Background : Wild-simulated ginseng (WSG, panax ginseng C. A. Meyer) is known to be grown in high altitudes with a climate of cool, well ventilated and fertile humid forests. In this study, the effects of altitude and soil characteristics on the growth of WSG were investigated. Methods and Results : The seeds were planted on designated line, and the altitude was set at 600 highland and 300 highland. At each highland soil characteristics and forest condition were investigated also Hobo (HOBO U30 Werther station) was set up to measure the microclimate in the site. Experiments were conducted in each highland to determine the difference in the growth of WSG according to the soil condition. Growth characteristics of undergrowth, such as root thickness and length also upper growth such as stem length and thickness were measured. As a result of the soil characteristics survey, the value of available phosphorus was found to be 256.7 ㎎/㎏ at 300 highlands. On the other hand, in 600 highland, available phosphorus was 29.59 ㎎/㎏, which showed big difference. Conclusion : Overall, the cation exchange capacity was higher than the 600 highland at 300 highland. The weight, stem length, root length and leaflet size of the wild-simulated ginseng were higher than those of 600 highland at 300 highland.

Background : Panax ginseng C. A. Meyer is a medicinal plant corresponding to 'raw materials for food use' in Korean food standards codex, and it is known that it has the same origin although it is classified into wild ginseng, cultured ginseng and red ginseng. It varies morphologically, molecularly, depending on the cultivation or habitat environment. In this study, we carried out experiments for the morphological differentiation of cultured ginseng (Hongcheon, Gunsan, Miryang) and adventitious roots culture of wild mountain ginseng (by company). Methods and Results : Morphological observations were made with resin sectioning method (RSM). The test method was sampled, preprocessed, embedded, segmented, slid and then observed with an optical microscope. The epidermis, cortex and vascular tissues were identified in the inner cross section of adventitious roots culture of wild mountain ginseng. Phloem was formed at the center, and xylem was formed at the side. In cultured ginseng, the organs of the vascular tissues were differentiated into two, whereas in the adventitious roots culture of wild mountain ginseng, one vascular tissue was formed due to the thin adventitious. Conclusion : Plant resources have the diversity of using the same or different origins depending on their distribution. The morphological differentiation method of ginseng and adventitious roots culture of wild mountain ginseng is expected to be based on domestic fair trade to prevent discrimination.

Background : Korea's agriculture has been forced to change due to the decline in agricultural population, the aging of the population and the conclusion of the Convention on Biological Diversity. Thus, agriculture is seeking to develop into a sixth industry. For this reason, it is necessary to find high-function alternative plant resources for health promotion to meet the changes in national needs for agricultural products. The adventitious roots culture of wild mountain ginseng is the raw material of food code that produced by the biotechnology technique. introduced in the culture of the plant, which is the only way to use a raw material of food. Therefore, in order to increase the yield of ginsenoside in adventitious roots culture of wild mountain ginseng, ginsenoside precursor was treated and ginsenoside contents were analyzed. Methods and Results : In order to investigate effect of precursor treatment on the production of ginsenoside in adventitious roots culture of wild mountain ginseng, 5 g of adventitious roots culture of mountain ginseng were cultured in liquid SH media supplemented with 2 ㎎/l IBA, 3% sucrose for 8 weeks, which was co-cultured with β-sitosterol and Squalene (0.0625 to 1 mM) or without. Determination of 17 ginsenosides contents of each extract was carried out by HPLC. Rb3 was accumulated by only β-sitosterol, also it increased production of total ginsenoside in the cultured ginseng roots at a concentration of 0.125 mM, which was 2.47-fold higher than that in the control (78.13 ㎎/g of extract). Conclusion : These results are an important to improve the production yield of ginsenoside in adventitious roots culture of wild mountain ginseng, and they provide an opportunity for development of adventitious roots culture of wild mountain ginseng in dietary health supplement and pharmaceutical industries.

