- Volume 66, Issue 9, 2016

A Gram-staining-negative, facultatively aerobic, white-colony-forming bacterium, designated strain SE-S21T, was isolated from forest soil of Jeju Island in Korea. Cells were motile rods with a single polar flagellum, showing catalase- and oxidase-positive reactions. Growth was observed at 10–40 °C (optimum, 30 °C), pH 4.0–10.0 (optimum, pH 7.0–7.5) and with 0–4.0 % (w/v) NaCl (optimum, 0–2 %). Only ubiquinone-8 was detected as the isoprenoid quinone, and C16 : 0, C17 : 0 cyclo, C19 : 1ω8c cyclo and summed feature 2 (comprising C12 : 0 aldehyde and/or unknown) were found to be the major fatty acids. Phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, an unknown aminophospholipid, an unknown aminolipid and an unknown lipid were detected as the major polar lipids. Putrescine and 2-hydroxyputrescine were the predominant polyamines. The DNA G+C content was 61.0 mol%. Phylogenetic analyses based on 16S rRNA and DNA gyrase B gene sequences revealed that strain SE-S21T formed a phyletic lineage within the genus Pandoraea . Strain SE-S21T was most closely related to Pandoraea faecigallinarum KOxT and Pandoraea pnomenusa CCUG 38742T with 98.8 % and 98.7 % 16S rRNA gene sequence similarities, respectively. However, the DNA–DNA relatedness values between strain SE-S21T and the type strains of P. faecigallinarum and P. pnomenusa were 26.6±5.7 % and 20.5±3.7 %, respectively. On the basis of phenotypic, chemotaxonomic and molecular features, strain SE-S21T clearly represents a novel species of the genus Pandoraea , for which the name Pandoraea terrae sp. nov. is proposed. The type strain is SE-S21T (=KACC 18127T=JCM 30137T). An emended description of the genus Pandoraea is also proposed.

Three Gram-stain-negative, rod-shaped bacteria, designated strains NH153T, F-2-11 and M-1-78, were isolated from surface seawater of the South China Sea and the East China Sea. The three isolates were able to grow at 15–45 °C (optimum 28–37 °C), but no growth occurred at 4 or 50 °C. The pH range for growth was pH 5.5–9.5 (optimum pH 7.5–8.5). The isolates required sea salts for growth and growth occurred in the presence of 0–10 % (w/v) NaCl (optimum 3–5 %); no growth occurred in the presence of 12.0, 15.0 or 20.0 % (w/v) NaCl. They were positive for hydrolysis of gelatin and Tween 80. The sole respiratory quinone was ubiquinone-8 (Q-8). The major cellular fatty acids (>10 %) were C16 : 0, C18 : 1 ω7c and summed feature 3 (C16 : 1 ω7c and/or iso-C15 : 0 2-OH). The major polar lipid components were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, one unidentified glycolipid, one unidentified phospholipid and one unidentified lipid. The genomic DNA G+C content of strain NH153T was 41.4 mol%. Based on 16S rRNA gene sequence analysis, the isolates were closely related to the type strain of Pseudoalteromonas shioyasakiensis (98.0–98.6 % sequence similarity). The 16S rRNA gene sequence similarities between the three isolates were 98.8–99.7 %. Phylogenetic analysis indicated that they formed a distinct lineage and clustered with P. shioyasakiensis and Pseudoalteromonas arabiensis. The level of DNA–DNA relatedness among the three isolates was 78.0–85.5 %. Strain NH153T exhibited average nucleotide identity values of 93.4 and 84.2 % with respect to P. shioyasakiensis JCM 18891T and P. arabiensis JCM 17292T, respectively. The genome-to-genome distance analysis revealed that strain NH153T shared 52.4 % DNA relatedness with P. shioyasakiensis JCM 18891T and 28.1 % with P. arabiensis JCM 17292T. On the basis of the phenotypic, genotypic and chemotaxonomic characterizations, as well as phylogenetic inference obtained in this study, strains NH153T, F-2-11 and M-1-78 represent a novel species within the genus Pseudoalteromonas , for which the name Pseudoalteromonas gelatinilytica sp. nov. is proposed. The type strain is NH153T (=CGMCC 1.15370T=DSM 100951T), and F-2-11 (=CGMCC 1.15364=DSM 100953) and M-1-78 (=CGMCC 1.15365=DSM 100952), are additional strains of the species.

