• Research in Plant Disease
• /
• v.24 no.3
• /
• pp.242-247
• /
• 2018

Pseudomonas chlororaphis O6 induces systemic tolerance in plants against drought stress. A volatile, 2R, 3R-butanediol, produced by the bacterium causes partial stomatal closure, thus, limiting water loss from the plant. In this study, we report that applications of extracellular polymeric substances (EPS) from P. chlororaphis O6 to epidermal peels of leaves of Arabidopsis thaliana also reduce the size of stomatal openings. Growth of A. thaliana seedlings with applications of the EPS from P. chlororaphis O6 reduced the extent of wilting when water was withheld from the plants. Fluorescence measurements showed photosystem II was protected in the A. thaliana leaves in the water stressed EPS-exposed plants. These findings indicate that P. chlororaphis O6 has redundancy in traits associated with induction of mechanisms to limit water stress in plants.

• The Plant Pathology Journal
• /
• v.34 no.1
• /
• pp.44-58
• /
• 2018

Pseudomonas chlororaphis 30-84 is a biological control agent selected for its ability to suppress diseases caused by fungal pathogens. P. chlororaphis 30-84 produces three phenazines: phenazine-1-carboxylic acid (PCA), 2-hydroxy-phenazine-1-carboxylic acid (2OHPCA) and a small amount of 2-hydroxy-phenazine (2OHPHZ), and these are required for fungal pathogen inhibition and wheat rhizosphere competence. The two, 2-hydroxy derivatives are produced from PCA via the activity of a phenazine-modifying enzyme encoded by phzO. In addition to the seven biosynthetic genes responsible for the production of PCA, many other Pseudomonas strains possess one or more modifying genes, which encode enzymes that act independently or together to convert PCA into other phenazine derivatives. In order to understand the fitness effects of producing different phenazines, we constructed isogenic derivatives of P. chlororaphis 30-84 that differed only in the type of phenazines produced. Altering the type of phenazines produced by P. chlororaphis 30-84 enhanced the spectrum of fungal pathogens inhibited and altered the degree of take-all disease suppression. These strains also differed in their ability to promote extracellular DNA release, which may contribute to the observed differences in the amount of biofilm produced. All derivatives were equally important for survival over repeated plant/harvest cycles, indicating that the type of phenazines produced is less important for persistence in the wheat rhizosphere than whether or not cells produce phenazines. These findings provide a better understanding of the effects of different phenazines on functions important for biological control activity with implications for applications that rely on introduced or native phenazine producing populations.

• The Plant Pathology Journal
• /
• v.29 no.3
• /
• pp.323-329
• /
• 2013

The stationary-phase sigma factor, RpoS, influences the expression of factors important in survival of Pseudomonas chlororaphis O6 in the rhizosphere. A partial proteomic profile of a rpoS mutant in P. chlororaphis O6 was conducted to identify proteins under RpoS regulation. Five of 14 differentially regulated proteins had unknown roles. Changes in levels of proteins in P. chlororaphis O6 rpoS mutant were associated with iron metabolism, and protection against oxidative stress. The P. chlororaphis O6 rpoS mutant showed increased production of a pyoverdine-like siderophore, indole acetic acid, and altered isozyme patterns for peroxidase, catalase and superoxide dismutase. Consequently, sensitivity to hydrogen peroxide exposure increased in the P. chlororaphis O6 rpoS mutant, compared with the wild type. Taken together, RpoS exerted regulatory control over factors important for the habitat of P. chlororaphis O6 in soil and on root surfaces. The properties of several of the proteins in the RpoS regulon are currently unknown.

• Research in Plant Disease
• /
• v.23 no.3
• /
• pp.212-227
• /
• 2017

Pseudomonas chlororaphis O6, isolated from the roots of dryland, field-grown commercial wheat in the USA, enhances plant health and therefore it is used in agriculture as a biofertilizer and biocontrol agent. The metabolites produced by this pseudomonad stimulate plant growth through direct antagonism of pathogens and by inducing systemic resistance in the plant. Studies upon P. chlororaphis O6 identify the pathways through which defined bacterial metabolites generate protection against pathogenic microbes, insects, and nematodes. P. chlororaphis O6 also triggers plant resistance to drought and salinity stresses. The beneficial determinants are produced from bacterial cells as they form biofilms during root colonization. Molecular control these processes in P. chlororaphis O6 involves the global regulatory Gac/Rsm signaling cascade with cross-talk between other global regulatory pathways. The Gac/Rsm regulon allows for coordinate phasing of expression of the genes that encode these beneficial traits among a community of cells. This review provides insights on the Gac/Rsm regulon in expression of beneficial traits of the P. chlororaphis O6 which can contribute to help yield enhancement and quality in agricultural production.

• The Plant Pathology Journal
• /
• v.27 no.3
• /
• pp.272-279
• /
• 2011

Root colonization of Arabidopsis thaliana with Pseudomonas chlororaphis O6 induces systemic tolerance against diverse pathogens, as well as drought and salt stresses. In this study, we demonstrated that 11 genes in the leaves were up-regulated, and 5 genes were down-regulated as the result of three- to five-days root colonization by P. chlororaphis O6. The identified priming genes were involved in cell signaling, transcription, protein synthesis, and degradation. In addition, expression of selected priming genes were induced in P. chlororaphis O6-colonized plants subjected to water withholding. Genes encoding defense proteins in signaling pathways regulated by jasmonic acid and ethylene, such as VSP1 and PDF1.2, were additional genes with enhanced expression in the P. chlororaphis O6-colonized plants. This study indicated that the expression of priming genes, as well as genes involved in jasmonic acid- and ethylene-regulated genes may play an important role in the systemic induction of both abiotic and biotic stress due to root colonization by P. chlororaphis O6.