Background : Recently, wild ginseng cultured ginseng cultured in a bioreactor is mass produced using biotechnological tissue culture technology. PgTRx1 gene which is involved in the production of useful substances in fermented wild ginseng cultured root was selected and introduced into a model plant (Nicotiana benthamiana) to investigate transformation useful gene expression and possible production of useful substances. Methods and Results : The PgTRx1 gene was amplified and isolated from fermented wild ginseng cultured root. Isolated PgTRx1 gene was ligated to the plant expression vector pMBP1. Overexpression genes were recombined and cloned into E. coli. Agrobacterium tumefaciens LBA4404 was transformed, cultured A. tumefaciens LBA4404 was agro-infiltrated into a model plant for transient assay. Agro-infiltration model plants were sampled on days 0, 1, 2, and 3, and cDNA synthesis was performed after total RNA extraction. The expression level of PgTRx1 gene increased with time, and NbNR, NbHSR, NbAPx, NbSIP, NbPAL, NbPR1a and NbNOA1 genes showed a decrease in the expression level. The samples were taken to determine antioxidant activity, acetylcholine hydrolase inhibitory activity and glutamate content at 0 h, 12 h, 14 h, and 36 h. The highest antioxidant activity was observed at 24 h of sample, acetylcholine hydrolase inhibitory activity at 12 h, and glutamate at 36 h. Conclusion : The possibility of introducing the model plant of the PgTRx1 gene derived from fermented wild ginseng cultured root was confirmed. The results showed that various activities were increased with time of agro-infiltration.

Background: To obtain useful cosmetic resources, this study aimed to determine the non-saponin fatty acid and inhibitory activities of collagenase and elastase by treatment of Saccharomyces cerevisiae in supercritical fluid extracted oil of the adventitious root culture of wild mountain ginseng. Methods and Results: We performed supercritical fluid extraction at various conditions such as pressure, temperature, time, and use of co-solvents, unlike the n-hexane extraction for the adventitious roots culture of wild mountain ginseng. The non-saponin-fatty acid obtained from the oil of the adventitious roots culture was incresed by treatment with S. cerevisiae. The supercritical fluid extraction was conducted using gas chromatography. Non-saponin-fatty acid content, in the oil of adventitious roots culture of wild mountain ginseng treated with S. cerevisiae for 2 days were three times higher than that in the control. In addition, the oil of the adventitious roots culture treated with S. cerevisiae was investigated for the anti-wrinkle effect by using collagenase and elastase. The oil of adventitious roots culture treated with S. cerevisiae exhibited higher collagenase and elastase inhibitory activities than those in the control. Conclusions: Supercritical fluid extracted oil of the adventitious roots culture of wild mountain ginseng treated with S. cerevisiae was found to have decreased ratio of saturated fatty acids and incresed ratio and content of unsaturated fatty acids increased. Furthermore, it showed anti-wrinkle effects in vitro.

Background: Hot steaming is known to be effective in improving the biological activities of plant extracts by breaking down useful compounds to low molecular weight ones. Methods and Results: This study aimed to develop an optimal extraction and steam processing method for enhancing the low molecular ginsenoside contents of the adventitious roots culture of wild mountain ginseng. The total ginsenoside was optimally extracted when 70% EtOH was used at 50℃, whereas low molecule ginsenoside such as Rg2, Rh1, Rh4 and Rk1 could be extracted using 70% EtOH at 70℃. The adventitious roots culture of wild mountain ginseng is known to contain four major ginsenosides, i.e., Rb2, Rb1, Rg1 and Rd, however new ginsenosides Rg6, Rh4, Rg3, Rk1 and Rg5 were new abundantly obtaind after steam processing method was applied. The contents of total ginsenosides were the highest when thermal steam processing was conducted at 120℃ for 120 min. Unlike ginsenosides such as Rg1, Re, Rb1, Rc, Rb2, and Rh1, which decreased after steam processing, Rg3, Rk1, and Rg5 increased after thermal processing. Steam processing significanltly reduced the content of Rb1, increased that of Rg6 by about ten times than that in the adventitious roots culture of wild mountain ginseng. Conclusions: Our study showed that the optimal extraction and steam processing method increased the content of total ginsenosides and allowed the extraction of minor ginsenosides from major ones.

