• Journal of Applied Biological Chemistry
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• v.64 no.4
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• pp.333-341
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• 2021

In this study, to select strains suitable as microbial agent from among rhizosphere microorganisms present in rhizosphere soil and roots, the mineral solubilization ability, antifungal activity against 10 types of plant pathogenic fungi, and plant growth-promoting activity of rhizosphere microorganisms were evaluated. As a result, DDP346 was selected because it has solubilization ability of phosphoric acid, calcium carbonate, silicon, and zinc; nitrogen fixing ability; production ability of siderophore, indole-3-acetic acid, and aminocyclopropane-1-carboxylate deaminase; and antifungal activity against seven types of plant pathogenic fungi. DDP346 showed a 99.9% homology with Acinetobacter pittii DSM 21653 (NR_117621.1); phylogenetic analysis also revealed a close relationship with Acinetobacter pittii based on the 16S rRNA base sequence. The growth conditions of DDP346 were identified as temperatures in the range of 10-40 ℃, pH in the range of 5-11, and salt concentrations in the range of 0-5%. In addition, a negative correlation coefficient (r² = -0.913, p <0.01) was shown between pH change and the solubilized phosphoric acid content of Acinetobacter sp. DDP346, and this is assumed to be due to the organic acid generated during culture. Consequently, through the evaluation of its mineral solubilization ability, antifungal activity against plant pathogenic fungi, and plant growth-promoting activity, the potential for the utilization of Acinetobacter sp. DDP346 as a multi-purpose microbial agent is presented.

• 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.18 no.4
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• pp.396-403
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• 2014

Ginseng (Panax ginseng C. A. Meyer) is an economically valuable pharmaceutical crop in Korea. In order to find promising biocontrol agents for soil-borne fungal pathogens which infect ginseng roots, we have isolated actinomycete, BK185 from soil. The isolate was investigated for the antifungal activity against to ginseng rot pathogens prior to testing genetic and chemical properties. The strain was identified as Streptomyces sp. using phylogenetic analysis based on 16S rRNA gene sequence. The most closely related species was S. sporoclivatus and S. geldanamycininus with high similarities (>99%). The isolate, BK185 showed positive reaction for PCR detection targeting biosynthetic gene clusters of PKS (Type-I polyketide synthase) and NRPS (Non-ribosomal polypeptide synthetase) genes. Major metabolite from the BK185 was analyzed by The LC/MS and identified to geldamycin, which was known to contained broad antibacterial, antifungal or anticancer activities. The results provide evidences that the strain, BK185 can be promising biocontrol agent for ginseng organic farming.

• The Korean Journal of Pesticide Science
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• v.19 no.4
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• pp.428-439
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• 2015

Pepper anthracnose caused by Collectotrichum acutaum has been known as one of the most damaging diseases of pepper, which has reduced not only yield but also quality of pepper produce almost every year. This study was conducted to develop control strategy against pepper anthracnose by alternate application of agricultural organic materials and chemical fungicides. The alternate application effect of agricultural organic materials and chemical fungicides for controlling pepper anthracnose were examined in vitro and in the field. First, thirteen microbial agents and twenty two agricultural organic materials were screened for antifungal activity against C. acutatum through the dual culture method and bioassay. As a result, one microbial agent (Bacillus subtilis QST-713) and three agricultural organic materials (sulfur, bordeaux mixture, marine algae extracts) were found to show high inhibition effect against C. acutatum. In the field test, when Iminoctadine tris+thiram, a chemical fungicide for controlling pepper anthracnose, was sprayed, it reduced disease incidence by 89.5%. Meanwhile Sulfur, bordeaux mixture, copper, marine algae extracts and Bacillus subtilis QST-713 showed low disease incidence at the range of 33.1~81.0%. However, when Iminoctadine tris+thiram and agricultural organic materials(bordeaux mixture, marine algae extracts) were applied to pepper fruits alternately two times at 7 days interval, there was a 81.7 and 87.1% reduction in disease, respectively. Consequently, the alternate spray of chemical fungicide (Iminoctadine tris+thiram) and agricultural organic materials (bordeaux mixture, marine algae extracts) could be recommended as a control method to reduce the using amount of chemical fungicide.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.5
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• pp.102-108
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• 2019

Melaleuca alternifolia contains terpineol-4, cineol, sesquiterpenes etc., and has a germicidal effect and skin moisturizing effect. It also has the characteristics of relieving acne inflammation, treating dandruff, relieving pain, and relieving depression. In this study, an antimicrobial substance extracted from tea tree using an organic solvent (methanol, dichloromethane, ethyl acetate) and hydrothermal extraction method. And confirmed the antimicrobial activity of each extract. In order to verify the antimicrobial activity, nine pathogenic bacteria (Bacillus cereus, Bacillus subtilis, Listeria monocytogenes, Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumoniae, Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa and Vibrio parahaemolyticus) were used. The antimicrobial activity of each extracts were confirmed by the commonly used disc diffusion method. The results showed that the fraction extracts of ethyl acetate and methanol had antimicrobial effects against V. parahaemolyticus and S. aureus. Using these results, we confirmed the antimicrobial activity of each fraction extracts and hot water extracts against V. parahaemolyticus. After the treat of samples, we confirmed at over 99.9 % of antimicrobial activity. In case of antifungal activities, we confirmed of preservation effect during over 45 hours. Based on the results of this research, further studies will be conducted to confirm the possibility of development as a new antimicrobial agent.

