• KSBB Journal
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• v.16 no.6
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• pp.568-572
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• 2001

Light is one of the most important environmental factors controlling plant physiology. The human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was produced from cell suspension cultures of transgenic tobacco under different light conditions (24 hr light, 18 hr light/dark cycle, dark). Under 24 hr light condition, cell growth was best and dry cell weight reached 14.4 g/L. Light did not influenced the secretion of total proteins. However, in the dark condition, the ratio of secreted total protein/dry cell weight was 1.5 fold higher than those of ethel conditions. Production of hGM-CSF was highest with 18 hr light condition and reached 496.5 ug/L. In addition, the content of hGM-CSf in secreted total proteins was 1.8 fold higher than that of 24 hr light condition, which is beneficial for the purificationof the protein.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.97.2-98
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• 2003

A new and effective formulation using antagonistic bacteria, Burkholderia cepacia YC5025 in vegetable oil was developed for the biocontrol of anthracnose. The bacterial population in the formulation was maintained to 5x10/sup7/ cfu/ml upto 60 days at room temperature. Control efficacy of the formulation for anthracnose was over 80％ by spraying of diluted suspension(x1,000) in growth chamber tests. On the contrary, the bacterial suspension in distilled water or bacterial culture broth containing same number of spores as the formulation had low control efficacy around 40％ even 2-weeks storage after preparation. The shelf-life of the formulation was longer than that of bacterial preparation using clay minerals such as talc or bentonite. The mechanisms of newly developed bacterial formulation are possibly the formation of water film on the surface of cucumber leaves and inactivation of the bacteria in the vegetable oils during storage. Further field tests and improvements with new liquid bacteiral formulation need to be done for practical application.

• Korean Journal of Plant Tissue Culture
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• v.25 no.3
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• pp.141-146
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• 1998

Feeding experiment of [$^3$H] 5-epi-aristolochene (5-EAS) demonstrated in suspension cultures of tobacco (Nicotiana tabacum L.) that 5-EAS hydroxylase activity was absent from control cells, but induced to a maximum level within 18 h after the addition of cellulase, and was very similar to induction pattern of sesquiterpene cyclase. This result suggest that the conversion of 5-EAS to capsidiol is catalyzed by at least one elicitor-inducible hydroxylase. Cytochrome P450 inhibitors, ancymidol and ketoconazole, suppressed the elicitor-induced capsidiol accumulation by inhibiting hydroxylase activity, suggesting that the hydroxylase may be a Cyt P450 type enzyme.

• Korean Journal of Pharmacognosy
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• v.21 no.3
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• pp.205-209
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• 1990

The study for the quantitative analysis of saikosaponin of Bupleurum falcatum root produced by tissue culture was attempted. The optimal concentration of 2,4-D was 0.1 mg/l for inducing the callus. The induction and growth of the adventitious root from the callus was obtained in the suspension culture containing MS basal medium supplemented with no plant growth regulators. The results of the quantitative analysis of saikosaponins by high performance liquid chromatography (HPLC) showed that the content of saikosaponin a and c was high in the one-year-old root from somatic embryos and that of saikosaponin d was remarkably high in three-months-old adventitious root from callus.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.3
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• pp.187-191
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• 2003

Proteolytic enzymes existing in plant cell cultured media are the major reason for the loss of secreted human granulocyte-macrophage colony-stimulating factor (hGM-CSF). The addition of pepstatin, aprotinin and PMSF relatively decreased the proteolytic degradation of hGM-CSF in a conditioned medium, but sufficient prevention against the proteolytic activity could not be obtained with chemical protease inhibitors. Gelatin, as a competitive substrate for protease, showed a stabilizing effect in a conditioned medium. Compared to the initial hGM-CSF concentration in a conditioned medium. with 10 g/L of gelatin, 68% of the hGM-CSF remained after 5 days. In a cell culture experiment, 5 g/L of gelatin significantly stimulated the hGM-CSF production and accumulation in culture media, with no growth inhibition. compared to the controls (4.72 $\mu\textrm{g}$/L), the extracellular hGM-CSF level could be increased to 39.78 $\mu\textrm{g}$/L with the addition of 5 g/L of gelatin.

• Korean Journal of Medicinal Crop Science
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• v.4 no.2
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• pp.132-138
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• 1996

This experiment was carried out to establish a year-round production system of pathogen-free stock through micropropagation in Fritillaria thunbergii as medicinal bulbous plant. The effect of different types of culture method and plant growth regulators, activated charcoal and mannitol on bulblet formation and subsequent growth were investigated. The MS solid medium containing 1. 0 mg/L kinetin and 0. 3 mg/L NAA was effective on bulblet formation and propagation rate compared to liquid and suspension culture. Addition of activated charcoal at 0. 01% to 0. 1% in the medium promoted bulbing of cultured bulblets and shoot formation. Addition of 1% to 2% mainnitol in MS medium was effective on the formation of bulblet and subsequent growth of bulblets compared to control. In addition of inhibitors, $10{\sim}100\;mg/L$ B-9 and Chloromequat had effective to stimulate bulblet growth but their effects were not so much as mannitol treatment. ABA treatment had detrimental effect on survival rate of explant and bulblet formation.