Background : Panax ginseng C.A. Meyer is a perennial herb belongs to the family Araliaceae. Wild-cultivated ginseng (WCG) is a specific type of ginseng in Korea which cultivated on artificial forest cultivation method. To obtain a WCG which is similar to wild ginseng (WG), this method usually performed in a mountain using seeds or seedlings of cultivated ginseng (CG) and WG. WCG is very expensive because it is difficult to cultivate. However, systematic cultivation method have not yet been developed compared to high added value. Furthermore, very high price of WCG caused the problem that Panax notoginseng or Panax quinquefolium are sold as WCG in Korean market. In this study, we analyzed the genetic diversity of WCG collected from five areas in Korea using SSR markers. Methods and Results : WCG samples were collected from five areas in Korea (Bucheon, Cheongju, Hoengseong, Judeok and Ulsan). DNA extraction was performed using CTAB method. SSR markers were collected from the published papers. After test PCR using the markers, one of the primer pair was labeled with fluorescence dye (FAM, NED, PET, or VIC) and GeneScan analysis were performed. DNA amplification was conducted using T-100 Thermal Cycler (Bio-Rad). PCR products were separated by capillary electrophoresis on the ABI 3730 DNA analyzer (Applied Biosystems). Conclusion : Eight SSR markers were collected from the published literature and used for the analysis. From the 8 tested SSR markers, 7 SSR markers showed polymorphism between varieties. GenScan analysis were performed using the selected SSR markers to analyze the phylogenetic relationship of WCG. From the results, WCG cultivated in Korea showed that they have a very diverse genetic background.

Background : Wild-cultivated ginseng (WCG) prices are very different according to root ages. Generally, two methods are used to discriminate the root ages of Panax ginseng C.A. Meyer. The first method is the yearly determination method by the ring dyeing method, and the second method is the confirmation the number of stem vestiges in the rhizome. In this study, we analyzed the agronomic and growth characteristics of the WCG cultivated in Korea. In this study, to determine the appropriate root ages discrimination method for the determination of the root ages of WCG, the root ages of WCG and cultivated ginseng was examined. Methods and Results : We examined the cultivated ginseng (CG) and WCG that was collected and sold by regional groups at the Korean market. WCG does not form annual rings, which are clear and regular in wild ginseng. Therefore, it is impossible to identify the age of WCG by using the annual growth rings staining method. However, the age can be estimated by determining the number of stem vestiges in the rhizome. Conclusion : From the results of the Study on identification of root age for quality evealuation in WCG in Korea. Appropriate root ages discrimination method of WCG was confirmation the number of stem vestiges in the rhizome.

Background : Panax ginseng C.A. Meyer is a representative medicinal plants and it has been used in traditional medicine because the plant have many effective component such as saponins. To obtain a wild-cultivated ginseng (WCG) which is similar to wild ginseng (WG), this method usually performed in a mountain using seeds or seedlings of cultivated ginseng (CG) and WG. WCG is very expensive because it is difficult to cultivate. However, systematic cultivation method have not been developed compared to their high added value. Furthermore, very high price of WCG caused the problem that Panax notoginseng or Panax quinquefolium are sold as WCG in Korean market. This is concerned as a serious problem to consumers. In this study, we analyzed the agronomic and growth characteristics of the WCG cultivated in Korea. Methods and Results : We examined the WCG that was collected and sold by regional groups at the Korean market. The root age, growth conditions, and quality level of the cultivated WCG were confirmed. WCG samples were collected from five areas in Korea (Bucheon, Cheongju, Hoengseong, Judeok and Ulsan). The main root diameter, root shape index, rhizome length, and root weight showed high level of variation and they did not form annual rings. Conclusion : Agronomic and growth characteristics of WCG showed high variations according to cultivating regions.

