Hymeniacidon perleve associated bioactive bacterium pseudomonas sp. NJ6-3-1

Among the marine bacteria isolated from cytotoxic sponge Hymeniacidon perleve, one strain NJ6-3-1 classified as Pseudomonas sp. showed both cytotoxic and antimicrobial activities. Fatty acid analysis indicated that the bacterial strain consists mainly of C16:1, C16:0, C18:1, C18:0, C15:0, C14:0. One unusual 9,10-cyclopropane-C17:0 fatty acid, and C26:0 also constitute as major component as well as the existence of squalene, the precursor of triterpenoids. The major metabolites in the culture broth were identified as alkaloids, including diketopiperazines and indole compounds, namely 3,6-diisopropylpiperazin-2,5-dione, 3-benzyl-3-isopropylpiperazin-2,5-dione, 3,6-bis-(2-methylpropyl)-piperazin-2,5-dione, indole-3-carboxaldehyde, indole-3-caroxylic acid methyl ester, indole-3-ethanol, and quinazoline-2,4-dione.     

<Emphasis Type="Italic">Sphingomonas jejuensis</Emphasis> sp. nov., isolated from marine sponge <Emphasis Type="Italic">Hymeniacidon flavia</Emphasis>

A Gram-negative, non-motile, rod shaped, and orange-pigmented chemoheterotrophic bacterium, strain MS-31^T was isolated from the marine sponge Hymeniacidon flavia, collected from near Jeju Island, Korea. The Strain MS-31^T was subjected to a polyphasic taxonomic study. The phylogenetic analysis based on the 16S rRNA gene sequences revealed that the novel isolate could be affiliated within the genus Sphingomonas. The strain MS-31^T showed 95.6% of 16S rRNA gene sequence similarity with the most closely related species Sphingomonas koreensis JSS26^T. The DNA G+C content of the strain MS-31^T was 69.4 mol%. The major isoprenoid quinone was ubiqunone 10 and predominant cellular fatty acids were summed feature 7 (comprising C_18:1 ω7c, C_18:1 Ω9t and/or C_18:1 ωl2t, 39.7%), C_16:0 (16.3%), C_14:0 2OH (15.9%) and summed feature 3 (comprising C_16:1 ω7c and/or C_15:0 iso 2OH, 11.7%). The polar lipids were sphingoglycolipid, phosphatidyletha-nolamine, phosphatidylglycerol, diphosphatidylglycerol and unidentified glycolipid. Based on the evidence from the polyphasic taxonomic study, the strain should be classified as a new species of the genus Sphingomonas. As a result, the name Sphingomonas jejuensis sp. nov. (type strain MS-31^T =KCTC 23321^T =NBRC 107775^T) is proposed.     

An evaluation of the diet of <Emphasis Type="Italic">Mysis relicta</Emphasis> using gut contents and fatty acid profiles in lakes with and without the invader <Emphasis Type="Italic">Bythotrephes longimanus</Emphasis> (<Emphasis Type="Italic">Onychopoda,Cercopagidae</Emphasis>)

Diets of Mysis relicta from four lakes in central Ontario that had been invaded by Bythotrephes longimanus and three lakes that had not been invaded were investigated using gut content analysis and fatty acid (FA) composition. Gut content analysis of M. relicta revealed a high incidence of cannibalism in all lakes, and consumption of B. longimanus and native zooplanktivorous midges in the genus Chaoborus in lakes where these were present. Cladocera other than B. longimanus were present in the guts of all M. relicta examined except those from Bernard Lake, the lake with the most B. longimanus. In that lake, B. longimanus was the most frequent diet item. Copepod remains were found in 60–100% of M. relicta guts with the lowest frequency occurring in Bernard Lake. Fatty acids (FA) that contributed strongly to the variation in FA composition in M. relicta, as revealed by a principal component analysis, were C16:0 (palmitic acid), C16:1n7 (palmitoleic acid), C18:1n9c (oleic acid), C20:4n6 (arachidonic acid), C20:5n3 (eicosapentaenoic acid), and C22:6n3 (docosahexaenoic acid). Significant differences in FA amount and composition of M. relicta were found between invaded and non-invaded lakes, and among lakes within these groups. Generally, M. relicta in non-invaded lakes had higher concentrations of C16:0, C18:1n9c, C18:2n6c (linoleic acid), C18:3n3 (α-linolenic acid) and C20:4n6, while M. relicta in invaded lakes had higher concentrations of C22:6n3. Two of the non-invaded lakes had lower water transparency, as measured by Secchi depth, which may be the reason why mysids and abundant populations of Chaoborus spp. could be found in the water column during the day. However, differences in FA profiles and gut contents of M. relicta between invaded and non-invaded lakes are consistent with competition for Cladocera in the presence of the invader rather than pre-existing differences among lakes. We conclude that the diet of M. relicta is affected by the invasion of B. longimanus.     

