The aim of this study was to compare the two in vitro culture systems callus and adventitious root by investigating the biomass and phenolic compounds in calli and adventitious roots induced from four different explants (leaf, root, petal, and ovary) in Camellia japonica. The biomass of calli and adventitious roots was examined after 4 and 8 weeks of cultivation, respectively, and 22 phenolic compounds were analyzed using high-performance liquid chromatography (HPLC). The biomass of the ovary-derived calli (2.0 g･mass-1) was 1.5-fold that of the leaf-derived calli. The dry weight (DW) was highest in ovary-derived calli; however, the highest dry matter content was obtained from leaf-derived calli. Differences in the investigated characteristics depended on the callus origin. In adventitious roots, the highest biomass was achieved in the leaf-derived adventitious root system; its fresh weight was 2.3-fold (89 mg･ea-1) higher, and its DW was 1.8-fold (16 mg･ea-1) higher than those of ovary-derived adventitious root system. Active cell division was detected in petal-derived lines in both the calli and adventitious roots. Results of the HPLC analysis revealed that the total content of 22 phenolic compounds was highest in ovary-derived calli and ovary-derived adventitious roots. Our experiments confirmed that the calli and adventitious roots of C japonica have different cytological characteristics and bioactive compounds depending on the explant origin. In addition, callus culture was a more suitable system than adventitious root for producing phenolic compounds when the duration of the culture period and biomass were considered.

The influence of the culture media on the growth and catechins and caffeine production was investigated in adventitious root cultures of tea tree. The growth rate of adventitious roots was higher than that of solid medium. Growth rate of adventitious roots was noted to be optimal in N6 liquid medium. Yields of (-)-epicatechin gallate (ECG) and caffeine were the highest when adventitious root cultures were maintained in Nitsch and B5 liquid medium, respectively. The production of (-)-epicatechin (EC) and (-)-epigallocatechin gallate (EGCG) from adventitious roots was maximal when cultured in 1/2MS liquid medium. The adventitious root extracts of tea tree produced on catechins as EC (20.77 mg/g) and EGCG (1.84 mg/g) in 1/2MS medium; EGC (24.39 mg/g) and caffeine (12.97 mg/g) in B5 medium and (-)-epigallocatechin (ECG) (2.16 mg/g) in Nitsch liquid medium.

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 : Mountain ginseng adventitious roots are being produced in large quantities by using plant tissue cultivation techniques to overcome the disadvantages of recent cultivation techniques. In this study, we intend to provide basic data on the development of cosmetic products by measuring the antioxidants of mountain ginseng adventitious roots extracted from each ethanol concentration and the activity of tyrosinase and elastase enzyme inhibition. Methods and Results : After drying, the mountain ginseng adventitious roots, it was crushed to less than 0.5 ㎜. for testing. It was then extracted at 70℃ for 180 minutes, adding 10 times its weight to each ethanol concentration (0, 20, 40, 60 and 80%) and filtered. The extract is depressurised and used in experiments after freeze-drying. The total phenols and flavonoids content measurement showed the most content in 80% ethanol extract. However, Trolox equivalent antioxidant capacity, ferric reducing antioxidant power showed the best ability in 60% ethanol extract. In addition, the active measurements of tyrosinase and elastase enzyme inhibition performed to check skin whitening and wrinkles improvement showed the highest activity in the 80% ethanol extract. Conclusion : As a result, the optimum conditions for the use of natural antioxidants and functional foods were maximized when it is extracted with 60% ethanol. It is also expected to be valuable as a natural cosmetics material when extracted with 80% ethanol as a solvent when combined with the results of enzyme inhibition activity.

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: 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 : 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 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.

