rRNA, gyrB, hsp60 and rpoB gene sequences placed strain MA5T in the genus Arcobacter in an independent phylogenetic line. Comparison of the 16S rRNA gene sequence of this strain with those of the type strains of the established Arcobacter species revealed A. nitrofigilis (95.1 %) as nearest neighbour. Strain MA5T grew optimally at 25 °C, pH 6.0 to 9.0 and in the presence of 2 to 5 % (w/v) NaCl under both aerobic and microaerobic conditions. The predominant fatty acids found were summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 1 ω7c), C12 : 0 3-OH and C18 : 1 ω7c. Menaquinone-6 (MK-6) and menaquinone-7 (MK-7) were found as the major respiratory quinones. The major polar lipids detected were phosphatidylethanolamine and phosphatidylglycerol. Strain MA5T could be differentiated phenotypically from the phylogenetic closest Arcobacter species by its ability to grow on 0.05 % safranin and 0.01 % 2,3,5-triphenyl tetrazolium chloride (TTC), but not on 0.5 % NaCl. The obtained DNA G+C content of strain MA5T was 27.9 mol%. Based on the phylogenetic, chemotaxonomic and phenotypic distinctiveness of MA5T, this strain is considered to represent a novel species of the genus Arcobacter , for which the name Arcobacter haliotis sp. nov. is proposed. The type strain is MA5T (=LMG 28652T=JCM 31147T).

A novel endophytic bacterium, strain J11-6T, was isolated from rice stems. Its taxonomic position was investigated using a polyphasic approach. The novel strain was Gram-staining-negative, facultatively anaerobic, motile and rod-shaped. Although the results of phylogenetic analysis based on 16S rRNA gene sequences indicated that J11-6T represented a member of the genus Rahnella , multilocus sequence analysis (MLSA) on the basis of concatenated partial atpD, gyrB, rpoB and infB gene sequences showed a clear distinction of J11-6T from the type strains of species of the genus Rahnella but indicated that it lay within the clade of the genus Serratia . The phylogenetically closest species were Serratia fonticola and Serratia aquatilis on the basis of the results of the MLSA phylogenetic analysis. The predominant cellular fatty acids were C16 : 1ω7c (38.7 %) and C16 : 0 (25.0 %). The DNA G+C content was 53.2 mol%. The DNA–DNA relatedness was 17.4 % between J11-6T and Rahnella aquatilis CIP 78.65T, and 29.2 % between J11-6T and S. fonticola LMG 7882T which indicates that this strain represents a novel species of the genus Serratia . Characterization by genotypic and phenotypic analysis indicated that J11-6T (=ACCC 19934T=KCTC 52529T) represents a novel species of the genus Serratia , for which the name Serratia oryzae sp. nov. is proposed.

A Gram-stain-negative, non-motile, aerobic, non-spore-forming, spherical bacterium (strain MIMtkB18T) was isolated from desert soil collected from part of a Mongolian Plateau, territory of Inner Mongolia, PR China. Cell growth could be observed at 20–45 °C (optimum at 40 °C), at a pH of 6–9 (optimum at pH 8.6) and in the presence of 0–1 % (w/v) NaCl (optimum 0 %). The genomic DNA G+C content was 69.6 mol%. 16S rRNA gene sequence analysis showed that strain MIMtkB18T was most closely related to Methylobrevis pamukkalensis PK2T (94.1 %), species of the genus Pleomorphomonas (93.4–94.0 %), and Hartmannibacter diazotrophicus E19T (93.9 %). The sole respiratory quinone was Q-10. The major fatty acids (>5 %) were C18 : 0 (5.7 %) and summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) (81.6 %). Polar lipids were mainly composed of phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, phosphatidylmonomethylethanolamine and unidentified phospholipids. Based on phenotypic, chemotaxonomic and phylogenetic characteristics, it is concluded that strain MIMtkB18T represents a novel genus and species, for which the name Mongoliimonas terrestris sp. nov. is proposed. The type strain is MIMtkB18T (=KCTC 42635T=MCCC 1K00571T).

