Regulation of the expression of prtW::gusA fusions in Erwinia carotovora subsp. carotovora

Marits, Reet; Tshuikina, Marina; Pirhonen, Minna; Laasik, Eve; Mäe, Andres

doi:10.1099/00221287-148-3-835

Cited by 19 publications

(16 citation statements)

References 27 publications

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“…Extracellular enzyme activities were measured in cell-free supernatant samples according to previously published protocols [31], [32]. To perform semi-quantitative agarose plate assays of extracellular protease and polygalacturonase production, cells were spotted on respective indicator plates described by Chatterjee and coworkers [23], with the exception that a milk-containing agar plate additionally contained 0.05% polygalacturonic acid as an inducer for proteases.…”

Section: Methodsmentioning

confidence: 99%

Lack of RsmA-Mediated Control Results in Constant Hypervirulence, Cell Elongation, and Hyperflagellation in Pectobacterium wasabiae

et al. 2013

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The posttranscriptional regulator RsmA controls the production of plant cell wall degrading enzymes (PCWDE) and cell motility in the Pectobacterium genus of plant pathogens. In this study the physiological role of gene regulation by RsmA is under investigation. Disruption of rsmA gene of the Pectobacterium wasabiae strain, SCC3193 resulted in 3-fold decrease in growth rate and increased virulence. The comparison of mRNA levels of the rsmA− mutant and wild-type using a genome-wide microarray showed, that genes responsible for successful infection, i.e. virulence factors, motility, butanediol fermentation, various secretion systems etc. were up-regulated in the rsmA− strain. The rsmA− strain exhibited a higher propensity to swarm and produce PCWDE compared to the wild-type strain. Virulence experiments in potato tubers demonstrated that in spite of its more efficient tissue maceration, the rsmA− strain's ability to survive within the host is reduced and the infection site is taken over by resident bacteria. Taken together, in the absence of RsmA, cells revert to a constitutively infective phenotype characterized by expression of virulence factors and swarming. We hypothesize that lack of control over these costly energetic processes results in decreased growth rate and fitness. In addition, our findings suggest a relationship between swarming and virulence in plant pathogens.

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Section: Methodsmentioning

confidence: 99%

Lack of RsmA-Mediated Control Results in Constant Hypervirulence, Cell Elongation, and Hyperflagellation in Pectobacterium wasabiae

et al. 2013

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“…Metalloproteases secreted through the Prt T1SS contribute to SRE virulence (59). In Pectobacterium, the T1SS is upregulated by plant extracts and acylhomoserine lactone (AHL), and controlled by GacAS (97,98). In Dickeya, this system is controlled by PecS (59,104) cand GacAS (85).…”

Section: Type I Secretion System: Metalloproteases and Adhesinsmentioning

confidence: 99%

The Role of Secretion Systems and Small Molecules in Soft-Rot Enterobacteriaceae Pathogenicity

Charkowski¹,

Blanco

Condemine

et al. 2012

Annu. Rev. Phytopathol.

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220

251

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Soft-rot Enterobacteriaceae (SRE), which belong to the genera Pectobacterium and Dickeya, consist mainly of broad host-range pathogens that cause wilt, rot, and blackleg diseases on a wide range of plants. They are found in plants, insects, soil, and water in agricultural regions worldwide. SRE encode all six known protein secretion systems present in gram-negative bacteria, and these systems are involved in attacking host plants and competing bacteria. They also produce and detect multiple types of small molecules to coordinate pathogenesis, modify the plant environment, attack competing microbes, and perhaps to attract insect vectors. This review integrates new information about the role protein secretion and detection and production of ions and small molecules play in soft-rot pathogenicity.