Background : Korean mountain ginseng (Panax ginseng C.A. Meyer) are difficult to industrially apply because of its scarcity and high cost. Advances in plant biotechnology have made it possible to produce mountain ginseng on a large scale using adventitious root cultures in bio-reactors. This study was conducted to develop a cosmetic emulsion using ginsenoside and physiological activity - enhanced raw materials by fermentation process. Methods and Results : Wild ginseng adventitious roots were fermented with Pediococcus pentosaceus HLJG 0702 (KACC 81017BP). ginsenoside contents was analysed by using HPLC. Antioxidant activity was measured by DPPH and ABTS radical scavenging activity and whitening effect was measured by tyrosinase inhibitory activity. After microfluidizer processing was performed to prepare emulsions with homogenized particles, particle size and distribution were measured through a transmission electron microscop e(TEM). Particle stability compares pH, viscosity, light and zeta potential. When fermented with Pediococcus pentosaceus HLJG 0702, the highest change rates of Rg3, Rk1 and Rg5 were shown and the antioxidant activity was increased. The whitening effect was 73.2 ± 0.9% when treated at 100 ㎍/㎖, 1.5 times higher than the control. The optimum particle size and distribution were shown to be 418.0 ± 14.9 ㎚ for 6 times treatment with 0 - 10 times microfluidizer treatment. Stability was about 3% in pH, viscosity and light test. the zeta potential was found to be homogeneous at –33.33 mV. Conclusion : Pediococcus pentosaceus HLJG 0702 Fermented Wild ginseng adventitious roots were found to have effective ingredients and improved physiological activity. We have also developed emulsions that exhibit optimal particle size and distribution

Background : The study about cultured wild ginseng root (Panax ginseng C. A. Meyer) have been reported mainly ginsenosides in saponins family. However metabolites of fermented wild ginseng roots by microorganisms was not reported yet. Methods and Results : Cultured wild ginseng roots were used for fermentation of ginseng roots using Pediococcus pentosaceus and other bacterial strains. We analyzed different types of ginsenoside contents, metabolite and enzyme contents, and gene expression by using microorganisms. Results showed considerable differences in ginseonoside contents specially Rk1 and Rg5. The highest enzyme activity level was by Glutathione reductase (GR) and Glutathione S transferase (GST) in fermented ginseng roots than control (non-fermented), whereas Glutathione peroxidase (GPX) and Peroxidase (POD) contents were reduced. Score plots and loading plots of principal components 1 of the PCA result obtained from the data on 43 metabolites in fermented wild ginseng root of five conditions. The concentration of metabolite such as β-alanin and 4-aminobutyric acid (GABA), which is used to improve memory were increased in fermented ginseng roots than control. We found functional gene in wild ginseng root related with metabolic process. The APX gene expression gradually increased in fermented ginseng root with respect to fermentation times. Conclusion : In this study, accumulation of functional metabolite in cultured ginseng r

Background : The minor saponins produced by the hydrolysis of a major saponins sugar. The minor saponins has high absorption and efficacy compared to major saponin. The acid treatment, heat treatment and fermentation with minor saponin research has been actively conducted. This study was performed in order to investigate the bioconversion of ginsenoside Rg5 of fermented wild ginseng adventitious roots by using lactic acid bacteria. Methods and Results : 20g adventitious roots of ginseng was added to water (10-fold v/w). 10% (v/v) of lactic acid bacteria (Pediococcus pentosaceus HLJG0702[KACC 81017BP]) were inoculated with wild ginseng adventitious roots. For the fermentation process the inoculated samples were transferred to culture room for 1, 3 and 5 days. The fermented samples were dried at room temperature and extracted with 70% ethanol. Extract was concentrated completely at 50 ℃ and Rg5 was analysed by using HPLC. Results showed no significant difference the dry weight of non-fermented and fermented wild ginseng adventitious roots. During the fermentation process, the pH changed from 5.7 to 4.2. HPLC analysis showed higher ginsenoside Rg5 (39.588 mg/g) at 3 days. Conclusion : The fermentation of ginseng root can increase the Rg5 contents and minor saponin composition. This process may be used to enhance the minor saponin thereby increasing in fermented property of wild ginseng adventitious roots.