An <Emphasis Type="Italic">ipdC</Emphasis> gene knock-out of <Emphasis Type="Italic">Azospirillum brasilense</Emphasis> strain SM and its implications on indole-3-acetic acid biosynthesis and plant growth promotion

The indole-3-pyruvate decarboxylase gene (ipdC), coding for a key enzyme of the indole-3-pyruvic acid pathway of IAA biosynthesis in Azospirillum brasilense SM was functionally disrupted in a site-specific manner. This disruption was brought about by group II intron-based Targetron gene knock-out system as other conventional methods were unsuccessful in generating an IAA-attenuated mutant. Intron insertion was targeted to position 568 on the sense strand of ipdC, resulting in the knock-out strain, SMIT568s10 which showed a significant (∼50%) decrease in the levels of indole-3-acetic acid, indole-3-acetaldehyde and tryptophol compared to the wild type strain SM. In addition, a significant decrease in indole-3-pyruvate decarboxylase enzyme activity by ∼50% was identified confirming a functional knock-out. Consequently, a reduction in the plant growth promoting response of strain SMIT568s10 was observed in terms of root length and lateral root proliferation as well as the total dry weight of the treated plants. Residual indole-3-pyruvate decarboxylase enzyme activity, and indole-3-acetic acid, tryptophol and indole-3-acetaldehyde formed along with the plant growth promoting response by strain SMIT568s10 in comparison with an untreated set suggest the presence of more than one copy of ipdC in the A. brasilense SM genome.     

<Emphasis Type="Italic">Sphingomonas hunanensis</Emphasis> sp. nov., isolated from forest soil

A novel Gram-negative, catalase- and oxidase-positive, strictly aerobic, non spore-forming, rod-shaped bacterium, designated strain JSM 083058^T, was isolated from non-saline forest soil in Hunan Province, China. Growth occurred with 0–8% (w/v) NaCl (optimum, 0.5–3%) at pH 6.0–10.0 (optimum, pH 7.0) and at 5–35°C (optimum, 25–30°C). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain JSM 083058^T fell within the cluster comprising species of the genus Sphingomonas, clustering with Sphingomonas aestuarii K4^T, with which it shared highest 16S rRNA gene sequence similarity (99.2%). The chemotaxonomic properties of strain JSM 083058^T were consistent with those of the genus Sphingomonas. The predominant respiratory quinone was ubiquinone Q-10, and the major cellular fatty acids were summed feature 8 (C18:1ω7c/C18:1ω6c), C16:0, summed feature 3 (C16:1ω7c/C16:1ω6c) and C17:1ω6c. The polar lipids consisted of diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and sphingoglycolipid. The genomic DNA G+C content of strain JSM 083058^T was 65.5 mol%. The combination of phylogenetic analysis, DNA–DNA relatedness, phenotypic characteristics and chemotaxonomic data supported the view that strain JSM 083058^T represents a novel species of the genus Sphingomonas, for which the name Sphingomonas hunanensis sp. nov. is proposed. The type strain is JSM 083058^T (=CCTCC AA 209011^T = DSM 22213^T).     

<Emphasis Type="Italic">Paenibacillus camelliae</Emphasis> sp. nov., isolated from fermented leaves of <Emphasis Type="Italic">Camellia sinensis</Emphasis>

A novel bacterium, strain blls-2^T was isolated from Pu’er tea. The isolate was Gram-positive, endospore-forming motile rod that grew at 15∼42°C and pH 6.0∼10.2. The DNA G+C content was 48.3 mol%, the predominant isoprenoid quinone was MK-7, and the predominant cellular fatty acid was anteiso-C15:0 (54.2%) followed by C16:0 (15.5%) and iso-C16:0 (8.2%). The polar lipid pattern of blls-2^T was characterized by the presence of diphosphatidylglycerol, phosphatidylethanolamine, and phosphatidylglycerol. Phy-logenetic analysis based on 16S rRNA gene sequence showed that the strain was affiliated within the Paenibacillaceae. The strain was most closely related to Paenibacillus granivorans A30^T, with a similarity of 97.1%. Based on the phylogenetic and phenotypic characteristics of strain blls-2^T, the isolate is thought to represent a novel taxon in the genus Paenibacillus. The name Paenibacillus camelliae sp. nov. is proposed for the fermented tea isolate; the type strain is blls-2^T (= KCTC 13220^T= CECT 7361^T).     

