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Classification of 'Nocardioides fulvus' IFO 14399 and Nocardioides sp. ATCC 39419 in Kribbella gen. nov., as Kribbella flavida sp. nov. and Kribbella sandramycini sp. nov.

By 16S rDNA sequence analysis, two strains identified previously as members of the genus Nocardioides, namely 'Nocardioides fulvus' IFO 14399 and Nocardioides sp. ATCC 39419, were found to form lines of descent distinct from the genus Nocardioides. Therefore, the two strains were studied taxonomically in detail by phenotypic, particularly chemotaxonomic, characterization, and by phylogenetic analysis including other related taxa. The genomic DNA G + C contents of 'N. fulvus' IFO 14399 and Nocardioides sp. ATCC 39419 are 70 and 68 mol%, respectively. The two strains contain LL-diaminopimelic acid as the diamino acid in their peptidoglycan, together with L-alanine at position 1 of the peptide subunit. The predominant menaquinone found in the two strains is MK-9(H4). The major cellular fatty acid found in the two strains is anteiso-C15:0' which is a minor component of members of the genus Nocardioides. The diagnostic polar lipid of the two strains is phosphatidylcholine. A phylogenetic tree was constructed which showed that the two strains from evolutionary lineages distinct from other LL-diaminopimelic-acid-containing taxa such as, for example, the genus Nocardioides. On the basis of our data, a new genus, Kribbella gen. nov., and two new species, Kribbella flavida gen. nov., sp. nov. and Kribbellla sandramycini gen. nov., sp. nov., are proposed. It is also proposed that the new genus be included in the family Nocardioidaceae.

Actinomycetales↗

Gracilibacillus gen. nov., with description of Gracilibacillus halotolerans gen. nov., sp. nov.; transfer of Bacillus dipsosauri to Gracilibacillus dipsosauri comb. nov., and Bacillus salexigens to the genus Salibacillus gen. nov., as Salibacillus salexigens comb. nov.

A Gram-positive, extremely halotolerant bacterium was isolated from the Great Salt Lake, Utah, USA. The strain, designated NNT (= DSM 11805T), was strictly aerobic, rod-shaped, motile by peritrichous flagella and spore-forming. Strain NNT grew at salinities of 0-20% (w/v) NaCl. A distinctive feature of strain NNT was its optimal growth in salt-free medium. The polar lipid pattern of strain NNT consisted of phosphatidyl glycerol, diphosphatidyl glycerol and two phospholipids of unknown structure. The G + C content of its DNA was 38 mol%. The morphological, physiological and, particularly, the 16S rDNA sequence data, showed that strain NNT was associated with 'Bacillus group 1'. However, the organisms showing the greatest degree of sequence similarity to strain NNT were members of the genus Halobacillus and the species Marinococcus albus, Virgibacillus pantothenticus, Bacillus salexigens and Bacillus dipsosauri. On the basis of chemotaxonomic data, strain NNT was shown to be chemically most similar to B. salexigens and B. dipsosauri, with the greatest degree of similarity being shown to the latter organism. This was consistent with the 16S rDNA sequence data. Members of the genus Halobacillus comprise a chemically distinct group and can easily be distinguished from all other organisms of 'Bacillus group 1'. On the basis of the 16S rDNA data, chemotaxonomy and the physiology of strain NNT, it is proposed that this organism is a member of a new species, within a new genus, for which the name Gracilibacillus halotolerans is proposed. It is also proposed that B. dipsosauri be transferred to this genus as Gracilibacillus dipsosauri comb. nov. and that B. salexigens be transferred to the genus Salibacillus gen. nov., as Salibacillus salexigens comb. nov. Finally, additional data is provided to support the transfer of Bacillus pantothenticus to the genus Virgibacillus, as Virgibacillus pantothenticus Heyndrickx et al. (1998).

Bacillus↗

Microbacterium kitamiense sp. nov., a new polysaccharide-producing bacterium isolated from the wastewater of a sugar-beet factory.

Two strictly aerobic, heterotrophic and mesophilic new organisms, strains Kitami A1 and Kitami C2T, were isolated from the wastewater of a sugar-beet factory in Kitami City, Hokkaido, Japan. In batch cultures, these organisms produced both insoluble and soluble exopolysaccharides (EPSs) utilizing sucrose as the sole carbon source. The G + C contents of the strains Kitami C2T and Kitami A1 were 69.2 mol%. Both strains had anteiso-C15:0 acid, anteiso-C17:0 acid and iso-C16:0 as major components. The major isoprenoid quinones from these strains included menaquinone-11 and menaquinone-12. Physiological and biochemical characterization, phylogenetic analysis and DNA-DNA relatedness indicated that these two organisms are new species of the genus Microbacterium, for which the name Microbacterium kitamiense is proposed. The type strain of M. kitamiense is strain Kitami C2T (= JCM 10270T).

