Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 19 de 19
Filtrar
1.
J Microbiol Biotechnol ; 34(3): 538-546, 2024 Mar 28.
Artículo en Inglés | MEDLINE | ID: mdl-38146216

RESUMEN

Cinnamaldehyde is a natural compound extracted from cinnamon bark essential oil, acclaimed for its versatile properties in both pharmaceutical and agricultural fields, including antimicrobial, antioxidant, and anticancer activities. Although potential of cinnamaldehyde against plant pathogenic bacteria like Agrobacterium tumefaciens and Pseudomonas syringae pv. actinidiae causative agents of crown gall and bacterial canker diseases, respectively has been documented, indepth studies into cinnamaldehyde's broader influence on plant pathogenic bacteria are relatively unexplored. Particularly, Pectobacterium spp., gram-negative soil-borne pathogens, notoriously cause soft rot damage across a spectrum of plant families, emphasizing the urgency for effective treatments. Our investigation established that the Minimum Inhibitory Concentrations (MICs) of cinnamaldehyde against strains P. odoriferum JK2, P. carotovorum BP201601, and P. versatile MYP201603 were 250 µg/ml, 125 µg/ml, and 125 µg/ml, respectively. Concurrently, their Minimum Bactericidal Concentrations (MBCs) were found to be 500 µg/ml, 250 µg/ml, and 500 µg/ml, respectively. Using RNA-sequencing analysis, we identified 1,907 differentially expressed genes in P. carotovorum BP201601 treated with 500 µg/ml cinnamaldehyde. Notably, our results indicate that cinnamaldehyde upregulated nitrate reductase pathways while downregulating the citrate cycle, suggesting a potential disruption in the aerobic respiration system of P. carotovorum during cinnamaldehyde exposure. This study serves as a pioneering exploration of the transcriptional response of P. carotovorum to cinnamaldehyde, providing insights into the bactericidal mechanisms employed by cinnamaldehyde against this bacterium.


Asunto(s)
Acroleína/análogos & derivados , Antiinfecciosos , Pectobacterium , Pectobacterium carotovorum , Pectobacterium/metabolismo , Antibacterianos/farmacología , Antibacterianos/metabolismo , Antiinfecciosos/farmacología , Bacterias/metabolismo , Plantas/metabolismo , Enfermedades de las Plantas/microbiología
2.
Environ Microbiol ; 25(11): 2564-2579, 2023 11.
Artículo en Inglés | MEDLINE | ID: mdl-37622480

RESUMEN

The type VI secretion system (T6SS) is a contractile nanomachine widespread in Gram-negative bacteria. The T6SS injects effectors into target cells including eukaryotic hosts and competitor microbial cells and thus participates in pathogenesis and intermicrobial competition. Pseudomonas fluorescens MFE01 possesses a single T6SS gene cluster that confers biocontrol properties by protecting potato tubers against the phytopathogen Pectobacterium atrosepticum (Pca). Here, we demonstrate that a functional T6SS is essential to protect potato tuber by reducing the pectobacteria population. Fluorescence microscopy experiments showed that MFE01 displays an aggressive behaviour with an offensive T6SS characterized by continuous and intense T6SS firing activity. Interestingly, we observed that T6SS firing is correlated with rounding of Pectobacterium cells, suggesting delivery of a potent cell wall targeting effector. Mutagenesis coupled with functional assays then revealed that a putative T6SS secreted amidase, Tae3Pf , is mainly responsible for MFE01 toxicity towards Pca. Further studies finally demonstrated that Tae3Pf is toxic when produced in the periplasm, and that its toxicity is counteracted by the Tai3Pf inner membrane immunity protein.


Asunto(s)
Pectobacterium , Pseudomonas fluorescens , Solanum tuberosum , Sistemas de Secreción Tipo VI , Pseudomonas fluorescens/genética , Pseudomonas fluorescens/metabolismo , Sistemas de Secreción Tipo VI/genética , Sistemas de Secreción Tipo VI/metabolismo , Mutagénesis , Pectobacterium/genética , Pectobacterium/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo
3.
World J Microbiol Biotechnol ; 38(11): 184, 2022 Aug 16.
Artículo en Inglés | MEDLINE | ID: mdl-35972587

