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  • 1
    Online Resource
    Online Resource
    Identification of low-temperature-regulated genes in the fire blight pathogen Erwinia amylovora
    Canadian Science Publishing ; 2006
    In:  Canadian Journal of Microbiology Vol. 52, No. 5 ( 2006-05-01), p. 468-475
    Details
    In: Canadian Journal of Microbiology, Canadian Science Publishing, Vol. 52, No. 5 ( 2006-05-01), p. 468-475
    Abstract: Genes involved in pathogenicity of several plant pathogens were shown to be induced at relatively cold temperatures. Loci from the fire blight pathogen Erwinia amylovora (Burrill) induced at 18 °C were identified using the miniTn5 transposon that contains the promoterless reporter gene gusA coding for β-glucuronidase (GUS). Certain mutants (2.7%) expressed GUS predominantly at 18 °C on minimal medium plates, indicating that the transposon had been inserted downstream of a putatively thermoregulated promoter. Those mutants were further screened with a quantitative GUS fluorometric assay. A total of 21 mutants were selected: 19 mutants had a transposon insertion in temperature-dependent genetic loci, with a 2.2- to 6.3-fold induction of gusA gene expression at 18 °C, and two mutants with impaired growth at 18 °C. Some of these genetic loci encoded (i) proteins implicated in flagella biosynthesis, biotin biosynthesis, multi-drug efflux, and type II secretion protein, and (ii) proteins of unknown function.Key words: fire blight, Erwinia amylovora, transposon mutagenesis, gene regulation, low temperature.
    Type of Medium: Online Resource
    ISSN: 0008-4166 , 1480-3275
    URL: Article
    DOI: 10.1139/w05-153
    RVK:
    WA 15000
    Language: English
    Publisher: Canadian Science Publishing
    Publication Date: 2006
    detail.hit.zdb_id: 280534-0
    detail.hit.zdb_id: 1481972-7
    SSG: 12
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  • 2
    Online Resource
    Online Resource
    Contribution of alginate and levan production to biofilm formation by Pseudomonas syringae
    Microbiology Society ; 2006
    In:  Microbiology Vol. 152, No. 10 ( 2006-10-01), p. 2909-2918
    Details
    In: Microbiology, Microbiology Society, Vol. 152, No. 10 ( 2006-10-01), p. 2909-2918
    Abstract: Exopolysaccharides (EPSs) play important roles in the attachment of bacterial cells to a surface and/or in building and maintaining the three-dimensional, complex structure of bacterial biofilms. To elucidate the spatial distribution and function of the EPSs levan and alginate during biofilm formation, biofilms of Pseudomonas syringae strains with different EPS patterns were compared. The mucoid strain PG4180.muc, which produces levan and alginate, and its levan- and/or alginate-deficient derivatives all formed biofilms in the wells of microtitre plates and in flow chambers. Confocal laser scanning microscopy with fluorescently labelled lectins was applied to investigate the spatial distribution of levan and an additional as yet unknown EPS in flow-chamber biofilms. Concanavalin A (ConA) bound specifically to levan and accumulated in cell-depleted voids in the centres of microcolonies and in blebs. No binding of ConA was observed in biofilms of the levan-deficient mutants or in wild-type biofilms grown in the absence of sucrose as confirmed by an enzyme-linked lectin-sorbent assay using peroxidase-linked ConA. Time-course studies revealed that expression of the levan-forming enzyme, levansucrase, occurred mainly during early exponential growth of both planktonic and sessile cells. Thus, accumulation of levan in biofilm voids hints to a function as a nutrient storage source for later stages of biofilm development. The presence of a third EPS besides levan and alginate was indicated by binding of the lectin from Naja mossambica to a fibrous structure in biofilms of all P. syringae derivatives. Production of the as yet uncharacterized additional EPS might be more important for biofilm formation than the syntheses of levan and alginate.
    Type of Medium: Online Resource
    ISSN: 1350-0872 , 1465-2080
    URL: Article
    DOI: 10.1099/mic.0.28875-0
    Language: English
    Publisher: Microbiology Society
    Publication Date: 2006
    detail.hit.zdb_id: 2008736-6
    SSG: 12
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  • 3
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    Online Resource
    The algT Gene of Pseudomonas syringae pv. glycinea andNew Insights into the Transcriptional Organization of the algT-muc Gene Cluster
    American Society for Microbiology ; 2006
    In:  Journal of Bacteriology Vol. 188, No. 23 ( 2006-12), p. 8013-8021
    Details
    In: Journal of Bacteriology, American Society for Microbiology, Vol. 188, No. 23 ( 2006-12), p. 8013-8021
    Abstract: The phytopathogenic bacterium Pseudomonas syringae pv. glycinea infects soybean plants and causes bacterial blight. In addition to P. syringae , the human pathogen Pseudomonas aeruginosa and the soil bacterium Azotobacter vinelandii produce the exopolysaccharide alginate, a copolymer of d -mannuronic and l -guluronic acids. Alginate production in P. syringae has been associated with increased fitness and virulence in planta. Alginate biosynthesis is tightly controlled by proteins encoded by the algT-muc regulatory gene cluster in P. aeruginosa and A. vinelandii . These genes encode the alternative sigma factor AlgT (σ 22 ), its anti-sigma factors MucA and MucB, MucC, a protein with a controversial function that is absent in P. syringae , and MucD, a periplasmic serine protease and homolog of HtrA in Escherichia coli . We compared an alginate-deficient algT mutant of P. syringae pv. glycinea with an alginate-producing derivative in which algT is intact. The alginate -producing derivative grew significantly slower in vitro growth but showed increased epiphytic fitness and better symptom development in planta. Evaluation of expression levels for algT , mucA , mucB , mucD , and algD , which encodes an alginate biosynthesis gene, showed that mucD transcription is not dependent on AlgT in P. syringae in vitro. Promoter mapping using primer extension experiments confirmed this finding. Results of reverse transcription-PCR demonstrated that algT , mucA , and mucB are cotranscribed as an operon in P. syringae . Northern blot analysis revealed that mucD was expressed as a 1.75-kb monocistronic mRNA in P. syringae .
    Type of Medium: Online Resource
    ISSN: 0021-9193 , 1098-5530
    URL: Article
    DOI: 10.1128/JB.01160-06
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2006
    detail.hit.zdb_id: 1481988-0
    SSG: 12
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  • 4
    Online Resource
    Online Resource
    Thermo-responsive expression and differential secretion of the extracellular enzyme levansucrase in the plant pathogenic bacterium Pseudomonas syringae pv. glycinea
    Oxford University Press (OUP) ; 2006
    In:  FEMS Microbiology Letters Vol. 265, No. 2 ( 2006-12), p. 178-185
    Details
    In: FEMS Microbiology Letters, Oxford University Press (OUP), Vol. 265, No. 2 ( 2006-12), p. 178-185
    Type of Medium: Online Resource
    ISSN: 0378-1097 , 1574-6968
    URL: Issue
    URL: Article
    DOI: 10.1111/fml.2006.265.issue-2
    DOI: 10.1111/j.1574-6968.2006.00486.x
    RVK:
    WA 15000
    Language: English
    Publisher: Oxford University Press (OUP)
    Publication Date: 2006
    detail.hit.zdb_id: 1501716-3
    SSG: 12
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