Strain NRRL B-41902T and three closely related strains were isolated from iceberg lettuce. The strain was found to consist of strictly aerobic, Gram-stain-negative rods that formed cocci in late stationary phase. 16S rRNA gene sequence analysis showed that strain NRRL B-41902T was most closely related to species within the genera Acinetobacter , and that a grouping of it and the three other closely related strains was most closely related to the type strain of Acinetobacter pittii, which was also confirmed through a phylogenomic analysis. Moreover, in silico DNA–DNA hybridization analysis revealed a substantial amount of genomic divergence (39.1 %) between strain NRRL B-41902T and the type strain of A. pittii , which is expected if the strains represent distinct species. Further phenotypic analysis revealed that strain NRRL B-41902T was able to utilize a combination of l-serine, citraconic acid and citramalic acid, which differentiated it from other, closely related Acinetobacter species. Therefore, strain NRRL B-41902T (=CCUG 68785T) is proposed as the type strain of a novel species, Acinetobacter lactucae sp. nov.

A mesophilic, facultatively anaerobic, facultatively chemolithoautotrophic bacterium, designated strain TC8T, was isolated from a sulfidic shallow-water marine gas vent located at Tor Caldara, in the Tyrrhenian Sea, Italy. Cells were Gram-stain-negative curved rods with one or more polar flagella. Cells were approximately 1–1.5 µm in length and 0.6 µm in width. Strain TC8T grew between 20 and 35 °C (optimum 30 °C), with between 5 and 45 g NaCl l−1 (optimum 15–20 g l−1) and between pH 4.5 and 8.5 (optimum pH 6.0–7.0). The generation time under optimal conditions was 8 h. Strain TC8T was a facultative chemolithoautotroph also capable of using organic substrates as electron donors and carbon sources. Chemolithoautotrophic growth occurred with sulfur and thiosulfate as the electron donors, CO2 as the carbon source, and nitrate, oxygen (5 %, v/v) and ferric iron as the electron acceptors. Chemoorganoheterotrophic growth occurred with tryptone, peptone, Casamino acids, pyruvate and glycerol as substrates, while chemolithoherotrophic growth occurred with d(+)-glucose, sucrose, yeast extract, acetate, lactate, citrate and l-glutamine. The G+C content of the genomic DNA was 59.9 mol%. Phylogenetic analysis of the 16S rRNA gene sequence of strain TC8T showed that this organism formed a lineage within the family Rhodospirillaceae , which branched separately from the two closest relatives, Magnetovibrio blakemorei MV1T (91.25 % similarity) and Magnetospira thiophila MMS-1T (90.13 %). Based on phylogenetic, physiological and chemotaxonomic characteristics, it is proposed that the organism represents a novel species of a new genus within the family Rhodospirillaceae,Varunaivibrio sulfuroxidans gen. nov., sp. nov. The type strain of Varunaivibrio sulfuroxidans is TC8T (=DSM 101688T=JCM 31027T).

Members of the genus Neisseria have been isolated from or detected in a wide range of animals, from non-human primates and felids to a rodent, the guinea pig. By means of selective culture, biochemical testing, Gram staining and PCR screening for the Neisseria -specific internal transcribed spacer region of the rRNA operon, we isolated four strains of the genus Neisseria from the oral cavity of the wild house mouse, Mus musculus subsp. domesticus. The isolates are highly related and form a separate clade in the genus, as judged by tree analyses using either multi-locus sequence typing of ribosomal genes or core genes. One isolate, provisionally named Neisseria musculi sp. nov. (type strain AP2031T=DSM 101846T=CCUG 68283T=LMG 29261T), was studied further. Strain AP2031T/ N. musculi grew well in vitro. It was naturally competent, taking up DNA in a DNA uptake sequence and pilT-dependent manner, and was amenable to genetic manipulation. These and other genomic attributes of N. musculi sp. nov. make it an ideal candidate for use in developing a mouse model for studying Neisseria –host interactions.