RESUMEN

Bacterial intercellular communication mediated by small diffusible molecules, known as quorum sensing (QS), is a common mechanism for regulating bacterial colonisation strategies and survival. Influence on QS by plant-derived molecules is proposed as a strategy for combating phytopathogens by modulating their virulence. This work builds upon other studies that have revealed plant-derived QS inhibitors extracted from oak bark (Quercus sp.). It was found that co-incubation of Pectobacterium carotovorum VKM-B-1247 with oak bark extract (OBE) reduced the production of acyl-HSL. This was accompanied by a dose-dependent decrease in the bacterial cellulolytic and protease activity. At the transcriptomic level, the OBE treatment suppressed the main QS-related genes expR/expI. Potato tubers pre-treated with OBE showed resistance to a manifestation of soft-rot symptoms. Analysis of the component composition of the OBE identified several biologically active molecules, such as n-hexadecanoic acid, 2,6-di-tert-butyl-4-methylphenol, butylated hydroxytoluene (BHT), gamma-sitosterol, lupeol, and others. Molecular docking of the binding energy between identified molecules and homology models of LuxR-LuxI type proteins allow to identify potential inhibitors. Collectively, obtained results figure out great potential of widely distributed oak-derived plant material for bacterial control during storage of potato.


Asunto(s)
Pectobacterium , Quercus , Solanum tuberosum , Proteínas Bacterianas/metabolismo , Simulación del Acoplamiento Molecular , Pectobacterium/genética , Pectobacterium/metabolismo , Pectobacterium carotovorum/metabolismo , Corteza de la Planta/metabolismo , Percepción de Quorum/genética , Solanum tuberosum/microbiología , Virulencia/genética
4.
Int J Mol Sci ; 22(14)2021 Jul 08.
Artículo en Inglés | MEDLINE | ID: mdl-34298965

RESUMEN

Pectobacterium parmentieri is a Gram-negative plant-pathogenic bacterium able to infect potato (Solanum tuberosum L.). Little is known about lytic bacteriophages infecting P. parmentieri and how phage-resistance influences the environmental fitness and virulence of this species. A lytic phage vB_Ppp_A38 (ϕA38) has been previously isolated and characterized as a potential biological control agent for the management of P. parmentieri. In this study, seven P. parmentieri SCC 3193 Tn5 mutants were identified that exhibited resistance to infection caused by vB_Ppp_A38 (ϕA38). The genes disrupted in these seven mutants encoded proteins involved in the assembly of O-antigen, sugar metabolism, and the production of bacterial capsule exopolysaccharides. The potential of A38-resistant P. parmentieri mutants for plant colonization and pathogenicity as well as other phenotypes expected to contribute to the ecological fitness of P. parmentieri, including growth rate, use of carbon and nitrogen sources, production of pectinolytic enzymes, proteases, cellulases, and siderophores, swimming and swarming motility, presence of capsule and flagella as well as the ability to form biofilm were assessed. Compared to the wild-type P. parmentieri strain, all phage-resistant mutants exhibited a reduced ability to colonize and to cause symptoms in growing potato (S. tuberosum L.) plants. The implications of bacteriophage resistance on the ecological fitness of P. parmentieri are discussed.


Asunto(s)
Bacteriófagos , Regulación Bacteriana de la Expresión Génica , Mutación , Pectobacterium , Enfermedades de las Plantas/microbiología , Polisacáridos Bacterianos , Solanum tuberosum/microbiología , Factores de Virulencia/biosíntesis , Bacteriófagos/genética , Bacteriófagos/metabolismo , Pectobacterium/genética , Pectobacterium/metabolismo , Pectobacterium/patogenicidad , Pectobacterium/virología , Polisacáridos Bacterianos/genética , Polisacáridos Bacterianos/metabolismo , Factores de Virulencia/genética
5.
Mol Plant Microbe Interact ; 34(1): 100-109, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-32960719

RESUMEN

Potato (Solanum tuberosum L.) is the primary vegetable crop consumed worldwide and is largely affected by bacterial pathogens that can cause soft rot and blackleg disease. Recently, resistance to these diseases has been identified in the wild potato S. chacoense, and the mechanism of resistance is unknown. Here, it was hypothesized that S. chacoense stems or tubers have unique chemistry that confers resistance to the pathogen Pectobacterium brasiliense through bactericidal, bacteriostatic, or antivirulence activity. Stem and tuber metabolite extracts were collected from S. chacoense and tested for effects on Pectobacterium bacterial multiplication rates, and activity and expression of known exoenzymes and virulence genes using S. tuberosum extracts as a comparative control. Comparatively, the S. chacoense extracts did not affect bacterial multiplication rate; however, they did reduce pectinase, cellulase, and protease activities. The chemical extracts were profiled using a bioassay-guided fractionation, and a nontargeted metabolomics comparison of S. chacoense and S. tuberosum stems and tubers was performed. The data showed that selected alkaloids, phenolic amines, phenols, amines, and peptides are integrative chemical sources of resistance against the bacteria.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Asunto(s)
Pectobacterium , Enfermedades de las Plantas , Solanum tuberosum , Factores de Virulencia , Metaboloma , Pectobacterium/metabolismo , Pectobacterium/patogenicidad , Enfermedades de las Plantas/microbiología , Tubérculos de la Planta/microbiología , Solanum tuberosum/microbiología , Factores de Virulencia/metabolismo
6.
Mol Plant Pathol ; 22(2): 271-283, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33301200

