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1.
J Food Sci ; 86(1): 95-102, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-33258157

RESUMEN

Pectobacterium carotovorum is a phytopathogenic bacteria that causes significant economic loses in food crops, such as bell pepper, which is of special significance in the value of production and trade in Mexico. Therefore, a solution for fruit conservation must be sought. Due to environmental concerns, it is necessary the use of environmentally-friendly active packaging. In this article, chitosan and chitosan-thyme essential oil nanocoatings were used for the preservation of green bell pepper. Different formulations based on chitosan nanoparticles (CSNPs) and chitosan-thyme essential oil nanoparticles (15, 30, and 45%) were prepared. For uncoated and coated bell peppers, the quality and physiological variables of inoculated and uninoculated fruit with P. carotovorum during 12-day storage period were assessed. According to the results, the weight loss of the fruit remained almost constant over the storage days for the different formulations. A decrease in fruit firmness and an increase in the respiration rate and ascorbic acid content until day 8 with a decrease at the end of the storage period were observed. Of all the evaluated nanocoatings, the fruit treated with the formulation containing 15% CSNPs showed the lowest colony-forming units and disease incidence. Also, the coated bell peppers with this formulation had lower CO2 production compared to the remaining treatments, and the weight loss and firmness were maintained. Therefore, the use of CSNP coatings could represent a good alternative for the protection of bell pepper against the pathogenic bacteria P. carotovorum. PRACTICAL APPLICATION: The results of the application of nanocoatings based on chitosan and chitosan-thyme essential oil as an antibacterial agent against P. carotovorum on green bell pepper during 12-day storage period suggest that nanoparticle-based coatings can be a natural option for the preservation of fruit quality during ripening.


Asunto(s)
Capsicum , Embalaje de Alimentos/instrumentación , Almacenamiento de Alimentos/métodos , Frutas/microbiología , Nanopartículas , Aceites Volátiles , Ácido Ascórbico , Quitosano/química , México , Pectobacterium carotovorum/efectos de los fármacos , Pectobacterium carotovorum/crecimiento & desarrollo , Thymus (Planta)/química
2.
Molecules ; 24(16)2019 Aug 10.
Artículo en Inglés | MEDLINE | ID: mdl-31405174

RESUMEN

Ornithogalum is an ornamental flowering species that grows from a bulb and is highly susceptible to soft-rot disease caused by Pectobacterium carotovorum (Pc). Interspecific hybridization between O. thyrsoides and O. dubium yielded hybrids with enhanced resistance to that pathogen. The hybrids displayed distinct phenolic-compound profiles with several peaks that were specifically heightened following Pc infection. Three of these compounds were isolated and identified as novel kaempferol O-tri-glycosides. The structures of these compounds were elucidated using reversed phase high-performance liquid chromatography (RP-LC), RP-LC coupled to high-resolution mass spectrometry (RP-LC-MS), and nuclear magnetic resonance (NMR) (1D 1H and 13C, DEPT, HMQC, HMBC, COSY, and NOE), in order to achieve pure and defined compounds data. The new compounds were finally identified as kaempferol 3-O-[4-O-α-l-(3-O-acetic)-rhamnopyranosyl-6-O-ß-d-xylopyranosyl]-ß-d-glucopyranoside, kaempferol 3-O-[4-O-α-l-(2-O-acetic)-rhamnopyranosyl-6-O-ß-d-xylopyranosyl]-ß-d-glucopyranoside and kaempferol 3-O-[4-O-α-l-(2,3-O-diacetic)-rhamnopyranosyl-6-O-ß-d-xylopyranosyl]-ß-d-glucopyranoside.


Asunto(s)
Glicósidos , Quempferoles , Ornithogalum/microbiología , Pectobacterium carotovorum/crecimiento & desarrollo , Enfermedades de las Plantas/microbiología , Cruzamientos Genéticos , Glicósidos/química , Glicósidos/farmacología , Quempferoles/química , Quempferoles/farmacología , Relación Estructura-Actividad
3.
Arch Microbiol ; 201(5): 615-622, 2019 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-30741323

RESUMEN

Pectobacterium carotovorum was incubated in formulations of chitosan nanoparticles or thyme essential oil-loaded chitosan nanoparticles for a maximum period of 48 h time. The cellular changes and viability were evaluated by transmission electron microscopy (TEM), and two colorimetric assays 3-(4,5 dimethylthiazol-2-yl)-2,5diphenyl tetrazolium bromide (MTT) and alamar blue (AMB), respectively. The incubation time and the addition of the secondary metabolite to the formulation were key factors to the cell damage and cell inhibitory effects on P. carotovorum, TEM observations overall demonstrated on the treated bacterium, cell surface alterations such as deforming and disappearance of the cell wall and the plasmatic membrane, with agglomeration of nanoparticles outside and inside of the cells, loss of cell content and lysis. Cell viability was reduced about 80% (MTT) and 100% (AMB) in the applied treatment of chitosan-loaded thyme essential oil nanoparticles after 48 h incubation, in addition, the total cell inhibition was shown from 6 h incubation onwards with the AMB assay. The TEM micrographs and the cell viability assays provided enough evidence of the antimicrobial activity of the nanostructured formulations compared with the control where no damage was observed.


