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  • 1
    In: Journal of Phytopathology, Wiley, Vol. 158, No. 4 ( 2010-04), p. 288-295
    Type of Medium: Online Resource
    ISSN: 0931-1785 , 1439-0434
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2010
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    detail.hit.zdb_id: 633487-8
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  • 2
    In: Microbial Biotechnology, Wiley, Vol. 2, No. 4 ( 2009-07), p. 465-475
    Abstract: Erwinia amylovora causes fire blight on several plant species such as apple and pear, which produce diverse phytoalexins as defence mechanisms. An evolutionary successful pathogen thus must develop resistance mechanisms towards these toxic compounds. The E. amylovora outer membrane protein, TolC, might mediate phytoalexin resistance through its interaction with the multidrug efflux pump, AcrAB. To prove this, a tolC mutant and an acrB/tolC double mutant were constructed. The minimal inhibitory concentrations of diverse antimicrobials and phytoalexins were determined for these mutants and compared with that of a previously generated acrB mutant. The tolC and arcB/tolC mutants were considerably more susceptible than the wild type but showed similar levels as the acrB mutant. The results clearly indicated that neither TolC nor AcrAB significantly interacted with other transport systems during the efflux of the tested toxic compounds. Survival and virulence assays on inoculated apple plants showed that pathogenicity and the ability of E. amylovora to colonize plant tissue were equally impaired by mutations of tolC and acrB/tolC . Our results allowed the conclusion that TolC plays an important role as a virulence and fitness factor of E. amylovora by mediating resistance towards phytoalexins through its exclusive interaction with AcrAB.
    Type of Medium: Online Resource
    ISSN: 1751-7915 , 1751-7915
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2009
    detail.hit.zdb_id: 2406063-X
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  • 3
    Online Resource
    Online Resource
    Scientific Societies ; 2004
    In:  Molecular Plant-Microbe Interactions® Vol. 17, No. 1 ( 2004-01), p. 43-54
    In: Molecular Plant-Microbe Interactions®, Scientific Societies, Vol. 17, No. 1 ( 2004-01), p. 43-54
    Abstract: The enterobacterium Erwinia amylovora causes fire blight on members of the family Rosaceae, with economic importance on apple and pear. During pathogenesis, the bacterium is exposed to a variety of plant-borne antimicrobial compounds. In plants of Rosaceae, many constitutively synthesized isoflavonoids affecting microorganisms were identified. Bacterial multidrug efflux transporters which mediate resistance toward structurally unrelated compounds might confer tolerance to these phytoalexins. To prove this hypothesis, we cloned the acrAB locus from E. amylovora encoding a resistance nodulation division-type transport system. In Escherichia coli, AcrAB of E. amylovora conferred resistance to hydrophobic and amphiphilic toxins. An acrB-deficient E. amylovora mutant was impaired in virulence on apple rootstock MM 106. Furthermore, it was susceptible toward extracts of leaves of MM 106 as well as to the apple phytoalexins phloretin, naringenin, quercetin, and (+)-catechin. The expression of acrAB was determined using the promoterless reporter gene egfp. The acrAB operon was up-regulated in vitro by the addition of phloretin and naringenin. The promoter activity of acrR, encoding a regulatory protein involved in acrAB expression, was increased by naringenin. In planta, an induction of acrAB was proved by confocal laser scanning microscopy. Our results strongly suggest that the AcrAB transport system plays an important role as a protein complex required for virulence of E. amylovora in resistance toward apple phytoalexins and that it is required for successful colonization of a host plant.