RESUMEN

Pectobacterium carotovorum has an incomplete Entner-Doudoroff (ED) pathway, including enzyme 2-keto-3-deoxy-6-phosphogluconate aldolase (Eda) but lacking phosphogluconate dehydratase (Edd), while P. atrosepticum (Pba) has a complete pathway. To understand the role of the ED pathway in Pectobacterium infection, mutants of these two key enzymes, Δeda and Δedd, were constructed in Pba SCRI1039. Δeda exhibited significant decreased virulence on potato tubers and colonization in planta and was greatly attenuated in pectinase activity and the ability to use pectin breakdown products, including polygalacturonic acid (PGA) and galacturonic acid. These reduced phenotypes were restored following complementation with an external vector expressing eda. Quantitative reverse transcription PCR analysis revealed that expression of the pectinase genes pelA, pelC, pehN, pelW, and pmeB in Δeda cultured in pyruvate, with or without PGA, was significantly reduced compared to the wild type, while genes for virulence regulators (kdgR, hexR, hexA, and rsmA) remained unchanged. However, Δedd showed similar phenotypes to the wild type. To our knowledge, this is the first demonstration that disruption of eda has a feedback effect on inhibiting pectin degradation and that Eda is involved in building the arsenal of pectinases needed during infection by Pectobacterium.


Asunto(s)
Aldehído-Liasas/metabolismo , Pectobacterium/metabolismo , Hidroliasas/metabolismo , Redes y Vías Metabólicas , Pectinas/metabolismo , Pectobacterium/enzimología , Pectobacterium/patogenicidad , Solanum tuberosum/microbiología , Virulencia
7.
PLoS One ; 15(4): e0230842, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32240203

RESUMEN

Globally, there is a high economic burden caused by pre- and post-harvest losses in vegetables, fruits and ornamentals due to soft rot diseases. At present, the control methods for these diseases are limited, but there is some promise in developing biological control products for use in Integrated Pest Management. This study sought to formulate a phage cocktail which would be effective against soft rot Pectobacteriaceae species affecting potato (Solanum tuberosum L.), with potential methods of application in agricultural systems, including vacuum-infiltration and soil drench, also tested. Six bacteriophages were isolated and characterized using transmission electron microscopy, and tested against a range of Pectobacterium species that cause soft rot/blackleg of potato. Isolated bacteriophages of the family Podoviridae and Myoviridae were able to control isolates of the Pectobacterium species: Pectobacterium atrosepticum and Pectobacterium carotovorum subsp. carotovorum. Genomic analysis of three Podoviridae phages did not indicate host genes transcripts or proteins encoding toxin or antibiotic resistance genes. These bacteriophages were formulated as a phage cocktail and further experiments showed high activity in vitro and in vivo to suppress Pectobacterium growth, potentially indicating their efficacy in formulation as a microbial pest control agent to use in planta.


Asunto(s)
Myoviridae/metabolismo , Pectobacterium/efectos de los fármacos , Podoviridae/metabolismo , Bacteriófagos/genética , Agentes de Control Biológico/metabolismo , Genómica , Myoviridae/genética , Pectobacterium/crecimiento & desarrollo , Pectobacterium/metabolismo , Pectobacterium carotovorum/genética , Control de Plagas/métodos , Filogenia , Enfermedades de las Plantas/microbiología , Podoviridae/genética , Solanum tuberosum/microbiología
8.
Environ Microbiol ; 17(11): 4730-44, 2015 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-26271942

RESUMEN

Integrative and conjugative elements (ICEs) contribute to the rapid evolution of bacterial pathogens via horizontal gene transfer of virulence determinants. ICEs have common mechanisms for transmission, yet the cues triggering this process under natural environmental or physiological conditions are largely unknown. In this study, mobilization of the putative ICE horizontally acquired island 2 (HAI2), present in the chromosome of the phytopathogen Pectobacterium atrosepticum SCRI1043, was examined during infection of the host plant potato. Under these conditions, mobilization of HAI2 increased markedly compared with in vitro cultures. In planta-induced mobilization of HAI2 was regulated by quorum sensing and involved the putative ICE-encoded relaxase ECA0613. Disruption of ECA0613 also reduced transcription of genes involved in production of coronafacic acid (Cfa), the major virulence factor harboured on HAI2, whereas their expression was unaffected in the quorum-sensing (expI) mutant. Thus, suppression of cfa gene expression was not regulated by the mobilization of the ICE per se, but was due directly to inactivation of the relaxase. The identification of genetic factors associated solely with in planta mobilization of an ICE demonstrates that this process is highly adapted to the natural environment of the bacterial host and can influence the expression of virulence determinants.


