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Volume 137,
Issue 8,
1991
Volume 137, Issue 8, 1991
- Physiology And Growth
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Bacterial metabolism of 3-chloroacrylic acid
More LessTwo bacterial strains were isolated with 3-chloroacrylic acid (CAA) as sole source of carbon and energy. Strain CAA1, a Pseudomonas cepacia sp., was capable of growth with only the cis-isomer of CAA. Strain CAA2, a coryneform bacterium, utilized both isomers of CAA as sole source of carbon and energy. Strain CAA1 contained cis-CAA hydratase and strain CAA2 contained two hydratases, one with cis-CAA hydratase activity and one with trans-CAA hydratase activity. The product of the hydratase activities with CAA was malonate semialdehyde. In both strains malonate semialdehyde was subsequently decarboxylated by a cofactor-independent decarboxylase yielding acetaldehyde and CO2.
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- Plant-Microbe Interactions
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Isolation and characterization of behavioural mutants and genes of Agrobacterium tumefaciens
Agrobacterium tumefaciens exhibits an unusual flagellation pattern in the electron microscope. The typical pattern is of two flagella in a polar or sub-polar tuft at one pole only, plus one or two lateral flagella on each side. Twenty independent behavioural mutants were isolated, after Tn5 mutagenesis, and divided into several classes. Fourteen non-motile mutants were differentiable into non-flagellate and flagellate subclasses, one non-chemotactic mutant appeared to have a defect in the intracellular signalling pathway, and one other mutant displayed an unusual pattern of polar cell pairing. Four mutants with impaired swarming ability were also isolated. Root colonization ability was impaired in the non-motile mutant for which this property was tested. From all but three of the mutants, the Tn5 insertion sites, including flanking sequences, have been cloned and used as probes to isolate a set of seven cosmids from a wild-type A. tumefaciens library. No hybridization to Escherichia coli or Pseudomonas DNA was detected for the A. tumefaciens mutant flanking sequences, but varying degrees of similarity to Rhizobium meliloti behavioural genes were detected. By complementation of A. tumefaciens behavioural mutants and hybridization of mutant flanking sequences, one cosmid insert was shown to contain at least nine behavioural loci, and another, linked to it on the chromosome, at least three. This suggests that, as in other organisms, behavioural genes are clustered in the A. tumefaciens genome.
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- Systematics
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A numerical classification of the genera Streptomyces and Streptoverticillium using miniaturized physiological tests
More LessEight hundred and twenty-one strains of the genera Streptomyces and Streptoverticillium were physiologically characterized using a total of 329 miniaturized tests. Overall similarities of all strains were determined by numerical taxonomic techniques using the UPGMA algorithm and the SSM and the S J coefficients as measures of similarity. Test error was within acceptable limits. Comparison of photometric and visual test reading revealed overall differences of 7·45%. A total of 15 major clusters (six or more strains), 34 minor clusters (less than six strains) and 40 single-member clusters were defined at the 81.5% similarity level (S SM). Two clusters containing physiologically, and in some cases morphologically and genetically, different groups could be further subdivided at the 84·0% similarity level (S SM). Generally, similar groupings were obtained with the Jaccard coefficient at similarity levels ranging from 59·6% to 64·6% similarity (S J), with changes in the definition of clusters and subclusters. The cophenetic correlation coefficients r CS for the UPGMA/S J and the UPGMA/S sm analysis were 0·6929 and 0·8239, respectively. Several phena showed significant overlap with others, indicating the physiological variability within the species. The phenetic data in most cases confirm the major phena of the study of Williams et al. (1983), Journal of General Microbiology 129, 1743-1813 (although the cluster-groups defined in that study could only be detected in part) and the results indicate that the genus Streptomyces is still overspeciated. Some of the major groupings obtained were very much in line with chemotaxonomic and genetical data. However, several clusters containing only a few strains should be regarded as preliminary ‘species’ until further information is available. The majority of Streptoverticillium strains presently assigned to different species formed a homogeneous subcluster defined at the 84·0% similarity level (S SM). Thus, on the basis of numerical phenetic and (published) molecular genetic and chemotaxonomic data, our study supports the suggestion that members of the genus Streptoverticillium be reclassified into the genus Streptomyces.