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“…Samples for GUS activity assay were taken at indicated time points and the growth was monitored by measuring the OD 600 . GUS activity was measured by using p-nitrophenyl β-D-glucuronide as substrate (Marits et al 2002;Novel et al 1974).…”

Section: Enzyme Assaysmentioning

confidence: 99%

A Novel Plant Ferredoxin-Like Protein and the Regulator Hor Are Quorum-Sensing Targets in the Plant Pathogen Erwinia carotovora

Sjöblom¹,

Harjunpää²,

Brader³

et al. 2008

MPMI

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Quorum sensing (QS), a population-density-sensing mechanism, controls the production of the main virulence determinants, the plant cell-wall-degrading enzymes (PCWDEs) of the soft-rot phytopathogen Erwinia carotovora subsp. carotovora. In this study, we used random transposon mutagenesis with a gusA reporter construct to identify two new QS-controlled genes encoding the regulator Hor and a plant ferredoxin-like protein, FerE. The QS control of the identified genes was executed by the QS regulators ExpR1 and ExpR2 and mediated by the global repressor RsmA. Hor was shown to contribute to bacterial virulence at least partly through its control of PCWDE production. Our results showed that FerE contributes to oxidative stress tolerance and in planta fitness of the bacteria and suggest that QS could be central to control of oxidative stress tolerance. The presence of the FerE protein appears to be rather unique in heterotrophic bacteria and suggests an acquisition of the corresponding gene from plant host by horizontal gene transfer.Additional keywords: acylhomoserine lactone, AHSL, ExpI, Pectobacterium carotovorum.Bacteria use a cell-to-cell communication system, referred to as quorum sensing (QS), to monitor their population density and to regulate a wide variety of functions accordingly (Fuqua et al. 2001;Waters and Bassler 2005). These include bioluminescence in Vibrio fischerii (Eberhard et al. 1981;Engebrecht and Silverman 1984), conjugal plasmid transfer in Agrobacterium tumefaciens (Zhang et al. 1993), biofilm formation and virulence in Pseudomonas aeruginosa (Parsek and Greenberg 2000;Winson et al. 1995), production of the exopolysaccharide stewartan in Pantoea stewartii (Beck von Bodman and Farrand, 1995), and carbapenem antibiotics and plant cellwall-degrading enzymes (PCWDEs) in Erwinia carotovora subsp. carotovora (also referred to as Pectobacterium carotovorum) (Bainton et al. 1992;Jones et al. 1993;Pirhonen et al. 1993).In contrast to the divergence of QS-controlled processes, the basic QS system is similar in many gram-negative bacteria. QS is achieved by the production and sensing of diffusible signal molecules, N-acylhomoserine lactones (AHSL), synthesized by an AHSL synthase, an enzyme of the LuxI family and recognized by a QS regulator, a transcription factor of the LuxR family that in turn controls the expression of downstream genes (Fuqua et al. 2001;Whitehead et al. 2001). Different bacteria produce AHSL with diverse acyl side chain lengths (4 to 16 carbons and with alterations in the oxidative status) that are recognized by a QS regulator specific for the cognate AHSL. Depending on the strain, E. carotovora subsp. carotovora produces mainly either 3-oxo-hexanoylhomoserine lactone (3-oxo-C6-HSL) or 3-oxo-octanoylhomoserine lactone (3-oxo-C8-HSL) as the major AHSL, with 3-oxo-C8-HSL as the cognate AHSL of E. carotovora subsp. carotovora SCC3193 (Brader et al. 2005). Normally, the QS regulator is unable to respond to noncognate AHSL. This confers a high degree of specificity to the QS system and s...

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Regulation of the expression of prtW::gusA fusions in Erwinia carotovora subsp. carotovora

Cited by 19 publications

References 27 publications

Lack of RsmA-Mediated Control Results in Constant Hypervirulence, Cell Elongation, and Hyperflagellation in Pectobacterium wasabiae

Lack of RsmA-Mediated Control Results in Constant Hypervirulence, Cell Elongation, and Hyperflagellation in Pectobacterium wasabiae

The Role of Secretion Systems and Small Molecules in Soft-Rot Enterobacteriaceae Pathogenicity

A Novel Plant Ferredoxin-Like Protein and the Regulator Hor Are Quorum-Sensing Targets in the Plant Pathogen Erwinia carotovora

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