• Journal of Life Science
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• v.17 no.2 s.82
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• pp.293-297
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• 2007

Green fluorescent protein (GFP) is an attractive reporter for bioprocess monitoring. A fluorescence-based method was developed to quantify GFP levels in transgenic plants and protein extracts. In this study, GFP was produced and secreted from suspension cells derived from transgenic rice. The RAmy3E promoter placed before the GFP gene controlled by sugars such as sucrose. The effects of sucrose concentration on the secretion of GFP and total protein into the medium were investigated in batch suspension culture. It was possible, therefore, to induce the expression of the GFP by removing sucrose from the cultured media or by allowing the rice suspension cells to deplete sucrose catabolically. The dry cell weight (7.06 g/L) and GFP level were detected as highest at 12%, 3% sucrose after 20 day culture, respectively. However secreted GFP fluorescence at the other sucrose concentrations (6%, 12%, 18% and 24%) were a little amount in media.

• Journal of Life Science
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• v.9 no.4
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• pp.408-413
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• 1999

Extracellular capsidiol, sesquiterpenoid phytoalexin, in the medium of pepper (Capsicum annuum L.) suspension cells was not identified from control cells, but highly accumulated in the elicitor-induced cells within 6 hours after the addition of 0.05$\mu\textrm{g}$/$m\ell$ cellulase. Capsidiol production in elicitor-induced cells was markedly suppressed by cytochrome P450 inhibitors, such as ancymidol and ketoconazole demonstrating that biosynthesis of capsidiol is catalyzed by at least on hydroxylation enzyme in the biochemical pathway. Based on protein electrophoresis, two bands, 23.0kDa and 27.5kDa, were identified as newly synthesized polypeptides in the elicitor-induced suspension cells, suggesting that pepper cells which were subjected to elicitor treatment activate specific gene(s) for capsidiol biosynthesis in cultured cells.

• KSBB Journal
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• v.18 no.2
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• pp.133-139
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• 2003

Established cell-line cultures of cultured and wild mountain ginseng were characterized and their abilities to produce ginsenoside were determined. Cell lines were made of calli induced from the roots of wild mountain ginseng and cultured ginseng(Panax ginseng). Suspension cultures of wild mountain ginseng and cultured ginseng showed different growth and ginsenoside production rate. Their specific growth rates were 0.067 and 0.0035 day-1 in spite of having the same sugar consumption rates, where cells from wild mountain ginseng grew almost twice as fast as those of cultured ginseng. Their respective abilities to produce ginsenoside, however, were 0.53 and 2.53 mg/L.day, which means cells from cultured ginseng produced around 5 times more than wild mountain ginseng.

• BMB Reports
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• v.32 no.5
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• pp.451-455
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• 1999

The concentrations of L-ascorbic acid (AsA, ascorbate, vitamin C) and its biosynthetic and metabolically-related enzymes such as L-galactono-1,4-lactone dehydrogenase (GLDase), ascorbate peroxidase (APX), and ascorbate oxidase (ASO) were investigated in suspension cultures of Scutellaria baicalensis. Cells growing from 4 days after subculture (DAS) to 9 DAS and from 16 DAS to 19 DAS showed a diauxic growth, and then growth rapidly decreased with further culturing. The AsA content slowly increased to 19 DAS, reached a maximum at 21 DAS (ca $120\;{\mu}g/g$ dry cell wt), and then rapidly decreased with further culturing. GLDase and ASO activity were well correlated with the cell growth curve, showing a maximum at 19 DAS, whereas APX activity showed a good correlation with the changes in AsA content, showing a maximum at 21 DAS. The total ascorbate contents (reduced form, AsA, and oxidized form, dehydroascorbate, DHA) were markedly enhanced at 10 DAS when L-galactose and L-galactono-1,4-lactone (25 mM) were added to SH medium supplemented with 20 g/l sucrose at 9 DAS, by 5.5 and 6.8 times, respectively. DHA composed more than 90% of the total ascorbate contents in suspension cultures of S. baicalensis, even though the ratio of reduced to oxidized form slightly varied with cell growth stage. The results indicate that L-galactose and L-galactono-1,4-lactone are effective precursors of AsA in cell cultures of S. baicalensis, and that in vitro cultured cells provide suitable biomaterials for the study of biosynthesis and metabolism of AsA.

• Korean Journal of Plant Tissue Culture
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• v.24 no.1
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• pp.57-61
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• 1997

We investigated changes in the superoxide dismutase (SOD) activity and SOD isoenzyme pattern in suspension cultures of tomato (Lycopersicon esculentum), which were compared with those of intact tomato plants. two grams (fr wt) of cells subcultured at 15-day intervals were inoculated into 50 mL MS medium containing l mg/L 2,4-D and 30 g/L sucrose in a 300 mL flask and maintained at $25^{\circ}C$ in the dark (100 rpm). The cell growth reached a maximum at 20 days after subculture (DAS), followed by a rapid decrease with further cultures. The cell colour changed from white to black from 23 DAS. The intracellular SOD activity (units/g cell dry wt) was significantly increased from 23 DAS and reached a maximum at 28 DAS (52,400 units), followed by a decrease with further cultures, whereas the extracellular SOD activity showed a maximum at 25 DAS (27,800 units/50 mL medium). The total SOD activity per flask showed a maximum at 25 DAS (35,700 units), in which the extracellular SOD activity occupied about 75%. The tomato cultured cells had four SOD isoenzymes and their patterns were well correlated with SOD activity without a qualitative change during the cell cultures. The intact tomato plants had an additional CuZnSOD isoenzyme, showing the different isoenzyme patterns from cultured cells.

