• 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.

• Journal of Plant Biotechnology
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• v.36 no.4
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• pp.352-359
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• 2009

Panax ginseng roots produce triterpene saponins called ginsenosides, which are high value secondary metabolites and has been used as drugs, detergents, sweeteners, and cosmetics. In the recent years plant cell, tissue and organ cultures have developed as important alternative sources for the saponin production in Panax ginseng. Adventitious roots and hairy roots have been successfully induced and cultured for the improvement of saponin contents. Genetic and metabolic engineering to regulate saponin biosynthesis in P. ginseng might be important way to improve the medicinal values of P. ginseng. Here we introduced the protocol of genetic transformation and recent progress of functional characterization of genes involved in saponin biosynthesis in P. ginseng.

• 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.

• KSBB Journal
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• v.21 no.2
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• pp.123-128
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• 2006

A transgenic hairy root clone AG-04 of Astragalus membranaceus was obtained following co-cultivation of leaf explants with Agrobacterium rhizogenes ATCC15834. This clone was examined for its growth and production of cyclolanostane-type saponins, astragalosides I, II, and III, under various culture conditions. Among the five basal media tested, Shenk and Hildebrandt(SH)(18) medium was best for roots growth and astragalosides production. The maximum root biomass was obtained at inoculum size of 500 mg FRW per flask, initial sucrose concentration of 3%, and shaking speeds of 90 rpm. The astagalosides production was promoted when the hairy root clone AG-04 was cultured at shaking speeds of 120 rpm and light irradiation of 18 h. Astragaloside contents was also stimulated with high initial sucrose concentration, and the maximum astargalosides contents of 6.21 %/g DRW was obtained at initial sucrose concentration of 6%. The addition of chitosan(100 mg/L) to the culture medium was significantly increased astragalosides production. This was 2.1 times higher than that obtained in a control culture without chitosan.

• Korean Journal of Medicinal Crop Science
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• v.11 no.5
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• pp.358-363
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• 2003

ScopoIamine and hyoscyamine which belong to tropane alkaloids are the pharmaceutically valuable anticholinergic drugs. In order to increase the productivities, the effects of elicitation were investigated during hairy root cultures of Scopolia. parviflora. Biotic elicitors originated from 3 fungi and 1 yeast were prepared as homogenate and supernatant and added to 3-week-old cultures. Both of homogenate and supernatant of Candida albicans elicitors increased the scopolamine production. The production of hyoscyamine was enhanced by homogenate of Fusarium solani and supernatant of C. albicans. Most of the other fungal elicitors were also improved on the tropane alkaloid production compared to non-treatment. Among the elicitors tested, C. albicans was proved the optimal biotic elicitor on tropane alkaloids production. These results will be served mass production of tropane alkaloids by large-scale production.

• 한국약용작물학회:학술대회논문집
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• 2011.09a
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• pp.324-325
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• 2011

• 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.

• Journal of Plant Biotechnology
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• v.5 no.1
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• pp.1-6
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• 2003

The adventitious root of Panax ginseng C.A. Meyer is regarded as an efficient alternative to cell culture or hairy root culture for biomass production due to its fast growth and stable metabolite production. To determine optimal culture conditions for the bioreactor culture of ginseng roots, experiments have been conducted on physical and chemical factors such as bioreactor type, dissolved oxygen, gas supply, aeration, medium type, macro- and micro-elements, medium supplement during culture period, sucrose concentration, osmotic agents, medium pH and light. Elicitation is a key step to increase ginsenoside accumulation in the adventitious roots but biomass growth is severely inhibited by elicitor treatment. To obtain high ginsenoside content with avoiding biomass decrease, we applied two-stage bioreactor culture system. Ginseng adventitious roots were cultured for 40 days to maximize biomass increase followed by elicitation for 7 days to enhance ginsenoside accumulation. We also experimented on types and concentrations of jasmonate to determine optimal elicitation methods. In this paper, we discussed several factors affecting the root propagation and ginsenoside accumulation. Based on the results obtained from previous experiments we have established large-scale bioreactor system (1 ton-10 ton) for the efficient production of ginseng adventitious roots and bioactive compounds including ginsenoside. Still, experiments are on going in our laboratory to determine other bioactive compounds having effects on diet, high blood pressure, DPPH elimination and increasing memories.

• KSBB Journal
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• v.9 no.1
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• pp.26-34
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• 1994

When effects of exogenous hormone on hairy root cultures of Raphanus sativs cv. Chungpihongsim examined, the highest anthocyanin synthesis and disorganization were observed when 2, 4-D was supplemented to the culture medium Cytokinins showed early weak induction after transfer and ABA showed inhibitory effect and GA3 showed no effects in anthocyanin synthesis. Hormones except for 2, 4-D in 1 mg/$\ell$ concentration did not induce disorganization of hairy root and retarded growth of hairy root. Time-course changes in anthocyanin synthesis, phenylalanine ammonia-lyase activity and chalcone synthase activity were examined in culture condition contalning 2, 4-D and kinetin. In a medium containing 2, 4-D, anthocyanin synthesis began to increase on the 9th day and reaching maxima on the 18th day after transfer. Maximum peak of PAL activity appeared on the 3-9th day and another minor peak appeared on the 18th day. CHS activity increased from 9th day, reaching maximum on the 18th day and remained at a relatively high level for culture period. In a medium containing kinetin, anthocyanin synthesis increased temporarily on the 6-9th days, early days after transfer and maintained at a low level for remaining culture period. Peak of PAL activity appeared on the 6th day and CHS activity increased from the 6th days, reaching maxima about 18th day and remained at a relatively high level. In particular, addition of kinetin after preculture in hormone free medium for 2 weeks which was thought of wound healing period showed no effects in anthocyanin synthesis. This results showed that stimulation of anthocyanin synthesis by 2, 4-D and kinetin was meaningfully connected with changes of PAL, CHS activity, and then suggested rate-limiting role of CHS on anthocyanin synthesis in that there is close correlation between anthocyanin synthesis and changes of CHS activity in time-course. Besides, it is considered that cytoklnins involving kinetin stimulated anthocyanin synthesis be due to "wound response" by cutting of young roots, and that difference in time-course peak and PAL, CHS activities expressed by 2, 4-D and kinetin result from occurrence of isozyme which have different regulatory mechanism.mechanism.