    Type of Medium: Online Resource
    ISSN: 0894-0282 , 1943-7706
    Language: English
    Publisher: Scientific Societies
    Publication Date: 2004
    detail.hit.zdb_id: 2037108-1
    detail.hit.zdb_id: 743331-1
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  • 4
    In: Microbiology, Microbiology Society, Vol. 154, No. 9 ( 2008-09-01), p. 2700-2708
    Type of Medium: Online Resource
    ISSN: 1350-0872 , 1465-2080
    Language: English
    Publisher: Microbiology Society
    Publication Date: 2008
    detail.hit.zdb_id: 2008736-6
    detail.hit.zdb_id: 1180712-X
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  • 5
    Online Resource
    Online Resource
    Microbiology Society ; 2008
    In:  Microbiology Vol. 154, No. 2 ( 2008-02-01), p. 413-421
    In: Microbiology, Microbiology Society, Vol. 154, No. 2 ( 2008-02-01), p. 413-421
    Type of Medium: Online Resource
    ISSN: 1350-0872 , 1465-2080
    Language: English
    Publisher: Microbiology Society
    Publication Date: 2008
    detail.hit.zdb_id: 2008736-6
    detail.hit.zdb_id: 1180712-X
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  • 6
    In: Applied and Environmental Microbiology, American Society for Microbiology, Vol. 76, No. 9 ( 2010-05), p. 2704-2711
    Abstract: The use of naturally occurring microbial antagonists to suppress plant diseases offers a favorable alternative to classical methods of plant protection. The soybean epiphyte Pseudomonas syringae pv. syringae strain 22d/93 shows great potential for controlling P. syringae pv. glycinea, the causal agent of bacterial blight of soybean. Its activity against P. syringae pv. glycinea is highly reproducible even in field trials, and the suppression mechanisms involved are of special interest. In this work we demonstrated that P . syringae pv. syringae 22d/93 produced a significantly larger amount of siderophores than the pathogen P. syringae pv. glycinea produced. While P . syringae pv. syringae 22d/93 and P. syringae pv. glycinea produce the same siderophores, achromobactin and pyoverdin, the regulation of siderophore biosynthesis in the former organism is very different from that in the latter organism. The epiphytic fitness of P . syringae pv. syringae 22d/93 mutants defective in siderophore biosynthesis was determined following spray inoculation of soybean leaves. The population size of the siderophore-negative mutant P . syringae pv. syringae strain 22d/93ΔSid was 2 orders of magnitude lower than that of the wild type 10 days after inoculation. The growth deficiency was compensated for when wound inoculation was used, indicating the availability of iron in the presence of small lesions on the leaves. Our results suggest that siderophore production has an indirect effect on the biocontrol activity of P . syringae pv. syringae 22d/93. Although siderophore-defective mutants of P . syringae pv. syringae 22d/93 still suppressed development of bacterial blight caused by P. syringae pv. glycinea, siderophore production enhanced the epiphytic fitness and thus the competitiveness of the antagonist.
    Type of Medium: Online Resource
    ISSN: 0099-2240 , 1098-5336
    RVK:
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2010
    detail.hit.zdb_id: 223011-2
    detail.hit.zdb_id: 1478346-0
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  • 7
    Online Resource
    Online Resource
    American Society for Microbiology ; 2004
    In:  Applied and Environmental Microbiology Vol. 70, No. 2 ( 2004-02), p. 693-703
    In: Applied and Environmental Microbiology, American Society for Microbiology, Vol. 70, No. 2 ( 2004-02), p. 693-703
    Abstract: Blossoms are important sites of infection for Erwinia amylovora , the causal agent of fire blight of rosaceous plants. Before entering the tissue, the pathogen colonizes the stigmatic surface and has to compete for space and nutrient resources within the epiphytic community. Several epiphytes are capable of synthesizing antibiotics with which they antagonize phytopathogenic bacteria. Here, we report that a multidrug efflux transporter, designated NorM, of E. amylovora confers tolerance to the toxin(s) produced by epiphytic bacteria cocolonizing plant blossoms. According to sequence comparisons, the single-component efflux pump NorM is a member of the multidrug and toxic compound extrusion protein family. The corresponding gene is widely distributed among E. amylovora strains and related plant-associated bacteria. NorM mediated resistance to the hydrophobic cationic compounds norfloxacin, ethidium bromide, and berberine. A norM mutant was constructed and exhibited full virulence on apple rootstock MM 106. However, it was susceptible to antibiotics produced by epiphytes isolated from apple and quince blossoms. The epiphytes were identified as Pantoea agglomerans by 16S rRNA analysis and were isolated from one-third of all trees examined. The promoter activity of norM was twofold greater at 18°C than at 28°C. The lower temperature seems to be beneficial for host infection because of the availability of moisture necessary for movement of the pathogen to the infection sites. Thus, E. amylovora might employ NorM for successful competition with other epiphytic microbes to reach high population densities, particularly at a lower temperature.
