• Journal of Life Science
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• v.23 no.8
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• pp.1010-1018
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• 2013

The greenhouse whitefly, Bemisia tabaci, is considered one of the most destructive pests of crops. In this study, we aimed to determine the optimal liquid culture conditions in shake flasks for maximal sporulation of Beauveria bassiana M130 using rice bran. The optimal initial pH for the spore production of B. bassiana using extracted rice bran medium was 5.2 and $28^{\circ}C$. The screening in shake flasks of carbon and nitrogen sources resulted in the identification of an optimal medium based on 0.5% $(NH_4)_2SO_4$, with extracted rice bran 8:1. Using this medium, a production level of $2.15{\times}10^9$ spores per ml was obtained after six days from culture inoculation at $28^{\circ}C$ in a rotary shaking incubator at 130 rpm. In addition, the specific activities of extracellular enzymes of chitinase and protease were $4,296{\mu}mol$ and $375{\mu}mol$, respectively. These results suggest that Beauveria bassiana M130 could be a bio-controller for the greenhouse whitefly.

• Korean Journal of Veterinary Research
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• v.46 no.3
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• pp.255-261
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• 2006

Experimental animals have been used to biological and medical purposes and the animals must be, for these purposes, healthy and clean to microbial infection. However, the animals can be easily exposed to pathogenic microorganism via several routes. Of the routes, environmental conditions are the most important factors to keep the animals healthy and clean, especially air condition. Monitoring of air-condition has been required to keep the animal healthy and clean. However, any guideline is not available for experimental conditions with pigs. Therefore, the sampling times and points were compared in different conditions to establish an optimal protocol for monitoring of air borne bacteria. Tryptic soy agar(TSA), blood agar containing 5% defibrinated sheep blood and Sabraud dextrose agar(SDA) were used as media to capture total bacteria, pathogenic bacteria and fungi, respectively. Two methods, compulsive capture using an air-sampler and capturing fall-down bacteria were used to capture the microorganisms in the air. The points and time of capturing were different at each experiment. Air borne microorganisms were captured at three and five points in the open and closed equipments, respectively. Air was collected using an air-sampler for 1 min and 5 min and the agar plates as open status were left from 30 min to 2hr. At first, we monitored an experimental laboratory which dealt with several pathogenic bacteria and then, a protocol obtained from the investigation was applied to open or close experimental conditions with pigs. Number of bacteria was high from 10:00 to 15:00, especially on 13:30-15:30 but sharply decreased after 17:00. The tendency of the number of bacteria was similar between two methods even though the absolute number was higher with air sampler. Critical difference in the number of cells was observed at 5 min with air sampler and 2 hr with fall-down capturing method. However, 1 min with air sampler and 1 hr with fall-down capturing were the best condition to identify bacterial species collected from the air. Number of bacteria were different depending on the sampling points in closed condition but not in opened condition. Based on our results, a guide-line was suggested for screening air-borne microorganism in the experimental conditions with pigs.

• Weed & Turfgrass Science
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• v.2 no.1
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• pp.70-75
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• 2013

Seven medicinal plant extracts were tested for antifungal activities against six species of the major turfgrass pathogenic fungi (Colletotrichum graminicola, Pythium spp., Rhizoctonia cerealis, Rhizoctonia solani AG1-1, Rhizoctonia solani AG2-2, and Sclerotinia homoeocarpa) using paper disk diffusion method. Three medicinal plant extracts, including Pinus densiflora showed antifungal activities. In suppression of mycelium growth test, on medium adding P. densiflora extract showed that inhibition rate of mycelium growth were above 80% in 10 mg/10 ml concentration of the extract. The inhibition rate of Pythium spp. was 100% and C. graminicola was 84.3% in 10 mg/10 ml concentrations of P. densiflora extract, respectively. In particularly, the inhibition rate of Pythium spp. was 89.5% in 2 mg/10 ml concentrations of P. densiflora extract. As a result, P. densiflora extract showed high antifungal activity to Pythium spp. and C. graminicola of the turfgrass pathogen in in vitro test.