A Gram-stain-negative, facultatively anaerobic, dark-yellow-pigmented bacterium, named SSR2A-4-2T, was isolated from coastal water in the East China Sea. Cells were ovoid or short rods with peritrichous flagella and contained carotenoid in addition to bacteriochlorophyll a pigment. A phylogenetic dendrogram based on 16S rRNA gene sequences showed that strain SSR2A-4-2T formed a distinct clade with members of the genus Blastomonas , with Blastomonas natatoria EY 4220T (=DSM 3183T) (similarity 95.6 %), Blastomonas ursincola KR-99T (=DSM 9006T) (95.5 %) and Blastomonas aquatica PE4-5T (=JCM 30179T) (94.8 %) as its closest phylogenetic relatives. Q-10 was the predominant respiratory quinone. The major fatty acids were summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c), summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c), C17 : 1ω6c and C18 : 1ω7c 11-methyl. The polar lipids contained diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, sphingoglycolipid, four unknown glycolipids and one unknown lipid. The DNA G+C content was 65.1 mol%. On the basis of the evidence presented in this study, strain SSR2A-4-2T represents a novel species of the genus Blastomonas , for which the name Blastomonas marina sp. nov. is proposed, with strain SSR2A-4-2T (=CGMCC 1.15297T=DSM 103453T) as the type strain.

A Gram-stain negative, rod-shaped, aerobic bacterial strain, BII-R7T, was isolated during a study targeting the culture-dependent microbial diversity occurring in bentonite formations from southern Spain. Comparative 16S rRNA gene sequence analysis showed that BII-R7T represented a member of the genus Stenotrophomonas (class Gammaproteobacteria ), and was related most closely to Stenotrophomonas rhizophila e-p10T (99.2 % sequence similarity), followed by Stenotrophomonas pavanii ICB 89T (98.5 %), Stenotrophomonas maltophilia IAM 12423T, Stenotrophomonas chelatiphaga LPM-5T and Stenotrophomonas tumulicola T5916-2-1bT (all 98.3 %). Pairwise sequence similarities to all other type strains of species of the genus Stenotrophomonas were below 98 %. Genome-based calculations (orthologous average nucleotide identity, original average nucleotide identity, genome-to-genome distance and DNA G+C percentage) indicated clearly that the isolate represents a novel species within this genus. Different phenotypic analyses, such as the detection of a quinone system composed of the major compound ubiquinone Q-8 and a fatty acid profile with iso-C15 : 0 and anteiso-C15 : 0 as major components, supported this finding at the same time as contributing to a comprehensive characterization of BII-R7T. Based on this polyphasic approach comprising phenotypic and genotypic/molecular characterization, BII-R7T can be differentiated clearly from its phylogenetic neighbours, establishing a novel species for which the name Stenotrophomonas bentonitica sp. nov. is proposed with BII-R7T as the type strain (=LMG 29893T=CECT 9180T=DSM 103927T).

A Gram-staining-negative, non-motile, aerobic bacterial strain, designated Y3L17T, was isolated from the saline–alkaline soil of a farmland, Hangjin Banner, Inner Mongolia, northern China. Y3L17T could grow at 15–45 °C (optimum 35 °C), pH 6.0–10.0 (optimum pH 8.0) and with 0–4 % (w/v) NaCl (optimum 0 %). The results of phylogenetic analysis based on the 16S rRNA gene and gyrB gene sequences revealed that Y3L17T tightly clustered with strains of members of the genus Arenimonas , sharing the highest 16S rRNA gene similarities with Arenimonas aestuarii S2-21T (99.5 %) and Arenimonas donghaensis HO3-R19T (98.2 %), and lower similarities (<97 %) with all the other type strains of species of this genus. However, Y3L17T shared only 92.62 % gyrB gene similarities with A. aestuarii S2-21T. The DNA–DNA hybridization values of Y3L17T with A. aestuarii S2-21T and A. donghaensis HO3-R19T were 20.1±2.5 and 18.2±3.2 %, respectively. Y3L17T contained phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, five unknown phospholipids and one unknown lipid as the major polar lipids. Ubiquinone-8 (Q-8) was the predominant respiratory quinone, while iso-C15 : 0, iso-C17 : 0ω9c and iso-C11 : 0 3-OH were the major cellular fatty acids. Its genomic DNA G+C content was 65.4 mol%. On the basis of its phenotypic, phylogenetic and genotypic characteristics, Y3L17T represents a novel species within the genus Arenimonas , for which the name Arenimonas soli sp. nov. is proposed, the type strain is Y3L17T (=CGMCC 1.15905T =KCTC 52420T).