• Journal of Plant Biotechnology
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• v.34 no.3
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• pp.207-212
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• 2007

This study describes a simple and rapid method for quantitative determination of $\gamma$-aminobutyric acid (GABA) using $^1H$ NMR spectroscopy from whole cell extracts of plant suspension cultures. When 9 cell lines derived from 8 species of higher plants maintained in liquid Marashige and Skoog (MS) medium supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) were subjected to $^1H$ NMR, a cell line of Coriandrum sativum L. exhibited the highest level of GABA. The level reached up to 16.9 mg/dry wt when cells were cultured in MS medium supplemented with 0.5 mg/L 2,4-D after 3 weeks of incubation. The method for quantitative determination of GABA using $^1H$ NMR established in this study could be applied to high-throughput screening of various plant resources for GABA production and the cell suspension culture system of C. sativum could be further developed for commercial production of GABA.

• Korean Journal of Medicinal Crop Science
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• v.12 no.6
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• pp.478-482
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• 2004

Suspension culture of Schisandra chinensis for production of gomisin J was perfomed in bioreactor. The inoculum size and initial sucrose concentration had significant effect on the cell growth and gomisin J accumulation. The maximum dry cell weight $(DCW;\;43.5\;g/{\ell})$ and gomisin J content $(0.71\;{\times}\;10^{-3}\;{\mu}g/g\;DCW)$ were obtained at inoculum size of 100 g fresh cell weight (FCW) per liter and MB5 medium containing 6% sucrose after 8 weeks of culture. The effect of oxygen supply on the cell growth and gomisin J accumulation was also investigated in an airlift-type bioreactor. The optimal cell growth and gomisin J content was obtained under 0.5 vvm. The productivity of gomisin J was 0.7 fold in bioreactor culture lower than that obtained in a flask cultivation.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.3
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• pp.306-316
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• 1997

Embryogenic calli were induced from mature seed scutella of anther culture-derived rice variety Zhonghua 8. Cell suspension cultures were initiated from friable embryogenic calli and utilized as source material for protoplast isolation. Generally, the older and finer cell suspensions gave higher protoplast yields than younger suspension cultures. Protoplasts exhibited sustained cell division and formed microcalli when cultured in KPR medium supplemented with 0.5 mg $l^{-1}$ 2,4-D, 1.0 mg $l^{-1}$ NAA and 0.5 mg $l^{-1}$ zeatin using the agarose embedding procedure without feeder cells. Protoplast plating efficiencies ranged from 0.20 to 0.54％. Microcalli were transferred to MS medium supplemented with 2.0 mg $l^{-1}$ kinetin and 0.5mg $l^{-1}$ NAA for plant regeneration. The regeneration frequencies were 2 to 12％, depending on the cell suspension lines of Zhonghua 8. The plants were transferred to the glasshouse and were fertile.

• Journal of Life Science
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• v.6 no.2
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• pp.94-103
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• 1996

To understand effect of inoculum size, cell density, sucrose concentration and concentrations of MS basal on suspension culture and protoplast isolation of wild viola(Viola patrinii DC.) callus from petiole segments this experiment was conducted. In the lot of 30 mesh inoculum size, two observations were; One was that a considerable increase in the fresh and dry weight of callus was determined. Another was that the callus mass was relatively compact compared with others. A recommendable cell density was 0.4g for 20ml culture medium and the higher sucrose concentration, the higher fresh and dry weight were obtained. The dilution of MS basal salt was differently affected on fresh and dry weight; the highest fresh weight was found in 1x MS salt, while the higest dry weight was in 1/3x dilution.The addition of casein hydrolysate(3g/L) was more effective to increase of both fresh and dry weight. THe contents of protein was great in the inoculum lots with larger inoculum sizeand higher concentration of MS basal salts contenting 3g/L of casein hydrolysate and higher sucrose compared with others. The greatest protoplasts were isolated from the lot of 10 mesh size treated with 1%pectinase SE-150 and 2% cellulase YC. In general, for optimal protoplast isolation the followingconditions were recommended; 1) Use of smaller cell size cultured for 2-5 weeks, and 2) more than 5 hours incubation using the combined mixture of the enzymes with proper concentrations.

• Journal of Plant Biotechnology
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• v.29 no.3
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• pp.185-188
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• 2002

Off-white, friable embryogenic calluses were formed on the internal integument of mature seeds of yuzu (Citrus junos) cultured on Murashige and Skoog's basal medium at a frequency of 1.2%. Embryogenic calluses were proliferated when cultured on medium with 1 mg/L 2,4-D. Upon transfer to medium with 0.1 mg/L kinetin, embryogenic calluses produced numerous somatic embryos. Embryogenic suspension cultures were established by placing embryogenic calluses into liquid medium with 1 mg/L 2,4-D. When plated onto medium with 0.5 mg/L ABA, embryogenic cells developed into somatic embryos at a high frequency, and then regenerated into plantlets. Plantlets were successfully transplanted to potting soil and grown in a greenhouse.