• The Korean Journal of Pesticide Science
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• v.17 no.4
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• pp.379-387
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• 2013

In order to improve practical utility of agro-microorganisms (AMs) which had been cultured and disseminated to promote plant growth and to control crop diseases, 51 isolates of AMs were collected from 18 agricultural extension centers in local government and screened for multi-functions such as antifungal activity, activities of phosphorus solubilization, IAA and siderophore production, nitrogen fixation, and hydrolytic enzyme activity. Finally we selected one isolate showing good antifungal activity and multi-functions related to plant growth and disease control. The selected isolate, Paenibacillus polymyxa CW, showed good inhibitory effect against plant pathogens, Pyricularia gresea, Colletotrichum acutatum, Fusarium oxysporum, Phomopsis sp., Aspergillus niger, Rhizoctonia solani and Phytophthora capsici. Suppressive effect of P. polymyxa CW against the used plant pathogens except for R. solani was much higher than that of P. polymyxa AC-1 storing in National Academy of Agricultural Science. We found P. polymyxa CW isolate showed good activity in siderophore and IAA formation, and nitrogen fixation. With P. polymyxa CW isolate, siderophore formation activity was similar to that of P. polymyxa AC-1, but IAA formation and nitrogen fixation activity was much higher than that of P. polymyxa AC-1. However neither P. polymyxa CW nor P. polymyxa AC-1 showed hydrolytic enzyme (chitinase, pectinase and cellulase) activity. The treatment of P. polymyxa CW with culture suspension of different cell density ($10^8$, $10^7$. $10^6$ cfu/ml) showed that the highest density reduced incidence of red pepper powdery mildew by 68.3% after 10 days of application. As application density of P. polymyxa CW was decreased, its control efficacy was proportionally decreased. In addition, when P. polymyxa CW was treated to control tomato powdery mildew at the same concentrations and their control effects were investigated after 7 days of inoculation, disease incidence was 0.03, 19.5, 45.7%, respectively, compared to 56.3% that of untreated check. Like red pepper powdery mildew, increase of application density of P. polymyxa CW resulted in increase of its control efficacy proportionally. P. polymyxa CW showed a density-dependent control efficacy against red pepper and tomato powdery mildews. Therefore we think that mode of action of the antagonist for suppressing two powdery mildew diseases might be antibiosis and density of more than $10^8cfu/ml$ was needed to control effectively the two diseases. On this basis, we think that P. polymyxa CW can be a promising control agent for suppressing powdery mildews of red pepper and tomato.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.5
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• pp.674-680
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• 2009

The objective of this study was to investigate antimicrobial activity, during the storage period, of animal feed and any effects on in vitro rumen digestion by supplementing different levels (5.55, 11.1, and 22.2 g/kg) of freeze dried citrus peel (FDCP) to the feed compared to untreated feed and feed treated with an antifungal agent (AA) at 0.05 g/kg. In a preservation test, feed supplemented with FDCP showed no deterioration over 21 days. Untreated feed and AA-treated feed, however, showed signs of deterioration after 16 days storage. Yellow colour and red colour, measured by spectro chromameter, decreased in the untreated and AA-treated feeds, but not in feed supplemented with FDCP. Aflatoxin was detected in untreated and AA-treated feeds at 16 days (8 ppb and 2 ppb) and 21 days (8 ppb and 4 ppb), but aflatoxin was not detected in the feed supplemented with FDCP. In a second experiment, fermentation by rumen microorganisms of FDCP (22.2 g/kg) and AA (0.05 g/kg) supplemented feeds was studied in vitro. Feeds were incubated with buffered rumen fluid for 3, 6, 9, 12, 24, and 48 h. Dry matter digestibility (DMD) and organic matter digestibility (OMD) were affected by treatment, but ammonia-N, total, and individual volatile fatty acids (VFA) were not adversely affected by treatment. In conclusion, the results indicated that FDCP might be useful for inhibiting microbial growth of animal feed during storage without disrupting rumen fermentation.

