Dataset Information

The Effect of Phenazine-1-Carboxylic Acid on Mycelial Growth of Botrytis cinerea Produced by Pseudomonas aeruginosa LV Strain.

ABSTRACT: One of the most important postharvest plant pathogens that affect strawberries, grapes and tomatoes is Botrytis cinerea, known as gray mold. The fungus remains in latent form until spore germination conditions are good, making infection control difficult, causing great losses in the whole production chain. This study aimed to purify and identify phenazine-1-carboxylic acid (PCA) produced by the Pseudomonas aeruginosa LV strain and to determine its antifungal activity against B. cinerea. The compounds produced were extracted with dichloromethane and passed through a chromatographic process. The purity level of PCA was determined by reversed-phase high-performance liquid chromatography semi-preparative. The structure of PCA was confirmed by nuclear magnetic resonance and electrospray ionization mass spectrometry. Antifungal activity was determined by the dry paper disk and minimum inhibitory concentration (MIC) methods and identified by scanning electron microscopy and confocal microscopy. The results showed that PCA inhibited mycelial growth, where MIC was 25 ?g mL-1. Microscopic analysis revealed a reduction in exopolysaccharide (EPS) formation, showing distorted and damaged hyphae of B. cinerea. The results suggested that PCA has a high potential in the control of B. cinerea and inhibition of EPS (important virulence factor). This natural compound is a potential alternative to postharvest control of gray mold disease.

SUBMITTER: Simionato AS

PROVIDER: S-EPMC5469906 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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The Effect of Phenazine-1-Carboxylic Acid on Mycelial Growth of Botrytis cinerea Produced by Pseudomonas aeruginosa LV Strain.

Simionato Ane S AS Navarro Miguel O P MOP de Jesus Maria L A MLA Barazetti André R AR da Silva Caroline S CS Simões Glenda C GC Balbi-Peña Maria I MI de Mello João C P JCP Panagio Luciano A LA de Almeida Ricardo S C RSC Andrade Galdino G de Oliveira Admilton G AG

Frontiers in microbiology 20170614

One of the most important postharvest plant pathogens that affect strawberries, grapes and tomatoes is Botrytis cinerea, known as gray mold. The fungus remains in latent form until spore germination conditions are good, making infection control difficult, causing great losses in the whole production chain. This study aimed to purify and identify phenazine-1-carboxylic acid (PCA) produced by the Pseudomonas aeruginosa LV strain and to determine its antifungal activity against B. ...[more]

PMID: 28659907

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Project description:Botrytis cinerea infection can be very devastating for tomato production, as it can result in a large-scale reduction in tomato fruit production and fruit quality after harvest. Thus, it negatively affects tomato yield and quality. In this study, a biocontrol bacteria CQ-4 was isolated and screened from the rhizosphere soil of tomato plants. Morphological, physiological, and biochemical characteristics and 16S rDNA sequence analysis revealed that it belongs to the species Pseudomonas aeruginosa, which has a strong antagonistic effect against Botrytis cinerea. In addition, the bacterium's antibacterial spectrum is relatively extensive, and antagonistic tests have shown that it also has varying degrees of inhibition on other 12 plant diseases. The growth promotion test showed that the strain has a clear promotion effect on tomato seed germination and seedling growth. The growth-promoting effect on plant height, stem thickness, dry and fresh weight and main root length of tomato seedlings was significantly improved after the seeds were soaked in a bacterial solution of 2.5 × 108 cfu mL-1 concentration. This did not only maintain the nutritional quality of tomato fruits, but also prevents them from rotting. In vitro and pot experiments showed that the strain CQ-4 can effectively control tomato gray mold, and the control effects on tomato leaves and fruits reached 74.4% and 66.0%, respectively. Strain CQ-4 induce plants to up-regulate the activities of four disease-resistant defense enzymes. The peak enzymatic activities of Phenylalanine Ammonia Lyase (PAL), polyphenol oxidase (PPO), peroxidase (POD), and Superoxide Dismutase (SOD) were increased by 35.6%, 37.6%, 46.1%, and 38.4%, respectively, as compared with the control group. This study found that the strain can solubilize phosphorus, fix nitrogen, and produce cellulase, protease, ferrophilin, and other antibacterial metabolites, but it does not produce chitinase, glucanase, and HCN (hydrocyanic acid). This research screened out an excellent Pseudomonas aeruginosa strain that can stably and effectively control tomato gray mold, and it provided theoretical basis for further development and the application of biological agents.