Asunto(s)
Antibacterianos/farmacología , Quitosano/farmacología , Aceites Volátiles/farmacología , Pectobacterium carotovorum/efectos de los fármacos , Pectobacterium carotovorum/crecimiento & desarrollo , Microscopía Electrónica de Transmisión , Nanopartículas , Thymus (Planta)/química
4.
Pak J Biol Sci ; 20(6): 278-288, 2017.
Artículo en Inglés | MEDLINE | ID: mdl-29023052

RESUMEN

BACKGROUND AND OBJECTIVE: Salicylic acid is a signal molecule which activates plant defense against plant pathogens such as the soft rot enterobacterium Pectobacterium carotovorum subsp. carotovorum. The objectives of study were to determine bactericidal effects of salicylic acid on the growth of P. carotovorum subsp. carotovorum and secondly, assess chemical responses of P. carotovorum subsp. carotovorum to salicylic acid. MATERIALS AND METHODS: Pectobacterium carotovorum subsp. carotovorum was grown in lysogeny broth amended with salicylic acid at concentrations of 0, 100, 200, 400, 800 and 1200 mg L-1. The P. carotovorum subsp. carotovorum cultures were incubated at 25°C and sampled at two time points, 0 h (sampled before incubation) and 24 h. Bacterial counts were done at the onset of the incubation (0 h) and after the 24 h incubation. The set which was incubated for 24 h was split into two, one subset was centrifuged and the other was not. From the centrifuged subset the supernatant was recovered and was, together with all the other samples (0 and 24 h not centrifuged), analyzed with1H nuclear magnetic resonance and gas chromatography. RESULTS: Bacterial counts done before and after incubation showed that the lower concentrations of salicylic acid, 0, 100, 200 and 400 mg L-1, supported the growth of P. carotovorum subsp. carotovorum whereas the higher concentrations of 800 and 1200 mg L-1 inhibited the growth of the bacterium completely. Nuclear magnetic resonance results showed either slight or no differences in the metabolite profiles and gas chromatography showed different responses without a clearly defined pattern among the experimental treatments. However, methanethiol was detected by both nuclear magnetic resonance and gas chromatography in all the treatments and was probably formed as a result of the breakdown of lysogeny broth. CONCLUSION: From the results obtained it was concluded that salicylic acid promotes the growth of P. carotovorum subsp. carotovorum at lower concentrations of 0-400 mg L-1 but higher concentrations of salicylic acid of 800 and 1200 mg L-1 inhibit bacterial growth. All the tested salicylic acid concentrations (0-1200 mg L-1) cause only slight chemical shifts in the bacterial culture. Methanethiol was detected in all treatments and it is probably formed from the breakdown of lysogeny broth.


Asunto(s)
Antibacterianos/farmacología , Pectobacterium carotovorum/efectos de los fármacos , Ácido Salicílico/farmacología , Cromatografía de Gases , Relación Dosis-Respuesta a Droga , Pectobacterium carotovorum/crecimiento & desarrollo , Pectobacterium carotovorum/metabolismo , Espectroscopía de Protones por Resonancia Magnética , Factores de Tiempo
5.
Plant Physiol Biochem ; 118: 479-493, 2017 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-28756346

RESUMEN

Plant Growth Promoting Rhizobacteria (PGPR) represent an alternative to improve plant growth and yield as well as to act as agents of biocontrol. This study characterized isolates of Streptomyces spp. (Stm) as PGPR, determined the antagonism of these isolates against Pectobacterium carotovorum subsp. brasiliensis (Pcb), evaluated the ability of Stm on promoting growth and modulating the defense-related metabolism of tomato plants, and the potential of Stm isolates on reducing soft rot disease in this species. The VOC profile of Stm was also verified. Promotion of plant growth was assessed indirectly through VOC emission and by direct interaction with Stm isolates in the roots. Evaluation of soft rot disease was performed in vitro on plants treated with Stm and challenged with Pcb. Enzymes related to plant defense were then analyzed in plants treated with three selected isolates of Stm, and PM1 was chosen for further Pcb-challenging experiment. Streptomyces spp. isolates displayed characteristics of PGPR. PM3 was the isolate with efficient antagonism against Pcb by dual-culture. Most of the isolates promoted growth of root and shoot of tomato plants by VOC, and PM5 was the isolate that most promoted growth by direct interaction with Stm. Soft rot disease and mortality of plants were significantly reduced when plants were treated with StmPM1. Modulation of secondary metabolism was observed with Stm treatment, and fast response of polyphenoloxidases was detected in plants pretreated with StmPM1 and challenged with Pcb. Peroxidase was significantly activated three days after infection with Pcb in plants pretreated with StmPM1. Results suggest that Streptomyces sp. PM1 and PM5 have the potential to act as PGPR.