Panax ginseng C.A. meyer (family: Araliaceae) is a perennial crop that has been widely used as a traditional medicine in Korea. Various P. ginseng cultivars exhibit a range of morphological and physiological traits as well as genetic diversity. To elucidate the differences of primary metabolism underlying such genetic diverstiy, we performed primary metabolite profiles in adventitious roots from five Panax ginseng cultivars using gas chromatography-mass spectrometry (GC-MS). The GC-MS analysis revealed eight primary metabolites as biomarkers and allowed us to classify the five cultivars into three groups. We selected three cultivars to represent each group and analyzed their transcriptomes by Illumina sequencing. We inspected 100 unigenes involved in seven primary metabolite biosynthesis pathways and found that 21 unigenes encoding 15 enzymes were differentially expressed among the three cultivars. Integrated analysis of transcriptomes and metabolomes revealed that the ginseng cultivars differ in primary metabolites as well as in the putative genes involved in the complex process of primary metabolic pathways. Our data derived from this integrated analysis provide insights into the underlying complexity of genes and metabolites that co-regulate flux through these pathways in ginseng.

Panax Ginseng is a perennial medicinal plant originated from North-east asia. Because of its well-known tonic effects mainly from ginsenosides, various types of processed ginseng products have been distributed around the world. Here, we analyzed secondary metabolite profiling of adventitious roots of 5 korean ginseng cultivars, Chunpoong (CP), Sunhyang (SH), Gopoong (GO), Sunun (SU), and Cheongsun (CS). At the same time, the profiles of relative gene expressions related to ginsenoside biosynthesis pathway were compared among ginseng cultivars. Secondary metabolite profiles were revealed by UPLC/Q-TOF-MS from extracts of bioreactor derived adventitious roots of five ginseng cultivars. Using principal component analysis, secondary metabolite profiles of ginseng cultivars were categorized into three groups. Metabolites with high VIP values were annotated using known database and standards compounds. Relative gene expression of ginsenoside related gene were analyzed using realtime PCR. The three groups had distinct metabolite contents. Furthermore, gene expression profiles related to ginsenoside were also different, which might contribute diverse secondary metabolite composition of ginseng cultivars. Further integrated analysis would provide a relationship between genetic background of ginseng cultivars and secondary metabolite profiles.

Korean Panax ginseng C. A. Meyer (P. ginseng) is a well-known and one of the most important tonic herbs used in traditional Korean medicine. The pharmacological effects of P. ginseng have been reported by many researchers. Nevertheless, little is known between the mechanism of action and the active compounds. In this study, we performed a comprehensive proteomic analysis and protein categorization in order to understand the physiological characteristics of the major components in the adventitious roots of P. ginseng. Whole proteins extracted from the cultured adventitious roots of P. ginseng were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE). Among the 1000 spots which were detected by silver staining, 113 spots were labeled and identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF MS). Our results showed that 40 proteins were identified among the 113 spots, with a hit ratio of 35.3%. A number of proteins identified on the 2-DE gels (30%; 16 spots) were involved in energy metabolism. These proteomic data will be helpful to better understand the physiological and pharmacological effects of P. ginseng.

We have established adventitious root culture systems of Codonopsis lanceolata, Codonopsis pilosula and Codonopsis ussuriensis. Root segments of C. lanceolata were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with 0.5mg/l NAA and produced 18.8±1.9 roots per explant. Root segments of C. pilosula were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with 1.0mg/l NAA and produced 8.5±1.8 roots per explant. Leaf segments of C. ussuriensis were the best explants for induction of adventitious roots and the number of adventitious root for explant was highest on solid medium with 0.5mg/l NAA and produced 7.8±0.4 roots per explant. In liquid culture, the best production of adventitious root (fresh weight) was obtained in 1/2 MS medium with 1.0mg/l NAA. This study demonstrated for the first time to produce adventitious roots in C. pilosula and C. ussuriensis.

The production of adventitious roots derived from root explant of Echinacea angustifolia and its secondary metabolite content were assessed in different types and levels of auxin. The induction of adventitious roots from root explant cultured in Murashige and Skoog solid medium supplemented with 1.0 mg/L indole -3-butyric acid (IBA) attained highest as 20.87 mg fresh weight and 3.07 mg dry weight per culture but root suspension culture at the same concentration of IBA enhanced biomass production as 3.07 g fresh weight and 0.38 g per culture after 4 weeks in culture. 3.0 mg/L α-naphthalene acetic acid (NAA) treatment had similar effect on root biomass production as 3.07 g fresh weight and 0.38 g per culture with liquid suspension culture, whereas adventitious roots exposed to over 3.0-5.0 mg/L IBA or 5.0 mg/L NAA were less responsive by reducing the number of adventitious roots and/or changing root morphology such as short and thick. The content of secondary metabolites such as phenolic, flavonoids and total caffeic acid in adventitious roots cultured on MS medium supplemented with 1.0 mg/L IBA were attained highest as 27.20, 9.60. 10.67 mg/g dry weight, respectively. Overall, the best production of root biomass and secondary metabolites were given by 1.0 mg/L IBA.

