• Microbiology and Biotechnology Letters
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• v.20 no.4
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• pp.384-389
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• 1992

Bacillus sp. YBL-7 which had been isolated from ginseng root-rot suppressive soil was able to antagonize Fusarium solani causing ginseng root-rot by their antibiotic substance. In order to develop multifunctional antagonist on Bacillus sp. YBL-7 as a biocontrol agent against Fusarium salam', optimal conditions for protoplast transformation system of Bacillus sp.YBL-7 by the vector plasmid pE194 were investigated. The protoplasts of Bacillus sp. YBL-7 were obtained at best efficiency by treatment with 200${\mu}g$/ml of lysozyme in the pH 7.0 of SMM buffer for 90 minutes at $40^{\circ}C$. The cell wall of the protoplast was regenerated on the agar plate containing 1.2% agar and 0.7 M mannitol. Under the best condition for protoplast formation and regeneration, the optimal transformation was achieved with 40% polyethylene glycol (M.W. 4000) treatment for 10minutes. The vector plasmid pE194 showed the best transformation frequency at 5$\mu$g/ml of final concentration. The pE194 was very stable over 80% in the transformants.

• Korean Journal of Microbiology
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• v.42 no.3
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• pp.223-229
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• 2006

To develop multifunctional microbial inoculant, an insluble phosphate-solubilizing bacterium with antifungal activity was isolated from plant rhizospheric soil. On the basis of its morphological, cultural and physiological characteristics and Biolog analysis, this bacterium was identified as Pseudomonas fluorescens RAF15. P. fluorescens RAF15 showed antifungal activities against phytopathogenic fungi Botrytis cinerea and Rhizoctonia solani. The optimal medium composition and cultural conditions for the solubilization of insoluble phosphate by P. fluorescens RAF15 were 1.5% of glucose, 0.005% of urea, 0.3% $MgCl_2{\cdot}6H_2\;0.01%\;of\;MgSO_4{\cdot}7H_2O\;0.01%,\;of\;CaCl_2{\cdot}2H_2O$, and 0.05% of NaCl along with initial pH 7.0 at $30^{\circ}C$. The soluble phosphate production under optimum condition was 863 mg/L after 5 days of cultivation. The solubilization of insoluble phosphates was associated with a drop in the pH of the culture medium. P. fluorescens RAF15 showed resistance against different environmental stresses like $10-35^{\circ}C$ temperature, 1-4% salt concentration and pH 2-11 range. The strain produced soluble phosphate to the culture broth with the concentrations of 971-1121 mg/L against $CaHPO_4$, 791-908 mg/L against $Ca_3(PO_4){_2}$, and 844 mg/L against hydroxyapatite, respectively. However, the strain produced soluble phosphate to the culture broth with the concentrations of 15 mg/L against $FePO_4$, and 5 mg/L against $AlPO_4$, respectively.

• Korean Journal of Soil Science and Fertilizer
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• v.21 no.1
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• pp.41-47
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• 1988

A laboratory experiment was conducted to find out the abilities of siderophore formation and competition of $Fe^{3+}$ and $Mn^{2+}$ absorption in synthetic medium with inoculation of fluorescence forming pseudomonas and soil saprophytic fungi as Stachybotrys chatarum, Fusarium solani, and F. oxysporum. The results obtained were summarized as follows; 1. The concentration of siderophore in synthetic medium with inoculation of Pseudomonas putida pt-II was increased (with progress of incubation time). 2. The negative correlation was obtained with the increase of $Fe^{3+}$ ion concentration and siderophore in synthetic medium with inoculation of fluorescence forming pseudomonas and soil saprophytic fungi. 3. The fresh weight of fungal hyphae was decreased with the increase of siderophore in synthetic medium. 4. There was insignificant relationship between the concentration of $Mn^{2+}$ and the concentration of siderophore while the positive correlation was obtained with the increase of fresh weight of fungal hyphae.