• Journal of the Korean Wood Science and Technology
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• v.27 no.2
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• pp.53-61
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• 1999

Effects of culture conditions and Mn(II) addition were investigated for production of extracellular manganese peroxidase by lignin-degrading fungus LSK-27, Nitrogen source was shown to more influence the production of extracellular manganese peroxidase by LSK-27 than carbon source. When peptone or yeast extract as nitrogen source was added, high MnP activity was obtained. Especially, nitrogen-sufficient culture condition was effective in MnP activity, showing significantly increase up to 1.0% peptone concentration, but carbon-sufficient was not. Mn(II) was shown to strongly induce the MnP production in culture fluids of LSK-27. Increase of MnP actiyity was obeserved up to addition of 100ppm Mn(II), and over this Mn(II) concentration appeared to be inhibitory. The highest level of MnP activity was attained when Mn(II) was added after 2 day incubation.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.16-16
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• 2015

Ectomycorrhizal (ECM) mushrooms play a major role in plant growth promotion through symbiotic association with roots of forest trees. They also provide an economically important food resource to us and therefore they have been studied for their artificial cultivation for decades in Korea. We have secured bio-resources of ECM mushrooms from Korean forests and performed their physiological studies. To investigate the cultural characteristics, the fungi were cultured under different conditions (medium, temperature, pH of the medium, inorganic nitrogen source). More than 90% of total 160 strains grew on three solid media (potato dextrose agar, PDA; sabouraud dextrose agar, SDA; modified Melin-Norkrans medium, MMN). The rate of mycelial growth on malt extract agar (MEA) was lower than those of three media (PDA, SDA, MMN). None of the Tricholomataceae strains grew on MEA. Many strains of ECM mushrooms were able to grow at the temperature range of $15{\sim}25^{\circ}C$ on PDA, while they showed poor growth at $10^{\circ}C$ or $30^{\circ}C$. In particular, the growth rates of both Gomphaceae and Tricholomataceae were significantly lower at $10^{\circ}C$ than at $30^{\circ}C$. The optimal pH of many strains was pH 5.0 when they cultured in potato dextrose broth (PDB). Fifty-seven percent of tested strains grew well on medium containing ammonium source than nitrate source. Many strains of Tricholomataceae showed a notable growth on ammonium medium than nitrate medium. Twenty-three percent of strains preferred nitrate source than ammonium source for their mycelial growth. The production and activity of two enzymes (cellulase and laccase) by ECM fungi were also assayed on the enzyme screening media containing CMC or ABTS. Each strains exhibited different levels of enzymatic activities as well as enzyme production. The number of laccase-producing strains was less than that of cellulase-producing strains. We found that 77% of tested strains produced both cellulase and laccase, whereas 2% of strains did not produce any enzymes. The morphological characteristics of mycelial colony were also examined on four different solid media. Yellow was a dominant color in mycelial colony and followed by white and brown on all culture media. ECM mushrooms formed mycelial colonies with a single or multiple colors within a culture medium depending on the strains and culture media. The most common shape of mycelial colony was a circular form on all media tested. Other families except for Amanitaceae formed an irregular colony on MMN than PDA. All strains of Tricholomataceae did not form a filamentous colony on all media. The pigmentation of culture media by mycelial colonies was observed in more than 50% of strains tested on both PDA and SDA. The degree of pigmentation on PDA or SDA was higher than MMN and brown color was dominant than yellow color. The production of exudates from mycelial colony was higher on PDA than MMN. Brown exudates were mainly produced by many strains on PDA or SDA, whereas transparent exudates were mainly produced by strains on MMN. We observed the mycelial colonies with a single or multiple textures in just one culture plate. Wrinkled or uneven colony surfaces were remarkably observed in many strains on PDA or SDA, while an even colony surface was observed in many strains on MMN. Sixty percent of Tricholomaceae strains formed wrinkled surface on PDA. However, they did not form any wrinkle on MMN plate. Cottony texture was observed in mycelia colonies of many strains. Velvety texture was often observed in the mycelial colonies on SDA than PDA and accounted for 60% of Suillaceae strains on SDA.

