• Journal of Ginseng Research
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• v.44 no.4
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• pp.563-569
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• 2020

Background: White ginseng consists of the roots and rhizomes of the Panax species, and red ginseng is made by steaming and drying white ginseng. While red ginseng has both polar and nonpolar ginsenosides, previous studies showed white ginseng to have only polar ginsenosides. Because nonpolar ginsenosides are formed through the manufacture of red ginseng from white ginseng, researchers have generally thought that nonpolar ginsenosides do not exist in white ginseng. Methods: We developed a simultaneous quantitative method for six nonpolar ginsenosides in white ginseng using reverse-phase high-performance liquid chromatography coupled with integrated pulsed amperometric detection. The nonpolar ginsenosides of white ginseng were extracted for 4 h under reflux with 50% methanol. Results: Using the gradient elution system, all target components were completely separated within 50 min. Nonpolar ginsenosides were determined in the rhizome head (RH), main root (MR), lateral root, and hairy root (HR) of 6-year-old white ginseng samples obtained from several regions (Geumsan, Punggi, and Kanghwa). The total content in the HR of white ginseng was 37.8-56.8% of that in the HR of red ginseng. The total content in the MR of white ginseng was 5.9-24.3% of that in the MR of red ginseng. In addition, the total content in the RH of white ginseng was 28.5-35.8% of that in the HR of red ginseng Conclusion: It was confirmed that nonpolar ginsenosides known to be specific components of red ginseng were present at substantial concentrations in the HR or RH of white ginseng.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.1
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• pp.43-47
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• 2006

Ginseng (Panax ginseng CA. Meyer) hairy root cultures, which are established via the infection of ginseng root discs with Rhizobium rhizogenes, have been used to construct profiles of both biomass growth and nutrient consumption in flask cultures. In a 250 mL shake flask culture, the maximum biomass was observed on the 59th day of the culture period, at 216.8 g (fresh wt) per liter or 11.4 g (dry wt) per liter. The hairy roots were determined to have a growth rate of 0.355 g-DW/g cells/day during the exponential growth phase and a maximum specific growth rate on day 7. Total ginseng saponin and phenolic compound contents were noted to have increased within the latter portion of the culture period. Linear correlations between increases in biomass weight and nutrient uptake were used to imply the conductivity yield $2.60g-DW/(L{\cdot}mS)$ and carbon yield 0.45 g-DW/(g sugar) in the 250 mL flask cultures. The biomass yield when two different nitrogen sources were used (ammonia and nitrate) was shown to remain approximately constant. at $0.47g-DW/(L{\cdot}mM\;NH_4$) and $0.33g-DW/(L{\cdot}mM\;NO_3$); it remained at these levels for 16 days with the ammonia. and for 24 days with the nitrate. The biomass yield when a phosphate source was used was also shown to remain approximately constant for 9 days, at $3.17g-DW/(L{\cdot}mM\;PO_4$), with an $R^2$ of 0.99.

• Proceedings of the Botanical Society of Korea Conference
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• 2000.04a
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• pp.20-20
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• 2000

• Journal of Plant Biotechnology
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• v.48 no.2
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• pp.100-105
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• 2021

Genetic engineering is a potential approach to improve secondary metabolism in plants. In order to elucidate the effect of production of anthocyanin pigment 1 (PAP1) overexpression on the bioactivity of ginseng, we analyzed its antioxidant, antimicrobial, and anti-elastase activities in this study. Our results showed that PAP1 overexpression increased the production of polyphenolic compounds including anthocyanins. The antioxidant, antimicrobial, and anti-elastase activities were stronger in anthocyanin-overproducing ginseng hairy roots (AOX) than in wild ginseng hairy roots. Using a different solvent system (0, 30, 70, and 100% (v/v) EtOH), we revealed that variations in the contents of the polyphenolic compounds were highly correlated with changes in the antioxidant and antimicrobial activities of AOX. The antioxidant, antimicrobial, and anti-elastase effects of AOX highlight genetic engineering as a powerful approach to enhance the therapeutic properties of plants. Our results show that AOX could potentially have various functional applications in the cosmetic and pharmaceutical industries.

• 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.

• Journal of Plant Biotechnology
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• v.34 no.4
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• pp.285-291
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• 2007

This study describes conditions for the mass production of activation-tagged mutant hairy root lines of ginseng by cocultivation with Agrobacterium rhizogenes. Because it is not currently possible to produce progeny from transgenic ginseng, a loss-of-function approach for functional genomics cannot be appliable to this species. A gain-of-function approach is alternatively the choice and hairy root production by cocultivation of A. rhizogenes would be most practical to obtain a large number of mutants. Various sources of explants were subjected to genetic transformation with various strains of A. rhizogenes harboring the activation-tagging vector pKH01 to determine optimum conditions for the highest frequency of hairy root formation on explants. Petiole explants cocultivated with A. rhizogenes R1000 produced hairy roots at a frequency of 85.9% after 4 weeks of culture. Conditions for maximum growth or branching rate of hairy roots were also investigated by using various culture media. Petiole explants cultured on half strength Schenk and Hildebrandt medium produced vigorously growing branched roots at a rate of 2.6 after 4 weeks of culture. A total of 1,989 lines of hairy root mutants were established in this study. These hairy root lines will be useful to determine functions of genes for biosynthesis of ginsenosides.

• Korean Journal of Plant Tissue Culture
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• v.27 no.1
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• pp.31-37
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• 2000

Proton extrusion by hairy roots of Panax ginseng C.A. Meyer was enhanced by iron-deficiency or single salt solutions of nitrate or ammonium salts. The enhancement effect of iron-deficiency was confined to the apical zones, whereas that of nitrate and ammonium was confined to the basal root zones. tinder iron-deficiency medium, the proton extrusion of ginseng hairy roots was higher at two times than that grown in the medium of iron-sufficiency. According to pH in the medium, uptake of inorganic elements showed various patterns for the kind of inorganic elements. $Ca^{2＋}$, $Mg^{2＋}$ and Fe$^{2＋}$ were highest uptake at the pH 4.8 and Mn$^{2＋}$, Zn$^{2＋}$ and Cu$^{2＋}$ were highest uptake at the pH 3.8. Free sugars and ginsenosides were analyzed by HPLC. Five kinds of free sugars were indentified in ginseng hairy roots. Glucose, fructose and sucrose were a major sugar in ginseng hairy roots and contents of free sugars were highest at pH 6.8 and pH 4.8, respectively. The contents of ginsenosides and phenolic compounds were highest at pH 5.8. This physiological responses in ginseng hairy root indicated that it may well be a good model system for the study of mechanism of mobilization and uptake of inorganic elements related with the red-colored phenomenon in ginseng roots.

• Journal of Ginseng Research
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• v.24 no.2
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• pp.74-78
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• 2000

To elucidate the effects of jasmonic acid and methyl jasmonate on the production of ginsenosides and growth, ginseng hairy root KGHR-8 clone was cultured on the 1/2 MS medium without growth regulators, which was supplemented with of various concentrations jasmonic acid and methyl jasmonate and culture period. The highest growth rate was obtained when 1$\mu\textrm{m}$ jasmonic acid and methyl jasmorlate were treated. However, the growth was inhibited at more than 30$\mu\textrm{m}$ of concentration. Treatment with high concen Dation of jasmonic acid (10$\mu\textrm{m}$) and methyl jasmonate (50$\mu\textrm{m}$) increased the contents and productivity of ginsenosides reversion of the growth inhibition. The highest contents and productivity of ginsenosides were appeared at 4 weeks after onset of the treatment of jasmonic acid and at 3 weeks in the case of methyl jasmonate.

• 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.

• Journal of Ginseng Research
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• v.21 no.1
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• pp.28-34
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• 1997

The effects of light on the pigment production and chloroplast development were examined on ginseng hairy roots cultured in 1/2MS liquid medium. The chlorophyll and carotenoid production were increased from 1,000 to 3,500 lux condition, but decreased drastically in 7,000 lux condition. The anthocyanin production was significantly increased with increment light intensity(1,000∼7,000 lux). The thylakoid membrane of chloroplast was proplastid in dark condition and it began to develop into thylakoid membrane in 1,000 lux condition and then intact thylakoid membrane was developed in 3,500 lux condition. However, the development of thylakoid membrane in 7,000 lux condition was inhibited comparing to 3,500 lux condition. The total chlorophyll production in blue light condition were high comparing to other wavelength and same as 40% of total chlorophyll on white light(3,500 lux) condition. The chlorophyll and carotenoid production by sucrose concentration were high in 3% sucrose condition and anthocyanin production was high in 4% condition. The production of chlorophyll and carotenoid by light periods was high when explants were cultured in dark condition for 1 week and then transferred to light condition for 4 weeks. Our results suggest that pigment production and chloroplast development could be accelerated by light Intensity of specific wavelength in cultures of ginseng hairy root.