Asunto(s)
Pectobacterium/genética , Pectobacterium/patogenicidad , Enfermedades de las Plantas/microbiología , Solanum tuberosum/microbiología , Factores de Virulencia/genética , Secuencia de Aminoácidos , Transferencia de Gen Horizontal , Indenos/metabolismo , Islas , Datos de Secuencia Molecular , Pectobacterium/metabolismo , Percepción de Quorum/genética , Alineación de Secuencia , Factores de Virulencia/metabolismo
9.
Heredity (Edinb) ; 114(5): 476-84, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-25585922

RESUMEN

Social bacteria use chemical communication to coordinate and synchronize gene expression via the quorum-sensing (QS) regulatory pathway. In Pectobacterium, a causative agent of the blackleg and soft-rot diseases on potato plants and tubers, expression of the virulence factors is collectively controlled by the QS-signals N-acylhomoserine lactones (NAHLs). Several soil bacteria, such as the actinobacterium Rhodococcus erythropolis, are able to degrade NAHLs, hence quench the chemical communication and virulence of Pectobacterium. Here, next-generation sequencing was used to investigate structural and functional genomics of the NAHL-degrading R. erythropolis strain R138. The R. erythropolis R138 genome (6.7 Mbp) contained a single circular chromosome, one linear (250 kbp) and one circular (84 kbp) plasmid. Growth of R. erythropolis and P. atrosepticum was not altered in mixed-cultures as compared with monocultures on potato tuber slices. HiSeq-transcriptomics revealed that no R. erythropolis genes were differentially expressed when R. erythropolis was cultivated in the presence vs absence of the avirulent P. atrosepticum mutant expI, which is defective for QS-signal synthesis. By contrast 50 genes (<1% of the R. erythropolis genome) were differentially expressed when R. erythropolis was cultivated in the presence vs absence of the NAHL-producing virulent P. atrosepticum. Among them, quantitative real-time reverse-transcriptase-PCR confirmed that the expression of some alkyl-sulfatase genes decreased in the presence of a virulent P. atrosepticum, as well as deprivation of organic sulfur such as methionine, which is a key precursor in the synthesis of NAHL by P. atrosepticum.


Asunto(s)
Genoma Bacteriano , Pectobacterium/patogenicidad , Percepción de Quorum , Rhodococcus/genética , Transcriptoma , Acil-Butirolactonas/metabolismo , Técnicas de Cocultivo , ADN Bacteriano/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Pectobacterium/metabolismo , Análisis de Secuencia de ADN , Solanum tuberosum/microbiología
10.
Plant Cell Rep ; 32(5): 579-89, 2013 May.
Artículo en Inglés | MEDLINE | ID: mdl-23479199

RESUMEN

KEY MESSAGE: Potato and tobacco cells are differentially suited to study oxylipin pathway and elicitor-induced responses. Synthesis of oxylipins via the lipoxygenase (LOX) pathway provides plant cells with an important class of signaling molecules, related to plant stress responses and innate immunity. The aim of this study was to evaluate the induction of LOX pathway in tobacco and potato cells induced by a concentrated culture filtrate (CCF) from Phytophthora infestans and lipopolysaccharide (LPS) from Pectobacterium atrosepticum. Oxylipin activation was evaluated by the measurement of LOX activity and metabolite quantification. The basal levels of oxylipins and fatty acids showed that potato cells contained higher amounts of linoleic (LA), linolenic (LnA) and stearic acids than tobacco cells. The major oxylipin in potato cells, 9(S),10(S),11(R)-trihydroxy-12(Z),15(Z)-octadecadienoic acid (9,10,11-THOD), was not detected in tobacco cells. CCF induced a sharp increase of LA and LnA at 8 h in tobacco cells. In contrast they decreased in potato cells. In CCF-treated tobacco cells, colneleic acid increased up to 24 h, colnelenic acid and 9(S)-hydroxyoctadecatrienoic acid (9(S)-HOT) increased up to 16 h. In potato cells, only colneleic acid increased slightly until 16 h. A differential induction of LOX activity was measured in both cells treated by CCF. With LPS treatment, only 9,10,11-THOD accumulation was significantly induced at 16 h in potato cells. Fatty acids were constant in tobacco but decreased in potato cells over the studied time period. These results showed that the two elicitors were differently perceived by the two Solanaceae and that oxylipin pathway is strongly induced in tobacco with the CCF. They also revealed that elicitor-induced responses depended on both cell culture and elicitor.