• Journal of Photoscience
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• v.7 no.2
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• pp.53-58
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• 2000

We performed the cloning of a chalcone synthase (CHS) gene, the key enzyme in the anthocyanin biosynthesis, from the cDNA library constructed with grape suspension cells irradiated UV-B. The PCR fragment was used to cloning the CHS gene. One CHS cDNA clone containing an open reading frame and a partial stilbene synthase (STS)cDNA, the stilbene-type phytoalexin, were isolated. The CHS cDNA clone (VCHS) showed 87% sequence homology with VvCHS (V.vinifea) and 72.3% identity with VSTSY(V.vinifea). its amino acid sequences were longer than any other CHS genes as 454 residues. Two genes were weakly expressed in white light irradiated cells, but highly induced in UV-B irradiated condition during 32 hours. Interestingly, the STS was quickly and abundantly expressed from 2 hours when supplemented with jasmonic acid (JA) and the maximum expression was observed at 4 hours and then gradually decreased. But, the additional UV-B or white light quickly degraded the STS expression than only JA treated grape suspension cells. The CHS also was rapidly induced with JA and the synergistical effect was observed at the addigional light treatment of UV-B or white light. These results are indicated that CHS and STS have different response mechanisms against the environmental stresses.

• Plant Resources
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• v.3 no.3
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• pp.219-222
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• 2000

For the purpose of establishing an efficient propagation through airlift bioreactor system of Zingiber of officinale Rosc. Korean native Seosanjong, the effect of different factors and bioreactor on cultured plantlets were investigated. The highest number of plantlets, fresh weight per plant was obtained from explants when cultured in MS liquid medium including 0.3 mg/L NAA and 2.0 mg/L kinetin for 40 days. A 10 L bottle type bubble bioreactor, compared with 250 mL Erlenmeyer flask, was more efficient, producing 4.7 plantlets or from 1.5 to 1.6 times more than did the Erlenmeyer flask. The results demonstrate the rapid mass propagation of airlift bioreactor to produce normal ginger.

• Journal of Plant Biotechnology
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• v.31 no.3
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• pp.239-248
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• 2004

This experiment was carried out to confirm that phytochrome regulates anthocyanin bio-synthesis during cell suspension culture system of grape or not. In suspension culture of grape, maximum accumulation of anthocyanin was observed at the stationary phase under continuous white light condition. From mono-chromatic light interruption for 24h at the 4th or 7th day on the suspension cultured cells, the anthocyanin accumulation was highly enhanced at the light interruption at 7th day than 4th day under all monochromatic light treatment. However, the cell growth patterns were not affected by any light treatment. In the darkness, the anthocyanin synthesis was very low but remarkably increased by blue light or red light irradiation. However, the increase of anthocyanin accumulation by blue or red light was suppressed by far-red light in the suspension cells of grape. This suppression by far-red light on the anthocyanin synthesis also observed on the cells treated red or far-red light alternatively. These results implied that phytochrome regulation system may be involved in the anthocyanin biosynthesis of the suspension grape cells. By RNA expression analysis, chalcone synthase (CHS) gene was expressed highly by blue and red light but low by far-red light. The synergistic increase of CHS gene expression was also observed at the treatment of blue light followed by red for 24h. This result may explain the increase of anthocyanin accumulation in B/R treatment. Although the expression of phytochrome gene (PHYA or PHYB) was not highly increased by all light treatment (blue, red, and far-red light) the expression of both PHYA gene and PHYB gene was increased a little in cells treated red or far-red light. In grape suspension cells, the red light enhanced the anthocyanin synthesis, whereas the far-red light was suppressed. Although it was not confirmed whether or not phytochrome gene is activated in anthocyanin accumulating grape cells, we believed that anthocyanin biosynthesis in grape cells may be regulated under phytochrome signal transduction system.

• The Plant Pathology Journal
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• v.24 no.3
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• pp.309-316
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• 2008

Mycelium of Ophiostoma quercus albino strain cultured in liquid culture media was harvested, lyophilized, and stored for examining biocontrol efficacy against wood discoloration by staining fungi in the laboratory and field conditions. Dry weight of mycelium grown in brown sugar yeast extract broth(BYB) showed 3.8 times higher than that grown in potato dextrose broth(PDB). The optimum culture period in BYB was 4 weeks. In vitality test of the albino strain, the lyophilized mycelium stored in liquid nitrogen($-196^{\circ}C$) or in a refrigerator($4^{\circ}C$) kept the vitality until 13 months after storage; however, the mycelium stored at room temperature lost the vitality completely after 13 months. The mycelium stored in liquid nitrogen or in a refrigerator protected wood chips from the discoloration by pretreating mycelial suspension on pine wood chips. The mycelium stored at room temperature for 7 months also showed complete protection. These results suggest that the lyophilized mycelium have a biocontrol efficacy only if it keeps the least vitality. In the field conditions, both albino strain and $Woodguard^{(R)}$(commercial chemical protectant) showed significant differences(p=0.05) in discoloration rate as compared to the non-treated control when these were treated on the wood logs of Pinus rigida. The albino strain showed better protection than $Woodguard^{(R)}$. Isolation frequency of blue stain fungi from the chips of wood logs treated with the albino strain was 0% at three months after treatment, while that treated with $Woodguard^{(R)}$ was 76.7%. In another experiment, pre-treatment of mycelial suspension on the cut surface of wood logs also showed significant protection from wood discoloration. Spraying of both albino strain on the cut surface and insecticides on the bark also showed relatively good control effects as compared to insecticide alone on the bark or nontreated control.