Actinomycetales↗

Ornithinicoccus hortensis gen. nov., sp. nov., a soil actinomycete which contains L-ornithine.

Two Gram-positive coccoid, non-motile bacteria with L-ornithine as diagnostic diamino acid of the peptidoglycan and an interpeptide bridge of L-Orn<--Gly(1,2)<--D-Glu were isolated from a sample of garden soil. The major menaquinone is MK-8(H4). 13-methyl and 12-methyl tetradecanoic acids are the predominant fatty acids. The polar lipids are phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylserine and two unknown phospholipids. Mycolic acids are absent. The DNA base composition is 72 mol% G + C. Recent comparative 16S rDNA studies revealed that strains HKI 0125T and HKI 0131 represent a novel lineage adjacent to the family Intrasporangiaceae of the order Actinomycetales but distinct from the previously described genera of this family. On the basis of the genotypic, chemotaxonomic, morphological and physiological characteristics of these two isolates it is proposed to classify HKI 0125T and HKI 0131 in a new genus and species for which the name Ornithinicoccus hortensis gen. nov., sp. nov. is proposed. The type strain is HKI 0125T (= DSM 12335T).

Actinomycetales↗

Nocardiopsis kunsanensis sp. nov., a moderately halophilic actinomycete isolated from a saltern.

A moderately halophilic actinomycete, designated HA-9T, was isolated from a saltern in Kunsan, Republic of Korea, and was the subject of polyphasic identification. Analysis of 16S rDNA indicated that the isolate belonged to the genus Nocardiopsis, but differed genetically from other Nocardiopsis species. Strain HA-9T contained meso-diaminopimelic acid, no diagnostic sugars, hexa- or octa-hydrogenated menaquinones with 10 isoprene units, straight-chain saturated or monounsaturated, iso-, anteiso-, 10-methyl branched fatty acids with 13-18 carbons and type III phospholipids. All of these characters consistently assign the isolate to the genus Nocardiopsis. All of the validly described Nocardiopsis species, including moderately halophilic Nocardiopsis halophila, can be differentiated from the saltern isolate using morphological and physiological traits. On the basis of polyphasic evidence, the name Nocardiopsis kunsanensis sp. nov. is proposed for strain HA-9T (= KCTC 9831T), which is designated the type strain.

Actinomycetales↗

Sphingomonas pituitosa sp. nov., an exopolysaccharide-producing bacterium that secretes an unusual type of sphingan.

Strain EDIVT, an exopolysaccharide-producing bacterium, was subjected to polyphasic characterization. The bacterium produced copious amounts of an extracellular polysaccharide, forming slimy, viscous, intensely yellow-pigmented colonies on Czapek-Dox (CZD) agar. The culture fluids of the liquid version of CZD medium were highly viscous after cultivation for 5 d. Cells of strain EDIVT were Gram-negative, catalase-positive, oxidase-negative, nonspore-forming, rod-shaped and motile. Comparisons of 16S rDNA gene sequences demonstrated that EDIVT clusters phylogenetically with the species of the genus Sphingomonas sensu stricto. The G+C content of the DNA (64.5 mol%), the presence of ubiquinone Q-10, the presence of 2-hydroxymyristic acid (14:0 2-OH) as the major hydroxylated fatty acid, the absence of 3-hydroxy fatty acids and the detection of sym-homospermidine as the major component in the polyamine pattern, together with the presence of sphingoglycolipid, supported this delineation. 16S rDNA sequence analysis indicated that strain EDIVT is most closely related (99.4% similarity) to Sphingomonas trueperi LMG 2142T. DNA-DNA hybridization showed that the level of relatedness to S. trueperi is only 45.5%. Further differences were apparent in the cellular fatty acid profile, the polar lipid pattern, the Fourier-transform infrared spectrum and whole-cell proteins and in a number of biochemical characteristics. On the basis of the estimated phylogenetic position derived from 16S rDNA sequence data, DNA-DNA reassociation and phenotypic differences, strain EDIVT (= CIP 106154T = DSM 13101T) was recognized as a new species of Sphingomonas, for which the name Sphingomonas pituitosa sp. nov. is proposed. A component analysis of the exopolysaccharide (named PS-EDIV) suggested that it represents a novel type of sphingan composed of glucose, rhamnose and an unidentified sugar. Glucuronic acid, which is commonly found in sphingans, was absent. The mean molecular mass of PS-EDIV was approximately 3 x 10(6) Da.

Base Composition↗

Pannonibacter phragmitetus gen. nov., sp. nov., a novel alkalitolerant bacterium isolated from decomposing reed rhizomes in a Hungarian soda lake.