Asunto(s)
Nicotiana/metabolismo , Oxilipinas/metabolismo , Solanum tuberosum/metabolismo , Ácidos Grasos Insaturados/metabolismo , Interacciones Huésped-Patógeno , Ácido Linoleico/metabolismo , Lipopolisacáridos/farmacología , Lipooxigenasa/metabolismo , Pectobacterium/metabolismo , Fenilanina Amoníaco-Liasa/metabolismo , Phytophthora infestans/metabolismo , Phytophthora infestans/patogenicidad , Solanum tuberosum/citología , Solanum tuberosum/efectos de los fármacos , Solanum tuberosum/microbiología , Nicotiana/citología , Nicotiana/efectos de los fármacos , Nicotiana/microbiología , Ácido alfa-Linolénico/metabolismo
11.
PLoS One ; 8(1): e54248, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23372695

RESUMEN

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.


Asunto(s)
Proteínas Bacterianas/genética , Flagelos/genética , Redes y Vías Metabólicas/genética , Pectobacterium/genética , Pectobacterium/patogenicidad , Proteínas Represoras/genética , Factores de Virulencia/genética , Proteínas Bacterianas/metabolismo , Quimiotaxis/fisiología , Flagelos/metabolismo , Eliminación de Gen , Regulación Bacteriana de la Expresión Génica , Pectobacterium/crecimiento & desarrollo , Pectobacterium/metabolismo , Enfermedades de las Plantas/microbiología , Proteínas Represoras/metabolismo , Solanum tuberosum/microbiología , Virulencia , Factores de Virulencia/metabolismo
12.
PLoS One ; 7(4): e35176, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22539957

RESUMEN

BACKGROUND: Several small diffusible molecules are involved in bacterial quorum sensing and virulence. The production of autoinducers-1 and -2, quinolone, indole and γ-amino butyrate signaling molecules was investigated in a set of soft-rot bacteria belonging to six Dickeya or Pectobacterium species including recent or emerging potato isolates. METHODOLOGY/PRINCIPAL FINDINGS: Using bacterial biosensors, immunoassay, and chromatographic analysis, we showed that soft-rot bacteria have the common ability to produce transiently during their exponential phase of growth the N-3-oxo-hexanoyl- or the N-3-oxo-octanoyl-l-homoserine lactones and a molecule of the autoinducer-2 family. Dickeya spp. produced in addition the indole-3-acetic acid in tryptophan-rich conditions. All these signaling molecules have been identified for the first time in the novel Dickeya solani species. In contrast, quinolone and γ-amino butyrate signals were not identified and the corresponding synthases are not present in the available genomes of soft-rot bacteria. To determine if the variations of signal production according to growth phase could result from expression modifications of the corresponding synthase gene, the respective mRNA levels were estimated by reverse transcriptase-PCR. While the N-acyl-homoserine lactone production is systematically correlated to the synthase expression, that of the autoinducer-2 follows the expression of an enzyme upstream in the activated methyl cycle and providing its precursor, rather than the expression of its own synthase. CONCLUSIONS/SIGNIFICANCE: Despite sharing the S-adenosylmethionine precursor, no strong link was detected between the production kinetics or metabolic pathways of autoinducers-1 and -2. In contrast, the signaling pathway of autoinducer-2 seems to be switched off by the indole-3-acetic acid pathway under tryptophan control. It therefore appears that the two genera of soft-rot bacteria have similarities but also differences in the mechanisms of communication via the diffusible molecules. Our results designate autoinducer-1 lactones as the main targets for a global biocontrol of soft-rot bacteria communications, including those of emerging isolates.