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Probabilistic identification of streptomycetes using miniaturized physiological tests
More LessA frequency matrix of positive results for phena defined in a previous phenetic classification was constructed. A total of 329 physiological characters from 782 strains was taken as the basis for this identification matrix. The minimum number of diagnostic characters for the matrix was selected using computer programs for calculation of different separation indices (CHARSEP) and selection of group diagnostic properties (DIACHAR). The resulting matrix consisted of 52 phena versus 50 characters. Overlap of phena was found to be relatively small (OVERMAT program). Identification scores for the most typical hypothetical organism of each phenon was satisfactory (MOSTTYP program). The matrix was evaluated theoretically and practically (MATIDEN program). For members of major clusters and subclusters, e.g. Streptomyces albidoflavus, S. halstedii, S. griseus, S. fradiae, S. rimosus and S. albus, identification scores were high. Organisms of phena containing only small numbers of strains could be identified correctly, but with lower accuracy. The identification rate of the matrix (Willcox probability > 0.90) in the theoretical evaluation was 84.39%, and in the practical evaluation 78.12%.
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Mass spectrometry as a tool for identifying group D2 corynebacteria by their fatty acid profiles
More LessCorynebacterium group D2 (CGD2) are lipophilic antibiotic-multiresistant bacteria involved in some infections of immunocompromised patients. The fatty acid composition and structure of different strains was established by several mass spectrometric methods, particularly negative ion tandem mass spectrometry coupled with capillary gas chromatography. Non-hydroxylated fatty acid profiles of three strains of CGD2 (ATCC 43042, ATCC 43043, ATCC 43044) were almost identical and revealed the presence of several straight chain unsaturated fatty acids from the ω-9 series, with even carbon numbers ranging from 14 to 24. Branched saturated fatty acids were mainly anteiso-heptadecanoic acid and tuberculostearic acid. Surprisingly, a relatively large quantity of 10-methylene octadecanoic acid was found. The non-hydroxylated fatty acid profile of one rare β-lactam susceptible strain (SC1) was different; 10-methylene octadecanoic acid was lacking whereas tuberculostearic acid was much more abundant. In contrast, the four CGD2 strains displayed highly similar mycolic acid patterns. The major mycolic acid species corresponded to C32, C30 and C28 bis-unsaturated with a double bond on each branch at the ω-9 position. The comparison of the mycolic acid composition and structure with those of other medically important corynebacteria strains, revealed a characteristic pattern for CGD2 strains, and CGD2 strains were easily distinguished from Corynebacterium jeikeium (CIP 82.51).
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Molecular and evolutionary relationships among enteric bacteria
More LessClassification of bacterial species into genera has traditionally relied upon variation in phenotypic characteristics. However, these phenotypes often have a multifactorial genetic basis, making unambiguous taxonomic placement of new species difficult. By designing evolutionarily conserved oligonucleotide primers, it is possible to amplify homologous regions of genes in diverse taxa using the polymerase chain reaction and determine their nucleotide sequences. We have constructed a phylogeny of some enteric bacteria, including five species classified as members of the genus Escherichia, based on nucleotide sequence variation at the loci encoding glyceraldehyde-3-phosphate dehydrogenase and outer membrane protein 3A, and compared this genealogy with the relationships inferred by biotyping. The DNA sequences of these genes defined congruent and robust phylogenetic trees indicating that they are an accurate reflection of the evolutionary history of the bacterial species. The five species of Escherichia were found to be distantly related and, contrary to their placement in the same genus, do not form a monophyletic group. These data provide a framework which allows the relationships of additional species of enteric bacteria to be inferred. These procedures have general applicability for analysis of the classification, evolution, and epidemiology of bacterial taxa.
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- Corrigendum
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