• Korean Journal of Plant Tissue Culture
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• v.22 no.5
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• pp.291-298
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• 1995

This study describes plant regeneration from leaf explant of Bupleurum falcatum through somatic embryogenesis, and the effect of 2,4-dichlorophenoxyacetic acid on somatic embryo abnormalities. The relationship between the cotyledon number of somatic embryo and its germinability is also described. Embryogenic calli were selected from calli formed on explants cultured on MS solid basal medium supplemented with 1 mg/L 2,4-D. Cotyledonary abnormalities were observed in somatic embryos which were developed from calli cultured on MS medium with 1 mg/L 2,4-D for 6-week and then subcultured on 2,4-D free MS medium for 3 weeks. The frequency of abnormalities was as follows: 7% of somatic embryos had one cotyledon, 65% of them had two cotyledons, 25% three cotyledons, 5% four cotyledons, 2% five cotyledons, and 3% trumpet-like cotyledons. The two cotyledon somatic embryos were germinated at a frequency of 80%. However the germination frequency of one cotyledon embryo and multicotyledonary embryo was lower than that of the two cotyledon somatic embryo. All of trumper-like somatic embryos did not germinate. Histological observations of multicotyledon embryo showed circular procambium in the root but pocambial strands in the cotyledonary node or upper hypocotyl. The number of the strands was equal to the cotyledon number.

• Korean Journal of Organic Agriculture
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• v.22 no.4
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• pp.743-760
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• 2014

This study aimed to isolate and identify freshwater algae from the organic agricultural ecosystems and investigate its biological characteristics to study the possibility of utilizing a biomass freshwater algae in organic farming. In the survey area, average water temperature was $12.4{\sim}28.2^{\circ}C$ and the pH ranges were from 6.1 to 8.5. The solid culture method is more suitable than liquid culture method for isolation of freshwater algae with lower contamination level and higher isolation frequency. A total of 115 strains were isolated from six freshwater algae habitats in nine regions in Korea. BGMM (BG11 Modified Medium) amended with NaNO3 and $KNO_3$ as a nitrogen, and $Na_2CO_3$ as carbon source was designed to isolate and culture freshwater algae. Absorbance of freshwater algae culture has increased dramatically to four days and decreased after eight days after inoculation. CHK008 of the seven isolates showed the highest absorbance in seven days after culturing in BGMM. The optimal pH of BGMM for culturing freshwater algae was pH 6-7. As light intensity increased, growth of freshwater algae increased. Among the five kinds of carbon sources, glucose and galactose promoted good growth of freshwater algae in BGMM. The colony color of purified 16 green algae isolates showed a separation of green, dark and light green, and of them, eleven algae strains showed a strong fluorescent light under fluorescence microscopy. Cell size of the green algae showed a wide range of variation depending on the species. General morphology of the green algae strains was spherical. Chlamydomonas sp. was elliptical, and Chlorella sorokiniana was ellipsoidal and cylindrical. All strains of the green algae except for Chlamydomonas sp. did not have flagella. One isolate of Chlamydomonas sp. and five isolates of C. sorokiniana secreted mucus. Sixteen isolates of 16 green algae were identified as two family and six species, Chlorella vulgalis, C. sorokiniana, C. pyrenoidosa, C. kessleri, C. emersonii, and Chlamydomonas sp. based on their morphological characteristics.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.5
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• pp.509-516
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• 2010

This study investigates characteristics of diesel degradation and variations of microbial community with the soil enrichment cultures. The cultures has yellow(YE-5) and transparent color's(WH-5) colony on solid plate medium. The bacillus type of YE-5 and WH-5 cultures showed diesel degradation at the rate of 99.07mg-Diesel/$L{\cdot}day$ and 57.82mg-Diesel/$L{\cdot}day$ in the presence of 1%(v/v) initial diesel concentration. Diesel degradation was 1.7 times faster than WH-5 culture. YE-5 or WH-5 culture could degrade a wide range of diesel compounds from $C_8$ to $C_24$. Microbial community analysis by PCR-DGGE technique shows that Psedomonas, Klebsiella, Escherichia and Stenotrophomonas as proteobacteria take role on the diesel degradation. uncultured Senotrophomonas sp. was only detected with YE-5 culture. It is concluded that proper combination of the microorganism should be present to stimulate the degradation of diesel and further studies are recommended for the effect of uncultured Senotrophomonas sp. or Escherichia hermannii on diesel degradation.

