• Korean Journal of Plant Tissue Culture
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• v.28 no.1
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• pp.7-13
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• 2001

Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.

• The Plant Pathology Journal
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• v.22 no.1
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• pp.75-89
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• 2006

The actinomycete strain JK-1 that showed strong inhibitory activity against some plant pathogenic fungi and oomycetes was isolated from Jung-bal Mountain in Ko-yang, Korea. The strain JK-1 produced spores singly borne on sporophores and the spores were spherical and 0.9-1.2 11m in diameter. The cell wall of the strain JK-1 contained meso-diaminopimelic acid. The actinomycete strain JK-1 was identified as the genus Micromonospora based on the morphological, physiological, biochemical and chemotaxonomic characteristics. From the 168 rDNA analysis, the strain JK-1 was assigned to M aurantiaca. The antibiotic MA-1 was purified from the culture broth of M aurantiaca JK-1 using various purification procedures, such as Diaion HP20 chromatography, C18 flash column chromatography, silica gel flash column chromatography and Sephadex LH-20 column chromatography. $^{1}H-$, $^{13}C-NMR$ and EI mass spectral analysis of the antibiotic MA-1 revealed that the antibiotic MA-1 is identical to phenylacetic acid. Phenylacetic acid showed in vitro inhibitory effects against fungal and oomycete pathogens Alternaria mali, Botrytis cinerea, Magnaporthe grisea, Phytophthora capsici and yeast Saccharomyces cerevisiae at < 100 $\mug$ $ml^{-1}$. In addition, phenylacetic, acid completely inhibited the growth of Sclerotinia sclerotiorum, Bacillus subtilis, Candida albicans, Xanthomonas campestris pv. vesicatoria at < $\mug$ $ml^{-1}$. Phenylacetic acid strongly inhibited conidial germination and hyphal growth of M grisea and C. orbiculare. Phenylacetic acid showed significantly high levels of inhibitory' effect against rice blast and cucumber anthracnose diseases at 250 $\mug$ $ml^{-1}$. The control efficacies of phenylacetic acid against the two diseases were similar to those of commercial compounds tricyclazole, iprobenfos and chlorothalonil .n the greenhouse.

• Applied Biological Chemistry
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• v.36 no.6
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• pp.562-566
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• 1993

The conditions required for plant transformation through the electroporation system and for the electrofusion of the prtoplasts were investigated for geranium (Pelargonium zonale hybrids). The optimum condition for electroporation was 1.77 kV/cm for $40\;{\mu}sec$ under which 70% of the protoplasts were viable and 58% of the viable protoplasts were stained with methylene blue. The pBin19 DNA plasmid used as a carrier vector was isolated from E.coli $DH5{\alpha}$ strain, purified, identified by the electrophoresis on agarose gel and electroporated into the protoplasts. The KM8 liquid medium gave better cell division than any other media. One MHz of AC frequency with 40 V/cm of amplitude for 15 sec followed by 0.5 kV/cm of DC amplitude for $60\;{\mu}sec$ was most efficient for the electrofusion of protoplasts.

• Journal of Bio-Environment Control
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• v.12 no.2
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• pp.101-105
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• 2003

This experiment was conducted to find out the effect of day length on the production of high quality plug seedlings in onion (Allium cepa L.). Two cultivars, ‘Changnyongdaego’ and‘Wolryun’, were grown to seedlings in 200-cell plug trays under 11.5, 12.5, 13.5 hours and natural day length. These seedlings were transplanted to the pot (16 cm In diameter) and grown under 16 hours day length. Number of leaves and neck diameter showed better growth in the longer than shorter day length treatments, but plant height old sheath length were retarded in the longer day length treatments. Growth such as no. of leaves, neck diameter, plant height and sheath length increased with the passage of day, but plant height and neck diameter decreased by treatment over 20 days with 13.5 hours day length. Bulbing and bulb size of onion after transplanting were enhanced in the seedlings cultured under longer day lengths. From the above results, treatment of long day length during seedling culture in plug tray can control the overgrowth and produce high quality plug seedlings.