    Type of Medium: Online Resource
    ISSN: 0099-2240 , 1098-5336
    RVK:
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2004
    detail.hit.zdb_id: 223011-2
    detail.hit.zdb_id: 1478346-0
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  • 8
    Online Resource
    Online Resource
    American Society for Microbiology ; 2008
    In:  Applied and Environmental Microbiology Vol. 74, No. 11 ( 2008-06), p. 3387-3393
    In: Applied and Environmental Microbiology, American Society for Microbiology, Vol. 74, No. 11 ( 2008-06), p. 3387-3393
    Abstract: In gram-negative bacteria, transporters belonging to the RND family are the transporters most relevant for resistance to antimicrobial compounds. In Pseudomonas aeruginosa , a clinically important pathogen, the RND-type pump MexAB-OprM has been recognized as one of the major multidrug efflux systems. Here, homologues of MexAB-OprM in the plant pathogens Pseudomonas syringae pv. phaseolicola 1448A, P. syringae pv. syringae B728a, and P. syringae pv. tomato DC3000 were identified, and mexAB-oprM -deficient mutants were generated. Determination of MICs revealed that mutation of MexAB-OprM dramatically reduced the tolerance to a broad range of antimicrobials. Moreover, the ability of the mexAB - oprM -deficient mutants to multiply in planta was reduced. RNA dot blot hybridization revealed growth-dependent regulation of the mexAB-oprM operon in P. syringae ; the expression of this operon was maximal in early exponential phase and decreased gradually during further growth.
    Type of Medium: Online Resource
    ISSN: 0099-2240 , 1098-5336
    RVK:
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2008
    detail.hit.zdb_id: 223011-2
    detail.hit.zdb_id: 1478346-0
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  • 9
    Online Resource
    Online Resource
    MDPI AG ; 2009
    In:  International Journal of Molecular Sciences Vol. 10, No. 2 ( 2009-02-19), p. 629-645
    In: International Journal of Molecular Sciences, MDPI AG, Vol. 10, No. 2 ( 2009-02-19), p. 629-645
    Abstract: AcrAB-TolC is the major multidrug efflux system in Enterobacteriaceae recognizing structurally unrelated molecules including antibiotics, dyes, and detergents. Additionally, in Escherichia coli it mediates resistance to bile salts. In the plant pathogen Erwinia amylovora AcrAB-TolC is required for virulence and phytoalexin resistance. Exchange analysis of AcrAB-TolC was conducted by complementing mutants of both species defective in acrB or tolC with alleles from either species. The acrB and tolC mutants exhibited increased susceptibility profiles for 24 different antibiotics. All mutants were complemented with acrAB or tolC, respectively, regardless of the taxonomic origin of the alleles. Importantly, complementation of E. amylovora mutants with respective E. coli genes restored virulence on apple plants. It was concluded that AcrAB and TolC of both species could interact and that these interactions did not yield in altered functions despite the divergent ecological niches, to which E. coli and E. amylovora have adopted.
    Type of Medium: Online Resource
    ISSN: 1422-0067
    Language: English
    Publisher: MDPI AG
    Publication Date: 2009
    detail.hit.zdb_id: 2019364-6
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  • 10
    Online Resource
    Online Resource
    Scientific Societies ; 2004
    In:  Molecular Plant-Microbe Interactions® Vol. 17, No. 10 ( 2004-10), p. 1095-1102
    In: Molecular Plant-Microbe Interactions®, Scientific Societies, Vol. 17, No. 10 ( 2004-10), p. 1095-1102
    Abstract: Coronatine (COR) is a chlorosis-inducing phytotoxin produced by the plant-pathogenic bacterium Pseudomonas syringae. Confocal laser scanning microscopy was used to investigate in vitro and in planta expression of COR genes by two model organisms, P. syringae pv. glycinea PG4180, a pathogen of soybean, and P. syringae pv. tomato DC3000, a pathogen of tomato and crucifers. Previously, it was shown in vitro that the cma operon involved in COR synthesis in PG4180 is expressed in a temperature-dependent manner, with maximal rates at 18°C and low activity at 28°C. However, nothing was known about the influence of temperature on the expression of COR biosynthetic genes in planta. Therefore, transcriptional fusions of the PG4180 and DC3000 cma promoter regions to a promoterless egfp gene were constructed and expressed in both P. syringae strains. The fluorescence patterns in response to temperature during growth of a strain in vitro were consistent with its COR production and the cma transcript abundance as revealed by RNA dot blot hybridization. Quantification of fluorescence indicated that cma promoter activity was dependent on the genetic background of the host strain. Expression of cma∷egfp in PG4180 was temperature-dependent in minimal medium as well as inside the plant tissue. In contrast, transcription of the cma operon was not significantly affected by temperature in DC3000. However, cells of DC3000 harboring the cma∷egfp fusions showed higher levels of fluorescence when recovered from infected host plants compared with cells grown in minimal medium. These results indicate that the signals for induction of COR biosynthesis differ significantly in PG4180 and DC3000.
    Type of Medium: Online Resource
    ISSN: 0894-0282 , 1943-7706
    Language: English
    Publisher: Scientific Societies
    Publication Date: 2004
    detail.hit.zdb_id: 2037108-1
    detail.hit.zdb_id: 743331-1
    SSG: 12
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