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Frontiers in Microbiology 2016inhibition of the fungal pathogen by PA23 is reliant upon a LysR-type transcriptional regulator (LTTR) called PtrA. In the current study, we show that Sclerotinia...
inhibition of the fungal pathogen by PA23 is reliant upon a LysR-type transcriptional regulator (LTTR) called PtrA. In the current study, we show that Sclerotinia stem rot and leaf infection are significantly increased in canola plants inoculated with the -mutant compared to the wild type, establishing PtrA as an essential regulator of PA23 biocontrol. LTTRs typically regulate targets that are upstream of and divergently transcribed from the LTTR locus. We identified a short chain dehydrogenase () gene immediately upstream of . Characterization of a mutant revealed that it is phenotypically identical to the wild type. Moreover, transcript abundance was unchanged in the mutant. These findings indicate that PtrA regulation does not involve , rather this LTTR controls genes located elsewhere on the chromosome. Employing a combination of complementation and transcriptional analysis we investigated whether connections exist between PtrA and other regulators of biocontrol. Besides was the only gene able to partially rescue the wild-type phenotype, establishing a connection between PtrA and the sensor kinase GacS. Transcriptomic analysis revealed decreased expression of biosynthetic () and regulatory genes () in the mutant; conversely, , and were markedly upregulated. The transcript abundance of was nine-fold higher in the mutant background indicating that this LTTR negatively autoregulates itself. In summary, PtrA is an essential regulator of genes required for PA23 biocontrol that is functionally intertwined with GacS.
PubMed: 27713742
DOI: 10.3389/fmicb.2016.01512 -
PloS One 2016Several bacterial species from the Burkholderia cepacia complex (Bcc) are feared opportunistic pathogens that lead to debilitating lung infections with a high risk of...
Several bacterial species from the Burkholderia cepacia complex (Bcc) are feared opportunistic pathogens that lead to debilitating lung infections with a high risk of developing fatal septicemia in cystic fibrosis (CF) patients. However, the pathogenic potential of other Bcc species is yet unknown. To elucidate clinical relevance of Burkholderia contaminans, a species frequently isolated from CF respiratory samples in Ibero-American countries, we aimed to identify its key virulence factors possibly linked with an unfavorable clinical outcome. We performed a genome-wide comparative analysis of two isolates of B. contaminans ST872 from sputum and blood culture of a female CF patient in Argentina. RNA-seq data showed significant changes in expression for quorum sensing-regulated virulence factors and motility and chemotaxis. Furthermore, we detected expression changes in a recently described low-oxygen-activated (lxa) locus which encodes stress-related proteins, and for two clusters responsible for the biosynthesis of antifungal and hemolytic compounds pyrrolnitrin and occidiofungin. Based on phenotypic assays that confirmed changes in motility and in proteolytic, hemolytic and antifungal activities, we were able to distinguish two phenotypes of B. contaminans that coexisted in the host and entered her bloodstream. Whole genome sequencing revealed that the sputum and bloodstream isolates (each representing a distinct phenotype) differed by over 1,400 mutations as a result of a mismatch repair-deficient hypermutable state of the sputum isolate. The inferred lack of purifying selection against nonsynonymous mutations and the high rate of pseudogenization in the derived isolate indicated limited evolutionary pressure during evolution in the nutrient-rich, stable CF sputum environment. The present study is the first to examine the genomic and transcriptomic differences between longitudinal isolates of B. contaminans. Detected activity of a number of putative virulence factors implies a genuine pathogenic nature of this novel Bcc species.
Topics: Bacterial Proteins; Burkholderia; Burkholderia Infections; Child; Communicable Diseases, Emerging; Cystic Fibrosis; Female; Gene Expression Profiling; Gene Expression Regulation, Bacterial; Genome, Bacterial; Humans; Opportunistic Infections; Quorum Sensing; Virulence; Virulence Factors
PubMed: 27512997
DOI: 10.1371/journal.pone.0160975 -
Scientific Reports Jul 2016We isolated Pseudomonas putida (P. putida) strain 1A00316 from Antarctica. This bacterium has a high efficiency against Meloidogyne incognita (M. incognita) in vitro and...
We isolated Pseudomonas putida (P. putida) strain 1A00316 from Antarctica. This bacterium has a high efficiency against Meloidogyne incognita (M. incognita) in vitro and under greenhouse conditions. The complete genome of P. putida 1A00316 was sequenced using PacBio single molecule real-time (SMRT) technology. A comparative genomic analysis of 16 Pseudomonas strains revealed that although P. putida 1A00316 belonged to P. putida, it was phenotypically more similar to nematicidal Pseudomonas fluorescens (P. fluorescens) strains. We characterized the diversity and specificity of nematicidal factors in P. putida 1A00316 with comparative genomics and functional analysis, and found that P. putida 1A00316 has diverse nematicidal factors including protein alkaline metalloproteinase AprA and two secondary metabolites, hydrogen cyanide and cyclo-(l-isoleucyl-l-proline). We show for the first time that cyclo-(l-isoleucyl-l-proline) exhibit nematicidal activity in P. putida. Interestingly, our study had not detected common nematicidal factors such as 2,4-diacetylphloroglucinol (2,4-DAPG) and pyrrolnitrin in P. putida 1A00316. The results of the present study reveal the diversity and specificity of nematicidal factors in P. putida strain 1A00316.
Topics: Antarctic Regions; Antinematodal Agents; Genome, Bacterial; Genomics; Phloroglucinol; Proline; Pseudomonas fluorescens; Pseudomonas putida; Pyrrolnitrin
PubMed: 27384076
DOI: 10.1038/srep29211 -
Microbiological Research 2016Pseudomonas fluorescens FD6 has been shown to possess many beneficial traits involved in the biocontrol of fungal plant pathogens, such as Botrytis cinerea and Monilinia...
Pseudomonas fluorescens FD6 has been shown to possess many beneficial traits involved in the biocontrol of fungal plant pathogens, such as Botrytis cinerea and Monilinia fructicola. Vfr (virulence factor regulator) a highly conserved global regulator of gram-negative bacteria, such as the human pathogen Pseudomonas aeruginosa, is required for the expression of many important virulence traits. The role of Vfr in the regulation of biocontrol traits, such as the production of antibiotics to control fungal pathogens by antagonistic bacteria, has not been elucidated. This study investigated the effect of a vfr mutant derived from P. fluorescens FD6 to better understand the regulation of some important biocontrol traits associated with the bacterium. Biochemical studies indicated that the production of the antibiotics 2,4-diacetylphloroglucinol, pyrrolnitrin and pyoluteorin, was markedly enhanced in the vfr mutant. The vfr mutation also increased biofilm production, swimming motility and the expression of exopolysaccharide-associated gene (pelA, pslA and pslB) transcripts, but reduced protease production. Wheat rhizosphere and root tip colonization by the vfr mutant was higher than that by the wild type at 7 and 21days after inoculation. These findings demonstrate that Vfr modulates the expression of several key traits and the production of important antibiotics involved in the biocontrol potential of P. fluorescens FD6.
Topics: Antibiosis; Antifungal Agents; Cyclic AMP Receptor Protein; Gene Expression Regulation, Bacterial; Gene Knockout Techniques; Humans; Pest Control, Biological; Phenols; Phloroglucinol; Pseudomonas fluorescens; Pyrroles; Pyrrolnitrin; Soil Microbiology; Triticum
PubMed: 27296968
DOI: 10.1016/j.micres.2016.04.013 -
Current Microbiology Sep 2016Tomato is one of the most economically attractive vegetable crops due to its high yields. Diseases cause significant losses in tomato production worldwide. We carried...
Tomato is one of the most economically attractive vegetable crops due to its high yields. Diseases cause significant losses in tomato production worldwide. We carried out Polymerase Chain Reaction studies to detect the presence of genes encoding antifungal compounds in the DNA of Pseudomonas putida strain PCI2. We also used liquid chromatography-electrospray tandem mass spectrometry to detect and quantify the production of compounds that increase the resistance of plants to diseases from culture supernatants of PCI2. In addition, we investigated the presence of 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase in PCI2. Finally, PCI2 was used for inoculation of tomato seeds to study its potential biocontrol activity against Fusarium oxysporum MR193. The obtained results showed that no fragments for the encoding genes of hydrogen cyanide, pyoluteorin, 2,4-diacetylphloroglucinol, pyrrolnitrin, or phenazine-1-carboxylic acid were amplified from the DNA of PCI2. On the other hand, PCI2 produced salicylic acid and jasmonic acid in Luria-Bertani medium and grew in a culture medium containing ACC as the sole nitrogen source. We observed a reduction in disease incidence from 53.33 % in the pathogen control to 30 % in tomato plants pre-inoculated with PCI2 as well as increases in shoot and root dry weights in inoculated plants, as compared to the pathogenicity control. This study suggests that inoculation of tomato seeds with P. putida PCI2 increases the resistance of plants to root rot caused by F. oxysporum and that PCI2 produces compounds that may be involved at different levels in increasing such resistance. Thus, PCI2 could represent a non-contaminating management strategy potentially applicable in vegetable crops such as tomato.
Topics: Amino Acids, Cyclic; Disease Resistance; Fusarium; Solanum lycopersicum; Plant Diseases; Pseudomonas putida
PubMed: 27246499
DOI: 10.1007/s00284-016-1068-y -
Archives of Microbiology Aug 2016In a previous study (Mikiciński et al. in Eur J Plant Pathol, doi: 10.1007/s10658-015-0837-y , 2015), we described the characterization of novel strain 49M of...
In a previous study (Mikiciński et al. in Eur J Plant Pathol, doi: 10.1007/s10658-015-0837-y , 2015), we described the characterization of novel strain 49M of Pseudomonas graminis, isolated from the phyllosphere of apple trees in Poland showing a good protective activity against fire blight on different organs of host plants. We now report investigations to clarify the basis for this activity. Strain 49M was found to produce siderophores on a medium containing complex CAS-Fe(3+) and HDTMA, but was not able to produce N-acyl homoserine lactones (AHLs). Moreover, it formed a biofilm on polystyrene and polyvinyl chloride (PVC) surfaces. Strain 49M gave a positive reaction in PCR with primers complementary to gacA, the regulatory gene influencing the production of several secondary metabolites including antibiotics. The genes prnD (encoding pyrrolnitrin), pltC, pltB (pyoluteorin), phlD (2,4-diacetyl-phloroglucinol) and phzC as well as phzD (and their homologs phzF and phzA encoding phenazine), described for antagonistic fluorescent pseudomonads, however, were not detected. Research into the biotic relationship between strain 49M and Erwinia amylovora strain Ea659 on five microbiological media showed that this strain clearly inhibited the growth of the pathogen on King's B and nutrient agar with glycerol media, to a very small extent on nutrient agar with sucrose, and not at all on Luria-Bertani agar. On medium 925, strain 49M even stimulated E. amylovora growth. The addition of ferric chloride to King's B resulted in the loss of its inhibitory ability. Testing the survival of 49M in vitro showed its resistance to drought, greater than that of E. amylovora.
Topics: Acyl-Butyrolactones; Anti-Bacterial Agents; Antibiosis; Bacterial Proteins; Biofilms; Biological Control Agents; DNA Primers; Erwinia amylovora; Malus; Plant Diseases; Poland; Polystyrenes; Polyvinyl Chloride; Pseudomonas; Siderophores
PubMed: 27002332
DOI: 10.1007/s00203-016-1207-7 -
Applied and Environmental Microbiology Dec 2015Pseudomonas protegens strain Pf-5 is a rhizosphere bacterium that suppresses soilborne plant diseases and produces at least seven different secondary metabolites with...
An Interspecies Signaling System Mediated by Fusaric Acid Has Parallel Effects on Antifungal Metabolite Production by Pseudomonas protegens Strain Pf-5 and Antibiosis of Fusarium spp.
Pseudomonas protegens strain Pf-5 is a rhizosphere bacterium that suppresses soilborne plant diseases and produces at least seven different secondary metabolites with antifungal properties. We derived mutants of Pf-5 with single and multiple mutations in biosynthesis genes for seven antifungal metabolites: 2,4-diacetylphoroglucinol (DAPG), pyrrolnitrin, pyoluteorin, hydrogen cyanide, rhizoxin, orfamide A, and toxoflavin. These mutants were tested for inhibition of the pathogens Fusarium verticillioides and Fusarium oxysporum f. sp. pisi. Rhizoxin, pyrrolnitrin, and DAPG were found to be primarily responsible for fungal antagonism by Pf-5. Previously, other workers showed that the mycotoxin fusaric acid, which is produced by many Fusarium species, including F. verticillioides, inhibited the production of DAPG by Pseudomonas spp. In this study, amendment of culture media with fusaric acid decreased DAPG production, increased pyoluteorin production, and had no consistent influence on pyrrolnitrin or orfamide A production by Pf-5. Fusaric acid also altered the transcription of biosynthetic genes, indicating that the mycotoxin influenced antibiotic production by Pf-5 at the transcriptional level. Addition of fusaric acid to the culture medium reduced antibiosis of F. verticillioides by Pf-5 and derivative strains that produce DAPG but had no effect on antibiosis by Pf-5 derivatives that suppressed F. verticillioides due to pyrrolnitrin or rhizoxin production. Our results demonstrated the importance of three compounds, rhizoxin, pyrrolnitrin, and DAPG, in suppression of Fusarium spp. by Pf-5 and confirmed that an interspecies signaling system mediated by fusaric acid had parallel effects on antifungal metabolite production and antibiosis by the bacterial biological control organism.
Topics: Antibiosis; Antifungal Agents; Culture Media; Fusaric Acid; Fusarium; Metabolic Networks and Pathways; Microbial Interactions; Pseudomonas; Signal Transduction; Transcription, Genetic
PubMed: 26655755
DOI: 10.1128/AEM.02574-15 -
Microbiological Research Nov 2015A hybrid sensor kinase termed RetS (regulator of exopolysaccharide and Type III secretion) controls expression of numerous genes in Pseudomonas aeruginosa. To...
A hybrid sensor kinase termed RetS (regulator of exopolysaccharide and Type III secretion) controls expression of numerous genes in Pseudomonas aeruginosa. To investigate the function of RetS in P. fluorescens FD6, the retS gene was disrupted. Genetic inactivation of retS resulted in enhanced production of 2, 4-diacetylphloroglucinol, pyrrolnitrin, and pyoluteorin. The retS mutant also exhibited significant increase in phlA-lacZ, prnA-lacZ, and pltA-lacZ transcription levels, influencing expression levels of the small regulatory RNAs RsmX and RsmZ. In the gacSretS double mutant, all the phenotypic changes caused by the retS deletion were reversed to the level of gacS single mutant. Furthermore, the retS mutation drastically elevated biofilm formation and improved the colonization ability of strain FD6 on wheat rhizospheres. Based on these results, we proposed that RetS negatively controlled the production of antibiotics through the Gac/Rsm pathway in P. fluorescens FD6.
Topics: Anti-Bacterial Agents; Bacterial Proteins; Biofilms; Galactosidases; Gene Silencing; Meristem; Mutation; Phenols; Phloroglucinol; Pseudomonas fluorescens; Pyrroles; Pyrrolnitrin; Rhizosphere; Triticum; Virulence Factors
PubMed: 26505308
DOI: 10.1016/j.micres.2015.07.005 -
Genomics Data Jun 2015Pseudomonas chlororaphis HT66, a plant growth-promoting rhizobacterium that produces phenazine-1-carboxamide with high yield, was compared with three genomic sequenced...
Pseudomonas chlororaphis HT66, a plant growth-promoting rhizobacterium that produces phenazine-1-carboxamide with high yield, was compared with three genomic sequenced P. chlororaphis strains, GP72, 30-84 and O6. The genome sizes of four strains vary from 6.66 to 7.30 Mb. Comparisons of predicted coding sequences indicated 4833 conserved genes in 5869-6455 protein-encoding genes. Phylogenetic analysis showed that the four strains are closely related to each other. Its competitive colonization indicates that P. chlororaphis can adapt well to its environment. No virulence or virulence-related factor was found in P. chlororaphis. All of the four strains could synthesize antimicrobial metabolites including different phenazines and insecticidal protein FitD. Some genes related to the regulation of phenazine biosynthesis were detected among the four strains. It was shown that P. chlororaphis is a safe PGPR in agricultural application and could also be used to produce some phenazine antibiotics with high-yield.
PubMed: 26484173
DOI: 10.1016/j.gdata.2015.01.006 -
Journal of Biotechnology Oct 2015Burkholderia pyrrocinia 2327(T) (=DSM 10685(T), having an origin history as a strain Fujisawa Pharm 2327(T) from Fujisawa Pharmaceutical Co., Ltd.) is the first...
Burkholderia pyrrocinia 2327(T) (=DSM 10685(T), having an origin history as a strain Fujisawa Pharm 2327(T) from Fujisawa Pharmaceutical Co., Ltd.) is the first industrial bacterium for the isolation of antifungal antibiotic pyrrolnitrin. Herein, we present the first complete genome sequence of strain 2327(T), which consists of three circular chromosomes with one plasmid for the total 7,961,346bp sized genome with a GC content of 66.5%. This information will provide better understanding of molecular mechanisms in strain 2327(T), leading the insight of whole-cell system for the practical application of strain with the virtue of antibiotic capacity.
Topics: Anti-Bacterial Agents; Antifungal Agents; Base Sequence; Burkholderia; Genome, Bacterial; Industrial Microbiology; Pyrrolnitrin
PubMed: 26150017
DOI: 10.1016/j.jbiotec.2015.06.420