• Korean Journal of Plant Tissue Culture
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• v.26 no.1
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• pp.53-58
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• 1999

The gamma radiation-induced changes of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in callus cultures of cassava (Manihot esculenta Crantz) selected as a high yield of cell line for SOD were investigated. In normal cultures, the cell growth reached a maximum at 30 days after subculture (DAS), followed by a rapid decrease with further cultures. The SOD and POD specific activities (units/mg protein) showed the highest at the immediately after subculture and subsequently decreased to 20 DAS, and then increased to 30 DAS, whereas the CAT activity showed the lowest at just after subculture, and it continuously increased from 15 DAS to 30 DAS, showing a good correlation with the cell growth. Irradiation of gamma-ray of 50 and 70 Gy on 7 DAS inhibited significantly the cell growth by 50% and 80% at 14 days after treatment (DAT), respectively. In the cells irradiated with 70 Gy, SOD and POD specific activities increased by 4 and 2.5 folds at 14 DAT, respectively, whereas CAT activity was not affected. The results indicate that SOD and POD may be involved in the antioxidative mechanism in relation to oxidative stresses induced by subcultures and by gamma radiation in callus cultures of cassava.

• KSBB Journal
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• v.14 no.3
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• pp.328-336
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• 1999

A green microalga, Chlorella vulgaris UTX 259, was cultivated in a bench-scale raceway pond. During the culture, 15%(v/v) $CO_2$ was supplied and industrial wastewater discharged from a steel-making plant was used as a culture medium. In a small scale culture bottle, the microalga grew up to 1.8 g $dm^{-3}$ of cell concentration and ammonia was completely removed from the wastewater with an yield coefficient of 25.7 g dry cell weight $g^{-1}\;NH_3-N$. During the bottle-culture, microalga was dominant over heterotrophic microorganisms in the culture medium. Therefore, the amount of carbon dioxide fixation could be estimated from the change of dry cell weight. In a semi-continuous operation of raceway pond with intermittent lighting (12 h light and 12 h dark), increase of dilution rate resulted in increase of the ammonia removal rate as well as the $CO_2$ fixation rate but the ammonia removal efficiency decreased. Ammonia was not completely removed from the medium (wastewater) of raceway pond which was operated in a batch mode under a light intensity up to 20 klux. The incomplete removal of ammonia was believed due to insufficient light supply. A mathematical model, capable of predicting experimental data, was developed in order to simulate the performance of the raceway pond under the light intensity of sun during a bright daytime. Simulation results showed that the rates of $CO_2$ fixation and ammonia removal could be enhanced by increasing light intensity. According to the simulation, 80 mg $dm^{-3}$ of ammonia in the medium could be completely removed if the light intensity was over 60 klux with a continuous lighting. Under the optimal operating condition determined by the simulation, the rates of carbon dioxide fixation and ammonia removal in the outdoor operation of raceway pond were estimated as high as $24.7 g m^{-2} day^{-1}$ and $0.52 g NH_3-N m^{-2} day^{-1}$, respectively.

• KSBB Journal
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• v.21 no.5
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• pp.331-335
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• 2006

Suspension cells of Angelica gigas Nakai were cultivated to produce extracellular polysaccharide(ECP) as immunostimulating agents. Effects of environmental conditions such as sucrose and 2,4-dichlorophenoxyacetic acid(2,4-D) concentrations on the growth and production of ECP were studied using suspension cultures of A. gigas Nakai. Final dry cell weight was increased with an increase of initial sucrose concentration from 30 to 60 g/L. The maximum production of ECP(1.2 g/L) was achieved at an initial sucrose concentration of 50 g/L on day 8. High 2,4-D concentration was effective for ECP production but not for cell growth. In addition, various fungal elicitors were investigated for the enhanced production of ECP in A. gigas suspension cultures. Among the tested fungal elicitors, Verticillium dahliae was the most effective for the production of ECP in A. gigas suspension culture.