Asunto(s)
Enterobacteriaceae/metabolismo , Pectobacterium/metabolismo , Percepción de Quorum , 4-Butirolactona/análogos & derivados , 4-Butirolactona/metabolismo , Acil-Butirolactonas/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Enterobacteriaceae/efectos de los fármacos , Enterobacteriaceae/crecimiento & desarrollo , Homoserina/análogos & derivados , Homoserina/metabolismo , Ácidos Indolacéticos/metabolismo , Cinética , Lactonas/metabolismo , Complejos Multienzimáticos/genética , Complejos Multienzimáticos/metabolismo , Pectobacterium/efectos de los fármacos , Pectobacterium/crecimiento & desarrollo , Quinolonas/metabolismo , ARN Mensajero/metabolismo , Transducción de Señal , Solanum tuberosum/microbiología , Triptófano/farmacología , Ácido gamma-Aminobutírico/metabolismo
13.
Nat Prod Commun ; 7(1): 65-9, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22428249

RESUMEN

This study was carried out to produce bioconverted products by microbial fermentation of tomato using a plant pathogenic bacterium Pectobacterium atrosepticum and to evaluate their in vitro antimycotic effect against pathogenic Candida species. The bioconverted products (500 microg/disc) provoked promising antimycotic effects against pathogenic isolates of Candida species as shown by the diameters of zones of inhibition (9 +/- 0.6 to 14 +/- 0.4 mm), along with their respective minimum inhibitory and minimum fungicidal concentration values, which increased from 250 to 1000 and 250 to 2000 microg/mL, respectively. With the viable counts of the tested fungal pathogens, exposure of the bioconverted products revealed a remarkable antimycotic effect. In addition, the morphology of a clinical isolate of C. glabrata KBN06P00368, visualized by scanning electron microscopy, showed a severe detrimental effect produced by the bioconverted products at the minimum inhibitory concentration (250 microg/mL). The bioconverted products significantly inhibited the in vitro growth of all the tested clinical and pathogenic laboratory isolates of Candida species. This study confirmed the potent antimycotic efficacy of the bioconverted products of tomato, hence justifying the therapeutic uses of bioconverted products in pharmaceutical preparations as an alternative approach to support the antifungal activity of conventional antimycotics.


Asunto(s)
Antifúngicos/metabolismo , Candida/efectos de los fármacos , Pectobacterium/metabolismo , Solanum lycopersicum/microbiología , Fermentación , Pruebas de Sensibilidad Microbiana , Microscopía Electrónica de Rastreo
14.
J Appl Microbiol ; 112(4): 782-92, 2012 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-22292528

RESUMEN

AIMS: The early molecular events underlying the elicitation of plant defence reactions by Gram-positive bacteria are relatively unknown. In plants, calcium and reactive oxygen species are commonly involved as cellular messengers of a wide range of biotic stimuli from pathogenic to symbiotic bacteria. In the present work, we checked whether nonpathogenic Streptomyces sp. strains could induce early signalling events leading to defence responses in BY2 tobacco cell suspensions. METHODS AND RESULTS: We have demonstrated that nonpathogenic Streptomyces sp. OE7 strain induced a cytosolic Ca(2+) increase and a biphasic oxidative burst in the upstream signalling events, leading to defence responses in BY2 tobacco cell suspensions. Streptomyces sp. OE7 also elicited delayed intracellular free scopoletin production and programmed cell death. In agreement with scopoletin production, OE7 induced accumulation of PAL transcripts and increased accumulation of transcripts of EREBP1 and AOX genes that are known to be regulated by the jasmonate/ethylene pathway. Transcript levels of PR1b and NIMIN2α, both salicylic acid pathway-linked genes, were not modified. Moreover, Streptomyces sp. OE7 culture filtrates could reduce Pectobacterium carotovorum- and Pectobacterium atrosepticum-induced death of BY2 cells and soft rot on potato slices. CONCLUSIONS: New insights are thus provided into the interaction mechanisms between Streptomyces sp. and plants; Streptomyces sp. could be sensed by plant cells, and through cytosolic Ca(2+) changes and the generation of reactive oxygen species, defence responses were induced. SIGNIFICANCE AND IMPACT OF THE STUDY: These induced defence responses appeared to participate in attenuating Pectobacterium-induced diseases in plants. Thus, Streptomyces sp. OE7 could be a biocontrol agent against Pectobacterium sp.


Asunto(s)
Calcio/metabolismo , Nicotiana/metabolismo , Nicotiana/microbiología , Especies Reactivas de Oxígeno/metabolismo , Apoptosis , Pectobacterium/metabolismo , Pectobacterium carotovorum/metabolismo , Células Vegetales/inmunología , Células Vegetales/metabolismo , Células Vegetales/microbiología , Escopoletina/metabolismo , Transducción de Señal , Solanum tuberosum/metabolismo , Streptomyces/metabolismo , Streptomyces/patogenicidad , Nicotiana/citología , Nicotiana/inmunología
15.
Mol Microbiol ; 82(3): 719-33, 2011 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-21992096