Asunto(s)
Pectobacterium carotovorum/crecimiento & desarrollo , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Raíces de Plantas/microbiología , Solanum lycopersicum/microbiología , Streptomyces/crecimiento & desarrollo , Streptomyces/aislamiento & purificación
6.
FEMS Microbiol Lett ; 363(23)2016 12.
Artículo en Inglés | MEDLINE | ID: mdl-27915254

RESUMEN

Carocin D is a bacteriocin produced by Pectobacterium carotovorum subsp. carotovorum Pcc21. Carocin D inhibits the growth of P carotovorum subsp. carotovorum and closely related strains. Pectobacterium carotovorum subsp. carotovorum is a causative bacterium for soft rot disease and leads to severe economic losses. Bacteriocins recognize and interact with a specific membrane protein of target bacteria as a receptor. To identify the receptor responsible for carocin D recognition, mutants that underwent a phenotypic change from carocin D sensitivity to carocin D insensitivity were screened. Based on Tn5 insertions, carocin D sensitivity was dependent on expression of the outer membrane protein OmpF. The insensitivity of the mutant (Pcc3MR) to carocin D was complemented with ompF from carocin D-sensitive strains, not from carocin D-resistant strains. The selectivity between sensitive and resistant strains could be attributed to variation in OmpFs in the cell-surface-exposed regions. Based on sequence analysis and complementation assays, it appears that carocin D uses OmpF as a receptor and is translocated by the TonB system. According to previously reported translocation mechanisms of colicins, OmpF works along with the TolA system rather than the TonB system. Therefore, the current findings suggest that carocin D is imported by a unique colicin-like bacteriocin translocation system.


Asunto(s)
Antibacterianos/metabolismo , Bacteriocinas/metabolismo , Pectobacterium carotovorum/crecimiento & desarrollo , Pectobacterium carotovorum/genética , Porinas/genética , Secuencia de Aminoácidos , Sitios de Unión/genética , Regulación Bacteriana de la Expresión Génica , Pruebas de Sensibilidad Microbiana , Mutagénesis , Enfermedades de las Plantas/microbiología , Análisis de Secuencia de Proteína , Transposasas/genética
7.
BMC Genomics ; 17(1): 614, 2016 08 11.
Artículo en Inglés | MEDLINE | ID: mdl-27515663

RESUMEN

BACKGROUND: Long noncoding RNAs (lncRNAs) represent a class of RNA molecules that are implicated in regulation of gene expression in both mammals and plants. While much progress has been made in determining the biological functions of lncRNAs in mammals, the functional roles of lncRNAs in plants are still poorly understood. Specifically, the roles of long intergenic nocoding RNAs (lincRNAs) in plant defence responses are yet to be fully explored. RESULTS: In this study, we used strand-specific RNA sequencing to identify 1113 lincRNAs in potato (Solanum tuberosum) from stem tissues. The lincRNAs are expressed from all 12 potato chromosomes and generally smaller in size compared to protein-coding genes. Like in other plants, most potato lincRNAs possess single exons. A time-course RNA-seq analysis between a tolerant and a susceptible potato cultivar showed that 559 lincRNAs are responsive to Pectobacterium carotovorum subsp. brasiliense challenge compared to mock-inoculated controls. Moreover, coexpression analysis revealed that 17 of these lincRNAs are highly associated with 12 potato defence-related genes. CONCLUSIONS: Together, these results suggest that lincRNAs have potential functional roles in potato defence responses. Furthermore, this work provides the first library of potato lincRNAs and a set of novel lincRNAs implicated in potato defences against P. carotovorum subsp. brasiliense, a member of the soft rot Enterobacteriaceae phytopathogens.


Asunto(s)
Resistencia a la Enfermedad/genética , Regulación de la Expresión Génica de las Plantas , Interacciones Huésped-Patógeno , Pectobacterium carotovorum/patogenicidad , ARN Largo no Codificante/genética , ARN de Planta/genética , Solanum tuberosum/genética , Cromosomas de las Plantas/química , Exones , Biblioteca de Genes , Ontología de Genes , Anotación de Secuencia Molecular , Pectobacterium carotovorum/crecimiento & desarrollo , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/microbiología , Inmunidad de la Planta/genética , Tallos de la Planta/genética , Tallos de la Planta/inmunología , Tallos de la Planta/microbiología , ARN Largo no Codificante/clasificación , ARN Largo no Codificante/inmunología , ARN de Planta/clasificación , ARN de Planta/inmunología , Solanum tuberosum/inmunología , Solanum tuberosum/microbiología
8.
Sci Rep ; 6: 27085, 2016 06 03.
Artículo en Inglés | MEDLINE | ID: mdl-27255932