• Journal of Practical Agriculture & Fisheries Research
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• v.14 no.1
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• pp.61-83
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• 2012

To identification of fungi that occurs round bale silages, 253 fungal contaminated samples were collected from 2009 to 2011. Total 253 silage samples from Italian ryegrass, sudan grass, rye, corn, barley and oat were analysed. Total 270 strains were purely isolated from contaminated round bale silages. The fungi were identified with morphological characteristics and rDNA sequence analysis. Nineteen species of fungi(Rhizopus sp., Fusarium spp., Coprinus sp., Blastomyces sp., Aureobasidium sp., Polypaecilum sp., Botryoderma sp., Mucor sp., Scytalidium sp., Sphaeropsis sp., Aspergillus spp., Trichocladium sp., Humicola sp., Staphylotrichum sp., Periconia sp., Verticillium sp., Diplococcium sp., Penicillium spp. and Trichoderma spp.) were identified by morphological characteristics. On the other hand, fungi isolated from silage were identified to Acremonium strictum, Aspergillus tubingensis, Bionectria ochroleuca, Dipodascaceae sp., Fusarium proliferatum, Fusarium oxysporum, Fusrium solani, Gelasinospora reticulata, Gibberella moniliformis, Gibberella zeae, Nectria mauritiicola, Penicillium paneum, Pseudallecheria boydii, Schizophyllum commune, Scopulariopsis brevicaulis and Simplicillium lamellicola by rDNA sequence analysis. Penicillium sp. and Trichoderma sp., were isolated 74 and 64 strains, respectively. Humicola sp., Aspergillus sp., Coprinus sp., and Fusarium spp. were identified 10 to 30 strains. Most fungi were isolated together with more than one species in a sample looked like one species with the naked eyes.

• The Plant Pathology Journal
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• v.38 no.1
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• pp.25-32
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• 2022

Plant pathogens pose major threats on agriculture and horticulture, causing significant economic loss worldwide. Due to the continuous and excessive use of synthetic pesticides, emergence of pesticide resistant pathogens has become more frequent. Thus, there is a growing needs for environmentally-friendly and selective antimicrobial agents with a novel mode of action, which may be used in combination with conventional pesticides to delay development of pesticide resistance. In this study, we evaluated the potentials of lichen substances as novel biopesticides against eight bacterial and twelve fungal plant pathogens that have historically caused significant phytopathological problems in South Korea. Eight lichen substances of diverse chemical origins were extracted from axenic culture or dried specimen, and further purified for comparative analysis of their antimicrobial properties. Usnic acid and vulpinic acid exhibited strong antibacterial activities against Clavibacter michiganensis subsp. michiganensis. In addition, usnic acid and vulpinic acid were highly effective in the growth inhibition of fungal pathogens, such as Diaporthe eres, D. actinidiae, and Sclerotinia sclerotiorum. Intriguingly, the growth of Rhizoctonia solani was specifically inhibited by lecanoric acid, indicating that lichen substances exhibit some degrees of selectivity to plant pathogens. These results suggested that lichen substance can be used as a selective biopesticide for controlling plant disease of agricultural and horticultural significance, minimizing possible emergence of pesticide resistant pathogens in fields.

• The Plant Pathology Journal
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• v.31 no.3
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• pp.278-289
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• 2015