A Gram-negative, motile, aerobic and coccoid rod-shaped bacterium, designated strain 2T4P-2-4T, was isolated from a piece of surface-sterilized bark of Aegiceras corniculatum collected from Cotai Ecological Zones in Macao, China, and tested by a polyphasic approach to clarify its taxonomic position. Strain 2T4P-2-4T grew optimally without NaCl at 28–30 °C, pH 7.0–8.0. The 16S rRNA gene sequence of strain 2T4P-2-4T had the highest similarity (96.2 %) to Aureimonas rubiginis CC-CFT034T. Phylogenetic analysis showed that the strain grouped with species of the genus Aureimonas . The predominant quinone system of strain 2T4P-2-4T was ubiquinone 10 (Q-10). The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, sulfoquinovosyldiacylglycerol, phosphatidylmethylethanolamine, two unidentified amino lipids, an unidentified aminophospholipid and five unidentified lipids. The predominant cellular fatty acid was C18 : 1ω7c (61.2 %). The DNA G+C content of strain 2T4P-2-4T was 69.8 mol%. Based on the phylogenetic, phenotypic and chemotaxonomic features, strain 2T4P-2-4T is a representative of a novel species of the genus Aureimonas , for which the name Aureimonas endophytica sp. nov. is proposed. The type strain of Aureimonas endophytica sp. nov. is 2T4P-2-4T (=KCTC 52217T=CGMCC 1.15367T).

A bacterial strain, designated gyp-1T, was isolated from a mangrove in Taiwan and characterized using the polyphasic taxonomic approach. Cells of gyp-1T were Gram-staining-negative, aerobic, poly-β-hydroxybutyrate-accumulating, non-motile, coccoid or short-rod-shaped and formed cream-coloured colonies. Growth occurred at 15–37 °C (optimum, 25–30 °C), at pH 5.5–7.0 (optimum, pH 6.0) and with 0–4 % NaCl (optimum, 1–2 %). Phylogenetic analyses based on 16S rRNA gene sequences indicated that gyp-1T represented a member of the genus Paracoccus and showed the highest levels of sequence similarity with respect to Paracoccus lutimaris HDM-25T (97.8 %) and Paracoccus aminovorans DM-82T (97.7 %). The major fatty acids (>10 %) of gyp-1T were C18 : 1ω7c and C16 : 0. The polar lipid profile consisted of phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, diphosphatidylglycerol, an unidentified glycolipid and two unidentified phospholipids. The major isoprenoid quinone was Q-10. The DNA G+C content was 64.6 mol%. The DNA–DNA hybridization value for gyp-1T with P. lutimaris HDM-25T and P. aminovorans DM-82T was less than 50 %. Differential phenotypic properties, together with the phylogenetic inference, demonstrate that gyp-1T should be classified as representing a novel species of the genus Paracoccus , for which the name Paracoccus mangrovi sp. nov. is presented. The type strain is gyp-1T (=BCRC 80920T=LMG 29172T=KCTC 42899T).

