• Journal of Ginseng Research
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• v.27 no.2
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• pp.78-85
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• 2003

By the Agrobacterium rhizogenes A$_4$ were induced a transformed callus of ginseng hairy root and examine to find the possibility whether it can produce certain ginsenoside. Investigations for a finding out to optimal culture medium showed that BA application is better than more factorial composition between auxins and cytokinins. For the induction of hairy root callus of ginseng, l/2 MS medium containing 1 to 3 mg of benzyladenine(BA) per liter gave the best result. The growth of ginseng hairy root callus(GHC) cultured with the 1/2MS medium supplemented with 2 mg BA/L was selected for best suspension cultures. The optimum concentration of BA for ginsenosides production was found to be 2 mg/L. Probably the inoculum size of callus plays a role with the ginsenoside production in suspension culture. AS for inoculum size of callus, 50 mg was superior to 150 mg for growth and ginsenoside production. Ginsenoside contents were highest in the suspension culture grown for four weeks under continuous light condition. In fact that continous light treatment promote strongly the synthesis of ginsenoside of the hairy root callus is first result in the world and the numerously induced root hairs of the callus leads a new method for ginsenoside production.

• Korean Journal of Plant Tissue Culture
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• v.22 no.2
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• pp.71-76
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• 1995

The effects of light on the activities of several antioxidative enzymes, catalase (CAT), superoxide dismutase(SOD), ascorbate oxidase(AO), and peroxidase(POD) were examined in the hairy root cultures of Phytolacca esculenta van Houtte induced by Agrobacterium tumefaciens $A_4$T. Activities of CAT, SOD, and AO were significantly decreased with incresing light intensity (500-2,000 lx). The activity of AO under high light condition (2,000 lx)was decreased by 92% compared to the dark condition. The activities of glutathoine peroxidase (GPO), ascorbate peroxidase (APO) and general POD were increased under lower light intensify below 500 lx. The activity of GPO under 2,000 lx was decreased by 85% compared to the dark condition. The activities of antioxidative enzymes were more decreased in blue light (400-500nm). The activities of antioxidative enzymes in blue light intensity were increased in lower light intensity below 30 lx, but decreased 21-70% under 200 lx. The activity of AO was decreased by 70% under 200 lx with increasing blue light intensity. Our results suggest that the activities of antioxidative enzymes in hairy roots might be inhibited by endogenous oxidants generated under the high blue light conditions.

• Korean Journal of Plant Tissue Culture
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• v.24 no.5
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• pp.273-277
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• 1997

The sesquiterpene cyclase (SC) was induced and its products were accumulated in the culture media of Hyoscyamus muticus hairy roots by addition of Rhizoctonia solani extracts. The cumulative production of solavetivone was nearly doubled by integrated extraction of the products from the media during the 24 h accumulation period. Western blots with monoclonal antibodies against SC show that the enzyme levels are the same for both extracted and non-extracted cultures. SC activities measured in vitro with radioactive substrate are not significantly different. These results suggest that productivity is controlled by substrate availability within the terpenoid pathway, and feedback regulation precedes the branch-point enzyme sesquiterpene cyclase.

• Horticultural Science & Technology
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• v.30 no.1
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• pp.79-84
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• 2012

An efficient protocol for cryopreservation of madder hairy root cultures has been developed using droplet-vitrification. In previous study, combining loading solution C4 (35% PVS3) and vitrification solution B5 (80% PVS3) was the most effective method. In this study, we tried three types of vitrification solution, B5, A3 (90% PVS2, on ice), and A5 (70% PVS2, on ice). Combining loading solution C4 and vitrification solution A5 (on ice) showed the best regeneration rate in this study. Histological changes of the cells within the hairy root of madder were also observed in different steps. The cells from the hairy roots of the control treatment were full and intact with different size of vacuoles and obvious cell nucleus having a dark nucleolus. After the stage of preparing for cryopreservation (after preculturing, loading, followed by dehydration solution A5 or B5), intercellular spaces had become distinct, and within cells, the cytoplasms had become denser and week plasmolyses had appeared. The cell plasmolyses were much more apparent and we measured the degree of plasmolysis by calculating, the area of cell/the area of cytoplasm. The value of plasmolysis degree was the highest in the combination of preculture, loading solution C4, and dehydration solution A5, 1.97. Because the highest regeneration rates appeared in the treatment of A5 for 20 min, we could assume that the optimal degree of plasmolysis for cryopreservation might be around 1.97. The changes in cell structure during cryopreservation might be a useful basis for the development of a proper long-term preservation method for madder germplasms.

• Journal of Ginseng Research
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• v.23 no.4
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• pp.205-210
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• 1999

The growth of hairy roots were increased $69\%$ by 0.1 mM DCPIP under light conditions. In these conditions, the contents of seven ginsenosides were none significant variation. The influence of electron inhibitors on growth and ginsenoside contents in ginseng hairy roots was tested. The growth was inhibited $71\%\;and\;22\%$ respectively by CCCP and methylamine. The ginsenoside contents were as decreased above $45\%$ in all treatment tested except triazine treatment. In antioxidants treatment, the growth of hairy roots was increased about $68\%$ by propylgallic acid, about $23\~25\%$ by ascorbic acid or 2,5-dimethylfuran, while the contents of seven ginsenosides were none significant variation. The ginsenoside productivity was high when hairy roots were cultured in $\circledR^{\wedge}MS$ medium for 4 weeks and then transferred to 1/2MS medium with ascorbic acid or 2,5-dimethylfuran for 1 weeks in light conditions. It is suggested that ginsenoside productivity could be accelerated by some antioxidants in hairy root cultures of ginseng.

• 한국약용작물학회:학술대회논문집
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• 2007.05a
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• pp.263-264
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• 2007

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.317-317
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• 2003

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2002.04b
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• pp.150-150
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• 2002

• KSBB Journal
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• v.8 no.2
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• pp.110-114
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• 1993

Vinblastine, an anticancer agent was produced by chemical coupling of two different monomeric indole alkaloids, vinblastine and catharanthine in the presence of ferric ion. Vindoline was efficiently extracted from the leaves of vinca (Cafharanthus roseus) by using supercritical carbon dioxide, whereas catharanthine was chemically extracted from the in vitro cultured hairy roots. The extracted crude monomeric precursors were purified by a two-step preparative TLC. The coupling reaction was carried out in the 0.1M glycine buffer(pH 2.0, 5ml) containing 40mM FeC13 with purified vindoline(0.3mg) and catharanthine(0.3mg) at 4$^{\circ}C$. The production yields (weight %) of vinblastine and 3', 4'-anhy-drovinblastine in the products were 23.2 and 26.0, respectively. The produced vinblastine was confirmed by FAB-MS.

• Journal of Plant Biology
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• v.39 no.1
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• pp.63-69
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• 1996

A hairy root clone of Panax ginseng C.A. Meyer, HRB-15 was cultured iu various conditions with 3 L bubble type bioreactor to enhance both growth and ginsenoside production. The hairy roots were more rapidly grown under the dark condition than under the light condition. However, total amount of ginsenoside of hairy roots cultured under the light for 30 days increased 2 folds as compared with the dark condition and was 1.10% based on 6 ginsenosides. Especially, ginsenoside-Re was significantly increased and some ginsenosides except for ginsenoside-Re was slightly reduced. Also, the growth of hairy roots decreased about 30% as compared with the dark condition. In contrast, addition of sodium acetate led to decreased production of ginsenoside and growth of hairy roots under light condition. The influence of potassium dihydrogenphosphate concentration was examined in MS medium and a 1.25 mM concentration was found to be the most appropriate for growth and ginsenoside production under light condition. Two-step process of hairy roots culture with yeast elicitation or without ammonia in culture medium was developed to enhance growth and giusenoside synthesis. $50\;\mu\textrm{g}$ of yeast elicitor per g of fresh weight showed a synergistic effect on the ginsenoside synthesis of hairy roots on 20 days after culture. At that time, the content of total ginsenoside was 1.15%, while the growth of hairy roots decreased 21 % as compared with the dark condition. In addition, when elimination of ammonia on 20 days after culture, the content of total ginsenoside was 1.26% with significant increment of ginsenoside-Rd (0.27%) in addition to ginsenoside-Re and the growth of hairy roots decreased 10% as compared with the dark condition. In this system, we have demonstrated a unique two-step process of hairy root cultures to maximize biomass and secondary metabolites. It has found possibility to enhance ginsenosides production by growing hairy roots in this method.