One beige-pigmented, Gram-stain-negative, rod-shaped bacterium, strain E3/4T, was isolated from a zebrafish, Danio rerio. Phylogenetic analysis based on nearly full-length 16S rRNA gene sequences showed that the isolate shared 98.5 % 16S rRNA gene sequence identity to the type strain of Undibacterium macrobrachii and 97.8 % to the type strain of Undibacterium seohonense . Lower 16S rRNA gene sequence similarities (<97.0 %) could be found in comparison with all other species of the genus Undibacterium . DNA–DNA hybridization with Undibacterium macrobrachii LMG 26891T showed a low level of relatedness, <35 %. The main cellular fatty acids of the strain were summed feature 3 fatty acids (C16 : 1ω7c/C16 : 1ω8c), C10 : 0 3-OH and C16 : 0. The polyamine pattern of strain E3/4T contained predominantly putrescine and 2-hydroxyputrescine. The major quinone was ubiquinone Q-8. Major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. On the basis of phylogenetic, chemotaxonomic, genomic and phenotypic analyses we propose a novel species of the genus Undibacterium , Undibacterium danionis , with strain E3/4T (=DSM 102221T=CCM 8677T=CIP 111017T) as the type strain.

A novel yellow bacterial strain, designated UCM-28T, was isolated from forest soil in Gyeonggi-Do, South Korea. The isolated strain was Gram-stain-negative, aerobic, non-spore-forming, non-motile and rod-shaped, and grew at 10–37 °C, pH 5.5–9 and with 0–1 % NaCl. It could reduce nitrate to nitrite and hydrolyse aesculin. We determined the taxonomic position of strain UCM-28T; based on the 16S rRNA gene sequence, the strain belongs to the genus Novosphingobium . The bacterium showed the highest similarity to Novosphingobium piscinae SLH-16T (98.9 %), Novosphingobium rhizosphaerae JM-1T (97.7 %), Novosphingobium taihuense T3-B9T (97.2 %), Novosphingobium subterraneum DSM 12447T (97.1 %), Novosphingobium aromaticivorans DSM 12444T (97.1 %) and Novosphingobium capsulatum GIFU 11526T (96.7 %). Phylogenic trees also confirmed that strain UCM-28T is most closely related to Novosphingobium piscinae SLH-16T and others, and is positioned within the genus Novosphingobium . The DNA relatedness of strain UCM-28T with its references was in the range of 20.9–35.2 %. The polar lipid profile revealed diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, sphingoglycolipid, phosphatidylcholine, phosphatidylmonomethylethanolamine, six unidentified polar lipids and two unknown glycolipids. The major quinone was ubiquinone Q-10, and the major polyamine was spermidine. The DNA G+C content was 63.5 mol%. The major fatty acids included (>10 %) summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) (46.3 %), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) (24.9 %) and C14 : 0 2-OH (11.8 %). Based on the phylogenetic and phenotypic data, strain UCM-28T should be classified within the genus Novosphingobium as a representative of a novel species, named Novosphingobium flavum sp. nov. The type strain is UCM-28T (=KACC 18571T=NBRC 111647T).