• Journal of Life Science
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• v.19 no.9
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• pp.1245-1250
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• 2009

Beimu (Fritillaria thunbergii Miquel), a bulbous plant of Liliaceae found in Korea, Japan and China, has been used as an antitussive and expectorant agent, and is also useful in alleviating stonsillitis and bronchiolitis. Most researches have been focused on micro-propagation and plant regeneration, component analysis, and dormancy relieving of beimu. Reports regarding the biological activity of beimu, such as anti-Helicobacter pyroli or platelet aggregation inhibition activity, are few and not widely available. In this study, methanol extract and its organic solvent fractions were prepared from Fritillaria thunbergii Miquel and their antimicrobial, antithrombin, and antioxidant activities were evaluated, respectively. The methanol extract contained lots of water-soluble materials (58.98%) and hexane-soluble oils (14.85%). The ethylacetate and butanol fraction at $500{\mu}g$/disc concentration showed strong antibacterial activity against tested bacteria, except Escherichia coli. Antifungal activity was not observed in methanol extract and its fractions. The hexane, ethylacetate and butanol fractions showed strong antithrombin activity at 4.8 mg/ml concentration. Especially, the ethylacetate fraction showed 95.4 sec of thrombin time at a concentration of 1.2 mg/ml, which is comparable to aspirin, a widely used antithrombosis agent. For antioxidation activity, the ethylacetate and butanol fraction showed good 1,1-diphenyl-2-picryl hydrazyl scavenging activity ($IC_{50}$ of $344{\sim}368{\mu}g$/ml). In superoxide dismutase-like activity and reducing power, the fractions showed $20{\sim}25%$ of vitamin C, and $51{\sim}54%$ of butyl hydroxytoluene, respectively.

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.133-139
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• 2009

Yam (Dioscorea batatas Decne) has been recognized as healthy food due to its various biological activities, such as anti-obesity, anti-constipation, anti-proliferation, anti-mutagenic activity, and decrease of blood glucose and cholesterol level. In this study, the methanol extract and its organic solvent fractions were prepared from dried Dioscorea batatas Decne, and their antimicrobial, antithrombin, and antioxidant activity were evaluated, respectively. The 66.7% of methanol extract was fractioned into water residue, and most of total-polyphenol/total-flavonoids were found in ethylacetate fraction. This ethylacetate fraction at $500\;{\mu}g/disc$ concentration showed strong antibacterial activity against gram-negative and gram-positive bacteria, except Staphylococcus aureus. Antifungal activity was not observed in methanol extract and its fractions. But, strong antithrombin activity was found in methanol extract. At 4.8 mg/mL concentration, thrombin time was 99.5 sec, which is coincides with 30% activity of aspirin, a commercial available antithrombosis agent. The ethylacetate fraction showed strong DPPH scavenging activity ($IC_{50}$ of $80.5\;{\mu}g/mL$), compared to vitamin C ($IC_{50}$ of $15.2\;{\mu}g/mL$). Also, the ethylacetate fraction showed strong SOD-like activity and reducing power, which are coincide with 43% of vitamin C and 82.7% of butylated hydroxytoluene activity, respectively.

• Journal of Life Science
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• v.17 no.11
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• pp.1562-1570
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• 2007

This experiment was conducted to develop fertilizer which promotes plant growth as well as suppressing pathogenic fungi. The fertilizer was made from the mixture of Ju-Back (Korean rice wine cake) and indigenous rhizosphere-bacterium. The main ingredients of Ju-Back were investigated as 6.04% total nitrogen, 42.59% total carbohydrate, 1.01% available phosphate, 73.42% organic matter, 7.72% potassium oxide, 1.35% calcium oxide, 0.53% magnesium oxide. The enzyme activities of Ju-Back were estimated to be 980 units/g for ${\alpha}-amylase$, 300 units/g for glucoamylase, and 1800 units/g for acid pretense. Indigenous rhizosphere bacteria which produced antifungal agent were isolated from soil, and was selected KMU-13 strain which can antagonize against various plant pathogenic fungi (Botrytis cinerea KACC 40573, Sclerotinia sclerotiorum KACC 41065, Fusairum oxysporum KACC 40052, Pythium aphanidermatum KACC 40156, Phytophthora capsici KACC 40476 and Glomerella cingulata KACC 40299). KMU-13 strain was identified as Bacillus subtilis KMU-13 by biochemical and 16s rDNA analysis. The organic fertilizer was made as prototype which was composed 20% Ju-Back, 70% carrier, 9.7% microorganism cultivated solution, 0.3% trace-element. We also investigated an application of fertilizer using Ju-Back for cultivating lettuce (Lactuca sativar) which were grown in three soil conditions that had chemical fertilizer, barnyard manure, lime power, urea, potassium chloride and superphosphate as a control, the whole quantity (80 kg/10a) of posted fertilizer with the control and the half quantity (40 kg/10a) with the control. The growth characteristics were examined and analysed with several weeks interval from 3 weeks to 8 weeks on head length (cm), head width (cm/head), number of leaf and fresh weight (g/plant). The results are summarized as follows. The head width and fresh weight of lettuce were the highest at posted fertilizer 1 (whole quantity) was applied chemical, organic matter (Ju-Back) and carrier. The head length was the highest at posted fertilizer 2 (whole quantity) was applied Ju-Back only.