• The Plant Pathology Journal
• /
• v.28 no.2
• /
• pp.202-206
• /
• 2012

Root colonization by the rhizobacterium Pseudomonas chlororaphis O6 in Arabidopsis thaliana Col-0 plants resulted in induced tolerance to drought and salinity caused by halide salt-generated ionic stress but not by osmotic stress caused by sorbitol. Stomatal apertures decreased following root colonization by P. chlororaphis O6 in both wild-type and ABA-insensitive Arabidopsis mutant plants. These results suggest that an ABA-independent stomatal closure mechanism in the guard cells of P. chlororaphis O6-colonized plants could be a key phenotype for induced systemic tolerance to drought and salt stress.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.10
• /
• pp.1659-1662
• /
• 2008

Pseudomonas chlororaphis O6 exhibits induced systemic resistance (ISR) against P. syringae pv. tabaci in tobacco. To identify one of the ISR metabolites, O6 cultures were extracted with organic solvents, and the organic extracts were subjected to column chromatography followed by spectroscopy analyses. The ISR bioassay-guided fractionation was carried out for isolation of the metabolite. High-resolution mass spectrometric analysis of the metabolite found $C_{9}H_{9}O_{3}N$ with an exact mass of 179.0582. LC/MS analysis in positive mode showed an $(M+H)^{+}$ peak at m/z 180. Nuclear magnetic resonance ($^{1}H,\;^{13}C$) analyses identified all protons and carbons of the metabolite. Based on the spectroscopy data, the metabolite was identified as 4-(aminocarbonyl) phenylacetate (4-ACPA). 4-ACPA applied at 68.0 mM exhibited ISR activity at a level similar to 1.0 mM salicylic acid. This is the first report to identify an ISR metabolite produced by P. chlororaphis O6 against the wildfire pathogen P. syringae pv. tabaci in tobacco.

• The Plant Pathology Journal
• /
• v.34 no.1
• /
• pp.35-43
• /
• 2018

Root-knot nematodes (Meloidogyne spp.) are parasites that attack many field crops and orchard trees, and affect both the quantity and quality of the products. A root-colonizing bacterium, Pseudomonas chlororaphis O6, possesses beneficial traits including strong nematicidal activity. To determine the molecular mechanisms involved in the nematicidal activity of P. chlororaphis O6, we constructed two mutants; one lacking hydrogen cyanide production, and a second lacking an insecticidal toxin, FitD. Root drenching with wild-type P. chlororaphis O6 cells caused juvenile mortality in vitro and in planta. Efficacy was not altered in the fitD mutant compared to the wild-type but was reduced in both bioassays for the mutant lacking hydrogen cyanide production. The reduced number of galls on tomato plants caused by the wild-type strain was comparable to that of a standard chemical nematicide. These findings suggest that hydrogen cyanide-producing root colonizers, such as P. chlororaphis O6, could be formulated as "green" nematicides that are compatible with many crops and offer agricultural sustainability.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.3
• /
• pp.480-491
• /
• 2017

Fluorescent pseudomonads have been isolated from halophytes, mesophytes, and xerophytes of Pakistan. Among these, eight isolates, GS-1, GS-3, GS-4, GS-6, GS-7, FS-2 (cactus), ARS-38 (cotton), and RP-4 (para grass), showed antifungal activity and were selected for detailed study. Based on biochemical tests and 16S rRNA gene sequences, these were identified as strains of P. chlororaphis subsp. chlororaphis and aurantiaca. Secondary metabolites of these strains were analyzed by LC-MS. Phenazine-1-carboxylic acid (PCA), 2-hydroxy-phenazine, Cyclic Lipopeptide (white line-inducing principle (WLIP)), and lahorenoic acid A were detected in variable amounts in these strains. P. aurantiaca PB-St2 was used as a reference as it is known for the production of these compounds. The phzO and PCA genes were amplified to assure that production of these compounds is not an artifact. Indole acetic acid production was confirmed and quantified by HPLC. HCN and siderophore production by all strains was observed by plate assays. These strains did not solubilize phosphate, but five strains were positive for zinc solubilization. Wheat seedlings were inoculated with these strains to observe their effect on plant growth. P. aurantiaca strains PB-St2 and GS-6 and P. chlororaphis RP-4 significantly increased both root and shoot dry weights, as compared with uninoculated plants. However, P. aurantiaca strains FS-2 and ARS-38 significantly increased root and shoot dry weights, respectively. All strains except PB-St2 and ARS-38 significantly increased the root length. This is the first report of the isolation of P. aurantiaca from cotton and cactus, P. chlororaphis from para grass, WLIP and lahorenoic acid A production by P. chlororaphis, and zinc solubilization by P. chlororaphis and P. aurantiaca.

• The Plant Pathology Journal
• /
• v.34 no.3
• /
• pp.241-249
• /
• 2018

Commercial biocontrol of microbial plant diseases and plant pests, such as nematodes, requires field-effective formulations. The isolate Pseudomonas chlororaphis O6 is a Gram-negative bacterium that controls microbial plant pathogens both directly and indirectly. This bacterium also has nematocidal activity. In this study, we report on the efficacy of a wettable powder-type formulation of P. chlororaphis O6. Culturable bacteria in the formulated product were retained at above $1{\times}10^8$ colony forming units/g after storage of the powder at $25^{\circ}C$ for six months. Foliar application of the diluted formulated product controlled leaf blight and gray mold in tomato. The product also displayed preventative and curative controls for root-knot nematode (Meloidogyne spp.) in tomato. Under laboratory conditions and for commercially grown melon, the control was at levels comparable to that of a standard commercial chemical nematicide. The results indicated that the wettable powder formulation product of P. chlororaphis O6 can be used for control of plant microbial pathogens and root-knot nematodes.