• KSBB Journal
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• v.30 no.6
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• pp.307-312
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• 2015

Transgenic rice cells using RAmy3D promoter can provide high productivity, and the production of recombinant protein is induced by sugar starvation. In this system, productivity was reduced during the scale-up processes. To ensure the influences of shear stress and oxygen transfer rate, working volume and mixing performances were investigated under various agitation speeds and working volumes. In addition, inoculation methods including suspended cells and filtered cells were compared. Working volumes and shaking speeds were 300, 450 mL and 80, 120 rpm, respectively. Hydrodynamic environment of each condition was measured numerically like mixing time and $k_La$. Good mixing performance and high shear stress were measured at high agitation speed and low volume. The highest level of hCTLA4Ig was 30.7 mg/L at 120 rpm, 300 mL. When conditioned medium was used for inoculation, increased cell growth was noticed during the day 0~4 and decreased slower than filtered cells. Compared with filtered cells, the maximum hCTLA4Ig level reached 37.8 mg/L at 120 rpm, 300 mL and lower protease activity level was observed. In conclusion mixing performance is critical factor for productivity and conditioned medium can have a positive effect on damaged cells caused by hydrodynamic shear stress.

• Natural Product Sciences
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• v.3 no.2
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• pp.81-88
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• 1997

NAD(P)H:quinone reductase (QR) is one of the protective phase II enzymes against toxicity that accomplishes the capacity of detoxification by modulating the effects of mutagens and carcinogens. The detoxification mechanism is that quinone reductase promotes the 2-electron reduction of quinones to hydroquinones which are less reactive. This study is to search new inducers of quinone reductase from natural products, which can be used as cancer chemopreventive agents. Plant extracts were evaluated by using quinone reductase generating system With Hepa 1c1c7 murine hepatoma cell lines for enzyme inducing properties and crystal violet staining method for the measurement of cytotoxicity provoked. We have tested approximately 106 kinds of natural products after partition into n-hexane, ethyl acetate and aqueous layers from 100% methanol extracts of natural products. The ethyl acetate fractions of Vitex rotundifolia $(fruits,\;2FC:\;12.7\;{\mu}g/ml)$, Cnidium officinale $(aerial\;parts,\;2FC:\;10.5\;{\mu}g/ml)$, Chrysanthemum sinese $(flowers,\;2FC:\;17.4{\mu}g/ml)$ and the hexane fractions of Angelica gigas $(roots,\;2FC:\;13.2\;{\mu}g/ml)$, Smilax china $(roots,\;2FC:\;l1.9\;{\mu}g/ml)$, Sophora flavescens $(roots,\;2FC:\;16.3\;{\mu}g/ml)$ revealed the significant induction of quinone reductase in a murine hepatic Hepa 1c1c7 cell culture system.

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.611-616
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• 2003

Korean ginseng(Panax Ginseng C.A. Meyer) known as a oriental miracle drug is an important medicinal plant. Ginseng has been used for geriatric, tonic, stomachic, and aphrodisiac treatments for thousands years. Also, it is an antibiotic and has therapeutic properties against stress and cancer. Ginseng is widely distributed all over the world. Among them, Korean mountain ginseng has the most valuable effect on pharmaceuticals. The roots of mountain ginseng contained several kinds of ginsenosides that have many active functions for the human body. However, the study of mountain ginseng has a limit because the mountain ginseng is very expensive and rare. So, we artificially cultured mountain ginseng adventitious roots using the bioreactor culture system. We induced callus from original mountain ginseng, directly dug up in mountain and aged about one hundred ten years. Separated adventitious roots were precultured in 500ml conical flasks and then, transferred in 20L bioreactors. The adventitious roots of mountain ginseng were harvested after culturing for 40days, dried and then, extracted with several solvents. In this study, we investigated the whitening effect, anti-wrinkle effect and the safety of tissue cultured adventitious roots extract of mountain ginseng in order to identify the merit as a cosmetic ingredient. Particularly, extract of mountain ginseng adventitious roots showed whitening and anti-wrinkle effects. The inhibitory effect of this extract on the melanogenesis was examined using B-16 melanoma cell. When B-16 melanoma cells were cultured with adventitious root extract, there was a dramatically decrease in melanin contents of 8-16 melanoma cell. And we identified this extract inhibited Dopa auto-oxidation significantly. Also, when transformed mouse fibroblast L929 cells were treated with this extract, there was a significant increase in collagen synthesis. The results show significant inhibited melanization and wrinkle without inhibiting cell viability.