Panax ginseng is a well-known herbal plant originated from North-east asia for its various tonic effects. However, production of ginseng roots takes long time in field condition, usually five through six years until harvest. Additionally, ginseng is very susceptible to many kinds of biotic and abiotic stresses, for example, Rhizoctonia solani, which causes damping-off, or high temperature. To overcome these limits, induction of adventitious roots has been studied for more than 10 years and also adventitious roots are widely used materials for genetic research of P.ginseng. In this study, we induced adventitious roots from registered Korean ginseng cultivars and cultured them in bioreactor condition. Induction rate of adventitious roots from nine Korean ginseng cultivars was evaluated and growth pattern of four cultivars in bioreactor scale was also studied. Furthermore, genes that are related to biosynthesis of saponins in ginseng, ginsenoside, were discovered in ginseng whole-genome shotgun sequences for genetic research.

일반적으로 쌍떡잎식물 국화과 허브의 한 종인 에키네시아는 미국 북미 대평원에 서식하는 야생 식물로 수세기동안 감기나 또 다른 바이러스 감염에 의한 질병에 대하여 면역 기능을 증진시키는 전통 식물 약제로 널리 사용되어 왔다. 본 연구에서는 생물반응장치를 이용하여 조직 배양된 에키네시아 식물체에 대하여 화장품 성분으로써 응용 가치를 평가하였다. 이미 몇몇 보고된 논문에서 에키네시아는 조직재생, 상처 치유, 그리고 염증 억제 등의 약리학적 활성에 대한 연구가 진행되었다. 본 연구에서는 생물반응장치를 이용하여 조직 배양된 에키네시아 추출물에 대하여 화장품 성분으로서 효능, 효과를 평가하였다. 실험 결과 조직배양된 에키네시아 추출물은 항산화 효과 및 단백분해효소 발현 억제 효과가 자연 상태의 에키네시아 추출물과 비교하여 우수한 결과를 보였다. 이러한 결과들은 피부보호를 위한 화장품 성분으로서 응용 가능성을 제공할 수 있을 것으로 사료된다.

본 실험은 여우구슬의 기내 부정근 유도 및 증식조건의 확립을 목적으로 수행되었다. 우선 여우구슬의 기내 발아체로부터 부위를 달리하여 부정근을 유도한 결과 줄기부위는 뿌리보다 양호한 부정근의 유도를 보였다. 또한 유도된 부정근을 이용하여 옥신의 종류(IAA, IBA, NAA와 2.4-D)에 따른 부정근 유도율을 조사한 결과 IBA와 NAA는 IAA와 2.4-D보다 높은 유도율을 보였다. IBA의 농도에 따른 유도율과 증식효율은 IBA가 0.5 mg/L첨가되었을 때 가장 높은 유도 및 증식효율을 보였다. 최적의 액체배지조건을 확인하고자 IBA의 농도는 0.5 mg/L로 첨가하고 sucrose의 농도를 달리하여 실험한 결과 sucrose는 30 g/L 첨가 되었을 때 가장 높은 생중량과 건중량을 나타냈다. 액체배양된 여우구슬의 부정근을 각각 MS, 1/2MS, 1/3MS배지에 30 g/L sucrose, 0.5 mg/L IBA가 첨가된 5 L 용량의 생물반응기에 4주간 배양한 결과 1/2MS 배지에서 양호한 생장을 보였다. 본 실험에서는 여우구슬의 종자발아체를 이용하여 부정근의 유도 및 증식조건에 필요한 기내배양조건과 2차적으로 유도된 부정근을 이용하여 플라스크와 생물반응기 배양을 통한 효율적인 증식조건을 확립하였다.