Two Gram-stain-negative, aerobic, non-motile, non-spore-forming and rod-shaped bacteria, designated strains T2T and T5, were isolated from a culture of Microcystis from Daejeon, Republic of Korea. Comparative 16S rRNA gene sequence studies placed the new isolates in the class Alphaproteobacteria and, notably, most closely related to Blastomonas aquatica PE 4-5T, Blastomonas natatoria DSM 3183T and Blastomonas ursincola KR-99T showing 99.4 %, 98.2 % and 97.9 % 16S rRNA gene sequence similarities, respectively. The two novel strains shared 100 % similarity with each other. The cells of strains T2T and T5 formed yellow colonies on R2A agar and contained Q-10 as the only ubiquinone, sphingoglycolipid, phosphatidylethanolamine, phosphatidylcholine, and phosphatidylglycerol as major polar lipids, and C17 : 1ω6c, summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), C17 : 1ω8c and C17 : 0 as the major fatty acids (>5 %). The DNA G+C content of the genomes was determined to be 64.2 mol% for strain T2T and 64.4 mol% for strain T5. The DNA–DNA hybridization values of strains T2T and T5 with B. aquatica PE 4-5T, B. natatoria DSM 3183T, and B. ursincola KR-99T were 19.7–42.4 %. Based on the combined genotypic and phenotypic data, we propose that strains T2T and T5 represent a novel species of the genus Blastomonas , for which the name Blastomonas fulvasp. nov. is proposed. The type strain is T2T (=KCTC 42354T=JCM 30467T).

Using a newly developed culture method for not yet cultured soil bacteria, three Gram-stain-negative, aerobic, non-spore-forming, motile, and rod-shaped bacteria (strain designated J18T, J11T and J9T) were isolated from forest soil at Kyonggi University, South Korea. Isolates were subjected to a taxonomic study by using a polyphasic approach. According to a phylogenetic tree based on 16S rRNA gene sequences, strains J18T, J11T and J9T belonged to the genus Massilia and clustered with Massilia haematophila CCUG 38318T (similarity range: 97.6~98.0 %). The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol, and the genomic DNA G+C contents of strains J18T, J11T and J9T were 63.4, 68.7 and 64.5 mol%, respectively. The major polyamines were putrescine and 2-hydroxyputescine, which were detected in all three strains. DNA–DNA between the three tested strains and the reference strains much lower than 70 %, the recommended threshold value for the delineation of genomic species. The predominant respiratory quinine was ubiquinone-8 (Q-8) and the major cellular fatty acids were Summed feature 3 (C16 : 1ω6c/C16  : 1ω7c) and C16 : 0. On the basis of phenotypic and genotypic data and DNA–DNA hybridization results, the three isolates are considered to represent three novel species of the genus Massilia , for which the names Massilia solisilvae sp. nov. for type strain J18T (=KEMB 9005-366T=JCM 31607T), Massilia terrae sp. nov. for type strain J11T (=KEMB 9005-360T=JCM 31606T) and Massilia agilis sp. nov. for type strain J9T (=KEMB 9005-359T=JCM 31605T) are proposed.

Two aerobic, Gram-stain-negative, gliding and yellow-pigmented bacteria, designated strains 03SUJ6T and WM95T were isolated from freshwater sediment of Juam reservoir and freshwater of Woopo wetland, Republic of Korea, respectively. Cells of the two strains are motile by gliding and catalase- and oxidase-positive. The 16S rRNA gene sequence similarity between 03SUJ6T and WM95T was 97.7 %, but their DNA–DNA relatedness was 55.1 %. A maximum-likelihood phylogenetic tree based on 16S rRNA gene sequences showed that 03SUJ6T and WM95T each form independent lineages within the genus Sphingomonas . 03SUJ6T was related distantly to Sphingomonas daechungensis CH15-11T (97.4 % 16S rRNA gene sequence similarity), Sphingomonas ginsengisoli Gsoil 634T (97.3 %) and Sphingomonas astaxanthinifaciens DMS 22298T (97.1 %). Closest relatives of strain WM95T were S . daechungensis CH15-11T (98.2 %), Sphingomonas jaspsi DSM 18422T (97.6 %), Sphingomonas sediminicola Dae 20T (97.5 %), Sphingomonas lutea JS5T (97.4 %) and S . ginsengisoli Gsoil 634T (97.2 %). The major fatty acids of the two isolates were summed feature 8 and C16 : 0. The predominant isoprenoid quinone was ubiquinone-10. The major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and sphingoglycolipid. sym-Homospermidine was the major polyamine of the isolates. Phenotypic characteristics distinguished 03SUJ6T and WM95T from the related species of the genus Sphingomonas . On the basis of the evidence presented in this study, the novel species, Sphingomonas limnosediminicola sp. nov. and Sphingomonas palustris sp. nov. are proposed for strain 03SUJ6T (=KCTC 23331T=JCM 17543T) and strain WM95T (=KACC 18738T=JCM 31399T), respectively.