<Emphasis Type="Italic">Oceaniovalibus guishaninsula</Emphasis> gen. nov., sp. nov., A Marine Bacterium of the Family <Emphasis Type="Italic">Rhodobacteraceae</Emphasis>

The alphaproteobacterial strain JLT2003^T was isolated from surface seawater off the coast of Guishan island, Taiwan. The strain was Gram negative, ovoid or coccoid, non-motile and formed pink colonies on marine agar 2216 (MA; DIFCO) medium. The dominant fatty acids were C_18:1ω7c, cyclo C_19:0ω8c, and C_16:0. The polar lipid profile consisted of diphosphatidylglycerol and phosphatidylglycerol. The major respiratory ubiquinone was Q-10. The DNA G+C content was 62.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain was most closely related to Pontibaca methylaminivorans GRP21^T with 94.8% similarity. The isolate was distinguishable from members of the family Rhodobacteraceae based on phenotypic and biochemical characteristics. On the basis of the taxonomic data presented, strain JLT2003^T is considered to represent a novel species of a new genus, for which the name Oceaniovalibus guishaninsula gen. nov., sp. nov. is proposed. The type strain of Oceaniovalibus guishaninsula is JLT2003^T (=JCM 17765^T = CGMCC 1.10827^T).     

Identification of the alga known as <Emphasis Type="Italic">Nannochloropsis</Emphasis> Z-1 isolated from a prawn farm in Hainan,China as <Emphasis Type="Italic">Chlorella</Emphasis>

An alga known as “Nannochloropsis”, isolated from a prawn farm in Hainan, China, has been critically investigated and identified as Chlorella, a member of the Chlorophyceae based on fatty acid composition, ultrastructure, and 18S rDNA. Cells of this alga were spherical, measured by 1–6 μm in diameter and were enclosed in thin walls of approximately 0.04 μm thickness. They contained several small mitochondria, two to three thylakoids and had no vacuoles. There were many pyrenoids in the algal cells and their thylakoid lamellae were sparse and not translucent. Many lipid droplets were present in the cytoplasm. The total lipid content of this alga was 3% per gram dry weight and its major fatty acids were C_16:0, C_18:0, C_18:1, C_18:2, C_18:3 and C_20:0. Eicosapentaenoic acid (C_20:5, EPA) was not detected. The length of its 18S rDNA sequence was 1,712 bp. 18S rDNA sequence analyses indicated that this alga was a species of Chlorella.     

Micropropagation of <Emphasis Type="Italic">Harpagophytum procumbens</Emphasis>

An efficient protocol for micropropagation of Harpagophytum procumbens DC., an endangered African medicinal plant, was developed. Maximum shoot multiplication without callus was obtained from nodal explants cultured on Murashige and Skoog (MS) basal salts plus Gamborg’s (B₅) vitamins supplemented with 0.1 mg dm⁻³ indole-3-acetic acid and 5.0 mg dm⁻³ kinetin. The shoots were subsequently subcultured every 3 weeks on the same medium. Detached axillary shoots were transferred to MS basal salts plus B₅ vitamins supplemented with various concentrations of α-naphthalene-acetic acid or indole-3-butyric acid (IBA), ranging from 0.5 to 2.5 mg dm⁻³ and 100 % rooting and optimal subsequent acclimatization was achieved on 1.0 mg dm⁻³ IBA. After 4 weeks of culture, the rooted shoots (>5 cm) were planted in pots containing peat, vermiculite and bark (2:1:1), covered with plastic domes and maintained at 25 °C for 2 weeks before being transferred to a glasshouse. Plant survival was about 40 %.     

