• Microbiology and Biotechnology Letters
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• v.25 no.1
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• pp.30-36
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• 1997

To genetically breed powerful multifunctional antagonistic bacteria, the urease gene of alkalophilic Bacillus pasteurii was transferred into Bacillus subtilis YB-70 which had been selected as a powerful biocontrol agent against root-rotting fungus Fusarium solani. Urease gene was inserted into the HindIII site of pGB215-110 and designated pGU266. The plasmid pGU266 containing urease gene was introduced into the B. subtilis YB-70 by alkali cation transformation system and the urease gene was very stably expressed in the transformant of B. subtilis YB-70(pGU266). The optimal conditions for the transfomation were also evaluated. From the in vitro antibiosis tests against F. solani, the antifungal activity of B. subtilis YB-70 containing urease gene was much efficient than that of the non-transformed strain. Genetic improvement of B. subtilis YB-70 by transfer of urease gene for the efficient control seemed to be responsible for enhanced plant growth and biocontrol efficacy by combining its astibiotic action and ammonia producing ability.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.97.2-98
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• 2003

A new and effective formulation using antagonistic bacteria, Burkholderia cepacia YC5025 in vegetable oil was developed for the biocontrol of anthracnose. The bacterial population in the formulation was maintained to 5x10/sup7/ cfu/ml upto 60 days at room temperature. Control efficacy of the formulation for anthracnose was over 80％ by spraying of diluted suspension(x1,000) in growth chamber tests. On the contrary, the bacterial suspension in distilled water or bacterial culture broth containing same number of spores as the formulation had low control efficacy around 40％ even 2-weeks storage after preparation. The shelf-life of the formulation was longer than that of bacterial preparation using clay minerals such as talc or bentonite. The mechanisms of newly developed bacterial formulation are possibly the formation of water film on the surface of cucumber leaves and inactivation of the bacteria in the vegetable oils during storage. Further field tests and improvements with new liquid bacteiral formulation need to be done for practical application.

• Korean Journal Plant Pathology
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• v.11 no.3
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• pp.252-257
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• 1995

Growth rates of the low temperature growing isolates, Pseudomonas fluorescens M45 and MC07, reached maximum stationary phase within 50 hrs at the low temperature, 4$^{\circ}C$. But an ordinary biocontrol agent P. putida Pf3 did not reach logarithmic growth phase until 80 hrs. The culture filtrates of M45 and MC07 significantly inhibited the mycelial growths of Pythium ultimum, Rhizoctonia solani and Phytophthora capsici. Detached cotyledons of cucumber grown on Murashige and Skoog agar medium were much enhanced in their growth, compared to those without the filtrates. Population densities of M45 and MC07 in the rhizosphere at 14$^{\circ}C$ were more stable than at 27$^{\circ}C$. When M45 and MC07 were treated into soil, the population density of MC07 continuously increased up to 9 days after treatment, and sustained the initial inoculum density up to 60 days. Cucumber damping-offs caused by P. ultimum and R. solani were significantly reduced by applying M45 as seed-inoculant and by soil treatment with MC07. The combined treatment of M45 and MC07 provided greater effect in reducing the disease incidence than that obtained by single treatments.

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

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1011-1021
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• 2005

Rhizobacteria are actively sought for the substitution of chemical fertilizers and pathogen control agents in environment-friendly sustainable agriculture. To be successfully commercialized in the current Korean market as agriculture biomaterials, microbial agents should exhibit both properties of plant growth promotion and pathogen control. That is, the organism must be a phytostimulator as well as a biocontrol agent. These criteria and the survival rate of a rhizobacterium, Bacillus amyloliquefaciens 7079, in the soil system were investigated to evaluate the suitability for future commercialization. B. amyloliquefaciens 7079-treated seedlings showed $22.8\%$ maximum increase in leaf-length growth, compared with water-treated controls, showing the phytostimulating property. The disease suppression rates of Phytophthora-blight of peppers and Fusarium-wilt of tomatoes by B. amyloliquefaciens 7079 were 1.5 and 2.2 times better, respectively, than by three popular chemical fungicides used in actual agricultural practices to control the respective pathogens. Survival of B. amyloliquefaciens 7079 on the rhizoplane and in the rhizosphere was favorable up to 50 days in the soil system employed. These positive properties show that B. amyloliquefaciens 7079 is likely to be a suitable candidate for commercialization to market as agricultural biomaterials.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.599-606
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• 2004

Gram-positive antagonistic bacilli were isolated from agricultural soils for possible use in biocontrol of plant pathogenic fungi, Fusarium oxysporum, Rhizoctonia solani, and/or Pythium ultimum. Among the 65 antagonistic Gram-positive soil isolates, 22 strains were identified as Bacillus species by 16S rDNA sequence analyses. Four strains, including DF14, especially exhibited multiple antagonistic properties against the three damping-off fungi. Genotypic properties of the Bacillus isolates were characterized by rapid molecular fingerprinting methods using repetitive extragenic palindromic-PCR (REP-PCR), ribosomal intergenic spacer-length polymorphisms (RIS-LP), 16S rDNA PCR-restriction fragment length polymorphisms (PCR-RFLP), and strain-specific PCR assays. The results indicated that the REP-PCR method was more valuable than the RIS-LP and 16S rDNA PCR-RFLP analyses as a rapid and reliable approach for bacilli typing and identification. The use of strain-specific primers designed based on 16S rDNA sequence comparisons enabled it to be possible to selectively detect a strain, DF14, which is being used as a biocontrol agent against damping-off fungi.