A yellow-pigmented, Gram-stain-negative, short-rod-shaped bacterial strain, MIMD3T, was isolated from biological soil crusts collected in Liangcheng, north-western China. Cell growth could be observed at 10–37 °C (optimum 25 °C), at pH 5–8 (optimum 6.6) and in the presence of 1 % (w/v) NaCl (optimum 0 %). The genomic DNA G+C content was 65.0 mol%. Analysis of 16S rRNA gene sequences showed that strain MIMD3T shared the highest similarity with Sphingomonas vulcanisoli KCTC 42454T (95.1 %), Sphingomonas oligophenolica JCM 12082T (94.8 %), Sphingomonas mali IFO 15500T (94.5 %), Sphingomonas . leidyi ATCC 15260T (94.4 %) and Sphingomonas formosensis CC-Nfb-2T (94.3 %). The strain had Q-10 as the predominant respiratory quinone, and sym-homospermidine as the major polyamine. The major fatty acids of the strain were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), C19 : 0 cyclo ω8c, C14 : 0 2-OH and C16 : 0. The main polar lipids of strain MIMD3T were phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine and sphingoglycolipid. Based on phenotypic, chemotaxonomic and phylogenetic characteristics, it is concluded that strain MIMD3T represents a novel species of the genus Sphingomonas , for which the name Sphingomonas crusticola sp. nov. is proposed. The type strain is MIMD3T (=KCTC 42801T=MCCC 1K01310T).

A Gram-stain-negative, aerobic, rod-shaped bacterium with a single polar flagellum, designated strain DCSW07T, was isolated from the surface water of the Bohai Sea, China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain DCSW07T shared highest similarity (96.97 %) with Phaeobacter gallaeciensis DSM 26640T, formed a lineage within the family Rhodobacteraceae and was distinct from the most closely related genera Phaeobacter and Pseudooceanicola (96.6–96.8 and 95.8–96.2 % 16S rRNA gene sequence similarity, respectively). Optimal growth occurred in the presence of 6 % (w/v) NaCl, at pH 6.0 and at 28 °C. Strain DCSW07T contained phosphatidylcholine, phosphatidylethanolamine, an unidentified phospholipid and two unidentified polar lipids as the major polar lipids, and C18 : 1ω7c as the main fatty acid (>10 % of the total). The DNA G+C content of strain DCSW07T was 64.8 mol%. On the basis of this polyphasic study, strain DCSW07T is considered to represent a novel species of a new genus in the Roseobacter clade of the family Rhodobacteraceae , for which the name Paraphaeobacter pallidus gen. nov., sp. nov. is proposed. The type strain of Paraphaeobacter pallidus is DCSW07T (=KCTC 52369T=MCCC 1K03197T=JCM 31458T=CGMCC 1.15762T).

Within the frame of a biotechnological screening, we isolated two Pseudomonas strains from forest soil. 16S rRNA gene sequence analysis indicated that strain CCOS 864T shared 99.8 % similarity with Pseudomonas donghuensis HYST, while strain CCOS 865T shared 99.0 % similarity with Pseudomonas putida DSM 291T and lower similarity with other P. putida group type strains. Based on multilocus sequence analysis, the two strains were genotypically distinct from each other, each forming a separate clade. Strains CCOS 864T and CCOS 865T were Gram-stain-negative, motile and rod-shaped, growing at a temperature range of 4–37 °C. Strain CCOS 864T could be phenotypically distinguished from P. putida group species by the combination of gelatinase-positive reaction and positive growth on N-acetyl-d-glucosamine, p-hydroxyphenylacetic acid and inosine but lack of fluorescein production on King’s B medium, while strain CCOS 865T could be distinguished from P. putida group species by the combination of positive growth with saccharic acid and negative growth with p-hydroxyphenylacetic acid and l-pyroglutamic acid. The major polar lipid for both strains was phosphatidylethanolamine; the major quinone was ubiquinone Q-9. DNA–DNA hybridization and average nucleotide identities confirmed the novel species status for the two strains. The DNA G+C contents of CCOS 864T and CCOS 865T were 62.1 and 63.8 mol%, respectively. The phenotypic, phylogenetic and DNA–DNA relatedness data support the suggestion that CCOS 864T and CCOS 865T represent two novel Pseudomonas species. The names Pseudomonas wadenswilerensis sp. nov. (type strain CCOS 864T=LMG 29327T) and Pseudomonas reidholzensis sp. nov. (type strain CCOS 865T=LMG 29328T) are proposed.