Three alkalitolerant bacterial strains were isolated from the surface of decomposing rhizomes of reed [Phragmites australia (Cav.) Trin. et Steudel] in Lake Fertö (Hungary). Cells of the novel isolates were Gram-negative, motile rods and formed star-shaped aggregates. They were facultatively anaerobic and chemo-organotrophic. Bacteriochlorophyll a was not synthesized under aerobic conditions. The strains were catalase and oxidase positive, produced acid from D-glucose under aerobic and anaerobic conditions and reduced nitrate to nitrogen. They tolerated pH values from 7.0 to 11.0 and grew in the absence of NaCl as well as in up to 5% (w/v) NaCl. The G + C content of the DNA was 64.6 mol% and the major isoprenoid quinone was Q-10. The dominant cellular fatty acid was C18 : 1omega7c. The cell membrane contained phosphatidyl glycerol, diphosphatidyl glycerol, phosphatidyl ethanolamine, phosphatidyl serine and one unknown phospholipid as polar lipids. Polyphasic taxonomic characterization revealed that strain C6/19T is most closely related to the Stappia-Roseibium cluster in the alpha-subclass of the Proteobacteria (showing 95.8-93.6% 16S rDNA sequence similarity). According to the phylogenetic and phenotypic evidence presented, a new genus and species is proposed, Pannonibacter phragmitetus gen. nov., sp. nov. The type strain is C6/19T (=DSM 14782T =NCAIM B02025T).

Alkalies↗

Cell-Wall Autohydrolysis in Isolated Endosperms of Lettuce (Lactuca sativa L.).

Cell walls prepared from the endosperm tissue of hydrated lettuce (Lactuca sativa L.) seeds undergo autohydrolysis. Release of carbohydrates is most rapid (0.4-0.6 [mu]g per endosperm) within the 1st h of incubation in buffer, but substantial autolysis is sustained for at least 10 h. Autolysis is temperature sensitive, and the optimum rate occurs at pH 5. The rate of autolysis increases markedly in the period just prior to radicle emergence. The cell-wall polysaccharide composition in micropylar and lateral endosperm regions differs significantly; the micropylar walls are rich in arabinose and glucose with substantially lower amounts of mannose. Although walls prepared from both micropylar and lateral regions undergo autolysis, micropylar walls release carbohydrates at a higher rate than lateral walls. Autolysis products elute as large polymers when subjected to size-exclusion chromatography, suggesting that endo-enzyme activity is responsible for release of fragments containing arabinose, galactose, mannose, and uronic acids. Arabinose, galactose, mannose, and glucose are also released as monomers. As a function of time, the ratio of polymers to monomers decreases, indicating that exo-enzyme activity is also present. Thermoinhibition or treatment with abscisic acid suppresses germination and reduces the rates of autolysis of walls isolated from the endosperm by about 25%. Treatments that alleviate thermoinhibition (kinetin and gibberellic acid) increase the rates of autolysis by 20 to 30% when compared to thermoinhibited controls.

Journal Article↗

Cell Wall and Extensin mRNA Changes during Cold Acclimation of Pea Seedlings.

During exposure to 2 degrees C, pea (Pisum sativum) seedlings cold acclimated to a killing temperature of -6 degrees C. Associated with this increase in freezing resistance was an increase in the weight of cell walls and changes in wall composition. Arabinosyl content increased by 100%, while other cell wall glycosyl residues and cellulose increased by about 20%. The cell wall hydroxyproline content increased by 80%. Arabinose and hydroxyproline are both major components of the structural cell wall glycoprotein, extensin. The increase in these components indicates that the level of extensin in the cell wall increases during cold acclimation. Northern blot analysis, using the pDC5A1 genomic clone as a probe, revealed a more than three-fold increase in total extensin mRNA during exposure to cold temperature. Specific extensin transcripts of 6.0, 4.5, 3.5, 2.6, 2.3, 1.8, and 1.5 kilobases were identified. Those at 6.0, 2.6, and 1.5 kilobases were especially promoted by low temperature treatment. The rise in extensin during cold acclimation may be regulated, at least in part, at the gene level. The possible structural role of this protein in freezing protection is discussed.

Journal Article↗

Rare, suppurative pulmonary infection caused by Nocardiopsis dassonvillei recognized by glycolipid markers.

An opportunistic actinomycete was isolated as the only etiological agent of a severe, suppurative pulmonary infection. The strain was rapidly recognised as Nocardiopsis by the taxonomically important and immunologically active glycolipid markers (G1 and G2). Identification of the clinical isolate, from a group of actinomycetes mainly known as soil habitants, was definitely proved by chemotaxonomic studies (cell wall/sugar, phospholipid and fatty acid types) as well as by genomic data (GC content, DNA-DNA reassociation). The level of DNA-DNA homology of the clinical actinomycete, in comparison with other reference members of this genus, revealed the highest (88%) relatedness to Nocardiopsis dassonvillei. The results confirmed the value and generic specificity of glycolipid markers from Nocardiopsis, the first time used for rapid recognition of a clinical strain causing a nocardiosis-like disease.