• Journal of Microbiology and Biotechnology
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• v.17 no.3
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• pp.474-480
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• 2007

Fusarium wilt of cucumbers was effectively controlled by Escherichia coli expressing an endochitinase gene (chiA), and the rate was as effective (60.0%) as the wild-type strain S. proteamaculans 3095 (55.0%) where the gene was cloned. However, live cells of soil inoculated E. coli host harboring the chiA gene did not proliferate but declined 100-fold from $10^8$ CFU during the first week and showed less than 10 cells after day 14, suggesting that E. coli was able to express and produce the chitinase enzyme to the soil even as the population was gradually decreasing. Because the majority of the strains was alive for only a short period of time and the Fusarium-affected seedlings showed symptoms of wilting within 7-10 days, it seems that the pathogen control was decided early after the introduction of the biocontrol agent, eliminating the survival of the antagonist. These results indicated that soil inoculated E. coli could sufficiently express and produce the recombinant protein to control the pathogen, and root or soil colonization of the antagonist might not be a significant factor in determining the efficacy of biological control.

• The Plant Pathology Journal
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• v.23 no.3
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• pp.180-186
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• 2007

The biocontrol agent, Serratia plymuthica A21-4, has been developed for controlling pepper Phytophthora blight. Serratia plymuthica A21-4 strongly inhibits the mycelial growth, zoospore formation, and cyst germination of Phytophthora capsici in vitro. The application of a cell suspension of strain A21-4 to pepper plants in pot experiments and in greenhouse successfully controlled the disease. The bacteria produced a potent antifungal substance which was a key factor in the suppression of Phytophthora capsici. The most active chemical com-pound was isolated and purified by antifungal activity-guided fractionation. The chemical structure was identified as a chlorinated macrolide $(C_{23}H_{31}O_8Cl)$ by spectroscopic (UV, IR, MS, and NMR) data, and was named macrocyclic lactone A21-4. The active compound significantly inhibited the formation of zoosporangia and zoospore and germination of cyst of P. capsici at concentrations lower than $0.0625{\mu}g/ml$. The effective concentrations of the macrocyclic lactone A21-4 for $ED_{50}$ of mycelial growth inhibition were $0.25{\mu}g/ml,\;0.25{\mu}g/ml,\;0.30{\mu}g/ml \;and\;0.75{\mu}g/ml$ against P. capsici, Pythium ultimum, Sclerotinia sclerotiorum and Botrytis cinerea, respectively.

• The Plant Pathology Journal
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• v.30 no.2
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• pp.220-227
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• 2014

The GacS/GacA system in the root colonizer Pseudomonas chlororaphis O6 is a key regulator of many traits relevant to the biocontrol function of this bacterium. Proteomic analysis revealed 12 proteins were down-regulated in a gacS mutant of P. chlororaphis O6. These GacS-regulated proteins functioned in combating oxidative stress, cell signaling, biosynthesis of secondary metabolism, and secretion. The extent of regulation was shown by real-time RT-PCR to vary between the genes. Mutants of P. chlororaphis O6 were generated in two GacS-regulated genes, trpE, encoding a protein involved in tryptophan synthesis, and prnA, required for conversion of tryptophan to the antimicrobial compound, pyrrolitrin. Failure of the trpE mutant to induce systemic resistance in tobacco against a foliar pathogen causing soft rot, Pectobacterium carotovorum SCCI, correlated with reduced colonization of root surfaces implying an inadequate supply of tryptophan to support growth. Although colonization was not affected by mutation in the prnA gene, induction of systemic resistance was reduced, suggesting that pyrrolnitrin was an activator of plant resistance as well as an antifungal agent. Study of mutants in the other GacS-regulated proteins will indicate further the features required for biocontrol-activity in this rhizobacterium.

• The Plant Pathology Journal
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• v.30 no.3
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• pp.279-287
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• 2014

KYC 3262 was selected as a biocontrol agent against anthracnose on hot pepper from 813 extracts of myxobacterial isolates. Dual culture with Colletotrichum acutatum and 813 myxobacterial extracts was conducted, and 19 extracts were selected that inhibited germination and mycelial growth of C. acutatum. All selections were Sorangium cellulosum, which are cellulolytic myxobacteria from soil. With the infection bioassay on detached fruits in airtight containers, KYC 3262, KYC 3512, KYC 3279, and KYC 3584 were selected. The listed four myxobacteria were cultured in CSG/1 liquid media, and harvested filtrates were sprayed on the infected fruits. KYC 3262 was selected from the studies of attached fruit in a greenhouse study. KYC 3262 filtrate was applied for 3 years (from 2011 to 2013) in a field study in Asan, Republic of Korea. Control values of the KYC 3262 in the field were 31%, 89%, and 82% in 2011, 2012, and 2013, whereas values of the fungicide spray treatment were 19%, 97%, and 91%, respectively. Yields (kg/20 plants) of the KYC 3262 were 2.66 kg and 18.6 kg in 2011 and 2013, respectively, and those of the fungicide treatment were 2.0 kg and 20.2 kg, in 2011 and 2013, respectively.