RESUMEN

Tunnelling nanotubes and cytonemes function as highways for the transport of organelles, cytosolic and membrane-bound molecules, and pathogens between cells. During viral infection in the model organism Drosophila melanogaster, a systemic RNAi antiviral response is established presumably through the transport of a silencing signal from one cell to another via an unknown mechanism. Because of their role in cell-cell communication, we investigated whether nanotube-like structures could be a mediator of the silencing signal. Here, we describe for the first time in the context of a viral infection the presence of nanotube-like structures in different Drosophila cell types. These tubules, made of actin and tubulin, were associated with components of the RNAi machinery, including Argonaute 2, double-stranded RNA, and CG4572. Moreover, they were more abundant during viral, but not bacterial, infection. Super resolution structured illumination microscopy showed that Argonaute 2 and tubulin reside inside the tubules. We propose that nanotube-like structures are one of the mechanisms by which Argonaute 2, as part of the antiviral RNAi machinery, is transported between infected and non-infected cells to trigger systemic antiviral immunity in Drosophila.


Asunto(s)
Proteínas Argonautas/genética , Proteínas de Drosophila/genética , Drosophila melanogaster/metabolismo , Orgánulos/metabolismo , ARN Bicatenario/genética , Proteínas Virales/antagonistas & inhibidores , Actinas/genética , Actinas/metabolismo , Animales , Proteínas Argonautas/metabolismo , Transporte Biológico , Comunicación Celular , Línea Celular , Dicistroviridae/genética , Dicistroviridae/crecimiento & desarrollo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/microbiología , Drosophila melanogaster/ultraestructura , Drosophila melanogaster/virología , Regulación de la Expresión Génica , Genes Reporteros , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Orgánulos/microbiología , Orgánulos/ultraestructura , Orgánulos/virología , Pectobacterium carotovorum/genética , Pectobacterium carotovorum/crecimiento & desarrollo , Interferencia de ARN , ARN Bicatenario/metabolismo , Tubulina (Proteína)/genética , Tubulina (Proteína)/metabolismo , Proteínas Virales/genética , Proteínas Virales/metabolismo , Proteínas de Unión al GTP rab/genética , Proteínas de Unión al GTP rab/metabolismo , Proteínas de Unión a GTP rab7
9.
Appl Environ Microbiol ; 82(4): 1353-1360, 2016 02 15.
Artículo en Inglés | MEDLINE | ID: mdl-26682862

RESUMEN

During their colonization of plants, human enteric pathogens, such as Salmonella enterica, are known to benefit from interactions with phytopathogens. At least in part, benefits derived by Salmonella from the association with a soft rot caused by Pectobacterium carotovorum were shown to be dependent on Salmonella KdgR, a regulator of genes involved in the uptake and utilization of carbon sources derived from the degradation of plant polymers. A Salmonella kdgR mutant was more fit in soft rots but not in the lesions caused by Xanthomonas spp. and Pseudomonas spp. Bioinformatic, phenotypic, and gene expression analyses demonstrated that the KdgR regulon included genes involved in uptake and metabolism of molecules resulting from pectin degradation as well as those central to the utilization of a number of other carbon sources. Mutant analyses indicated that the Entner-Doudoroff pathway, in part controlled by KdgR, was critical for the persistence within soft rots and likely was responsible for the kdgR phenotype.


Asunto(s)
Interacciones Microbianas , Viabilidad Microbiana , Pectobacterium carotovorum/crecimiento & desarrollo , Enfermedades de las Plantas/microbiología , Regulón , Salmonella enterica/crecimiento & desarrollo , Verduras/microbiología , Metabolismo de los Hidratos de Carbono , Carbono/metabolismo , Hidrólisis , Redes y Vías Metabólicas/genética , Mutación , Pectinas/metabolismo , Pseudomonas/crecimiento & desarrollo , Salmonella enterica/genética , Xanthomonas/crecimiento & desarrollo
11.
Proc Natl Acad Sci U S A ; 112(17): 5533-8, 2015 Apr 28.
Artículo en Inglés | MEDLINE | ID: mdl-25870275

RESUMEN

Oligogalacturonides (OGs) are fragments of pectin that activate plant innate immunity by functioning as damage-associated molecular patterns (DAMPs). We set out to test the hypothesis that OGs are generated in planta by partial inhibition of pathogen-encoded polygalacturonases (PGs). A gene encoding a fungal PG was fused with a gene encoding a plant polygalacturonase-inhibiting protein (PGIP) and expressed in transgenic Arabidopsis plants. We show that expression of the PGIP-PG chimera results in the in vivo production of OGs that can be detected by mass spectrometric analysis. Transgenic plants expressing the chimera under control of a pathogen-inducible promoter are more resistant to the phytopathogens Botrytis cinerea, Pectobacterium carotovorum, and Pseudomonas syringae. These data provide strong evidence for the hypothesis that OGs released in vivo act as a DAMP signal to trigger plant immunity and suggest that controlled release of these molecules upon infection may be a valuable tool to protect plants against infectious diseases. On the other hand, elevated levels of expression of the chimera cause the accumulation of salicylic acid, reduced growth, and eventually lead to plant death, consistent with the current notion that trade-off occurs between growth and defense.


