• Korean Journal Plant Pathology
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• v.10 no.1
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• pp.39-46
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• 1994

Pseudomonas fluorescens Biovar III strains S-2 antagonistic to Rhizoctonia solani was subjected to Tn5 mutagenesis by the transposon vector pGS9. Ampicillin and kanamycin resistant (Ampr, Kmr) transconjugants were recovered at a frequency of 1.3$\times$10-7 per initial recipient cell, when recipient cells were washed twice in TE buffer before conjugation. Of the ca. 3000 transconjugants, a frequency of noninhibitory (Inh-), nonfluorescent (Flu-) and auxotorphic (Pro-) mutants were 0.27%, 0.47% and 0.40%, respectively. In these mutants, all Inh- mutants showed the same colony morphology as wild type, whereas all Flu- and Pro- mutants inhibited the growth of R. solani. These mutants were also susceptible to chloramphenicol, indicating only the Tn5 element, except for parts of pGS9, was integrated into the recipient genome. In a Southern blot analysis, the Tn5 element inserted into one site on the chromosome for each of the chosen mutants. However, Tn5 insertion sites of Inh-, and Pro- mutants were differed in each other. These indicate that the genes essential for R. solani inhibition, fluorescent production and auxotrophic are chromosomally located, but not linked to each other.

• KSBB Journal
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• v.7 no.4
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• pp.296-301
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• 1992

A Pseudomonas fluorescens was selected from mushrooms and studied in both batch and fed-batch cultures in order to get maximal biomass concentration. P. fluorescens is an aerobic bacterium and antagonistic to Pseudomonas tolaasii which causes blotch disease on the mushroom cap. P fluarescens and P. tolaasii were identified by Gram staining, gelatin liquefaction, oxidase test, etc. and were characterized by pigment production, temperature sensitivity, salt tolerance and rapid pitting test, etc., Celts of P. fluorescens well in medium containing 30g/L of glucose, whereas the growth was inhibited at the glucose levels at higher than 30g/L. The highest values of specific growth rate and productivity were obtained when using 10g/1 of yeast extract. Optimum concentrations of $NH_4Cl$ and ${(NH_4Cl)}_2SO_4$ for culture were found to be 1.0g/L and 0.1g/L respectively. Optimum concentration of $MgSO_4{\cdot}7H_2O$ used as a sulfursource was 1.0g/L. It was also found that the cell concentrations reached the maximum level when grown on the medium containing 1.0g/L of $KH_2PO_4$ and 0.1g/L of $CaCl_2$. Also, the optimum culture conditions were $30^{\circ}C$ and pH 6.0. Cultivation of P. fluarescens at high dissolved oxygen (DO) concentration led to a decrease of bacterial productivity in batch culture. Maximum productivity was achieved at 40% DO concentration.

• The Plant Pathology Journal
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• v.25 no.1
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• pp.77-85
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• 2009

Pseudomonas fluorescens strain Gpf01, isolated from ginseng rhizosphere showed antiviral activity against Cucumber mosaic virus, when tested in a local host of CMV, Chenopodium amaranticolor. Transposon mutant library of Gpf01 was prepared using pGS9::Tn5 and the mutant Gpf01-RS19 was found to loose antiviral production. We developed primers from the flanking region of Tn5 and found a cosmid clone pAV1123, harboring 1.2 kb antiviral compound producing (avcf01) locus. When a sub-clone pPH9, which carried 9.3 kb region of pAV1123, was introduced into antivirus deficient P. fluorescens wild type strain B16, it exhibited antiviral activity. Using Tn3-gus mutagenesis and complementation analysis, it was found that the genes related to antiviral activity production resided in a 9.3 kb HindIII-HindIII fragment of pAV1123, indicating that the plasmid carries an essential genes promoting antiviral activity.