Background : Korean mountain ginseng(Panax ginseng C.A. Meyer) are difficult to clinically apply because of its scarcity and high cost. Advances in plant biotechnology have made it possible to produce mountain ginseng extracts on a large scale using adventitious root cultures in bio-reactors. This study investigated the variations of ginsenoside compounds composition and biological activities of wild ginseng adventitious roots by fermentation process. Methods and Results : Wild ginseng adventitious roots with five days fermentation using four strain of bacteria(Leuconostoc mesenteroides(KACC 15744), Bacillus circulans(KACC 15822), Bacillus licheniformis(KACC 15823), Bacillus subtilis subsp. inaquosorum(KACC 17047)). Ginsenoside contents was analysed by using HPLC. To examine the antioxidant activity associated with biological functions, radical scavenging analyses DPPH, ABTS and SOD-like activity analyses were conducted. The total phenolic and flavonoid contents were evaluated to determine the antioxidant activity increment. The result showed increased total ginsenoside contents by fermentation process. In particular, B. licheniformis showed the highest ginsenoside contents. Regarding ginseng fermented with B. licheniformis, values of 70.6 ± 1.4%, 44.3 ± 1.7%, and 88.4 ± 1.3% were measured using DPPH, ABTS, and SOD-like antioxdiant activity analyses, respectively. The total phenolic contents in ginseng fermented with B. licheniformis was 184.5 ± 0.9 ㎍·GAE/㎖, and the total flavonoid contents was 108.5 ± 1.8 ㎍·QE/㎖ in ginseng fermented with L. mesenteroides. Conclusion : The high activity of β-glucosidase was selected bacteria. The four types of lactic acid bacteria examined, the use of B. licheniformis to ferment ginseng resulted in greatest increase in biological activities and ginsenoside contents.

The aim of the present study was to determine the effect of dietary cultured wild ginseng root (CWGR) supplementation on goat milk composition and ginsenoside profiles. Sixteen Saanen dairy goats were allocated to two balanced groups based on lactation period, body weight (38.6 ± 3.2 kg), and dairy milk yield (2.85 ± 1.2 kg), and were kept in separate pens. Goats were fed a total mixed ration (TMR) feed (2.3 kg/d, dry matter basis) and 1.5 g of CWGR powder was supplemented in the experimental diet. The total feeding period was 3 weeks, and milk and blood samples were collected on the last three days of the experimental period. There was no effect of CWGR on daily milk yield and milk composition (fat, protein, lactose, and solid-not-fat). However, the CWGR-treatment group had significantly higher plasma IgG and protein contents than the control group (P < 0.05). Significant amounts of ginsenosides were observed in the milk of the CWGR-treatment group, whereas ginsenosides were not detected in the milk of the control group. In conclusion, dietary CWGR was a useful regimen to produce functional goat milk enriched in ginsenosides.

피부는 스모그, 담배연기 및 UV와 같은 외부환경으로부터 신체를 보호하는 가장 큰 기관이며, 보호 기작 으로서 각질세포와 그 사이를 메우고 있는 세라마이드, 콜레스테롤, 지방산 등의 세포간지질이 라멜라 액정 구조 로 피부 장벽을 이루고 있다. 본 연구에서는 세포간지질 중 세라마이드 생합성과 관련되어 있는 세린-팔미토일 전이효소(serine-palmitoyltransferase, SPT) 발현을 western blot으로 확인한 결과, 제주산양산삼 추출물이 농도의존적으로 SPT 단백질 발현을 증가시킴을 확인하였다. 또한 제주산양산삼 추출물을 5% 함유한 제형을 2주간 피부에 도포 후 TEWL을 측정하였을 때, 제주산양산삼 추출물을 함유한 에멀젼 도포부위의 TEWL이 대 조군에 비해 유의적으로 감소하는 것을 확인하였다. 이 연구결과는 제주산양산삼 추출물이 SPT의 발현 증가를 통해 세포간 지질의 핵심성분인 세라마이드의 생합성을 증가시켰음을 보여준다. 따라서 제주산양산삼 추출물은 피부장벽기능을 개선시켜 TEWL 감소 효과를 나타내며, 이를 통해 화장품 분야에서 피부장벽 강화 및 보습소재 로서 사용될 수 있다고 사료된다.