A taxonomic study was carried out on strain 19-m-6T, which was isolated from deep sea sediment of the South China Sea during the screening of alkane-degrading bacteria. The isolate was Gram-reaction-negative, and oxidase- and catalase- positive. On the basis of 16S rRNA gene sequence similarity, strain 19-m-6T was shown to belong to the genus Alcanivorax , related to Alcanivorax jadensis T9T (97.5 %), Alcanivorax hongdengensis A-11-3T (97.3  %), A. lcanivorax borkumensis SK2T (96.6 %) and seven other species of the genus Alcanivorax (93.9–95.4 %). Average nucleotide identity values between strain 19-m-6T and A. jadensis T9T, A. hongdengensis A-11-3T and A. borkumensis SK2T were 85.12, 85.87 and 84.35 %, respectively. The estimated DNA–DNA hybridization values between strain 19-m-6T and these three type strains were 22.0, 22.6 and 21.2 %, respectively. Four alkane hydroxylase (alkB) genes were obtained from the draft genome sequence. The G+C content of the chromosomal DNA was 56.44 mol%. The major fatty acids were C16 : 0, C18 : 1 ω7c and summed feature 3 (C16 : 1 ω6c and/or C16 : 1 ω7c). The polar lipids were phosphatidylglycerol, phosphatidylethanolamine, one aminolipid, three phospholipids, two glycolipids and two aminophospholipids. According to its phenotypic features, fatty acid composition and 16S rRNA gene sequence, the novel strain fitted well into the genus Alcanivorax , but could be clearly distinguished from all other known Alcanivorax species described to date. The nameAlcanivorax nanhaiticus sp. nov. is thus proposed, with 19-m-6T (=MCCC 1A05629T=KCTC 52137T) as the type strain.

A Gram-stain-negative, aerobic, motile with a single polar flagellum, non-spore-forming and rod-shaped bacterial strain named T33T was isolated from forest soil collected at Kyonggi University, South Korea. The strain was catalase- and oxidase-positive, colonies grew on R2A agar at 32 °C. Sequencing of the 16S rRNA gene and phylogenetic analysis revealed that T33T represented a member of the genus Massilia and is closely related to Massilia niastensis KACC 12599T (98.7 % sequence similarity), Massilia aerilata KACC 12505T (98.5 %), Massilia tieshanensis KACC 14940T (98.4 %), Massilia kyonggiensis KACC 17471T (98.1 %), Massilia norwichensis LMG 28164T (97.7 %), Massilia haematophila CCUG 38318T (97.4 %), Massilia consociata CCUG 58010T (97.3 %), and Massilia niabensis KACC 12632T (97.0 %). Ubiquinone Q-8 is the predominant respiratory quinone, and phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol are the major polar lipids. The major fatty acids are summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c), summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c), and C16 : 0. DNA–DNA hybridization revealed <70 % relatedness between strain T33T and the most closely related type strains. The DNA G+C content of strain T33Tis 69.4 mol%. Based on physiological and biochemical test results, Massilia pinisoli T33T is proposed as a novel species of the genus Massilia . The type strain is T33T (=KACC 18748T=KEMB 9005-368T=JCM 31316T).

A taxonomic study was carried out on a Gram-staining-negative bacterium, strain SF-12T, isolated from an unidentified starfish living in Sanya, PR China. Cells of SF-12T were non-spore-forming rods, 0.5–0.8 µm wide, 2.2–2.5 µm long and motile by means of flagella. SF-12T was facultatively anaerobic, heterotrophic, oxidase- and catalase-positive. Growth of SF-12T occurred at 15–38 °C (optimum, 30 °C), at pH 6.5–8.5 (optimum, pH 7.0), and in the presence of 2.0–7.0 % (w/v) NaCl (optimum, 3.0–4.0 %). The predominant fatty acids of SF-12T were C18 : 1ω7c and/or C18 : 1ω6c. Ubiquinone 10 was the sole respiratory quinone of SF-12T. The major polar lipids of SF-12T were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, three unknown aminolipids, and seven unknown phospholipids. The DNA G+C content was 61 mol%. SF-12T showed the highest 16S rRNA gene sequence similarity to Lutimaribacter pacificus W11-2BT (96.06 %), followed by Cribrihabitans neustonicus CC-AMHB-3T (96.02 %), Lutimaribacter saemankumensis SMK-117T (96.0 %), Cribrihabitans marinus CZ-AM5T (95.92 %), Lutimaribacter litoralis KU5D5T (95.92 %) and other species of the family Rhodobacteraceae (<95.9 %). However, phylogenetic trees based on 16S rRNA gene sequences showed that SF-12T formed a lineage with members of the genus Lutimaribacter in the trees. On the basis of phenotypic, chemotaxonomic and phylogenetic analyses, SF-12T is considered to represent a novel species of the genus Lutimaribacter , for which the name Lutimaribacter marinistellae sp. nov. is proposed. The type strain is SF-12T (=MCCC 1K01154T=KCTC 42911T).