• Journal of Microbiology and Biotechnology
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• v.13 no.2
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• pp.292-300
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• 2003

Pseudomonas fluorescens B16 is a plant glowth-prornoting rhizobacterium, which produces an antibacterial compound that is effective against plant root pathogens, such as Agrobacrerium tumefaciens and Raistonia solanacearum. We mutagenized the strain B16 with Omegon-Km and isolated six antibacterial-activity-deficient mutants. Two cosmid clones that hybridized with the mutant clones also were isolated from a genomic library of tile parent strain. Using deletion and complementation analyses, it was found that the biosynthesis genes resided in a 4.3-kb SalI-NarI fragment. When a plasmid clone carrying the fragment was introduced into P. fluorescens strain 1855.344, which does not exhibit any antibacterial activity, the transconjugants exhibited antibacterial activity, indicating that the plasmid clone carried all the genes essential for production of the antibacterial compound. DNA sequence analysis of the fragment identified four putative open reading frames (ORFs): orf1 through orf4 The deduced amino acid sequences of ORF1, ORF2, and ORF4 were similar to cystathionine gamma lyase, pyruvate formate-lyase activating enzyme, and transcriptional regulator, respectively, yet the amino acid sequence of ORF3 showed no similarities to any known proteins. It was also demonstrated that the antibacterial activity was responsible for biological control of the bacterial wilt caused by R. solanacearum.

• Korean Journal Plant Pathology
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• v.10 no.3
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• pp.197-210
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• 1994

This experiment was carried out to study the cause of degeneration and rot of cultivated mushroom. Among 597 bacterial isolates derived from the rots of Button mushroom (Agaricus bisporus), Oyster mushroom (Pleurotus ostreatus) and Oak mushroom (Lentinus edodes) collected from markets of 5 cities (Seoul, Suwon, Taegu, Pohang and Pusan) in Korea (1991~1993), 111 bacterial isolates (18.5%) were proved as pathogenic bacteria. These pathogenic bacteria causing bacterial rots of cultivated mushrooms were identified as Pseudomonas tolasii, P. agarici, and Eriwinia sp., and the main causal bacteria were P. tolaasii. P. fluorescens and Klebsiella plenticola were confirmed as saprophytic non-pathogenic bacteria. One hundred fifty nine isolates (Group No. 39) of the 486 saprophytic bacterial isolates were classified as P. fluorescens, and this species was most often found rot area of cultivated mushrooms. P. tolaasii, the causal organism of bacterial blotch, was classified into two groups; One group can be differentiated from the other by the formation of white precipitation band by white line reacting organisms of Pseudomonas Agar F media. P. tolaasii attacked the cultivated mushrooms relatively well at lower incubation temperature such as 5$^{\circ}C$, but P. agarici rarely attack at below 1$0^{\circ}C$. The temperature for the infection commercial cultivated mushrooms by P. agarici was higher than that of P. tolaasii. Optimum temperature for the infection of mushrooms by P. tolaasii and P. agarici were 2$0^{\circ}C$ and $25^{\circ}C$, respectively.

• Korean Journal of Microbiology
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• v.21 no.4
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• pp.207-212
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• 1983

Isolated Gram-negative bacteria, being capable of degrading toxic, recalcitrant, and pigment-producing furfural, were tentatively identified as Pseudomonas testosteroni, Pseudomonas maltophilia, Klebsiella Pneumoniae, and Pseudomonas fluorescens. They exhibited synergistic effects between P. testosteroni and the others in the degradation of colourproducing furfural. Synergistic effects and possible sequence of its degradation were attempted by manometric technique. P. testosteroni could degrade furfural to decolourize it and produce ninhydrin-reaction postive substance (NPS) which could be utilized by P. maltophilia and K. pneumoniae and the latter two bacteria could ,degrade furfural to 2-furoic acid as an oxidized form. Finally 2-furoic acid was further oxidized by P. fluorescens. Once NPS and 2-furoic acid were produced, the degradation efficiency was enhanced by competing four bacteria against furfural and 2-furoic acid.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.281-286
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• 2010

