• Journal of Microbiology and Biotechnology
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• v.34 no.5
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• pp.1040-1050
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• 2024

To isolate and analyze bacteria with Verticillium wilt-resistant properties from the fermentation residue of kitchen wastes, as well as explore their potential for new applications of the residue. A total of six bacterial strains exhibiting Verticillium wilt-resistant capabilities were isolated from the biogas residue of kitchen waste fermentation. Using a polyphasic approach, strain ZL6, which displayed the highest antagonistic activity against cotton Verticillium wilt, was identified as belonging to the Pseudomonas aeruginosa. Bioassay results demonstrated that this strain possessed robust antagonistic abilities, effectively inhibiting V. dahliae spore germination and mycelial growth. Furthermore, P. aeruginosa ZL6 exhibited high temperature resistance (42℃), nitrogen fixation, and phosphorus removal activities. Pot experiments revealed that P. aeruginosa ZL6 fermentation broth treatment achieved a 47.72% biological control effect compared to the control group. Through activity tracking and protein mass spectrometry identification, a neutral metalloproteinase (Nml) was hypothesized as the main virulence factor. The mutant strain ZL6ߡNml exhibited a significant reduction in its ability to inhibit cotton Verticillium wilt compared to the strain P. aeruginosa ZL6. While the inhibitory activities could be partially restored by a complementation of nml gene in the mutant strain ZL6CMߡNml. This research provides a theoretical foundation for the future development and application of biogas residue as biocontrol agents against Verticillium wilt and as biological preservatives for agricultural products. Additionally, this study presents a novel approach for mitigating the substantial amount of biogas residue generated from kitchen waste fermentation.

• Korean Journal of Environmental Biology
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• v.39 no.4
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• pp.479-485
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• 2021

A semi-selective agar medium was developed for detecting and enumerating Colletotrichum scovillei in chili (Capsicum annum) plant material. Potato-dextrose-agar(PDA) was used as the basic medium. The composition of the semi-selective agar medium was established after several attempts to favor the development of C. scovillei and inhibit the growth of other fungi and bacteria. The semi-selective agar medium contained PDA amended with pyribencarb and pydiflumetofen at 40 ㎍ mL^-1 each and streptomycin at 100㎍ mL^-1 for preventing bacterial growth. The pH was adjusted to 4.8 with 85% lactic acid. The inhibition of the mycelial growth of C. scovillei was significantly less than that of most other fungi including Fusarium species when grown on the semi-selective medium. C. scovillei was detected from naturally infected chili plants by plating fruit and stem tissue suspensions on the semi-selective medium, which was found to be reliable and quantifiable. This was the first report of a semi-selective agar medium to detect the presence of C. scovillei in naturally infected chili tissue.

• Journal of Life Science
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• v.18 no.7
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• pp.974-979
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• 2008

The powdery mildew, a fungal plant disease found in varieties of plant cultures, is occurred by attack with Fusarium sp., Sphaerotheca sp., Leveilluna sp., and Eryshipe sp.. In this study we investigated the control of Fusarium graminearum, a barley powdery mildew fungus, by natural plant extracts. Among the 900 plant extracts tested, Zanthoxylum schinifolium, Ligusticum acutilobum, Bidens frondosa L., Dictamnus dasycarpus, Evodia officinalis, Disporum sessile, Scopolia japonica Max., Styrax japonica S. et Z., Dictamnus dasycarpus Turcz., Sinomenium acutum Rehder et Wils., Eugenia aromaticum, Rubus parvifolius L., Reynoutria elliptica, Coptis chinensis, Paeonia lactiflora Pall., Rheum undalatum, Paeonia suffruticosa, Oenothera odorata Jacq., Euphorbia pekinensis Rupr., and Nepeta cataria were selected based on spore germination inhibition assay. Further mycelial growth inhibition assay with economical and safety considerations led us to finally select Z. schinifolium (sancho) for control of F. graminearum. To produce antifungal sancho extract, methanol was suitable for extraction and subsequent fractionations of the extract showed that the water residue mainly had antifungal activity. The sancho extract and its fractions showed minor antibacterial activity against different pathogenic or food spoilage bacteria, but they did not show any harmful effects against young tomato plant by treatment of $1,000\;{\mu}g/ml$ in green chamber test. These results suggested that the extract of sancho has high potentials on control of a powdery mildew fungus, F. graminearum.