LasR Receptor for Detection of Long-Chain Quorum-Sensing Signals: Identification of <Emphasis Type="Italic">N</Emphasis>-Acyl-homoserine Lactones Encoded by the <Emphasis Type="Italic">avsI</Emphasis> Locus of <Emphasis Type="Italic">Agrobacterium vitis</Emphasis>

Bacterial biosensor strains have greatly facilitated the rapid discovery, isolation, and study of quorum-sensing systems. In this study, we determined the relative sensitivity of a LasR-based E. coli bacterial bioluminescence biosensor JM109 (pSB1075) for 13 diverse long-chain N-acyl-homoserine lactones (AHLs) including oxygen-substituted and -unsubstituted AHLs containing 14, 16, and 18 carbons and with and without double bonds. Furthermore, we show by bioassay, HPLC, and GC/MS that four long-chain AHLs of the C16-HSL family are encoded by the avsI gene of Agrobacterium vitis strain F2/5, a non-tumorigenic strain that inhibits pathogenic strains of A. vitis from causing crown gall on grape. The four C16-HSLs include: C16-HSL, N-hexadecanoyl homoserine lactone; 3-oxo-C16-HSL, N-(3-oxohexadecanoyl)homoserine lactone; C16:1-HSL, N-(cis-9-octadecenoyl)homoserine lactone; and 3-oxo-C16:1-HSL, N-(3-oxo-cis-11-hexadecenoyl)homoserine lactone. Thus, the LasR-based bioluminescent biosensor tested in this study should serve as a useful tool for the detection of various long-chain AHLs with and without double bonds as well as those oxylated at the third carbon from uninvestigated species.     

<Emphasis Type="Italic">Henriciella marina</Emphasis> gen. nov., sp. nov., a novel member of the family <Emphasis Type="Italic">Hyphomonadaceae</Emphasis> isolated from the East Sea

A bacterial strain, designated Iso4^T, was isolated from the East Sea of Korea and was subjected to a poly-phasic taxonomy study including phenotypic and chemotaxonomic characteristics as well as 16S rRNA gene sequence analysis. Cells of the strain were Gram-negative, motile, non-budding, non-stalked, and strictly aerobic. Strain Iso4^T grew optimally at 20°C in the presence of 1∼2% (w/v) NaCl and at pH 6.9∼7.6. The major respiratory quinone was Q-10 and the major cellular fatty acids were C_18:1 ω7c (53.5%), C_17:1 ω5c (11.7%), C_17:1 ω6c (8.1%), C_16:0 (7.8%), C_17:0 (4.8%), C_15:0 (2.9%), and C_16:1 ω5c (2.2%). The DNA G+C content of strain Iso4^T was 56.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain Iso4^T formed a monophyletic clade in the family Hyphomonadaceae, supported by high bootstrap value and was most closely related to the genus Hyphomonas (92∼94%), a member of marine bacteria in the family. The phenotypic, genotypic, and chemotaxonomic evidences also suggest strain Iso4^T represents a novel genus and species in the family Hyphomonadaceae, for which the name Henriciella gen. nov., sp. nov. is proposed. The type strain is Iso4^T (=KCTC 12513^T =DSM 19595^T =JCM 15116^T).     

Morphological evaluation and antioxidant activity of <Emphasis Type="Italic">in vitro</Emphasis>- and <Emphasis Type="Italic">in vivo</Emphasis>-derived <Emphasis Type="Italic">Echinacea purpurea</Emphasis> plants

An effective in vitro protocol for rapid clonal propagation of Echinacea purpurea (L.) Moench through tissue culture was described. The in vitro propagation procedure consisted of four stages: 1) an initial stage - obtaining seedlings on Murashige and Skoog (MS) basal medium with 0.1 mg L⁻¹ 6-benzylaminopurine, 0.1 mg L⁻¹ α-naphthalene acetic acid and 0.2 mg L⁻¹ gibberellic acid; 2) a propagation stage — shoot formation on MS medium supplemented with 1 mg L⁻¹ 6-benzylaminopurine alone resulted in 9.8 shoots per explant and in combination with 0.1 mg L⁻¹ α-naphthalene acetic acid resulted in 16.2 shoots per explant; 3) rooting stage — shoot rooting on half strength MS medium with 0.1 mg L⁻¹ indole-3-butyric acid resulted in 90% rooted microplants; 4) ex vitro acclimatization of plants. The mix of peat and perlite was the most suitable planting substrate for hardening and ensured high survival frequency of propagated plants. Significant higher levels were observed regarding water-soluble and lipid-soluble antioxidant capacities (expressed as equivalents of ascorbate and α-tocopherol) and total pnenols content in extracts of Echinaceae flowers derived from in vitro propagated plants and adapted to field conditions in comparison with traditionally cultivated plants.     