Indigenous strains of Trichoderma species isolated from rhizosphere soils of Tea gardens of Assam, north eastern state of India were assessed for in vitro antagonism against two important tea fungal pathogens namely Pestalotia theae and Fusarium solani. A potent antagonist against both tea pathogenic fungi, designated as SDRLIN1, was selected and identified as Trichoderma viride. The strain also showed substantial antifungal activity against five standard phytopathogenic fungi. Culture filtrate collected from stationary growth phase of the antagonist demonstrated a significantly higher degree of inhibitory activity against all the test fungi, demonstrating the presence of an optimal blend of extracellular antifungal metabolites. Moreover, quantitative enzyme assay of exponential and stationary culture filtrates revealed that the activity of cellulase, ${\beta}$-1,3-glucanase, pectinase, and amylase was highest in the exponential phase, whereas the activity of proteases and chitinase was noted highest in the stationary phase. Morphological changes such as hyphal swelling and distortion were also observed in the fungal pathogen grown on potato dextrose agar containing stationary phase culture filtrate. Moreover, the antifungal activity of the filtrate was significantly reduced but not entirely after heat or proteinase K treatment, demonstrating substantial role of certain unknown thermostable antifungal compound(s) in the inhibitory activity.

• Applied Biological Chemistry
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• v.41 no.8
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• pp.600-604
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• 1998

Methanol extracts from 53 species of oriental medicinal plants in 34 families were tested for their fungicidal activities against Pyricularia grisea, Rhizoctonia solani, Phytophthora capsici, Phytophthora infestans, Collectotrichum dematium, Botryospaeria dothidea, Fusarium oxysporum f. sp. cucumerinum, Botrytis cinerea, Puccinia recondita, and Erysiphe graminis. In in vitro study using impregnated paper disc method, the efficacy varied with both plant pathogen and plant species tested. Methanol extracts of Asarum sieboldii roots, Sinomenium acutum roots, Pinus densiflora leaves, Rheum undulatum root barks, Coptis japonica roots, and Phellodendron amurense barks showed potent fungicidal activities against the various pathogens when treated with 10 mg/disc. In a whole plant test, methanol extracts of P. densiflora leaves and roots and C. japonica roots were highly effective against a variety of plant pathogens. As a naturally occurring fungicide, P. densiflora- and C. japonica-derived materials could be useful as new fungicidal products against various plant diseases induced by plant pathogenic fungi.

• The Plant Pathology Journal
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• v.23 no.2
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• pp.97-102
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• 2007

A total of 39 essential oils were tested for antifungal activities as volatile compounds against five phytopathogenic fungi at a dose of 1 ${\mu}l$ per plate. Five essential oils showed inhibitory activities against mycelial growth of at least one phytopathogenic fungus. Origanum vulgare essential oil inhibited mycelial growth of all of the five fungi tested. Both Cuminum cyminum and Eucalyptus citriodora oils displayed in vitro antifungal activities against four phytopathogenic fungi except for Colletotrichum gloeosporioides. The essential oil of Thymus vulgaris suppressed the mycelial growth of C. gloeosporioides, Fusarium oxysporum and Rhizoctonia solani and that of Cymbopogon citratus was active to only F. oxysporum. The chemical compositions of the five active essential oils were determined by gas chromatography-mass spectrometry. This study suggests that both E. citriodora and C. cyminum oils have a potential as antifungal preservatives for the control of storage diseases of various crops.

• 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.

• Mycobiology
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• v.33 no.2
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• pp.97-103
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• 2005

Crude extracts and active principles from medicinal plants have shown potential role in controlling plant diseases in glasshouses as well as in fields as one of the safest and ecofriendly methods. The effect of nor-securinine (an alkaloid) isolated from Phyllanthus amarus has been seen against spore germination of some fungi (Alternaria brassicae, A. solani, Curvularia pennisetti, Curvularia sp., Erysiphe pisi, Helminthosporium frumentacei) as well as pea powdery mildew (Erysiphe pisi) under glasshouse conditions. The sensitivity of fungi to nor-securinine varied considerably. Nor-securinine was effective against most of the fungi. H. frumentacei was more sensitive even at the lowest concentration ($1,000\;{\mu}g/ml$). Likewise conidia of E. pisi were also inhibited in partially or completely appressorium formation. Pre-inoculation treatment showed greater efficacy than post-inoculation in inhibiting powdery mildew development on pea plants in a glasshouse. Maximum inhibition occurred at $2000\;{\mu}g/ml$.