A moderately psychrophilic, aerobic, hydrogen- and sulfur-oxidizing bacterium, designated strain MAS2T, was isolated from a tank containing coastal seawater from Tokyo Bay and a block of beef tallow added as organic material. Growth occurred under aerobic chemolithoautotrophic conditions in the presence of molecular hydrogen, thiosulfate, tetrathionate, elemental sulfur or sulfide as the sole energy source and bicarbonate as a carbon source. The isolate represented a Gram-staining-negative rod with a single polar flagellum and grew in artificial seawater medium with thiosulfate at 2–40 °C (optimum 30 °C). The isolate grew in media with thiosulfate at Na+ concentrations between 30 and 1380 mM (optimum 270 mM). MAS2T possessed C16 : 0, C16 : 1 and C18 : 1 as the major fatty acids. The G+C content of the genomic DNA was 39.6 mol%. The 16S rRNA gene sequence similarity analysis showed that the isolate represented a member of the genus Thiomicrospira within the class Gammaproteobacteria and was most closely related to Thiomicrospira frisia JB-A2T. On the basis of phenotypic and molecular properties, the isolate represents a novel species of the genus Thiomicrospira , for which the name Thiomicrospira hydrogeniphila sp. nov. is proposed (type strain, MAS2T=JCM 30760T=DSM 100274T).

The phenotypic and genotypic characteristics of a historical collection of strains identified as Achromobacter denitrificans were examined. Sequence analysis of a 765 bp nrdA gene fragment revealed that eight of these strains belonged to the recently described Achromobacter aegrifaciens, Achromobacter mucicolens , and Achromobacter insolitus , and that one strain belonged to Achromobacter xylosoxidans . The analysis also suggested the presence of four novel species of the genus Achromobacter among the remaining strains. The latter was confirmed by multilocus sequence analysis of concatenated nusA, eno, rpoB, gltB, lepA, nuoL andnrdA gene fragments and extensive genotypic and phenotypic characterization. We propose to name these novel species as Achromobacter agilis sp. nov., nom. rev. (type strain LMG 3411T=CCUG 62454T), Achromobacter pestifer sp. nov., nom. rev. (type strain LMG 3431T=CCUG 61959T) , Achromobacter kerstersii sp. nov. (type strain LMG 3441T=CCUG 62449T) and Achromobacter deleyi sp. nov. (type strain LMG 3458T=CCUG 62433T).

A new aerobic marine bacterium, strain S3431, was isolated from swab samples of an unidentified polychaete near Canal Concepción, Chile. This strain was thought to represent a new taxon within the genus Pseudoalteromonas . Although DNA–DNA reassociation values showed less than 70 % genomic DNA relatedness to established Pseudoalteromonas type strains, it shared 78 % DNA–DNA relatedness with Alteromonas fuliginea DSM 15748 (=KMM 216) (Romanenko et al., 1994). A. fuliginea has later been considered a heterotypic synonym of Pseudoalteromonas citrea (Ivanova et al., 1998). Relatedness between strains S3431, A. fuliginea DSM 15748 and the type strain P. citrea LMG 12323T was therefore studied. Physiological traits and genomic information were shared at a high level by strains S3431 and DSM 15748, but not between these and P. citrea LMG 12323T. There was only approximately 20 % DNA–DNA relatedness between P. citrea LMG 12323T and strains S3431 and DSM 15748. Based on the available phylogenetic and phenotypic data, the reclassification of A. fuliginea DSM 15748 (Romanenko et al., 1995) → Pseudoalteromonas citrea (Ivanova et al., 1998) as Pseudoalteromonas fuligineacomb. nov. is proposed, and strain S3431 should be assigned to this new species. The name Pseudoalteromonas fuliginea is proposed with KMM 216T (=DSM 15748T=CIP 105339T) as the type strain.