<Emphasis Type="Italic">In vitro</Emphasis> culture studies on <Emphasis Type="Italic">Stevia rebaudiana</Emphasis>

Summary Shoot apex, nodal, and leaf explants of Stevia rebaudiana Bertoni can regenerate shoots when cultured on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA; 8.87 μM) and indole-3-acetic acid (5.71 μM). Rooting of the in vitro-derived shoots could be achieved following subculture onto auxin-containing medium. A survival rate of 70% was recorded at the hardening phase on the substrate cocopeat. The presence of the sweet diterpene glycosides, viz. stevioside and rebaudioside, was confirmed in the in vitro-derived tissues of Stevia using HPTLC techniques. Callus cultured on agar-solidified MS medium supplemented with BA (8.87 μM) and indole-3-butyric acid (9.80 μM) showed the highest sweetener content.     

<Emphasis Type="Italic">Joostella atrarenae</Emphasis> sp. nov., a novel member of the <Emphasis Type="Italic">Flavobacteriaceae</Emphasis> originating from the black sea sand of Jeju Island

Strain M1-2^T was isolated from the black sand from the seashore of Jeju Island, Republic of Korea and was classified using a polyphasic taxonomic approach. Strain M1-2^T appeared as Gram-negative, motile rods that could grow in the presence of 1–10% (w/v) NaCl and at temperatures ranging from 4 to 37°C. This isolate has catalase and oxidase activity and hydrolyses aesculin, DNA and l-tyrosine. Based on phylogenetic analysis using 16S rRNA gene sequences, strain M1-2^T belongs to the genus Joostella and is clearly distinct from the other described species of this genus, Joostella marina (type strain En5^T). The 16S rRNA gene sequence similarity level between M1-2^T and J. marina En5^T is 97.2%, and the DNA–DNA relatedness value between the two strains is 23.9%. Strain M1-2^T contains MK-6 as the major menaquinone and iso-C_15:0, summed feature 3 (C_16:1 ω7c and/or iso-C_15:0 2OH) and iso-C_17:0 3OH as major cellular fatty acids. The DNA G + C content is 32.3 mol%. These data suggest that strain M1-2^T should be classified as a novel species, for which the name Joostella atrarenae sp. nov. is proposed. The type strain for the novel species is M1-2^T (= KCTC 23194^T = NCAIM B.002413^T).     

<Emphasis Type="Italic">Pseudonocardia serianimatus</Emphasis> sp. nov., a novel actinomycete isolated from the surface-sterilized leaves of <Emphasis Type="Italic">Artemisia annua</Emphasis> L.

An aerobic, non-motile, catalase-positive, Gram-stain positive actinomycete designated YIM 63233^T was isolated from the surface-sterilized leaves of Artemisia annua L. and characterized using a polyphasic taxonomic approach. Optimal growth occurred at 20–28°C, pH 6.0–7.0 and in the presence of 0–3% (w/v) NaCl. 16S rRNA gene sequence-based phylogenetic analysis showed that strain YIM 63233^T clustered with species of the genus Pseudonocardia, displaying ≥1.2% sequence divergence with recognized species of this genus (from 98.8 to 94.0%). Relatively low levels of DNA–DNA relatedness were found between strain YIM 63233^T and Pseudonocardia petroleophila IMSNU 22072^T, which supported the classification of strain YIM 63233^T within a novel species of the genus Pseudonocardia. The G + C content of genomic DNA was 72.0 mol%. Strain YIM 63233^T possessed chemotaxonomic markers that were consistent with classification in the genus Pseudonocardia, i.e. the predominant fatty acids were iso-C16:0 (32.27%), C16:0 10-methyl (8.73%) and C17:1ω8c (8.30%), whilst the predominant menaquinone was MK-8(H₄). The diagnostic diamino acid of the cell-wall peptidoglycan was meso-diaminopimelic acid. The major cell wall sugars were glucose, arabinose, galactose, mannose and rhamnose. The results of physiological and biochemical tests and DNA–DNA hybridization allowed the phenotypic and genotypic differentiation of strain YIM 63233^T from its closest phylogenetic neighbours. Therefore, the new isolate YIM 63233^T represents a novel species of the genus Pseudonocardia, for which the name Pseudonocardia serianimatus sp. nov. is proposed. The type strain is YIM 63233^T (=DSM 45302^T = CCTCC AA 208079^T).     