A light yellow-coloured, Gram-stain-negative, non-motile and rod-shaped bacterium, designated strain K-3-6T, capable of degrading aliphatic hydrocarbons was isolated from oil-contaminated soil of Biratnagar, Morang, Nepal. It was able to grow at 15–45 °C, at pH 5.0–9.5 and with 0–6 % (w/v) NaCl. Based on 16S rRNA gene sequence analysis, strain K-3-6T belongs to the genus Sphingobium and is closely related to Sphingobium olei IMMIB HF-1T (98.4 % similarity), Sphingobium abikonense NBRC 16140T (98.3 %), Sphingobium rhizovicinum CC-FH12-1T (97.9 %), Sphingobium lactosutens DS20T (97.9 %), Sphingobium amiense NBRC 102518T (97.2 %), Sphingobium phenoxybenzoativorans SC_3T (97.2 %) and Sphingobium fontiphilum Chen16-4T (97.0 %). The predominant respiratory quinone was ubiquinone-10 and the major polyamine was spermidine. The polar lipid profile revealed the presence of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine, phosphatidyldimethylethanolamine, sphingoglycolipid and phosphatidylmonomethylethanolamine. The predominant fatty acids of strain K-3-6T were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), C14 : 0, C16 : 0 and C14 : 0 2-OH. The genomic DNA G+C content was 65.6 mol%. Levels of DNA–DNA relatedness between strain K-3-6T and S. olei IMMIB HF-1T, S. abikonense NBRC 16140T, S. lactosutens DS20T, S. rhizovicinum CC-FH12-1T, S. amiense NBRC 102518T and S. fontiphilum Chen16-4T were 34.0, 33.3, 28.7, 26.3, 29.0 and 22.3 %, respectively. The morphological, physiological, chemotaxonomic and phylogenetic analyses clearly distinguished this strain from its closest phylogenetic neighbours. Thus, strain K-3-6T represents a novel species of the genus Sphingobium , for which the name Sphingobium naphthae sp. nov. is proposed. The type strain is K-3-6T (=KEMB 9005-449T=KACC 19001T=JCM 31713T).

A novel Gram-stain-negative, aerobic, rod-shaped strain designated 166T was isolated from surface-sterilized root tissue of maize planted in the Fangshan District of Beijing, PR China. The 16S rRNA gene sequence analysis indicated that strain 166T belongs to the genus Rhizobium and is closely related to Rhizobium cellulosilyticum ALA10B2T and Rhizobium yantingense H66T with sequence similarities of 98.8 and 98.3 %, respectively. According to atp D and rec A sequence analysis, the highest sequence similarity between strain 166T and R. cellulosilyticum ALA10B2T is 93.8 and 84.7 %, respectively. However, the new isolate exhibited relatively low levels of DNA–DNA relatedness with respect to R. cellulosilyticum DSM 18291T (20.8±2.3 %) and Rhizobium yantingense CCTCC AB 2014007T (47.2±1.4 %). The DNA G+C content of strain 166T was 59.8 mol%. The main polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, diphosphatidylglycerol, an unidentified aminophospholipid and an unidentified aminolipid. The major fatty acids of strain 166T were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c). The results of the physiological and biochemical tests and minor differences in the fatty acid profiles allowed a clear phenotypic differentiation of strain 166T from the type strains of closely related species, R. cellulosilyticum DSM 18291T and R. yantingense CCTCC AB 2014007T. Strain 166T represents a novel species within the genus Rhizobium , for which the name Rhizobium wenxiniae sp. nov. is proposed, with the type strain 166T (=CGMCC 1.15279T=DSM 100734T).