• Journal of Plant Biotechnology
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• v.30 no.2
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• pp.173-177
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• 2003

The effect of IBA and NAA on adventitious root cultures of Panax ginseng C.A. Mater were investigated. Results indicated differences in growth and development of the roots according to 5mg/L IBA and 2mg/L NAA. IBA resulted in a normal root development and a higher growth compared to NAA. The roots formed on NAA-containing media were shorter and thicker than those in IBA, showing a hypertrophy of the root tip. NAA induced more than 1.6 times higher ethylene production compared to IBA, which caused inhibition of the root growth. Under the ventilation, in the other hand, on difference was observed in ethylene concentration and the root growth between IBA and NAA treatments. Under ventilation ethylene production was not detected until 10 days of culture, while detected from the initial stage under on ventilation. The results suggested the importance of ventilation during the culture for the growth and development of ginseng adventitious roots.

• Plant Resources
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• v.6 no.1
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• pp.1-6
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• 2003

Ginseng(Panax ginseng C. A. Meyer) is important medicinal plant but requires 4-year cultivation for root harvest because of slow growth. In contrast, ginseng hairy roots induced by introducing Ri-plasmid of Agrobacterium rhizogenes into genomic DNA of plant cells show vigorous growth, and the hairy roots produce the same or more saponins than natural ginseng roots. Therefore, hairy roots can be used for commercial purposes. The present study was carried out to induce hairy roots with both active growth and high saponin contents. Numerous hairy roots of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes R1000 A4T in dark condition after one month of culture. About 3 hundred lines of hairy roots were selected according as morphological characters on medium with carbenicillin. After pre-selection of fifteen lines of hairy roots with active growth, KGHR-l and KGHR-8 lines were finally selected which had characters of high content of ginsenoside-Rd and ginsenoside-Re, respectively. The optimum growth of hairy roots was achieved in the culture of 1/2 MS liquid medium in dark (22 $^{\circ}C$) under 60 rpm gyratory shaking. Hairy roots grew well in 5L Erlenmeyer flasks, lL roller drums, 10L jar-fermenters, and especially in 20L air-lift culture vessels.

• Korean Journal of Plant Tissue Culture
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• v.27 no.4
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• pp.309-315
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• 2000

Panax ginseng is an important medicinal plant that has been used worldwide for geriatric, tonic, stomachic, and aphrodisiac treatments. Ginsenosides contained in the ginseng root are the main substances having active functions for human body. The price of ginseng is very expensive due to a complex process of cultivation, and the yield of ginseng is limited, which cannot meet the demand of the increasing market. Researchers have applied plant biotechnology to solve the problems but there are still things to be determined towards ginsenoside production by large-scale adventitious root culture. In this experiment, 5 to 20 liter bioreactors were employed to determine optimal conditions for adventitious root culture and ginsenoside production of Panax gineng. Callus was induced from the ginseng root on MS agar medium containing 1.0 mg. $L^{-1}$ 2,4-D and 0.1 mg. $L^{-1}$ kinetin. Then the callus was cultured on MS agar medium supplemented with 2.0 mg. $L^{-1}$ IBA, 0.1 mg. $L^{-1}$ kinetin, and 30 g. $L^{-1}$ to induce adventitious roots. The maximum root growth and ginsenoside production were obtained in 1/2 MS medium. 2.0 mg. $L^{-1}$ naphthalene acetic acid resulted in greater root growth than 2.0 mg $L^{-1}$ indole-3-butyric acid. Ginsenoside content increased with 2.0 mg. $L^{-1}$ benzyl adenin or kinetin. High concentrations of benzyl adenin (above 3.0 mg. $L^{-1}$ ) decreased the adventitious root growth and ginsenoside productivity. N $H_{4}$$^{+}$ inhibited the ginsenoside accumulation, while high concentrations of $K^{+}$, $Mg_{2}$$^{+}$, and $Ca_{2}$$^{+}$ increased it. N $H_{4}$$^{+}$ at 0.5 and 1.0 times of the normal amount in 3/4 SH medium resulted in the greatest biomass increase, but the highest ginsenoside productivity was obtained when N $O_{3}$$^{-}$ was used as the sole nitrogen source in the medium. Most microelements at high concentrations in the medium inhibited the root growth, but high concentrations of MnS $O_4$enhanced the root growth. Root dry weight increased with increasing sucrose concentrations up to 50 g. $L^{-1}$ , but decreased from 70 g $L^{-1}$ Ginsenoside productivity was maximized at the range of 20 to 30 g. $L^{-1}$ sucrose. In the experiment on bioreactor types, cone and balloon types were determined to be favorable for both adventitious root growth and ginsenoside production. Jasmonic acid was effective for increasing ginsenoside contents and Rb group ginsenosides mainly increased. These results could be employed in commercial scale bioreactor cultures of Panax ginseng.x ginseng.

• 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.

• Korean Journal of Plant Tissue Culture
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• v.28 no.6
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• pp.347-351
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• 2001

Panax ginseng C.A. Meyer is a well-known Korean traditional medicine. Until now, even though major research of ginseng has been focused on the pharmacological effect, clinical application and chemical analysis of extracted secondary metabolite for several years, the physiology and gene functions of ginseng were not well known. In this research, we have developed the protein extraction methods of ginseng root and hairy root for proteome analysis in order to elucidate the gene(s) function of ginseng. Using the liquid nitrogen (equation omitted) TCA method as protein extraction method, about 660 protein spots were detected on the 2-DE gel of hairy root. Additionally, comparative analysis result of 2-DEs of ginseng root (equation omitted) hairy root suggested that proteomes of same organism could be changeable according to the culture condition, growth stages and other stimulus.

• Korean Journal Plant Pathology
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• v.14 no.3
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• pp.268-277
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• 1998

The production of Korean ginseng, one of the most important medicinal root crops, is limited by many factors including soil sickness, root rots in fields as well as during storage prior to consumption. Although much research has been conducted on the diseases in field condition, little information is available on the control of postharvest roots rots. To obtain better management strategy of postharvest root rots in ginseng, biological control using antagonistic bacteria was attempted. Of 208 bacteria obtained form suppressive soil samples, 4 were selected based on the inhibitory effect on mycelial growth of two major causal fungi for postharvest root rots in ginseng, Botrytis cinerea and Fusarium solani. The culture filtrates of these bacterial antagonists greatly inhibited the conidial germination of both pathogenic fungi and produced abnormal morphology such as swollen germ tubes in F. solani and vacuolation of nongerminated conidia in B. cinerea. The population levels of bacterial antagonists on the ginseng roots were gradually increased up to 8 days of incubation. Postharvest root rots of ginseng caused by f. solani and B. cinerea were controlled in dipping tests in the ranges of 60∼80% by antagonistic Bacillus spp. obtained from suppressive soil. These results suggest that biological control using these antagonistic bacteria would be an alternative strategy to control postharvest root rots in ginseng.

• Journal of Ginseng Research
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• v.28 no.1
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• pp.67-70
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• 2004

Bacillus subtilis B-4228 selected from ginseng field soil for prevention of rusty root was tested for the control of ginseng root rot. In petri-plate dual culture, mycelial growth of Cylindrocarpon destructans was inhibited by B-4228 and hyphal swelling of C. destructans was occurred. In pot experiment with C. destructans-contaminated soil B-4228 dipping of ginseng seedling showed significant preventive effect of root rot (p=0.01), percent healthy root 82% and 20% for treatment and control, root rot rate 6% and 50.4%, respectively.

• Journal of Ginseng Research
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• v.31 no.1
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• pp.14-22
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• 2007

As the zygotic embryo of North American ginseng (Panax quinquefolius L.) matured during stratification over 203 days it grew from 0.75 to 5.2 mm. Embryo excision and culturing on media containing different concentrations of two growth regulators, gibberellic acid ($GA_3$, 1 to 10 ${\mu}M$) and benzyladenine (BA, 1 to 5 ${\mu}M$), during stratification, showed that shoot and root number and the shoot, root and cotyledon length increased with increased stratification time. Gibberellic acid was the more effective growth regulator for increasing shoot and root number and shoot, root and cotyledon lengths. Immature embryos (stratified for up to 63 days) needed growth regulators for further development. Cultures on $GA_3$ at the last culture date (stratified for 203 days) when embryos were mature, produced multiple shoots but there was no effect of $GA_3$ concentration. Benzyladenine inhibited shoot and root growth regardless of embryo stratification. Growth regulators had little effect on cotyledon length of mature embryos. Embryos cultured on $GA_3$ combined with BA were green on all culture dates whereas greening in the control and BA treatments increased with culture date. The BA treatments induced 100% swelling of the embryos on the final culture date while in the control and $GA_3$ treatments there was no swelling. There was little or no curling in the control and BA treatments and a linear decrease in curling with culture date in the $GA_3$ and $GA_3$ + BA treatments.

• Journal of Bio-Environment Control
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• v.11 no.4
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• pp.199-204
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• 2002

To overcome a decrease of Korean ginseng production caused by successive cropping, we have tried to develop a nutrient culture system for Korean ginseng production. For determining the optimal substrate, mixture of sand and TKS-2 (S＋T), peatmoss (P), reused rockwool (RR), and granular rockwool (GR) were investigated. The overall physico-chemical properties of RR fell into the reported optimal range for the ginseng cultivation. However, bulk density of S＋T was a little higher than that of soil in Korean ginseng fields. The top fresh weight of the ginseng was high in RR and S＋T substrates. The root fresh and dry weights in the RR were remarkably greater than that in the conventional soil (CS) of Korean ginseng fields. In terms of ginseng quality, the vitamin C content of ginseng root in nutrient culture was higher than that in CS. However, the contents of crude saponin and total ginsenosides in ginseng between in the nutrient culture and in the soil culture did not show any significant differences.

• KSBB Journal
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• v.5 no.3
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• pp.263-268
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• 1990

New methods have been developed to transform Panax ginseng with Ri plasmids of Agrobacterium rhizogenes 15834 and A. rhizogenes A4. Modified leaf disc method was made feasible to establish hairy root culture even when an axonic plantlet was not available as in the case of P. ginseng. The contents of ginsenosides (Rgl, Rf, Rc, Rbl, and Rb2) in hairy roots. were determined by HPLC. Hairy root cultures, established as liquid culture in MS medium, was produced 0.34~1.19% ginsenosides on dry weight basis, and this result is significantly higher level than that of normal P. ginseng.