Effects of plant genotype and bacterial strain on <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>-mediated transformation of <Emphasis Type="Italic">Brassica oleracea</Emphasis> L. var. <Emphasis Type="Italic">capitata</Emphasis>

Two inbred lines of Brassica oleracea L. var. capitata were transformed with two Agrobacterium tumefaciens strains harboring resistance to herbicide Basta: AGL1/pDM805 and LBA4404/pGKB5 (LB5-1). Inoculated cotyledons and hypocotyls provided equally good explants and manifested a high percentage of shoot regeneration on MS medium supplemented with 1 mg/l benzyladenine and 0.5 mg/l indole-3-butyric acid. The P₃₄I₅ genotype was superior to P₂₂I₅ in shoot regeneration (48.1 vs. 26.9%), multiplication, and acclimation in the greenhouse (76 vs. 40%). A. tumefaciens AGL1/pDM805 provided more regenerated shoots per explant, especially in the case of cotyledon explants, and the higher transformation rate (up to 35% vs. up to 12%) as compared to LB5-1. Putative transformants survived spraying with 10–30 mg/l phosphinothricin. Transformation was confirmed by GUS assay and PCR analysis in T0 and T1 generations. Published in Russian in Fiziologiya Rastenii, 2007, vol. 54, No. 5, pp. 738–743. The text was submitted by the authors in English.     

High-frequency transformation of<Emphasis Type="Italic"> Lobelia erinus</Emphasis> L. by<Emphasis Type="Italic"> Agrobacterium</Emphasis>-mediated gene transfer

A highly efficient transformation procedure was developed for Lobelia erinus. Leaf or cotyledon discs were inoculated with Agrobacterium tumefaciens strain EHA105 harboring the binary vector plasmid pIG121Hm, which contains a -glucuronidase gene with an intron as a reporter gene and both the neomycin phosphotransferase II and hygromycin phosphotransferase genes as selectable markers. The hygromycin-resistant calli produced on the selection medium were transferred to MS medium supplemented with 0.5 mg/l benzyladenine and 0.2 mg/l indole-3-acetic acid for regeneration of adventitious shoots. Transgenic plants were obtained as a result of the high regeneration rate of the transformed calli, which was as high as 83%. In contrast, no transgenic plant was obtained by the procedure of direct shoot formation following inoculation with A. tumefaciens. Transgenic plants flowered 3–4 months after transformation. Integration of the transgenes was detected using PCR and Southern blot analysis, which revealed that one to several copies were integrated into the genomes of the host plants. The transformation frequency at the stage of whole plants was very high—45% per inoculated disc.Abbreviations BA: 6-Benzyladenine - 2,4-D: 2,4-Dichlorophenoxyacetic acid - GUS: -Glucuronidase - IAA: Indole-3-acetic acidCommunicated by G.C. Phillips     

<Emphasis Type="Italic">Methylophaga murata</Emphasis> sp. nov.: a Haloalkaliphilic Aerobic Methylotroph from Deteriorating Marble