A Pseudomonas fluorescens strain was isolated and found to show antagonistic activity against phytopathogenic fungi and to possess a gene responsible for production of antibiotic 2,4-diacetylphloroglucinol. For the extension of biocontrol range, a gene for an Androetonus australis Hector insect toxin 1 (AaHIT1), one of the most known toxic insect-selective peptides, was designed and synthesized according to the preferred codon usage of Pseudomonas fluorescens, cloned, and transformed into the strain by pSUP106 vector, a broad-host-range plasmid. Bioassays indicated that the engineered strain was able to produce AaHIT1 with insecticidal activity, and at the same time retain the activity against plant pathogen. The experiments for nonplanted soil and rhizosphere colonization showed that, similar to the population of the wild-type strain, that of the engineered strain remained relatively constant in the first 10 days, and the subsequent 50 days, suggesting that AaHIT1 expression in the bacterial cell does not substantially impair its long-term colonization. It is first reported that a Pseudomonas fluorescens strain expressing an active scorpion neurotoxin has dual activity against phytopathogenic fungi and insects, making at attractive for agronomic applications.

• Journal of Environmental Science International
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• v.17 no.9
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• pp.1033-1037
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• 2008

The effects of inorganic salts, inoculum concentration, aeration rate and shaking speed on insoluble phosphate solubilization by Pseudomonas fluorescens RAF15 were investigated. Soluble phosphate production was dependent on the presence of $MgCl_2{\cdot}6H_2O$ and $MgSO_4{\cdot}7H_2O$ in the medium. Supplementation of medium with 0.01% $CaCl_2{\cdot}2H_2O$ and 0.01% NaCl slightly increased soluble phosphate production. The optimal medium compositions for the solubilization of insoluble phosphate by P. fluorescens RAF15 were 1.5% glucose, 0.005% urea, 0.3% $MgCl_2{\cdot}6H_2O$, 0.01% $MgSO_4{\cdot}7H_2O$, 0.01% $CaCl_2{\cdot}2H_2O$ and 0.01% NaCl, respectively. Optimal inoculum concentration was 2.0%(v/v). Maximum soluble phosphate production was obtained with 20-50 ml/250-ml flask and 200 rpm of shaking speed, respectively. The addition of EDTA decreased cell growth and soluble phosphate production.

• Clean Technology
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• v.8 no.4
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• pp.199-203
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• 2002

To investigate the optimal conditions for biodegradation of toluene by Pseudomonas fluorescens KCTC 1767 in a batch soil-bioreactor, the effects of rpm change from 60 to 180, and temperature change from $15^{\circ}C$ to $30^{\circ}C$ in a batch culture and the flow rate change from 55 mL/min to 85 mL/Min in soil-bioreactor on the biodegradation of toluene were studied. In a batch culture the optimal operating conditons were 60 rpm, and $30^{\circ}C$ at initial pH 7, In a soil-bioreactor the optimal flow rate was 55 mL/min in the flow rate of circulation. The lower flow rate of circulation may help to biodegrade toluene adsorped in soil and dissolved in underground water.

• Korean Journal Plant Pathology
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• v.2 no.3
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• pp.165-173
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• 1986

Accumulation of Pseudomonas sp., P. fluorescens and Erwinia carotovora in 60 min treatment was greater in extracts from soil, exudate from ginseng root and solutions than distilled water. In bacterial movement toward rubber tube soil from chamber, accumulation of P. fluorescens in response to soil supplemented with soil extracts, exudate and solutions was generally greater in soil extracts compared to control and other solutions, but Pseudomonas sp. and E. carotovora were not much response to supplemented extracts, exudate and solutions. Accumulation of the bacteria in capillaries containing various exudates from fungal propagules with not attracted to the exudates. For an accumulation of bacteria in rubber tubes containing soil inoculated with fungal propagules, the Pseudomonas sp. was not attracted in soil inoculated by the organisms as attractant but P. fluorescens and E. carotovora to fungi were attracted to F. solani, F. oxysporum and mixed organism Alternaria panax did not affect on bacterial movement except E. carotovora. The organic matter conten in Kangwha and Kimpo soil were low in diseased and healthy soil. The K content was especially high in Kimpo healthy soil. Bacterial population in Goesan and Kangwha were more abundant than other soil. The number actinomycetes was populated abundant in healthy soil of Goesan and diseased soil of Poonggi.