• KSBB Journal
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• v.22 no.5
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• pp.297-304
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• 2007

For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

• Journal of Bio-Environment Control
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• v.22 no.2
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• pp.100-107
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• 2013

Unlike general microwave, QRD (Quadratic Residue Diffusor) Microwave used in this study is known as a new technology that enhances the sterilization effect with low power because it is possible to induce the average sterilization by changing wavelength phase difference. Therefore, basic research was conducted on the function that could sterilize culture media for plant factory by using environmentally friendly and low energy consuming QRD Microwave. The results are as follows: It was confirmed that there was no external deformation in the polyurethane foam and rock wool medium when changing the microwave level between 2 and 8 kW in different water content of culture media. However, PDA solid media at 2 kW were not dissolved in 60 and 180 seconds. All of the media were dissolved in other processing. There was little difference in the microwave irradiation level and surface temperature of the strain according to the processing time between Bacillus sp. and Burkholderia sp. In the sterility test according to the microwave irradiation level and processing time, it was confirmed that both Bacillus sp. and Burkholderia sp. grew in the microwave level 2 kW regardless of time. In the microwave level 6 kW, all experimental groups except the processing of Burkholderia sp. for 60 seconds were sterilized, and all of Bacillus sp. was killed in the all experimental groups. In the microwave level 8 kW, it was confirmed that both Bacillus sp. and Burkholderia sp. were sterilized regardless of time. The temperature in microwave-processed media after contaminating strains to each medium was maintained at more than 100 in polyurethane foam and rock wool medium after 60 seconds. In general, it was shown that it was possible to sterilize after 60 seconds. Therefore, it is considered that Bacillus sp. and Burkholderia sp. which are the biggest problems in the plant factory can be adequately sterilized by QRD Microwave used in this study.

• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.21-28
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• 2001

Enzyme linked-immunosorbent assay (ELISA) for a rapid detection of fungi, Aspergillus and Penicillium genera in food, were developed and their efficiencies were approved by detecting artificially contaminated agricultural commodities. Mice were immunized with partially purified Aspergillus flavus extracellualr polysaccharide (EPS) and lymph node cells of the mice were fused with the myeloma cells for production of monoclonal antibodies. Mab 1G11, one of the antibodies, was selected and purified. A sandwich ELISA was established and its detection limit toward A. flavus EPS was 1mg/ml. Among the 59 strains tested (including 18 species of Aspergillus, 16 of Penicillium, 11 of Fusarium, 1 of Absidia, 2 of Alternaria, 2 of Candida, 2 of Cladosporium, 2 of Geotrichum, 2 of Mucor, 2 of Rhizopus, 1 of Trichoderma), species of Aspergillus and penicillium had a high reactivity with Mab 1G11 even up to 10,000 times dilution of culture broths. The other genera except Cladosporium resinae showed no reactivity, thus Mab 1G11 was specific to the genera of Aspergillus and Penicillium. The epitope of A. flavus EPS against monoclonal Mab 1G11 was on the carbohydrate moiety when 1 to 100$\mu g/g$ A. flavus EPS were put into rice, potato, and mandarin orange, the average recoveries detected by sandwich ELIA were 123, 59, and 76%, respectively. Correlation was found to be linear between the EPS, and mycelium of A. flavus and Penicillium citrinum grown in a liquid medium (r=0.87 and 0.96), and also between the EPS and colony forming unit in solid media of rice of potato (r=0.91-0.99).