A novel Gram-staining-negative, aerobic and rod-shaped strain designated 541T was isolated from surface-sterilized root tissue of maize, collected from the Fangshan District of Beijing, People’s Republic of China, and was subjected to a taxonomic study using a polyphasic approach. According to a phylogenetic tree based on 16S rRNA gene sequences, strain 541T represented a member of the genus Sphingomonas and clustered with Sphingomonas sanxanigenens DSM 19645T, with which it shared the highest 16S rRNA gene sequence similarity (98.8 %). The predominant respiratory quinone was ubiquinone-10 (Q-10), the major polyamine was sym-homospermidine and the major cellular fatty acids were C18 : 1ω7c (50.9 %), C16 : 0 (22.0 %) and C14 : 0 2-OH (11.4 %). The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine and sphingoglycolipid. The DNA G+C content was 64.7 mol%. DNA–DNA relatedness between strain 541T and its closest phylogenetic relative Sphingomonas sanxanigenens DSM 19645T was 50.8 %. The results of physiological and biochemical tests and the differences in the fatty acid profiles allowed a clear phenotypic differentiation of strain 541T from closely related species of the genus Sphingomonas . Strain 541T represents a novel species within the genus Sphingomonas , for which the nameSphingomonas zeicaulis sp. nov. is proposed, with the type strain 541T (=CGMCC 1.15008T=DSM 100587T).

Two yeast strains that are members of the same species were isolated from different marine habitats, i.e. one from Mid-Atlantic Ridge ocean water samples located in the direct vicinity of black smokers near the Rainbow deep-sea hydrothermal vent and one from Brazilian marine water samples off the Ipanema beach. Strains CLIB 1964T and CLIB 1965 are anamorphic ascomycetous yeasts affiliated to the Yamadazyma clade of Saccharomycetales. Interestingly, these strains were phylogenetically and distinctly positioned into a group of species comprising all species of the genus Yamadazyma isolated from marine habitats including deep-sea hydrothermal vents, i.e.Candida atmosphaerica,C. spencermartinsiae,C. atlantica,C. oceani and C. taylorii. These strains differed significantly in their D1/D2 domain sequences of the LSU rRNA gene from the closely related species mentioned above, by 2.6, 3.0, 3.4, 3.8 and 6.0 %, respectively. Internal transcribed spacer region sequence divergence was also significant and corresponded to 4.6, 4.7, 4.7, 12.0 and 24.7 % with C. atlantica,C. atmosphaerica, C. spencermartinsiae,C. oceani and C. taylorii, respectively. Phenotypically, strains CLIB 1964T and CLIB 1965 could be distinguished from closely related species by their inability to assimilate l-sorbose. CLIB 1964T (=CBS 14301T=UBOCC-A-214001T) is the designated type strain for Yamadazyma barbieri sp. nov. The MycoBank number is MB 815884.

Papaya is an important fruit crop cultivated in tropical and subtropical regions. Papaya little leaf (PLL) disease was observed in China. The phytoplasma 16S rRNA gene was detected from symptomatic papaya trees via PCR using phytoplasma universal primers P1/P7 followed by R16F2n/R16R2. No amplification products were obtained from templates of asymptomatic papaya trees. These results indicated a direct association between phytoplasma infection and PLL disease. Comparative and phylogenetic analyses of 16S rRNA gene sequences indicated that the papaya-infecting phytoplasmas under study belonged to the peanut witches’ broom phytoplasma group (16SrII). Genotyping through use of computer-simulated RFLP analysis of 16S rRNA genes and coefficients of RFLP pattern similarities (0.97) reveal that the PLL phytoplasma was placed in a new subgroup. In this article, we describe the molecular characterization of a new phytoplasma associated with PLL disease and propose that the PLL phytoplasma be considered as a novel subgroup, 16SrII-U.

A number of recent Requests for an Opinion have cited Rule 18c of the International Code of Nomenclature of Bacteria as the Rule governing setting a time limit on the search for a neotype strain. This Rule only governs what happens when a neotype is proposed and not what happens when the original type appears to have been lost and a neotype is not found. It is appropriate to emphasize this issue and to examine what other alternatives are available.