• The Plant Pathology Journal
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• v.36 no.3
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• pp.303-303
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• 2020

• Journal of Plant Biotechnology
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• v.1 no.1
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• pp.1-7
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• 1999

Large-scale cultures of plant cell, tissue, and organ have been achieved by using BTBB. When different sized BTBBs (5 L, 20 L, 100 L, 300 L, and 500 L) were tested for the culture of yew cells (Taxus cuspidata Sieb. et Zucc.), cell growth increment reached to 94.5% in SCV after 24 days of culture with 30% of inoculation cell density. However, there were some variations in the production of taxol and its derivatives among the BTBBs of different size. Approximate 4 ㎎/l of taxol and 84 ㎎/l of total taxanes were obtained by using a 500L BTBB after 6 weeks of culture. With a 20L BTBB, about 20,000 cuttings of virus-free potatoes (cv. Dejima) could be obtained by inoculating 128 explants and maintaining 8 weeks under 16 hr light illumination. The frequency of ex vitro rooting of the cuttings revealed as more than 99% under 30% shade. By incorporating two-stage culture process consisting of multiple bulblet formation in solid medium and bulblet development in liquid medium, mass propagation of lily through bioreactor seemed to be possible. In the case of 'Marcopolo', the growth of mini-bulblets in BTBB was nearly 10 folds faster than that of the solid medium. Time course study revealed that maximum MAR yield of ginseng (Panax ginseng C. A. Meyer) in a 5 L and 20 L BTBB after 8 weeks of culture was 500 g and 2.2 ㎏, respectively. By cutting the MAR once and/or twice during the culture, the yield of root biomass could be increased more than 50% in fresh weight at the time of harvest. With initial inoculum of 500 g of sliced MAR in a 500 L BTBB, 74.8 ㎏ of adventitious root mass was obtained after 8 weeks of culture. The average content of total ginseng saponin obtained from small-scale and/or pilotscale BTBBs was approximately 1% per gram dry weight. Based on our results, we suggest that large-scale cultures of plant cell, tissue, and organ using BTBB system should be quite a feasible approach when compared with conventional method of tissue culture.

• Korean Journal of Plant Resources
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• v.34 no.5
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• pp.470-477
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• 2021

Amsonia ellipotica (Thumb.) Roem. & Schult. has less than 10 native places nationwide and is evaluated as an endangered species (EN) on the Korean Red List, so systematic breeding and cultivation methods are required. This study was carried out to establish an effective method for seedling production in A. elliopotica that can be developed as a genetic resource conservation and new ornamental material. This experiment was performed by varying the seedling production container (72, 105 and 128 cell trays), number of seeds sown in a cell (1, 2, and 4 per cell), soil type [horticultural soil and neutralized peat-moss: perlite mixed in 3:1, 4:1 (v:v)], additional fertilizer concentrations (0, 250, 500 and 1000 mg/L) and shading degrees (0, 55 and 75%). As a result of the study, growth increased with the increased in measuring capacity per cell of sowing container. According to number of seeding growth, sowing one seed per cell was the best. Seedling growth increased as the treatment concentrations increased, higher growth was in 1000 mg/L. On the other hand, during the shading treatment, growth significantly decreased regardless of shading degrees.

• Korean Journal of Plant Resources
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• v.36 no.6
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• pp.588-596
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• 2023

Long-term culture of cell lines is an important issue in in vitro culture and in plant science. In this study, the regeneration ability and ex vitro acclimatization of regenerants were evaluated. The ploidy level of regenerants derived from long-term cultured cell lines was measured in Imperata cylindrica 'Rubra', Poaceae. Adventitious buds (shoots) were successfully induced from five-year-cultured calli on MS medium containing 0.1 mg/L BA or 2.0 mg/L TDZ, combined with 0.01 mg/L auxins (IAA, IBA, NAA and 2,4-D), respectively. Adventitious roots were also induced on MS medium containing 0.01 mg/L auxins (IBA, NAA and 2,4-D), respectively. Interestingly, regenerants with both red and green leaf were successfully obtained when regenerants were cultured on MS medium with 9% sucrose. Regenerants derived from long-term cultured calli were transferred to pots using an optimal acclimatization process and successfully adapted to both pot and soil conditions. Moreover, the ploidy level was measured using calli and regenerants that had been kept on MS medium containing various kinds of plant growth regulators (PGRs).

• Journal of Plant Biotechnology
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• v.32 no.4
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• pp.313-319
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• 2005

The effects of various carbon sources on the secretion of hGM-CSF, total protein and protease into the medium were investigated in transgenic tobacco cells. The dry cell weight (11.2 g/L) and wet cell weight (310.8 g/L) were highest at 30 g/L glucose after 5-day culture but, the dry cell weight (13.4 g/L) and wet cell weight (480 g/L) were highest at 30 g/L sucrose after 10-day culture. The total protein (110.3 mg/L), protease activity (3950 U/L) and total secreted hGM-CSF (56 mg/L) were highest at 30 g/L sucrose after 10-day culture. Stabilization of the total secreted protein and hGM-CSF in various carbon source concentrations was determined. Total secreted protein was most stabilized in the medium containing sucrose. However, the loss of the total protein was increased with the concentrations of high level in medium containing sorbitol, mannitol, fructose, and glucose. hGM-CSF was more stabilized in the medium containing sucrose than in the medium containing sorbitol, mannitol, fructose, glucose.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2005.11a
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• pp.433-433
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• 2005

• Journal of Plant Biotechnology
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• v.32 no.3
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• pp.209-215
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• 2005

Lactoferrin is an iron-binding glycoprotein with many biological roles, including the protection against microbial and virus infection, stimulation of the immune system. We developed the transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing the human lactoferrin (hLf) protein following Agrobacterium tumefaciens-mediated transformation. A construct containing a targeting signal peptide from tobacco endoplasmic reticulum fused to hLf cDNA under the control of an oxidative stress-inducible SWPA2 promoter was engineered. Transgenic Siberian ginseng cultured cells to produce a recombinant hLf protein were successfully generated and confirmed by PCR and Southern blot analysis. ELISA and western blot analysis showed that full length-hLf protein was synthesized in the transgenic cells. The production of hLf increased proportionally to cell growth and reached a maximal (up to 3% of total soluble proteins) at the stationary phase. These results suggest that the transgenic Siberian ginseng cultured cells in this study will be biotechnologically useful for the commercial production of medicinal plant cell cultures to produce hLf protein.

• The Plant Pathology Journal
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• v.17 no.6
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• pp.315-324
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• 2001

Tobacco (Nicotiana tabacum cvs.Xanthi-nc and NC 82) plants infected with Tobacco mosaic virus (TMV) were examined ultrastructurally. Local lesions produced by TMV were sunken and withered. The plants were subjected to temperature shift (TS), a method to produce programmed cell death (PCD), by placing the infected plants initially at high temperature (35$^{\circ}C$) for 2 days and then shifting them to greenhouse temperature (22-27$^{\circ}C$). As a result, expanded lesions around the original necrotic lesions were produced. The expanded area initially had no symptoms, but it withered and became necrotic 15 h after TS. No ultrastructural changes related to PCD were noted at 0 h after TS in Xanthi-nc tobacco tissues as well as in healthy and susceptible tobacco tissues infected with TMV, At 6 h after TS, chloroplasts were convoluted and cytoplasm began to be depleted; however no necrotic cells were found. At 17 h after TS, ground cytoplasm of affected cells was completely depleted and chloroplasts were stacked together with bent cell wall or dispersed in the intracellular space. Necrotic cells were also observed, containing virus particles in the necrotic cytoplasm. There were initially two types of symptoms in the expanded lesions: chlorosis and non-chlorosis (green). Abundant TMV particles and X-bodies were only found in the chlorotic tissue areas. These results suggest that PCD by TMV infection may start with the wilting of cells and tissues before necrotic lesion formation.

• 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.

• BMB Reports
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• v.31 no.4
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• pp.413-417
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• 1998

Transgenic mice containing a bovine ${\beta}-Casein/Human$ lysozyme fusion gene (pBZ) were generated in order to produce human lysozyme in their milk. The expression vector was a quadripartite fusion consisting of a 2 kb upstream DNA of the bovine ${\beta}-casein$ gene, human lysozyme gene, intron II of the rabbit ${\beta}-globin$ gene, and the polyadenylation/termination signals of SV40 DNA. Fertilized mouse zygotes were microinjected with pBZ, then transferred into the oviduct of foster mothers. Out of 20 mice born, 11 survived until postweaning and three were identified as positivetransgenic by Southern blot analysis (one male and two females). The founder mice were mated to BCFl mice to produce transgenic progeny. It was confirmed by RT-PCR and Northern blot analyses that the transgene was specifically expressed in the mammary gland of the founder mice. Furthermore, the artificial introns within the transgenic RNA was proven to be correctly spliced out as judged by RT-PCR analysis. These results indicated that transgenic mice generated in this study properly expressed the human lysozyme RNA in their mammary gland.