• The Korean Journal of Mycology
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• v.22 no.2
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• pp.153-159
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• 1994

Effects of some sources on the vegetative growth of Naematoloma sublateritium (Fr.) Karst. were investigated using liquid and solid culture media. Temperature, pH, carbohydrates as carbon sources, amino acids as nitrogen sources, the optimal carbon/nitrogen ratio, mineral element and organic acids were studied for good mycelial growth. We could improve a new semisynthetic medium for mycelial growth in N. sublateritium.

• Korean Journal of Agricultural Science
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• v.3 no.2
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• pp.235-243
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• 1976

As an investigation on the catabolite repression system in cellulase production by Cellulomons sp. CS1-1, the organism was tested on the avicel overlay plates containing glucose or cellobiose at a range of concentration and was grown in continuous culture vessel, supplied by cellobiose medium, aiming the enhanced production of extracellular CM-cellulase at low dilution rates. Product inhibition of cellulase action by cellobiose was also tested. The results obtained are: i) no inhibition of CM-cellulase was observed up to 10 mM(3.4mg/ml) cellobiose in the reaction mixture, however 30% inhibition was observed at 20mM and 55% at 50mM, ii) the tests of catabolite repression on the solid media were successful, and avicel degradation was markedly repressed by glucose or cellobiose, iii) at low concentrations of cellobiose, dilution rate 0.05 and $1.0hour^{-1}$, no significant increase was observed in the production of either intra or extracellular CM-cellulase.

• Korean Journal of Plant Tissue Culture
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• v.25 no.6
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• pp.525-530
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• 1998

These studies were carried out to select the active grow hairy root lines induced from various ginseng(Panax ginseng C. A. Meyer) parts. Hairy roots were induced in root explants, stem and petiole in vitro by A. rhizogenes R1000 or A. rhizogenes $A_4$. These hairy roots could be grown on the phytohormone free medium, and PCR analysis of rol C and vir C gene fragments confirmed that hairy roots were transgenic tissues. We have selected 11 hairy root lines with active growing characters among 300 hairy root lines selected based on growth and morphological characteristics on 1/2MS solid media with 250 mg/L carbenicillin. Morphological characteristics of selected 11 hairy root lines were thickness and thiness of main roots, and many projection for lateral roots, active grow of lateral roots. Among selected 11 hair root lines prominent characteristics of hairy roots with active growing characters were thiness of main roots and active grow of lateral roots. But characteristics of low growing hairy roots were thickness of main roots and low grow of lateral roots. Finally we have selected actively growing hairy roots, KGHR-1, KGHR-5, KGHR-8 among 11 hairy root lines.

• Membrane and Water Treatment
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• v.4 no.2
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• pp.143-155
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• 2013

Four membranes were used to separate Chlorella sp. from their culture medium in cross-flow microfiltration (MF) experiments: cellulose acetate (CA), cellulose nitrate (CN), polypropylene (PP) and polyvinylidenefluoride (PVDF). It was found that the hydrophilic CA and CN membranes with a pore size of 1.2 ${\mu}m$ exhibited the best performances among all the membranes in terms of permeation flux. The hydrophobicity of each membrane material was determined by measuring the angle between the water (liquid) and membrane (solid). Contact angle measurements showed that deionized (DI) water had almost adsorbed onto the surfaces of the CA and CN membranes, which gave $0.00^{\circ}$ contact angle values. The PP and PVDF membranes were more hydrophobic, giving contact angle values of $95.97^{\circ}$ and $126.63^{\circ}$, respectively. Although the pure water flux increased with increasing pore diameter (0.8 < 1.2 < 3.0 ${\mu}m$) in hydrophilic CA and CN membranes, the best performance in term of filtration rate for filtering a microalgae suspension was attained by membranes with a pore size of 1.2 ${\mu}m$. The fouled membrane pore sizes and pore blocking were inspected using a scanning electron microscope (SEM). MF with large pore diameters was more sensitive to fouling that contributed to intermediate blocking, where the size of the membrane pores is almost equivalent to that of cells.