Actinomycetales Infections↗

Taxonomic relationships among strains of the anaerobic bacterium Bacteroides ruminicola determined by DNA and extracellular polysaccharide analysis.

DNA and extracellular polysaccharide (EPS) analyses were performed on 14 strains of Bacteroides ruminicola. The guanine-plus-cytosine (G+C) base contents, determined from the buoyant densities of chromosomal DNAs, showed a broad range of values, from 37.6 to 50.9 mol%. DNA hybridization showed generally low DNA relatedness among the strains. Seven strains formed two groups of closely related bacteria consisting of five (group 1) and two (group 2) strains, and another strain, E42g, showed moderate relatedness to group 1 strains. However, the remaining six strains were not related to any of the other strains. DNA reassociation indicates that the strains constitute a genetically diverse group representing as many as nine separate species. EPS analysis showed that the strains produced EPS with rather uniform sugar compositions, which did not correlate with strain relationships determined by DNA analysis. Four strains had EPS with acidic sugars or unknown compounds. The EPS of strain 20-63 contained the unusual acidic sugar 4-O-(1-carboxyethyl)-rhamnose. This monosaccharide has been shown to occur in nature in only one other bacterial species.

Animals↗

Analysis of mannans of two relatively avirulent mutant strains of Candida albicans.

We previously reported the isolation of two cerulenin-resistant mutant strains of Candida albicans 4918 that differ in adherence properties and are less virulent than the parental strain. In addition, biochemical characterization demonstrated significant differences in both protein and polysaccharide composition of cell wall material between the mutant and wild-type strains. These observations prompted studies concerning the chemical structure of mannans in these strains. After extraction and subsequent purification by ion-exchange chromatography, mannan fractions were subjected to either mild acid hydrolysis, alkali hydrolysis, or acetylation followed by acetolysis. Acid- and alkali-modified mannans were studied by proton magnetic resonance spectroscopy, and released products were analyzed by high-performance liquid chromatography on an Aminex HPX-42A column. The results demonstrated quantitative and qualitative differences between mannooligosaccharides of the wild-type and mutant strains in the identity of released oligosaccharides as well as in linkage of the oligosaccharides to the protein backbone.

Acetylation↗

Cloning, characterization, and inactivation of the gene pbpC, encoding penicillin-binding protein 3 of Staphylococcus aureus.

The gene pbpC from Staphylococcus aureus was sequenced: it encodes a 691-amino-acid protein with all of the conserved motifs of a class B high-molecular-weight penicillin-binding protein (PBP), including the transpeptidase conserved motifs SXXK, SXN, and KTG. Insertional inactivation of pbpC and introduction of the intact gene in a laboratory mutant missing PBP 3 showed that the pbpC gene encodes the staphylococcal PBP 3. Inactivation of pbpC caused no detectable change in the muropeptide composition of cell wall peptidoglycan and had only minimum, if any, effect on growth rates, but caused a small but significant decrease in rates of autolysis. Cells of abnormal size and shape and disoriented septa were produced when bacteria with inactivated pbpC were grown in the presence of a sub-MIC of methicillin.

Amino Acid Sequence↗

Characteristics of CDC group 1 and group 1-like coryneform bacteria isolated from clinical specimens.

Fifteen strains of CDC group 1 coryneform and biochemically similar bacteria were isolated from clinical specimens. Of the 15 strains isolated, 11 were derived from abscesses and purulent lesions, mostly from the upper part of the body, and 3 were grown from blood cultures. Nine strains were associated with mixed anaerobic but no other aerobic flora. Seven strains exhibited the classical biochemical profile of CDC coryneform group 1; however, eight strains were unable to reduce nitrate and were called "group 1-like." Other reactions to differentiate CDC group 1 and group 1-like coryneform rods include alpha-hemolysis on human blood agar, fermentation of adonitol, and the presence of alkaline phosphatase. Fifteen strains showed marked CAMP reactions on different erythrocyte agars. Gas-liquid chromatography of volatile and nonvolatile fatty acids as well as cellular fatty acid patterns and the composition of cell wall components suggest that CDC group 1 and group 1-like coryneform bacteria do not belong to the genus Corynebacterium but possibly to the genus Actinomyces or Arcanobacterium. DNA-DNA hybridization studies revealed that group 1 and group 1-like strains represent different species.

Actinomycetales↗