RESUMEN

Cyclic diguanylate (c-di-GMP) is a second messenger controlling many important bacterial processes. The phytopathogen Pectobacterium atrosepticum SCRI1043 (Pba1043) possesses a Type I secretion system (T1SS) essential for the secretion of a proteinaceous multi-repeat adhesin (MRP) required for binding to the host plant. The genes encoding the MRP and the T1SS are tightly linked to genes encoding several putative c-di-GMP regulatory components. We show that c-di-GMP regulates secreted MRP levels in Pba1043 through the action of two genes encoding predicted diguanylate cyclase (DGC) and phosphodiesterase proteins (ECA3270 and ECA3271). Phenotypic analyses and quantification of c-di-GMP levels demonstrated that ECA3270 and ECA3271 regulate secreted MRP levels by increasing and decreasing, respectively, the intracellular levels of c-di-GMP. Moreover, ECA3270 represents the first active DGC reported to have an alternative active-site motif from the 'canonical' GG[D/E]EF. ECA3270 has an A-site motif of SGDEF and analysis of single amino acid replacements demonstrated that the first position of this motif can tolerate functional substitution. Serine in position one of the A-site is also observed in many other DGCs. Finally, another T1SS-linked regulator (ECA3265) also plays an important role in regulating secreted MRP, with an altered localization of MRP observed in an ECA3265 mutant background. Mutants defective in these three T1SS-linked regulators exhibit a reduction in root binding and virulence, confirming that this complex, finely tuned regulation system is crucial in the interaction with host plants.


Asunto(s)
Adhesinas Bacterianas/metabolismo , Proteínas de Escherichia coli/metabolismo , Regulación Bacteriana de la Expresión Génica , Pectobacterium/metabolismo , Hidrolasas Diéster Fosfóricas/metabolismo , Liasas de Fósforo-Oxígeno/metabolismo , Secuencias de Aminoácidos , Sustitución de Aminoácidos , GMP Cíclico/análogos & derivados , GMP Cíclico/metabolismo , Proteínas de Escherichia coli/genética , Eliminación de Gen , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/metabolismo , Mutagénesis Sitio-Dirigida , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Pectobacterium/patogenicidad , Hidrolasas Diéster Fosfóricas/genética , Liasas de Fósforo-Oxígeno/genética , Raíces de Plantas/microbiología , Solanum tuberosum/microbiología , Virulencia
16.
Res Microbiol ; 162(9): 945-50, 2011 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-21288487

RESUMEN

Bacteria degrading quorum sensing (QS) signals have been proposed as biocontrol agents able to quench QS-dependent expression of virulence symptoms caused by Pectobacterium on potato plants. We report here that gamma-caprolactone (GCL) treatment stimulated growth of the native QS-degrading bacterial community in an industrial plant hydroponic system for culturing Solanum tuberosum. Post-GCL treatment, QS-degrading bacteria were mainly identified as Rhodococcus isolates, while Agrobacterium isolates dominated under similar untreated conditions. Most of the assayed Rhodococcus isolates exhibited efficient biocontrol activity for protecting potato tubers. Analytical chemistry approach revealed the rapid degradation of GCL introduced in the plant cultures.


Asunto(s)
Agrobacterium/metabolismo , Agentes de Control Biológico , Pectobacterium/metabolismo , Enfermedades de las Plantas/microbiología , Rhodococcus/metabolismo , Solanum tuberosum/microbiología , Agrobacterium/crecimiento & desarrollo , Caproatos/farmacología , Cromatografía Líquida de Alta Presión , Hidroponía , Lactonas/farmacología , Espectrometría de Masas , Pectobacterium/efectos de los fármacos , Pectobacterium/patogenicidad , Percepción de Quorum/efectos de los fármacos , Rhodococcus/crecimiento & desarrollo , Virulencia
17.
Mol Plant Microbe Interact ; 21(5): 547-54, 2008 May.
Artículo en Inglés | MEDLINE | ID: mdl-18393614

RESUMEN

To analyze whether metabolite import into Pectobacterium atrosepticum cells affects bacterial virulence, we investigated the function of a carrier which exhibits significant structural homology to characterized carboxylic-acid transport proteins. The corresponding gene, ECA3984, previously annotated as coding for a Na(+)/sulphate carrier, in fact encodes a highly specific citrate transporter (Cit1) which is energized by the proton-motive force. Expression of the cit1 gene is stimulated by the presence of citrate in the growth medium and is substantial during growth of P. atrosepticum on potato tuber tissue. Infection of tuber tissue with P. atrosepticum leads to reduced citrate levels. P. atrosepticum insertion mutants, lacking the functional Cit1 protein, did not grow in medium containing citrate as the sole carbon source, showed a substantially reduced ability to macerate potato tuber tissue, and did not provoke reduced citrate levels in the plant tissue upon infection. We propose that citrate uptake into P. atrosepticum is critical for full bacterial virulence.