Asunto(s)
Proteínas de Arabidopsis/biosíntesis , Arabidopsis/metabolismo , Proteínas Fúngicas/biosíntesis , Ácidos Hexurónicos/metabolismo , Enfermedades de las Plantas/inmunología , Inmunidad de la Planta , Proteínas de Plantas/biosíntesis , Poligalacturonasa/biosíntesis , Animales , Arabidopsis/genética , Arabidopsis/inmunología , Arabidopsis/microbiología , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/inmunología , Botrytis/crecimiento & desarrollo , Botrytis/inmunología , Proteínas Fúngicas/genética , Proteínas Fúngicas/inmunología , Ácidos Hexurónicos/inmunología , Ratones Transgénicos , Pectobacterium carotovorum/crecimiento & desarrollo , Pectobacterium carotovorum/inmunología , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/genética , Proteínas de Plantas/inmunología , Poligalacturonasa/genética , Poligalacturonasa/inmunología , Pseudomonas syringae/crecimiento & desarrollo , Pseudomonas syringae/inmunología , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología
12.
Dev Comp Immunol ; 49(1): 96-102, 2015 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-25445903

RESUMEN

Hemocyanins present in invertebrate hemolymph are multifunctional proteins, responsible for oxygen transport and contributing to innate immunity through phenoloxidase-like activity. In arthropods, hemocyanin has been identified as a source of broad-spectrum antimicrobial peptides during infection. Conversely, no hemocyanin-derived antimicrobial peptides have been reported for molluscs. The present study describes a putative antimicrobial region, termed haliotisin, located within the linking sequence between the α-helical domain and ß-sheet domain of abalone (Haliotis tuberculata) hemocyanin functional unit E. A series of synthetic peptides based on overlapping fragments of the haliotisin region were tested for their bactericidal potential. Incubating Gram-positive and Gram-negative bacteria in the presence of certain haliotisin peptides, notably peptides 3-4-5 (DTFDYKKFGYRYDSLELEGRSISRIDELIQQRQEKDRTFAGFLLKGFGTSAS) led to reductions in microbial growth. Furthermore, transmission electron micrographs of haliotisin-treated bacteria revealed damages to the microbial cell wall. Data discussed here provides the first evidence to suggest that molluscan hemocyanin may act as a source of anti-infective peptides.


Asunto(s)
Péptidos Catiónicos Antimicrobianos/metabolismo , Péptidos Catiónicos Antimicrobianos/farmacología , Bacterias/efectos de los fármacos , Gastrópodos/metabolismo , Hemocianinas/metabolismo , Secuencia de Aminoácidos , Animales , Péptidos Catiónicos Antimicrobianos/química , Bacillus subtilis/efectos de los fármacos , Bacillus subtilis/crecimiento & desarrollo , Bacillus subtilis/ultraestructura , Bacterias/crecimiento & desarrollo , Bacterias/ultraestructura , Pruebas de Sensibilidad Microbiana , Microscopía Electrónica de Transmisión , Modelos Moleculares , Datos de Secuencia Molecular , Pectobacterium carotovorum/efectos de los fármacos , Pectobacterium carotovorum/crecimiento & desarrollo , Pectobacterium carotovorum/ultraestructura , Péptidos/síntesis química , Péptidos/farmacología , Estructura Terciaria de Proteína , Homología de Secuencia de Aminoácido
13.
Mikrobiol Z ; 77(6): 47-54, 2015.
Artículo en Ruso | MEDLINE | ID: mdl-26829839

RESUMEN

Antagonistic activity 100 strains of Bacillus bacteria towards to museum and actual strains of phytopathogenic bacteria and fungy was defined. Relation between level of antagonistic activity to phytopathogenic bacteria and genus accessory of the last was shown. The medium level of antagonism to fungal phytopathogens at 30% of the studied strains of Bacillus bacteria was shown. 5 strains of Bacillus sp. with high and medium levels of antagonism to phytopathogens bacterial and fungy nature was selected and considered as perspective for creation of biological preparations for plant protection.