Project description:To develop new agents against strawberry grey mould and to aid in the development of biological pesticides, we investigated the inhibitory effect of a natural compound, phenazine-1-carboxamide (PCN), against Botrytis cinerea using a growth rate assay. Additionally, indoor toxicity and the in vitro control effect of PCN were further studied to determine its potential mechanisms of action on B. cinerea. PCN was inhibitory against B. cinerea with a 50% effective concentration (EC50) of 108.12 μg/mL; the toxicity of PCN was equivalent to that of carbendazim (CBM). The best in vitro control effect of PCN against grey mould in strawberry (fruit) reached 75.32%, which was slightly higher than that of CBM. The field control effect of PCN against grey mould reached a maximum of 72.31% at a PCN concentration of 700 μg/mL, which was 1.02 times higher than that of CBM. Fungistatic activity was observed at low concentrations of PCN, while high concentrations of PCN resulted in fungicidal activity against B. cinerea. This natural compound strongly inhibited both spore and sclerotium germination of B. cinerea, with the best relative inhibition rates of 77.03% and 82.11%, respectively. The inhibitory effect of PCN on mycelial growth of B. cinerea was significant and reached levels of 87.32%. Scanning electron microscopy observations revealed that after 48 h of PCN treatment, the mycelia appeared loose, locally twisted, and folded, with exudation of contents; the mycelia was withered and twisted, with edge burrs, deformations, ruptures and a sheet-like structure. Transmission electron microscopy observations revealed that after 48 h of PCN treatment, the structure of the cell nucleus was unclear and the vacuoles had ruptured; additionally, various organelles exhibited disordered structures, there were substantial non-membrane transparent inclusions, the cells were plasmolysed, the cell walls were collapsed in some cases, and the hyphal tissue was essentially necrotic. A PCN dosage of 35-140 μg/mL had no effect on the cell membrane permeability of the mycelia, while a PCN dosage of 700 μg/mL resulted in significant permeability. PCN inhibited B. cinerea toxin; the mycotoxin level was approximately 0.41 of the value recorded for the control at a PCN dosage of 700 μg/mL. PCN affected the activity of pectin methylgalacturonase (PMG), polygalacturonase (PG), cellulase (Cx) and β-glucosidase (BG); the lowest activities of PMG, PG, BG and Cx reached 0.3 U/mg, 0.62 U/mg, 0.64 U/mg, and 0.79 U/mg, respectively, after treatment with 700 μg/mL PCN.

Project description:BACKGROUND:Phytophthora infestans is responsible for late blight, one of the most important potato diseases. Phenazine-1-carboxylic acid (PCA)-producing Pseudomonas fluorescens strain LBUM223 isolated in our laboratory shows biocontrol potential against various plant pathogens. To characterize the effect of LBUM223 on the transcriptome of P. infestans, we conducted an in vitro time-course study. Confrontational assay was performed using P. infestans inoculated alone (control) or with LBUM223, its phzC- isogenic mutant (not producing PCA), or exogenically applied PCA. Destructive sampling was performed at 6, 9 and 12 days and the transcriptome of P. infestans was analysed using RNA-Seq. The expression of a subset of differentially expressed genes was validated by RT-qPCR. RESULTS:Both LBUM223 and exogenically applied PCA significantly repressed P. infestans' growth at all times. Compared to the control treatment, transcriptomic analyses showed that the percentages of all P. infestans' genes significantly altered by LBUM223 and exogenically applied PCA increased as time progressed, from 50 to 61% and from to 32 to 46%, respectively. When applying an absolute cut-off value of 3 fold change or more for all three harvesting times, 207 genes were found significantly differentially expressed by PCA, either produced by LBUM223 or exogenically applied. Gene ontology analysis revealed that both treatments altered the expression of key functional genes involved in major functions like phosphorylation mechanisms, transmembrane transport and oxidoreduction activities. Interestingly, even though no host plant tissue was present in the in vitro system, PCA also led to the overexpression of several genes encoding effectors. The mutant only slightly repressed P. infestans' growth and barely altered its transcriptome. CONCLUSIONS:Our study suggests that PCA is involved in P. infestans' growth repression and led to important transcriptomic changes by both up- and down-regulating gene expression in P. infestans over time. Different metabolic functions were altered and many effectors were found to be upregulated, suggesting their implication in biocontrol.

Dataset Information

The Effect of Phenazine-1-Carboxylic Acid on Mycelial Growth of Botrytis cinerea Produced by Pseudomonas aeruginosa LV Strain.

Publications

The Effect of Phenazine-1-Carboxylic Acid on Mycelial Growth of <i>Botrytis cinerea</i> Produced by <i>Pseudomonas aeruginosa</i> LV Strain.

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