• Asian Journal of Turfgrass Science
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• v.24 no.2
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• pp.176-181
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• 2010

Brown patch (Rhizoctonia solani AG1-1), dollar spot (Sclerotinia homoeocarpa), pythium blight (Pythium spp.), anthracnose (Colletotrichum graminicola), yellow patch (Rhizoctonia cerealis) and Zoysia patch (Rhizoctonia solani AG2-2) are the major turfgrasses diseases in Korea. In this study, 23 medicinal plant extracts were tested for antifungal activities against turfgrass pathogenic fungi. In paper disk diffusion method, 12 medicinal plant extracts, including Sophora flavescens, showed antifungal activity. Also, in the test of antifungal activity on media contained the extracts of S. flavescens, Curcuma longa, Rheum undulatum, Coptis chinensis and Asiasarum sieboldi showed above 80% inhibitory effects on the mycelial growth in 110 mg/10 ml concentration of the extracts. S. flavescens, in particularly, showed antifungal activity against the six turfgrass pathogenic fungi. The inhibition rate of S. homoeocarpa was 100% in 10 mg/10 ml, 5 mg/10 ml and 2 mg/10 ml concentrations of C. longa extract. In case of Pythium spp., the extracts of S. flavescens, R. undulatum and C. chinensis showed 100 % inhibition rate on the test media.

• The Korean Journal of Pesticide Science
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• v.12 no.3
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• pp.291-294
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• 2008

The Streptomyces padanus isolate TH04, isolated from mummified peaches, showed strong antifungal activity to Monilinia fructicola. The inhibition activity of the isolate TH04 to mycelial growth and spore germination at 1% concentration of sub-antifungal powder made from culture suspension (CS) was ranged from 79.8% to 81.0% and from 73.9% to 75.8% to M. fructicola four strains, respectively. In the test of antifungal activity in mixed culture of the isolate and M. fructicola, inhibition rate was 7.5%, 86.8% and 94.0% in 0.01, 0.1, and 1% concentration of CS containing bacterial cell of the isolate, respectively. On apples (cultivar; Fuji), the control values of the isolate TH04 crude filtrates (0.1 and 1%) were 85.9% and 100%, respectively. The results suggest that the isolate TH04 indicate development possibility as biocontrol agent of brown rot caused by M. fructicola with the study on delivery method and fermentation condition to produce an antifungal compound.

• Microbiology and Biotechnology Letters
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• v.39 no.2
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• pp.168-174
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• 2011

In order to develop safe and economic novel antifungal agents, we prepared 73 methanol extracts from medicinal and edible herbs and examined their 365 solvent fractions using n-hexane, methylene chloride, ethylacetate, butanol and water residue based on the sequential organic solvent fraction method. When using the various fractions in the screening step for antifungal activity, we discovered ethylacetate fraction of Morus alba L., methylene chloride fraction of Pimpinella brachycarpa (MCPB), and n-hexane fraction of Salvia miltiorrhiza Bunge, which all have activities in methanol extracts, as potential sources of antifungal agents. Amongst these, the antifungal activity of P. brachycarpa has not to date been reported on. In addition, the mycelial growth inhibition and spore germination inhibition activities of MCPB against A. niger were confirmed by disc-diffusion assay in a 10 day culture. The MIC and MFC of MCPB were determined as 0.25 and 0.5 mg/ml, respectively. The MCPB has no hemolytic activity against human RBC at 0.5 mg/ml and glycoside-flavonoids are theorized to be active constituents. These results suggest that MCPB has a prominent antifungal activity and that the application of sequential organic solvent fractions, instead of simple natural product extracts, is useful in the screening process of novel bioactive substances.

• Horticultural Science & Technology
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• v.31 no.6
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• pp.657-665
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• 2013

A large patch disease caused by Rhizoctonia solani AG2-2 (IV) is a serious problem in Korean lawngrass (Zoysia japonica) sites including golf courses and sports fields in Korea. Antagonistic microorganisms against R. solani AG2-2 (IV) were isolated from various forest and crop soil sources in Southern Korea. Among the 61 isolates, I-009, FRIN-001-1, and YPIN-022 strains showing dramatic inhibition of the mycelial growth of R. solani AG2-2 (IV) in the pairing culture were selected as the most potential antagonistic microorganisms for this study. Based on the 16s RNA sequence comparison, I-009 and FRIN-001-1 isolates were identified as Bacillus spp., while YPIN-022 isolate belongs to the genus Pseudomonas. The greater inhibition (clear) zone between two edges of the selected and pathogenic microbes ranged from 11 to 15 mm in three selections, but the others averaged to 7 mm out of 30 mm distance. In another antifungal test using culture filtrate, those three isolates represented a range of 51.7 to 63.5% suppression potential. The selected isolates also inhibited significantly the stem-segment colonization by R. solani AG2-2 (IV) in vivo test by 28.1%, 43.0%, and 23.7% when inoculated with I-009, FRIN-001-1, and YPIN-022, respectively. The highest antagonistic activity for the large patch disease was demonstrated by the isolate FRIN-001-1, which will be useful for developing a bio-pesticide against Rhizoctonia.