Asunto(s)
Antibiosis , Bacillus/patogenicidad , Agrobacterium tumefaciens/crecimiento & desarrollo , Bacillus/fisiología , Recuento de Colonia Microbiana , Fusarium/crecimiento & desarrollo , Pectobacterium carotovorum/crecimiento & desarrollo , Pseudomonas fluorescens/crecimiento & desarrollo , Pseudomonas syringae/crecimiento & desarrollo , Xanthomonas campestris/crecimiento & desarrollo
14.
FEMS Microbiol Lett ; 354(1): 75-82, 2014 May.
Artículo en Inglés | MEDLINE | ID: mdl-24655217

RESUMEN

Peptaibols, mainly produced by Trichoderma, play a pivotal role in controlling plant disease caused by fungi, virus, and Gram-positive bacteria. In the current study, we evaluated the control effect of Trichokonins, antimicrobial peptaibols from Trichoderma pseudokoningii SMF2, on soft rot disease of Chinese cabbage caused by a Gram-negative bacterium Pectobacterium carotovorum subsp. carotovorum and analyzed the mechanism involved. Trichokonins treatment (0.3 mg L(-1) ) enhanced the resistance of Chinese cabbage against Pcc infection. However, Trichokonins could hardly inhibit the growth of Pcc in vitro, even at high concentration (500 mg L(-1) ). Therefore, the direct effect of Trichokonins on Pcc may not the main reason why Trichokonins could control soft rot of Chinese cabbage. Trichokonin treatment led to an obvious increase in the production of reactive oxygen species hydrogen peroxide and superoxide radical, a significant enhance of the activities of pathogenesis-related enzymes catalase, polyphenoloxidase and peroxidase, and upregulation of the expression of salicylic acid - responsive pathogenesis-related protein gene acidic PR-1a in Chinese cabbage. These results indicate that Trichokonins induce resistance in Chinese cabbage against Pcc infection through the activation of salicylic acid signaling pathway, which imply the potential of Trichoderma and peptaibols in controlling plant disease caused by Gram-negative bacteria.


Asunto(s)
Alameticina/farmacología , Brassica/inmunología , Pectobacterium carotovorum/fisiología , Enfermedades de las Plantas/microbiología , Trichoderma/metabolismo , Alameticina/metabolismo , Brassica/efectos de los fármacos , Brassica/genética , Brassica/microbiología , Datos de Secuencia Molecular , Pectobacterium carotovorum/efectos de los fármacos , Pectobacterium carotovorum/crecimiento & desarrollo , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/inmunología , Ácido Salicílico/inmunología
15.
Biotechnol Lett ; 36(2): 371-81, 2014 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-24126536

RESUMEN

Non-coding, small RNAs (sRNAs) have been identified in a wide spectrum of organisms ranging from bacteria to humans; however, the role and mechanisms of these sRNA in plant immunity is largely unknown. To determine possible roles of sRNA in plant-pathogen interaction, we carried out a high-throughput sRNA sequencing of Brassica campestris using non-infected plants and plants infected with Erwinia carotovora. Consistent with our hypothesis that distinct classes of host sRNAs alerts their expression levels in response to infection, we found that: (1) host 28-nt sRNAs were strongly increased under pathogen infection; and (2) a group of host sRNAs homologous to the pathogen genome also accumulated at significantly higher level. Our data thus suggest several distinct classes of the host sRNAs may enhance their function by up-regulation of their expression/stability in response to bacterial pathogen challenges.


Asunto(s)
Brassica/microbiología , Interacciones Huésped-Patógeno , Pectobacterium carotovorum/crecimiento & desarrollo , ARN Pequeño no Traducido/biosíntesis , Transcriptoma , Brassica/genética
16.
Genetika ; 49(5): 609-16, 2013 May.
Artículo en Ruso | MEDLINE | ID: mdl-24159802

RESUMEN

Transgenic plants of camelina (Camelina sativa (L.) Crantz) with the synthetic gene of antimicrobial peptide cecropin P1 (cecP1) were obtained. Agrobacterium-mediated transformation is performed using the binary vector pGA482::cecP1 by vacuum infiltration of flower buds. The presence of the cecP1 gene in the genome of plants was confirmed by PCR. cecP1 gene expression in transgenic plants was shown by Western blot analysis and by antimicrobial activity of plant extracts against the bacterial phytopathogene Erwinia carotovora. The plants of F0 and F1 generations had the normal phenotype and retained the ability to form viable seeds in self-pollination. cecP1 plants exhibit enhanced resistance to bacterial and fungal phytopathogens: Erwinia carotovora and Fusarium sporotrichioides. The increased sustainability of cecropin P1-expressing plants against salt stress is shown. The possibility of the integration of the cecP1 gene into the overall protective system of plants against biotic and abiotic stresses is discussed.