• Journal of Food Hygiene and Safety
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• v.3 no.2
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• pp.65-73
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• 1988

T-2 toxin is one of mycotoxins produced by fungi such as Fusarium spp. and possesses a potent cytotoxicity to eukaryotic cell. The contamination of mycotoxins in cereals and feedstuffs is one of the great concerns in health authorities. Therefore, the development of the specific, sensitive and simplified analysis method for T -2 toxin is required. During more than ten years, several chemical and biological analysis methods were proposed and applied for the detection and quantification of T-2 toxin. TLC, GLC-FID and GC-MS are widely employed, but these methods required numerous clean-up procedures before analysis, and the detection limit for T-2 toxin is more than 10 ppb. Biological analysis methods with dermal tissues and cultured cells are not specific to T-2 toxin, since T-2 toxin and other related derivatives possess a similar toxicological activity although their relative activity is different each otber. Based on tbe specific reaction between antibody and antigen, the authors tried to introduce the immunochemical methods for determination of T-2 toxin. The enzyme-linked immunosorbent assay method using monoclonal antibody for T-2 toxin was applied to analyse T-2 toxin. The detection limit of T-2 toxin by ELISA method was 0.1 ppb. The correlation between ELISA and GC-MS method on these samples was very high. ELISA method developed for the detection and quantification of T -2 toxin in this paper possesses simplicity, high sensitivity and specific for T-2 toxin. Furthermore, the ELISA method with T-2 toxin monoclonal antibody was an excellent tool for the screening of Fusarium spp. which was suspected to produce T-2 toxin.

• Korean Journal of Microbiology
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• v.44 no.1
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• pp.56-62
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• 2008

Two hundred and sixty-eight bacterial strains were isolated from pine mushroom (Tricholoma matsutake) basal soil. In the course of screening for antifungal activity against seven plant pathogenic fungi (Alternaria panax, Botrytis cinerea, Colletotrichum gloeosprioides, Fusarium oxysporum, Phytopthora capsici, Pythium ultimum, Rizoctonia solani) of isolates, strain KM1-15 showed strong antibiotic activity against Alternaria panax and Colletotrichum gloeosprioides. In determining its relationship on the basis of 16S rDNA sequence, KM1-15 strain was most closely related to Bacillus $koguryoae^T$ (AY904033) (99.62%). When assayed with the API 50CHE Kit, unlike Bacillus koguryoae, it is positive for utilization of L-arabinose, cellobiose, inulin, and D-turanose. Results of cellular fatty acid analysis showed that major cellular fatty acids were 15:0 anteiso (35.78%) and 17:0 anteiso (17.97%). In particular, hydroxyl fatty acids such as 13:0 iso 3-OH, 14:0 iso 3-OH, 15:0 iso 3-OH, and 17:0 iso 3-OH were only restricted to strain KM1-15. DNA G+C content was 43.7 mol% and quinone system was MK-7 (100%) in strain KM1-15.

• Korean Journal of Soil Science and Fertilizer
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• v.40 no.6
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• pp.447-453
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• 2007

With a broad objective for the development of microbial based fertilizers, a total of 373 strains were isolated from rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass. The efficacy of the isolates to augument overall plant growth was evaluated. After screening for their plant growth promotion and antagonistic properties in vitro efficient strains were further selected. The most efficient strains was characterized by 16S rRNA gene sequences and biochemical techniques and was designated as Bacillus subtilis S37-2. The strains facilitated plant growth and inhibited the plant phathogenic fungi such as Fusarium oxysporum (KACC 40037, Rhizoctonia solani (KACC 40140), and Sclerotinia sclerotiorum (KACC 40457). Pot based bioassay using lettuce as test plant was conducted by inoculating suspension ($10^5$ to $10^8cells\;mL^{-1}$) of B. subtilis S37-2 to the rhizosphere of lettuce cultivated in soil pots. Compared with non-inoculated pots, marked increase in leaf (42.3%) and root mass (48.7%) was observed in the inoculation group where the 50ml of cell mixture ($8.7{\times}10^8cells\;ml^{-1}$) was applied to the rhizosphere of letuce either once or twice. Antagonistic effects of B. subtilis S37-2 strain on S. sclerotiorum (KACC 40457) were tested. All the tested lettuce plants perished after 9 days in treatment containing only S. sclerotiorum, but only 17% of lettuce was perished in the inoculation plot. B. subtilis grew well in the TSB culture medium. The isolates grew better in yeast extracts than peptone and tryptone as nitrogen source. The growth rate was 2~4 times greater at $37^{\circ}C$ as compared with $30^{\circ}C$ incubation temperature. B. subitlis S37-2 produced $0.1{\mu}g\;ml^{-1}$ of IAA (indole 3-acetic acid) in the TSB medium containing L-tryptophan($20mg\;L^{-1}$) in 24 hours.