• The Plant Pathology Journal
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• v.33 no.5
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• pp.499-507
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• 2017

In an attempt to develop a biological control agent against mycotoxigenic Fusarium species, we isolated Bacillus amyloliquefaciens strain DA12 from soil and explored its antimicrobial activities. DA12 was active against the growth of mycotoxigenic F. asiaticum, F. graminearum, F. proliferatum, and F. verticillioides both in vitro and in planta (maize). Further screening using dual culture extended the activity range of strain DA12 against other fungal pathogens including Botrytis cinerea, Colletotrichum coccodes, Endothia parasitica, Fusarium oxysporum, Raffaelea quercus-mongolicae, and Rhizoctonia solani. The butanol extract of the culture filtrate of B. amyloliquefaciens DA12 highly inhibited the germination of F. graminearum macroconidia with inhibition rate 83% at a concentration of $31.3{\mu}g/ml$ and 100% at a concentration of $250{\mu}g/ml$. The antifungal metabolite from the butanol extract was identified as iturin A by thin layer chromatography-bioautography. In addition, volatile organic compounds produced by DA12 were able to inhibit mycelial growth of various phytopathogenic fungi. The volatile compounds were identified as 2-heptanone, 5-methyl heptanone and 6-methyl heptanone by gas chromatography-mass spectrometry (GC-MS) analysis. These results indicate that the antagonistic activity of Bacillus amyloliquefaciens DA12 was attributable to iturin A and volatile heptanones, and the strain could be used as a biocontrol agent to reduce the development of Fusarium diseases and mycotoxin contamination of crops.

• The Korean Journal of Pesticide Science
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• v.14 no.1
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• pp.65-71
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• 2010

This study was performed to investigate the inhibiting activity of etridiazole and thiophanate-mthyl on mycelial growth of Rhizoctonia solani and Pythium ultimum and the effect of inoculum density of each pathogen on the control efficacy of the mixture of etridiazole and thiophanate-methyl in a seedling assay test. In mycelial growth inhibition test, $EC_{50}$ values of etridiazole and thiophanate-methyl against R. solani were 15.87 and 9.34 ${\mu}g\;mL^{-1}$, while those were 0.2 and more than $500\;{\mu}g\;mL^{-1}$ against P. ultimum, respectively. Controlling activity of the mixture of etridiazole and thiophanate-methyl against damping-off of pepper and cucumber, caused by R. solani and P. ultimum was tested in a greenhouse. With the inoculation of R. solani, disease incidences of seedling in pepper were 73% and 95% at 0.5% and 1.0% of inoculum concentration, while in cucumber those were 55% and 62% at 1.0% and 2.0%. When P. ultimum was inoculated into soil by 2.0% of inoculum concentration, those in pepper and cucumber were 66.7% and 96.8%, respectively. The efficacy of the mixture was somehow affected by the concentration of R. solani. While each control value of the mixture was 94.4% and 90.7% in pepper and cucumber at low inoculum concentration (0.05%), the efficacy of the fungicide decreased in pepper and cucumber by 70.7% and 72.9% at high concentration of R. solani (0.1% in pepper and 0.5% in cucumber). However, the control value of the mixture was 100% in pepper and cucumber, irrespective of the inoculum concentration of P. ultimum, however, the increase of inoculum concentration in soil did not result in the decrease of the fungicide efficacy.

• Research in Plant Disease
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• v.18 no.4
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• pp.354-360
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• 2012

This study tested the antifungal compound obtained from a medicinal plant, Dioscorea quinqueloba Thunb., in order to search the possibility of practical application of this product in agriculture through evaluating its activity using the citrus fruits. The extract of D. quinqueloba Thunb., which has the strongest antifungal activity, was selected as a candidate among 101 plant extracts. Based on this examination concerning antifungal activity of the product on Penicillium digitatum in vitro, it was confirmed its effect of mycelial growth inhibition showed over 87% at 0.5 mg/ml concentration. This natural product showed the stability of the substance, as it was not significantly influenced by pH, temperature, or ultraviolet radiation. While citrus fruits were stored at room temperature, P. digitatum was inoculated into them in order to prepare a similar environmental conditions with epidemic occurrence of the mold. As the result of our investigation, the disease preventive effects of the active antifungal substance evidenced a 100% at 0.5 mg/ml. When the phytotoxicity of the selected natural product on citrus at 2 mg/ml was assessed, we noted no toxic effects. Based on the superior preventive effects from this natural product extracted from the plant, it is presumed to be very useful in agricultural applications for the control of green mold, P. digitatum, which has been occurred often the biggest problem in the storage of citrus fruits.