• Proceedings of the Botanical Society of Korea Conference
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• 1987.07a
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• pp.213-237
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• 1987

In vitro flowering system may minimize the confounded influence of non-floral meristem parts of plants in studying the relationship of a given treatment and flowering responses. We have induced flower buds from plantlets regenerated from zygotic embryo-derived somatic embryos of ginseng, which circumvented the normal 2-year juvenile period before flowering. The result suggests that the adulthood of ginseng root explants in the experiment previously conducted by Chang and Hsing (1980; Nature 284: 341-342) is not prerequired to flowering of plantlets regenerated through somatic embryogenesis. We have also induced flower buds from elongated axillary brandches from cotyledonary nodes by culturing ginseng zygotic embryos, seedlings, and excised cotyledonary nodes. It was found that 6-benzyladenine (BA) supplemented to the medium was essential for flowering, whereas abscisic acid (ABA) was inhibitory. Gibberellic acid(GA3) was also required for flowering when ABA was present with BA in the medium. The results suggest that cytokinins, gibberellins, and inhibitors play primary, permissive, and preventive roles, respective-ly, in the induction of flowering of ginseng. Tran Thanh Van (1980; Int. Rev. Cytol., Suppl. IIA: 175-194) has developed the "thin cell layer system" in which the induction of shoots, roots, or flower buds from epidermal layer explants were controlled by culture conditions and exogenous growth regulators in the medium, Utilizing the thin cell layer system, Meeks-Wagner et al. (1989; The Plant Cell 1: 25-35) have cloned genes specifically expressed during floral evocation. However, the system is too tedious for obtaining a sufficient amount of plant materials for biochmical and molecular biological studies of flowering. We have developed a garlic callus culture system and one obvious advantaging over the thin cell layer system is that an abundant cells committed to develope into flower buds proliferate. When the above cells were compared by two-dimensional gel electrophoresis with those which have just lost the competence for developing into flower buds, a few putative proteins specific to floral evocation were detected. The garlic callus culture system can be further explored for elucidation of the molecular biological mechanism of floral evocation and morphogenesis.hogenesis.

• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.346-355
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• 2021

Ngoc Linh ginseng is one of the most valuable endemic medicinal herbs in Vietnam. In this study, Ngoc Linh ginseng callus was fermented by Lactobacillus plantarum ATCC 8014 (at 6, 7, and 8 log CFU/ml) to evaluate the extraction efficiency of bioactive compounds. The post-fermentation solution was spray-dried using maltodextrin with or without Stevia rebaudiana (3% and 6% v/v) as the wall material. Bioactive compounds such as polyphenols, polysaccharides, and total saponins, and L. plantarum viability during fermentation and after spray-drying, as well as under simulated gastric digestion, were evaluated in this study. The results showed that probiotic density had a significant effect on bioactive compounds, and L. plantarum at 8 log CFU/ml showed the best results with a short fermentation time compared to other tests. The total content of polyphenols, polysaccharides, and saponins reached 5.16 ± 0.18 mg GAE/g sample, 277.2 ± 6.12 mg Glu/g sample, and 4.17 ± 0.15 mg/g sample, respectively after 20 h of fermentation at the initial density of L. plantarum (8 log CFU/ml). Although there was no difference in the particle structure of the preparation, the microencapsulation efficiency of the bioactive compound in the samples containing S. rebaudiana was higher than that with only maltodextrin. The study also indicated that adding S. rebaudiana improved the viability of L. plantarum in gastric digestion. These results showed that S. rebaudiana, a component stimulating probiotic growth, combined with maltodextrin as a co-prebiotic, improved the survival rate of L. plantarum in simulated gastric digestion.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2005.04a
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• pp.140-140
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• 2005

• Proceedings of the Ginseng society Conference
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• 1974.09a
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• pp.101-113
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• 1974

The radioactive compound sodium $acetate-U-C^{14}$ (C-14 acetate) was administered to two- and four-year-old July and September American ginseng (Panax quinquefolium L.) plants and cuttings. The C-14 acetate uptake was approximately $99\%.$ The autoradiochromatograms suggest that the saponins(panaquilins) isolated by preparative thin-layer chromatography contained impurities, especially those isolated from the leaf and stem extracts. The root and fruit methanol extracts yielded relatively pure saponins. The large amounts of panaquilin B and its proximity to panaquilin C on preparative thin-layer plates resulted in some admixing. The average concentration $(\%$ plant dry weight) of semipurified saponins were high in the leaves $(13.8\%),$ compared to fruits $(9.8\%),\;stems\;(7.9\%)\;and\;roots\;(6.3\%).$ The average percentage of C-14 acetate incorporation into panaquilins was $4.8\%.$ The average percentage of C-14 acetate incorporation into panaquilins B and C was higher $(1.40\%\;and\;1.13\%,$ respectively) than that into panaquilin C, (d), G-1 and G-2 $(0.75\%,\;0.65\%,\;0.13\%\;and\;0.53\%,$ respectively). Panaquilin synthesis may be depending upon the part collection period and age of the plant. The average percentage of C-14 acetate incorporation into panaquilin B is high in roots $(0.58\%)\;and\;stems\;(0.48\%);$ that into panaquilins C and (d) high in leaves $(0.40\%\;and\;0.45\%,$ respectively); and that into panaquilin E high in roots and leaves $(0.55\%\and\;0.50\%,$ respectively). Panaquilin G-2 was synthesized in all parts of plants. The panaquilins appear to be biosynthesized more actively in July than September (exception-panaquilin G-l). Panaquilins B, C and G-1 may be biosynthesized more actively in four-year-old plants and panaquilins (d) and E more actively in two-year-old plants. The results from expectance with cuttings suggest that the panaquilins are synthesized de novo in the above-ground parts of ginseng plants, and that panaquilin G-l may be synthesized de novo in the leaf. It is known from the tissue culture studies that panaquilins are produced by leaf, stem and root callus tissues and callus-root cultures of American and Korean ginseng plants. Panaquilins may actively be synthesized de novo in most any cell or organ of the ginseng plants. It was verified that C-14 acetate was incorporated into the panaxadiol portions of the panaquilins of two-year-old plants (sp. act., 0.56 $m{\mu}Ci/mg$) and four-year-old plants (sp. act., 0.54 $m{\mu}Ci/mg$).

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.611-616
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• 2003

Korean ginseng(Panax Ginseng C.A. Meyer) known as a oriental miracle drug is an important medicinal plant. Ginseng has been used for geriatric, tonic, stomachic, and aphrodisiac treatments for thousands years. Also, it is an antibiotic and has therapeutic properties against stress and cancer. Ginseng is widely distributed all over the world. Among them, Korean mountain ginseng has the most valuable effect on pharmaceuticals. The roots of mountain ginseng contained several kinds of ginsenosides that have many active functions for the human body. However, the study of mountain ginseng has a limit because the mountain ginseng is very expensive and rare. So, we artificially cultured mountain ginseng adventitious roots using the bioreactor culture system. We induced callus from original mountain ginseng, directly dug up in mountain and aged about one hundred ten years. Separated adventitious roots were precultured in 500ml conical flasks and then, transferred in 20L bioreactors. The adventitious roots of mountain ginseng were harvested after culturing for 40days, dried and then, extracted with several solvents. In this study, we investigated the whitening effect, anti-wrinkle effect and the safety of tissue cultured adventitious roots extract of mountain ginseng in order to identify the merit as a cosmetic ingredient. Particularly, extract of mountain ginseng adventitious roots showed whitening and anti-wrinkle effects. The inhibitory effect of this extract on the melanogenesis was examined using B-16 melanoma cell. When B-16 melanoma cells were cultured with adventitious root extract, there was a dramatically decrease in melanin contents of 8-16 melanoma cell. And we identified this extract inhibited Dopa auto-oxidation significantly. Also, when transformed mouse fibroblast L929 cells were treated with this extract, there was a significant increase in collagen synthesis. The results show significant inhibited melanization and wrinkle without inhibiting cell viability.

• 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.30 no.3
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• pp.233-239
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• 2003

We have developed a reliable and high-frequency genetic transformation and regeneration system via somatic embryogensis of Eleutherococcus sessiliflorus. Embryogenic callus obtained from seed were co- cultivated with Agrobacterium tumefaciens strain EHA101/pIG121Hm harboring genes for intron-$\beta$-glucoronidase(GUS), kanamycin and hygromycin resistance. Following co-cultivation, two types of samples(fine embrogenic calli and early globular embryo clusters) were cultivated on Murashige and Skoog(MS) medium containing 1 mg/L2.4-D for 3day in dark. Transient expression of GUS gene was found to be higher in the early globular embryo clusters than in the embryogenic calli. Also, co-cultivated period affected expression of GUS gene; the best result was obtained when globular embryo clusters were co-cultivated with Agrobacterium for 3 days. Subsequently, this callus transferred to selective MS medium containing 1mg/L2.4-D, 50mg/L kanamycin or/and 30mg/L hygromycin and 300mg/L cefortaxime. These embryogenic calls were subcultured to the same selection medium at every 2 weeks intervals. Approximately 24.5% of the early globular embryos co-cultivated with Agrobacterium for 3days produced kanamycin or/and hygromycin-resistant calli. Transgenic somatic embryos were converted into plantlets in half strength MS medium supplemented with 3mg/L GA$_3$ kanamycin and were confirmed by GUS histochemical assay and polymerase chain reaction analysis. Genomic Southem blot hybridization confirmed the incorporation of NPT II gene into the host genome.

• Korean Journal of Plant Tissue Culture
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• v.28 no.6
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• pp.353-357
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• 2001

Transformation of ginseng plants was achieved by biolistic system with cotyledon explants and callus using phosphinothricin acetyl-transferase (PAT) gene resisting to a herbicide of Bialaphos. The binary vector for transformation was constructed with disarmed Ti-plasmid and with double 355 promoter. The introduced NPT II and PAT genes of the transgenic ginseng plants were successfully identified by the PCR, and the survival test on the medium with basta. The transgenic ginseng plants were propagated using repetitive secondary embryogenesis. The transgenic ginseng plantlets had normal structures of roots and shoots, and dormant buds for new year sprouting. We transferred the transgenic plants to greenhouse and observed the continuing growth until a new year.

• Proceedings of the Ginseng society Conference
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• 1990.06a
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• pp.175-189
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• 1990

Nitrogen compounds of Panax ginseng and their biological activities in plant and animal were reviewed. Major nitrogen compounds found in P. ginseng are free amino acids. Water solilble proteins, indouble proteins and peptides. Minor nitrogen compounds are dencichine. Glycolyroteins, amines, alkaloides, methoxy or alkyl pyrazine derivatives, free nucleosides and nucleic acid bases. 4-methyl-i-thiazoltethanol and pyroglutamic acid the contents of total nitrogen and protein in root Increased until 13 years old which was the highest age tinder investigation. Soluble protein content increased with the root weight and was higher in xylem pith than cortex-epidermis indicating the close relation with root growth. Arginine, which covered 58% of total free amino acids, may serve as storage nitrogen. Arginine seems to be changed into proline in rhizome. threonine in stem and again threonine and arginine in leaf. The greater the root weight the higher the polyamine stimulated Polyamine stimlllated the growth of root callus. Physiological roles of other minor nitrogen compounds are unknown although content is relatively high ((1.if) 6.w). Biochemical and pharmacological activities of some nitrogen compounds for animal were more investigated than physiological role there plant itself. Radiation and U.V protective function (heat stable protein). insulin-like activity in lipogenesis and livolysis (adenosine and pyroglutamic acid), depression of blood sugar content (glycopevtide). htmostatic and nellrotoxic activity (dencichine) and, sedative and hypnotic activity (4-methyl-i-thiazoleethanol) are reported. Heat stable protein increased with root age. The traditional quality criteria appear to be well in accordance with biological activities of nitrogen compounds. Chemical studies of nitrogen compounds seem relatively rare, probably due to difficulty of isolation, subsequently the investigations of biological activities are little.

• Journal of Ginseng Research
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• v.14 no.2
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• pp.317-331
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• 1990

Nitrogen compounds of Panax ginseng and their biological activities in plant and animal were reviewed. Major nitrogen combounds found in P. ginseng are free amino acids, Water soluble teins, insoluble proteins and peptides. Minor nitrogen compounds are dencichine. glycol)roteins. amines, alkaloides, methoxy or alkyl pyrazine derivatives. free nucleosides and nllrleir arid bases. 4-me- thymi-5-thiazoleethanol and pyroglutamic acid. The contents of total nitrogen and protein in root increased until 13 years old rvhich was the highest age tinder investigation. Soluble protein content increased With the root weight and was higher in xylem pith than cortex-epidermis indicating the rlosc relation with root growth. Arginine which covered 58% of total free amino aroids may serve as a storage nitrogen. Arginine seems to be changed into proline in rhizome, threonine in stem and again threoning and arginine in leaf. The greater the root weight the higher the polyaminc content. Polyamine stimulated the growth of root callus. Physiological roles of other minor nitrogen compounds are unknown although dencichine content is relatively high (0.5% d.w.). biochemical and pharmatological activities of some nitrogen compounds for animal were more investigated than physiological roll iota plant itself. Radiation and U.V. protective function (heat stable protein), insulin-like activity in lipogenesis and lipolysis (adenosine and pyroglutamic acid), depression of blood sugar content (glycopeptide). hemostatir and nellrotoxic activity (denrichine) and. sedative and hypnotic activity (4-methyl-5-thiazoleethilnol) are reported. Heat stable protein increased with root age. The traditional quality critsria appear to be well in accordance with biological activities of nitrogen compounds. Chemical stlldies of nitrogen compounds seem relatively rare, probably dole to difficulty of isolation, subsequently the investigations of biological activities are little.