Asunto(s)
Brassicaceae/metabolismo , Resistencia a la Enfermedad , Genoma de Planta , Pectobacterium carotovorum/crecimiento & desarrollo , Péptidos/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Brassicaceae/genética , Brassicaceae/microbiología , Péptidos/genética , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/microbiología
17.
J Microbiol Biotechnol ; 23(8): 1147-53, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23727798

RESUMEN

Pectobacterium carotovorum subsp. carotovorum (formerly Erwinia carotovora subsp. carotovora) is a plant pathogen that causes soft rot and stem rot diseases in several crops, including Chinese cabbage, potato, and tomato. To control this bacterium, we isolated a bacteriophage, PP1, with lytic activity against P. carotovorum subsp. carotovorum. Transmission electron microscopy revealed that the PP1 phage belongs to the Podoviridae family of the order Caudovirales, which exhibit icosahedral heads and short non-contractile tails. PP1 phage showed high specificity for P. carotovorum subsp. carotovorum, and several bacteria belonging to different species and phyla were resistant to PP1. This phage showed rapid and strong lytic activity against its host bacteria in liquid medium and was stable over a broad range of pH values. Disease caused by P. carotovorum subsp. carotovorum was significantly reduced by PP1 treatment. Overall, PP1 bacteriophage effectively controls P. carotovorum subsp. carotovorum.


Asunto(s)
Bacteriófagos/crecimiento & desarrollo , Pectobacterium carotovorum/crecimiento & desarrollo , Pectobacterium carotovorum/virología , Control Biológico de Vectores/métodos , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Podoviridae/crecimiento & desarrollo , Bacteriólisis , Bacteriófagos/clasificación , Bacteriófagos/aislamiento & purificación , Bacteriófagos/ultraestructura , Microscopía Electrónica de Transmisión , Podoviridae/clasificación , Podoviridae/aislamiento & purificación , Podoviridae/ultraestructura , Verduras/microbiología
18.
Phytopathology ; 103(12): 1268-79, 2013 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-23758294

RESUMEN

Pectobacterium carotovorum subsp. brasiliense is a newly identified member of the potato soft rot enterobacteriaceae. The pathogenesis of this pathogen is still poorly understood. In this study, an mCherry-P. carotovorum subsp. brasiliense-tagged strain was generated to study P. carotovorum subsp. brasiliense-potato plant interactions. Prior to use, the tagged strain was evaluated for in vitro growth, plasmid stability, and virulence on potato tubers and shown to be similar to the wild type. Four potato cultivars were evaluated for stem-based resistance against P. carotovorum subsp. brasiliense. Confocal laser-scanning microscopy and in vitro viable cell counts showed that P. carotovorum subsp. brasiliense is able to penetrate roots of a susceptible potato cultivar as early as 12 h postinoculation and migrate upward into aerial stem parts. Due to the phenotypic differences observed between tolerant and susceptible cultivars, a comparison of P. carotovorum subsp. brasiliense colonization patterns in these cultivars was undertaken. In the susceptible cultivar, P. carotovorum subsp. brasiliense cells colonized the xylem tissue, forming "biofilm-like" aggregates that led to occlusion of some of the vessels. In contrast, in the tolerant cultivar, P. carotovorum subsp. brasiliense appeared as free-swimming planktonic cells with no specific tissue localization. This suggests that there are resistance mechanisms in the tolerant cultivar that limit aggregation of P. carotovorum subsp. brasiliense in planta and, hence, the lack of symptom development in this cultivar.


Asunto(s)
Resistencia a la Enfermedad , Susceptibilidad a Enfermedades , Pectobacterium carotovorum/patogenicidad , Enfermedades de las Plantas/microbiología , Solanum tuberosum/microbiología , Proteínas Luminiscentes , Microscopía Confocal , Pectobacterium carotovorum/genética , Pectobacterium carotovorum/crecimiento & desarrollo , Fenotipo , Enfermedades de las Plantas/inmunología , Raíces de Plantas/inmunología , Raíces de Plantas/microbiología , Tallos de la Planta/inmunología , Tallos de la Planta/microbiología , Tubérculos de la Planta/inmunología , Tubérculos de la Planta/microbiología , Plásmidos , Proteínas Recombinantes de Fusión , Solanum tuberosum/inmunología , Virulencia , Proteína Fluorescente Roja
19.
Int J Biol Macromol ; 57: 185-92, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23511055

RESUMEN

The current study focuses on the preparation of new N-(cinnamyl) chitosan derivatives as antimicrobial agents against nine types of crop-threatening pathogens. Chitosan was reacted with a set of aromatic cinnamaldehyde analogs by reductive amination involving formation of the corresponding imines, followed by reduction with sodium borohydride to produce N-(cinnamyl) chitosan derivatives. The structural characterization was confirmed by (1)H and (13)C NMR spectroscopy and the degrees of substitution ranged from 0.08 to 0.28. The antibacterial activity was evaluated in vitro by minimum inhibitory concentration (MIC) against Agrobacterium tumefaciens and Erwinia carotovora. A higher inhibition activity was obtained by N-(α-methylcinnamyl) chitosan with MIC 1275 and 1025 mg/L against A. tumefaciens and E. carotovora, respectively followed by N-(o-methoxycinnamyl) chitosan (MIC=1925 and 1550 mg/L, respectively). The antifungal assessment was evaluated in vitro by mycelial radial growth technique against Alternaria alternata, Botrytis cinerea, Botryodiplodia theobromae, Fusarium oxysporum, Fusarium solani, Pythium debaryanum and Phytophthora infestans. N-(o-methoxycinnamyl) chitosan showed the highest antifungal activity among the tested compounds against the airborne fungi A. alternata, B. cinerea, Bd. theobromae and Ph. infestans with EC50 of 672, 796, 980 and 636 mg/L, respectively. However, N-(p-N-dimethylaminocinnamyl) chitosan was the most active against the soil born fungi F. oxysporum, F. solani and P. debaryanum (EC50=411, 566 and 404 mg/L, respectively). On the other hand, the chitosan derivatives caused significant reduction in spore germination of A. alternata, B. cinerea, F. oxysporum and F. solani compared to chitosan and the reduction in spore germination was higher than that of the mycelia inhibition. The synthesis and characterization of new chitosan derivatives are ongoing in our laboratory aiming to obtain derivatives with higher antimicrobial activities and used as safe alternatives to harmful microbicides.


Asunto(s)
Acroleína/análogos & derivados , Agrobacterium tumefaciens/crecimiento & desarrollo , Antiinfecciosos , Quitosano , Hongos/fisiología , Pectobacterium carotovorum/crecimiento & desarrollo , Acroleína/química , Antiinfecciosos/síntesis química , Antiinfecciosos/química , Antiinfecciosos/farmacología , Quitosano/análogos & derivados , Quitosano/síntesis química , Quitosano/química , Quitosano/farmacología , Micelio/crecimiento & desarrollo , Esporas Fúngicas/crecimiento & desarrollo
20.
mBio ; 4(1): e00557-12, 2013 Feb 12.
Artículo en Inglés | MEDLINE | ID: mdl-23404399

RESUMEN

UNLABELLED: Although enteric human pathogens are usually studied in the context of their animal hosts, a significant portion of their life cycle occurs on plants. Plant disease alters the phyllosphere, leading to enhanced growth of human pathogens; however, the impact of human pathogens on phytopathogen biology and plant health is largely unknown. To characterize the interaction between human pathogens and phytobacterial pathogens in the phyllosphere, we examined the interactions between Pectobacterium carotovorum subsp. carotovorum and Salmonella enterica or Escherichia coli O157:H7 with regard to bacterial populations, soft rot progression, and changes in local pH. The presence of P. carotovorum subsp. carotovorum enhanced the growth of both S. enterica and E. coli O157:H7 on leaves. However, in a microaerophilic environment, S. enterica reduced P. carotovorum subsp. carotovorum populations and soft rot progression by moderating local environmental pH. Reduced soft rot was not due to S. enterica proteolytic activity. Limitations on P. carotovorum subsp. carotovorum growth, disease progression, and pH elevation were not observed on leaves coinoculated with E. coli O157:H7 or when leaves were coinoculated with S. enterica in an aerobic environment. S. enterica also severely undermined the relationship between the phytobacterial population and disease progression of a P. carotovorum subsp. carotovorum budB mutant defective in the 2,3-butanediol pathway for acid neutralization. Our results show that S. enterica and E. coli O157:H7 interact differently with the enteric phytobacterial pathogen P. carotovorum subsp. carotovorum. S. enterica inhibition of soft rot progression may conceal a rapidly growing human pathogen population. Whereas soft rotted produce can alert consumers to the possibility of food-borne pathogens, healthy-looking produce may entice consumption of contaminated vegetables. IMPORTANCE: Salmonella enterica and Escherichia coli O157:H7 may use plants to move between animal and human hosts. Their populations are higher on plants cocolonized with the common bacterial soft rot pathogen Pectobacterium carotovorum subsp. carotovorum, turning edible plants into a risk factor for human disease. We inoculated leaves with P. carotovorum subsp. carotovorum and S. enterica or E. coli O157:H7 to study the interactions between these bacteria. While P. carotovorum subsp. carotovorum enhanced the growth of both S. enterica and E. coli O157:H7, these human pathogens affected P. carotovorum subsp. carotovorum fundamentally differently. S. enterica reduced P. carotovorum subsp. carotovorum growth and acidified the environment, leading to less soft rot on leaves; E. coli O157:H7 had no such effects. As soft rot signals a food safety risk, the reduction of soft rot symptoms in the presence of S. enterica may lead consumers to eat healthy-looking but S. enterica-contaminated produce.


Asunto(s)
Antibacterianos/metabolismo , Antibiosis , Ácidos Carboxílicos/metabolismo , Pectobacterium carotovorum/crecimiento & desarrollo , Salmonella enterica/crecimiento & desarrollo , Salmonella enterica/metabolismo , Bacterias , Ambiente , Escherichia coli , Escherichia coli O157/crecimiento & desarrollo , Escherichia coli O157/metabolismo , Concentración de Iones de Hidrógeno , Lactuca , Enfermedades de las Plantas/microbiología , Hojas de la Planta/microbiología , Plantas
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