Background : Wild-cultivated P. ginseng (WCG) is a specific ginseng in Korea which depends on artificial forest growth method. To obtain a WCG which is similar to wild ginseng (WG), this method usually performed in a mountain using seeds or seedlings of cultivated ginseng (CG) and WG. Recently, very high price of WCG caused the problem that Panax notoginseng or Panax quinquefolium are sold as WCG in Korean market. This is concerned as a serious problem to consumers. In this study, we tried to develop a method to discriminate WCG, CG or WG using simple sequence repeat (SSR) markers and phylogenetic analysis. Methods and Results : WCG samples (3, 5, or 6-years old) were collected in Hoengseong, Gangwondo. DNA extraction was performed using CTAB method. SSR markers were collected from the published papers. After test PCR using the markers, one of the primer pair was labeled with fluorescence dye (FAM, NED, PET, or VIC) and Gene Scan analysis were performed. NTsys-PC program was used for the phylogenetic analysis of the data. Eight SSR markers were collected from the published literature and used for the analysis. From the 8 tested SSR markers, 7 SSR markers showed polymorphism between varieties. GenScan analysis were performed using the selected SSR markers to analyze the phylogenetic relationship of WCG. Conclusion : Phylogenetic analysis showed the relationship between WCG and P. ginseng cultivars and the seven SSR markers used in this study are able to distinguish Wild-cultivated P. ginseng.

Background : Ginsenosides, the main ingredient of ginseng roots can be confirmed various physiological activity such as anticancer, antioxidant, a natural ginsenosides is there a structure to be absorbed into the body does not work well absorbed through this process biologically active thus a high conversion ginsenosides. β-glucosidase enzyme is observed in several of the microorganism with an enzyme that serves to convert a ginsenoside prosper that is absorbed into the body. Methods and Results : To view a primary β-glucosidase activity, the bacteria were innoculated in esculin agar medium and the color change of the media were measured by the time and degree of changing color. In the other method, 5 mM of p-nitrophenyl-β-D-glucopyranoside (pNPG) containing 25 mM phosphate buffer solution (pH 7.0) was added to 50 ul enzyme solution. Then the solution was added to 50 ul reaction for 5 min at 30°C. The amount of p-nitrophenol liberated measured at 405 nm absorbance. The experimental results showed higher β-glucosidase activity in Pediococcus pentosaceus, Leuconostoc mesenteroide, Leuconostoc mesenteroides subsp. cremoris, and Paenibacillus polymyxa by using esculin agar medium method. Similarly in second method, β-glucosidase activity was higher in P. pentosaceus 402.32±11.43 unit/l, L. mesenteroide 353.73±14.64 unit/l, Lactobacillus sakei 198.4±15.47 unit/l Lactobacillus plantarum subsp. plantarum 164.1±8.12 unit/l. Conclusion : The result that the β-glucosidase activity was higher in P. pentosaceus, L. mesenteroide, and L. plantarum subsp. plantarum as compared to tested microbes. Therefore selected bacteria can be used in the industry of functioned foods and beverage to improve human healths.

With the purpose of improving ginsenoside production in Korean wild ginseng (Panax ginseng Meyer) mutant adventitious root lines, a synthetic gene encoding squalene synthase (PgSS2) was placed under the control of 35S promoter and transferred to Panax ginseng. Embryogenic callus obtained from ginseng adventitious root lines were transformed by infection with A. tumefaciens strain EHA105 containing the PgSS2 gene. Ten phosphinothricin-resistant plants were generated on selection medium, and the transgene integration and expression in these plants were confirmed by PAT test strip, RT-PCR and Southern hybridization. Ginsenoside analysis by HPLC revealed that the total contents of the 8 ginsenoside types (Rg1, Re, Rf, Rh1, Rb1, Rc, Rb2, Rd) in transgenic adventitious root lines were about 1.6-fold higher than that of the mutant control line (MCL1). This transformation method may facilitate the improvement of Panax ginseng in terms of the accumulation levels of ginsenoside.