Asunto(s)
Ácido Cítrico/metabolismo , Pectobacterium/metabolismo , Solanum tuberosum/microbiología , Secuencia de Aminoácidos , Proteínas Bacterianas/genética , Proteínas Bacterianas/fisiología , Prueba de Complementación Genética , Datos de Secuencia Molecular , Mutación , Pectobacterium/genética , Pectobacterium/patogenicidad , Homología de Secuencia de Aminoácido , Virulencia/genética
18.
Trends Microbiol ; 15(8): 363-71, 2007 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-17627825

RESUMEN

Phytopathogens coordinate multifaceted life histories and deploy stratified virulence determinants via complex, global regulation networks. We dissect the global regulation of four distantly related model phytopathogens to evaluate large-scale events and mechanisms that determine successful pathogenesis. Overarching themes include dependence on centralized cell-to-cell communication systems, pervasive two-component signal-transduction systems, post-transcriptional regulation systems, AraC-like regulators and sigma factors. Although these common regulatory systems control virulence, each functions in different capacities, and to differing ends, in the diverse species. Hence, the virulence regulation network of each species determines its survival and success in various life histories and niches.


Asunto(s)
Proteínas Bacterianas/metabolismo , Regulación Bacteriana de la Expresión Génica , Pectobacterium/patogenicidad , Enfermedades de las Plantas/microbiología , Pseudomonas syringae/patogenicidad , Ralstonia solanacearum/patogenicidad , Xanthomonas campestris/patogenicidad , Proteínas Bacterianas/genética , Solanum lycopersicum/microbiología , Pectobacterium/genética , Pectobacterium/metabolismo , Pseudomonas syringae/genética , Pseudomonas syringae/metabolismo , Ralstonia solanacearum/genética , Ralstonia solanacearum/metabolismo , Transducción de Señal , Solanum tuberosum/microbiología , Virulencia , Xanthomonas campestris/genética , Xanthomonas campestris/metabolismo
19.
Plant Biol (Stuttg) ; 8(5): 636-45, 2006 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-16755465

RESUMEN

Lipopolysaccharides (LPS), ubiquitous cell surface components of Gram-negative bacteria, are directly implicated in plant/pathogen interactions. However, their perception by the plant, the subsequent signal transduction in both compatible and incompatible interactions, as well as the defence reactions induced in compatible interactions are as yet poorly understood. We focused on biochemical and physiological reactions induced in cell suspensions of three Solanaceae species (tobacco, tomato, and potato) by purified lipopolysaccharides from PECTOBACTERIUM ATROSEPTICUM (PA), a pathogen of potato, and PSEUDOMONAS CORRUGATA (PSC), a pathogen of tomato. LPS PA and LPS PSC caused a significant acidification of potato, tomato, and tobacco extracellular media, whereas laminarin (a linear beta-1,3 oligosaccharide elicitor) induced an alkalinisation in tobacco and tomato, but not in potato cell suspensions. None of the two LPS induced the formation of active oxygen species in any of the hosts, while laminarin induced H (2)O (2) production in cells of tobacco but not of tomato and potato. In tomato cells, LPS PA and LPS PSC induced a strong but transitory stimulation of lipoxygenase activity, whereas laminarin induced a stable or slightly increasing LOX activity over the first 24 h of contact. In tobacco, LOX activity was not triggered by either LPS, but significantly increased following treatment with laminarin. In potato, neither LPS nor laminarin induced LOX activity, in contrast with concentrated culture filtrate of PHYTOPHTHORA INFESTANS (CCF). These results demonstrate that LPS, as well as laminarin, are perceived in different ways by SOLANACEAE species, and possibly cultivars. They also suggest that defence responses modulated by LPS depend on plant genotypes rather than on the type of interaction.


Asunto(s)
Lipopolisacáridos/farmacología , Nicotiana/efectos de los fármacos , Pectobacterium/metabolismo , Pseudomonas/metabolismo , Solanum lycopersicum/efectos de los fármacos , Solanum tuberosum/efectos de los fármacos , Células Cultivadas , Relación Dosis-Respuesta a Droga , Peróxido de Hidrógeno/metabolismo , Concentración de Iones de Hidrógeno , Lipopolisacáridos/metabolismo , Lipooxigenasa/metabolismo , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Solanum tuberosum/genética , Solanum tuberosum/metabolismo , Especificidad de la Especie , Factores de Tiempo , Nicotiana/genética , Nicotiana/metabolismo
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA