Biocontrol Bacterium Pseudomonas Fluorescens Research Articles

Transcriptional Regulator PhlH Modulates 2,4-Diacetylphloroglucinol Biosynthesis in Response to the Biosynthetic Intermediate and End Product.

Certain strains of biocontrol bacterium Pseudomonas fluorescens produce the secondary metabolite 2,4-diacetylphloroglucinol (2,4-DAPG) to antagonize soilborne phytopathogens in the rhizosphere. The gene cluster responsible for the biosynthesis of 2,4-DAPG is named phlACBDEFGH and it is still unclear how the pathway-specific regulator phlH within this gene cluster regulates the metabolism of 2,4-DAPG. Here, we found that PhlH in Pseudomonas fluorescens strain 2P24 represses the expression of the phlG gene encoding the 2,4-DAPG hydrolase by binding to a sequence motif overlapping with the -35 site recognized by σ70 factors. Through biochemical screening of PhlH ligands we identified the end product 2,4-DAPG and its biosynthetic intermediate monoacetylphloroglucinol (MAPG), which can act as signaling molecules to modulate the binding of PhlH to the target sequence and activate the expression of phlG Comparison of 2,4-DAPG production between the ΔphlH, ΔphlG, and ΔphlHG mutants confirmed that phlH and phlG impose negative feedback regulation over 2,4-DAPG biosynthesis. It was further demonstrated that the 2,4-DAPG degradation catalyzed by PhlG plays an insignificant role in 2,4-DAPG tolerance but contributes to bacterial growth advantages under carbon/nitrogen starvation conditions. Taken together, our data suggest that by monitoring and down-tuning in situ levels of 2,4-DAPG, the phlHG genes could dynamically modulate the metabolic loads attributed to 2,4-DAPG production and potentially contribute to rhizosphere adaptation.IMPORTANCE 2,4-DAPG, which is synthesized by biocontrol pseudomonad bacteria, is a broad-spectrum antibiotic against bacteria, fungi, oomycetes, and nematodes and plays an important role in suppressing soilborne plant pathogens. Although most of the genes in the 2,4-DAPG biosynthetic gene cluster (phl) have been characterized, it is still not clear how the pathway-specific regulator phlH is involved in 2,4-DAPG metabolism. This work revealed the role of PhlH in modulating 2,4-DAPG levels by controlling the expression of 2,4-DAPG hydrolase PhlG in response to 2,4-DAPG and MAPG. Since 2,4-DAPG biosynthesis imposes a metabolic burden on biocontrol pseudomonads, it is expected that the fine regulation of phlG by PhlH offers a way to dynamically modulate the metabolic loads attributed to 2,4-DAPG production.

Effect of Wheat Roots Infected with the Pathogenic Fungus Gaeumannomyces graminis var. tritici on Gene Expression of the Biocontrol Bacterium Pseudomonas fluorescens Pf29Arp

Traits contributing to the competence of biocontrol bacteria to colonize plant roots are often induced in the rhizosphere in response to plant components. These interactions have been studied using the two partners in gnotobiotic systems. However, in nature, beneficial or pathogenic fungi often colonize roots. Influence of these plant-fungus interactions on bacterial behavior remains to be investigated. Here, we have examined the influence of colonization of wheat roots by the take-all fungus Gaeumannomyces graminis var. tritici on gene expression of the biocontrol bacterium Pseudomonas fluorescens Pf29Arp. Bacteria were inoculated onto healthy, early G. graminis var. tritici-colonized and necrotic roots and transcriptomes were compared by shotgun DNA microarray. Pf29Arp decreased disease severity when inoculated before the onset of necrosis. Necrotic roots exerted a broader effect on gene expression compared with early G. graminis var. tritici-colonized and healthy roots. A gene encoding a putative type VI secretion system effector was only induced in necrotic conditions. A common pool of Pf29Arp genes differentially expressed on G. graminis var. tritici-colonized roots was related to carbon metabolism and oxidative stress, with a highest fold-change with necrosis. Overall, the data showed that the association of the pathogenic fungus with the roots strongly altered Pf29Arp adaptation with differences between early and late G. graminis var. tritici infection steps.

Biocontrol strain Pseudomonas fluorescens WCS365 inhibits germination of Fusarium oxysporum spores in tomato root exudate as well as subsequent formation of new spores

Fusarium oxysporum f.sp.radicis-licopersici (Forl) is a soilborne pathogenic fungus which can cause tomato foot and root rot (TFRR). Tomato root exudate is a good source of nutrients for both Forl and the TFRR-suppressing biocontrol bacterium Pseudomonas fluorescens strain WCS365. Incubation of Forl microconidia in tomato root exudate stimulates their germination. This phenomenon is observed, to a lesser extent, upon incubation in plant nutrient solution supplemented with citrate or glucose, the major organic acid and sugar components, respectively, of tomato root exudate. Here we show that induction of germination of microconidia is significantly reduced in the presence of P. fluorescens WCS365 in all tested media. Scanning electron microscopy revealed that P. fluorescens WCS365 colonizes developing hyphae. Efficient colonization correlates with low nutrient availability. Eventually, new microconidia are formed. The presence of P. fluorescens WCS365 reduces the number of newly formed microconidia. This reduction does not depend on physical contact between bacteria and hyphae. We discuss that the ability of P. fluorescens WCS365 to slow down the processes of microconidia germination and development of new microconidia of the phytopathogen, and therefore the ability to reduce fungal dissemination, is likely to contribute to the biocontrol efficacy of this strain.

Secondary metabolite production facilitates establishment of rhizobacteria by reducing both protozoan predation and the competitive effects of indigenous bacteria

Summary 1. Rhizosphere bacteria live in close contact to plant roots feeding on root exudates and rhizodeposits. By producing toxic exoproducts rhizobacteria may inhibit plant pathogens thereby functioning as biocontrol agents and increasing plant fitness. However, the evolutionary basis why rhizobacteria protect plants is little understood. To persist toxin production needs to improve the competitiveness of the bacteria themselves. 2. We investigated the importance of secondary metabolite production for the establishment of the model soil biocontrol bacterium Pseudomonas fluorescens CHAO in the rhizosphere of rice. We compared the performance of this toxin-producing strain and its isogenic gacS deficient mutant defective in secondary metabolite production. The bacteria were added to the rhizosphere of rice, where they had to compete with the indigenous flora for resources and to resist pr?dation by the protist Acanthamoeba castellana. 3. Secondary metabolite production strongly enhanced the establishment of the inoculated bacteria by improving competitive strength and predator resistance. The fitness gain due to attenuation of pr?dation exceeded that due to competition by a factor of 2-3, confirming the importance of grazing resistance for rhizosphere bacteria. 4. Biocontrol properties of Plant Growth Promoting Bacteria such as P. fluor escens therefore gain a new dimension. Toxicity primary plays a role in the interaction with competitors and especially predators, and not in the protection of the host plant. Thus, establishment and efficiency of bio control bacteria may be improved by fostering predator defence via toxin production.

Genome-wide search reveals a novel GacA-regulated small RNA in Pseudomonas species

BackgroundSmall RNAs (sRNAs) are widespread among bacteria and have diverse regulatory roles. Most of these sRNAs have been discovered by a combination of computational and experimental methods. In Pseudomonas aeruginosa, a ubiquitous Gram-negative bacterium and opportunistic human pathogen, the GacS/GacA two-component system positively controls the transcription of two sRNAs (RsmY, RsmZ), which are crucial for the expression of genes involved in virulence. In the biocontrol bacterium Pseudomonas fluorescens CHA0, three GacA-controlled sRNAs (RsmX, RsmY, RsmZ) regulate the response to oxidative stress and the expression of extracellular products including biocontrol factors. RsmX, RsmY and RsmZ contain multiple unpaired GGA motifs and control the expression of target mRNAs at the translational level, by sequestration of translational repressor proteins of the RsmA family.ResultsA combined computational and experimental approach enabled us to identify 14 intergenic regions encoding sRNAs in P. aeruginosa. Eight of these regions encode newly identified sRNAs. The intergenic region 1698 was found to specify a novel GacA-controlled sRNA termed RgsA. GacA regulation appeared to be indirect. In P. fluorescens CHA0, an RgsA homolog was also expressed under positive GacA control. This 120-nt sRNA contained a single GGA motif and, unlike RsmX, RsmY and RsmZ, was unable to derepress translation of the hcnA gene (involved in the biosynthesis of the biocontrol factor hydrogen cyanide), but contributed to the bacterium's resistance to hydrogen peroxide. In both P. aeruginosa and P. fluorescens the stress sigma factor RpoS was essential for RgsA expression.ConclusionThe discovery of an additional sRNA expressed under GacA control in two Pseudomonas species highlights the complexity of this global regulatory system and suggests that the mode of action of GacA control may be more elaborate than previously suspected. Our results also confirm that several GGA motifs are required in an sRNA for sequestration of the RsmA protein.

Encapsulation of cabbage seeds in alginate polymer containing the biocontrol bacterium Pseudomonas fluorescens strain LRB3W1 for the control of cabbage soilborne diseases

Encapsulation of cabbage seeds in alginate polymer containing the biocontrol bacterium Pseudomonas fluorescens strain LRB3W1 for the control of cabbage soilborne diseases

Combined Use of the Biocontrol Bacterium Pseudomonas fluorescens Strain LRB3W1 with Reduced Fungicide Application for the Control of Tomato Fusarium Wilt

Pseudomonas fluorescens strain LRB3W1 inhibited the mycelial growth of Fusarium oxysporum f. sp. lycopersici and suppressed the Fusarium wilt of tomato. The chemical fungicide, benomyl, did not suppress the disease incidence at low concentrations. However, the disease incidence was decreased by the combined application of benomyl at low concentrations with strain LRB3W1. Combined application of benomyl with the bacterium was more effective than treatment with the bacterium alone. The survival of strain LRB3W1 was not influenced by the presence of benomyl. This combined use of the biocontrol bacterium, strain LRB3W1, and a fungicide, benomyl, should be an attractive approach for suppressing tomato wilt.

Structure, Chemistry, and Biological Activity of Pseudophomins A and B, New Cyclic Lipodepsipeptides Isolated from the Biocontrol Bacterium Pseudomonas fluorescens.

ChemInformVolume 34, Issue 28 Natural Products Structure, Chemistry, and Biological Activity of Pseudophomins A and B, New Cyclic Lipodepsipeptides Isolated from the Biocontrol Bacterium Pseudomonas fluorescens. M. Soledade C. Pedras, M. Soledade C. Pedras Dep. Chem., Univ. Saskatchewan, Saskatoon, Sask. S7N 5C9, Can.Search for more papers by this authorNargis Ismail, Nargis Ismail Dep. Chem., Univ. Saskatchewan, Saskatoon, Sask. S7N 5C9, Can.Search for more papers by this authorJ. Wilson Quail, J. Wilson Quail Dep. Chem., Univ. Saskatchewan, Saskatoon, Sask. S7N 5C9, Can.Search for more papers by this authorSusan M. Boyetchko, Susan M. Boyetchko Dep. Chem., Univ. Saskatchewan, Saskatoon, Sask. S7N 5C9, Can.Search for more papers by this author M. Soledade C. Pedras, M. Soledade C. Pedras Dep. Chem., Univ. Saskatchewan, Saskatoon, Sask. S7N 5C9, Can.Search for more papers by this authorNargis Ismail, Nargis Ismail Dep. Chem., Univ. Saskatchewan, Saskatoon, Sask. S7N 5C9, Can.Search for more papers by this authorJ. Wilson Quail, J. Wilson Quail Dep. Chem., Univ. Saskatchewan, Saskatoon, Sask. S7N 5C9, Can.Search for more papers by this authorSusan M. Boyetchko, Susan M. Boyetchko Dep. Chem., Univ. Saskatchewan, Saskatoon, Sask. S7N 5C9, Can.Search for more papers by this author First published: 08 July 2003 https://doi.org/10.1002/chin.200328207Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume34, Issue28July 15, 2003 RelatedInformation

Structure, chemistry, and biological activity of pseudophomins A and B, new cyclic lipodepsipeptides isolated from the biocontrol bacterium Pseudomonas fluorescens

Pseudophomins A and B are cyclic lipodepsipeptides isolated from Pseudomonas fluorescens strain BRG100, a bacterium with potential application for biocontrol of plant pathogens and weeds. Their chemical structures were established by a combination of spectroscopic data, X-ray crystallography, and selective chemical degradation. This unique chemical degradation allowed the unambiguous determination of the absolute configuration of the amino acid residue Leu-1, due to γ-lactam formation followed by selective cleavage of the adjacent N(8)–C(7) bond. To the best of our knowledge this is the first application of γ-lactam formation to the determination of absolute configuration of an adjacent amino acid. Pseudophomin B showed higher antifungal activity against the phytopathogens Phoma lingam/Leptosphaeria maculans and Sclerotinia sclerotiorum than pseudophomin A, and is likely to be the main component responsible for the antifungal activity of EtOAc extracts of strain BRG100. By contrast, pseudophomin A showed stronger inhibition of green foxtail ( Setaria viridis) root germination than pseudophomin B.

Deleterious impact of a virulent bacteriophage on survival and biocontrol activity of Pseudomonas fluorescens strain CHAO in natural soil.

Many biotic and abiotic factors affect the persistence and activity of beneficial pseudomonads introduced into soil to suppress plant diseases. One such factor may be the presence of virulent bacteriophages that decimate the population of the introduced bacteria, thereby reducing their beneficial effect. We have isolated a lytic bacteriophage (phi)GP100) that specifically infects the biocontrol bacterium Pseudomonas fluorescens CHA0 and some closely related Pseudomonas strains. phiGP100 was found to be a double-stranded-DNA phage with an icosahedral head, a stubby tail, and a genome size of approximately 50 kb. Replication of phiGP100 was negatively affected at temperatures higher than 25 degrees C. phiGP100 had a negative impact on the population size and the biocontrol activity of P. fluorescens strain CHA0-Rif (a rifampicin-resistant variant of CHA0) in natural soil microcosms. In the presence of phiGP100, the population size of strain CHA0-Rif in soil and on cucumber roots was reduced more than 100-fold. As a consequence, the bacterium's capacity to protect cucumber against a root disease caused by the pathogenic oomycete Pythium ultimum was entirely abolished. In contrast, the phage affected neither root colonization and nor the disease suppressive effect of a phiDGP100-resistant variant of strain CHA0-Rif. To our knowledge, this study is the first to illustrate the potential of phages to impair biocontrol performance of beneficial bacteria released into the natural soil environment.

Systemic resistance in Arabidopsis induced by rhizobacteria requires ethylene-dependent signaling at the site of application.

Root colonization of Arabidopsis thaliana by the nonpathogenic, rhizosphere-colonizing, biocontrol bacterium Pseudomonas fluorescens WCS417r has been shown to elicit induced systemic resistance (ISR) against Pseudomonas syringae pv. tomato (Pst). The ISR response differs from the pathogen-inducible systemic acquired resistance (SAR) response in that ISR is independent of salicylic acid and not associated with pathogenesis-related proteins. Several ethylene-response mutants were tested and showed essentially normal symptoms of Pst infection. ISR was abolished in the ethylene-insensitive mutant etr1-1, whereas SAR was unaffected. Similar results were obtained with the ethylene-insensitive mutants ein2 through ein7, indicating that the expression of ISR requires the complete signal-transduction pathway of ethylene known so far. The induction of ISR by WCS417r was not accompanied by increased ethylene production in roots or leaves, nor by increases in the expression of the genes encoding the ethylene biosynthetic enzymes 1-aminocyclopropane-1-carboxylic (ACC) synthase and ACC oxidase. The eir1 mutant, displaying ethylene insensitivity in the roots only, did not express ISR upon application of WCS417r to the roots, but did exhibit ISR when the inducing bacteria were infiltrated into the leaves. These results demonstrate that, for the induction of ISR, ethylene responsiveness is required at the site of application of inducing rhizobacteria.