The haloalkaliphilic methylotrophic bacterium (strain Kr3) isolated from material scraped off the deteriorating marble of the Moscow Kremlin masonry has been found to be able to utilize methanol, methylamine, trimethylamine, and fructose as carbon and energy sources. Its cells are gram-negative motile rods multiplying by binary fission. Spores are not produced. The isolate is strictly aerobic and requires vitamin B₁₂ and Na⁺ ions for growth. It is oxidase- and catalase-positive and reduces nitrates to nitrites. Growth occurs at temperatures between 0 and 40°C (with the optimum temperatures being 20–32°C), pH values between 6 and 11 (with the optimum at 8–9), and NaCl concentrations between 0.05 and 3 M (with the optimum at 0.5–1.5 M). The dominant cellular phospholipids are phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin. The major cellular fatty acids are palmitic (C_16:0), palmitoleic (C_16:1), and octadecenoic (C_18:1) acids. The major ubiquinone is Q₈. It accumulates ectoine and glutamate, as well as a certain amount of sucrose, to function as osmoprotectants and synthesizes an exopolysaccharide composed of carbohydrate and protein components. It is resistant to heating at 70°C, freezing, and drying; utilizes methanol, with the resulting production of formic acid, which is responsible for the marble-degrading activity of the isolate; and implements the 2-keto-3-deoxy-6-phosphogluconate variant of the ribulose monophosphate pathway. The G+C content of its DNA is 44.6 mol %. Based on 16S rRNA gene sequencing and DNA-DNA homology levels (23–41%) with neutrophilic and alkaliphilic methylobacteria from the genus Methylophaga, the isolate has been identified as a new species, Methylophaga murata (VKM B-2303^T = NCIMB 13993^T).__________Translated from Mikrobiologiya, Vol. 74, No. 4, 2005, pp. 511–519.Original Russian Text Copyright © 2005 by Doronina, Lee, Ivanova, Trotsenko.     

<Emphasis Type="Italic">Shewanella upenei</Emphasis> sp. nov., a lipolytic bacterium isolated from bensasi goatfish <Emphasis Type="Italic">Upeneus bensasi</Emphasis>

A Gram-staining-negative, motile, non-spore-forming and rod-shaped bacterial strain, 20-23R^T, was isolated from intestine of bensasi goatfish, Upeneus bensasi, and its taxonomic position was investigated by using a polyphasic study. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain 20-23R^T belonged to the genus Shewanella. Strain 20-23R^T exhibited 16S rRNA gene sequence similarity values of 99.5, 99.2, and 97.5% to Shewanella algae ATCC 51192^T, Shewanella haliotis DW01^T, and Shewanella chilikensis JC5^T, respectively. Strain 20-23R^T exhibited 93.1–96.0% 16S rRNA gene sequence similarity to the other Shewanella species. It also exhibited 98.3–98.4% gyrB sequence similarity to the type strains of S. algae and S. haliotis. Strain 20-23R^T contained simultaneously both menaquinones and ubiquinones; the predominant menaquinone was MK-7 and the predominant ubiquinones were Q-8 and Q-7. The fatty acid profiles of strain 20–23R^T, S. algae KCTC 22552^T and S. haliotis KCTC 12896^T were similar; major components were iso-C_15:0, C_16:0, C_16:1 ω7c and/or iso-C_15:0 2-OH and C_17:1 ω8c. The DNA G+C content of strain 20-23R^T was 53.9 mol%. Differential phenotypic properties and genetic distinctiveness of strain 20–23R^T, together with the phylogenetic distinctiveness, revealed that this strain is distinguishable from recognized Shewanella species. On the basis of the data presented, strain 20-23R^T represents a novel species of the genus Shewanella, for which the name Shewanella upenei sp. nov. is proposed. The type strain is 20–23R^T (=KCTC 22806^T =CCUG 58400^T).     

<Emphasis Type="Italic">Gaetbulibacter jejuensis</Emphasis> sp. nov., isolated from seawater

A novel marine bacterium, designated strain CNURIC014^T was isolated from coastal seawater of Jeju Island in Korea. Strain CNURIC014^T formed yellow colonies on marine agar 2216 and the cells were Gram-negative, non-motile, strictly aerobic, rod-shaped. The temperature, pH and NaCl ranges for growth were 15–37°C, pH 6.0–9.0 and 1.0–7.0% NaCl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain CNURIC014^T was most closely related to Gaetbulibacter marinus and Gaetbulibacter saemankumensis, with a sequence similarity of 95.1% and 94.6%, respectively. The DNA G+C content of the strain was 33.1 mol% and the major respiratory quinone was menaquinone-6. The major cellular fatty acids were iso-C_15:1 (22.8%), iso-C_15:0 (18.8%), summed feature 3 (iso-C_15:0 2-OH/C_16:1 ω7c, 12.9%) and iso-C_17:0 3-OH (11.5%). On the basis of phenotypic, phylogenetic, and genotypic data, strain CNURIC014^T represents a novel species within the genus Geatbulibacter, for which the name Gaetbulibacter jejuensis sp. nov. is proposed. The type strain is CNURIC014^T(=KCTC 22615^T =JCM 15976^T).