Background : Exposure to Ultraviolet B (UVB) causes oxidative stress, inflammation, pigmentation and severe skin damage. Astragalus membranaceus (AG) has been used as a traditional medicine and have been studied various physiological activities. During the roasting process, bioactive substances is change including antioxidant substances. The aim is study the antioxidant effects and reactive oxygen species (ROS) inhibitory effect of the roasted A. membranaceus (R-AG). on Human dermal fibroblast (HDF) cells. Methods and Results : To prepare of R-AG samples, roasting machine was used. AG and R-AG were extracted to water and 70% ethanol. AG samples were evaluated the antioxidant potential by measuring the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2`-azino-bis 3-ethylbenzothiazoline-6-sulphonic acid (ABTS) scavenging activities. Additionally, total phenolic contents and total flavonoid contents was compared with antioxidant ingredients. AG and R-AGs were analyzed with HPLC determine the major compounds such as calycosin, mononetin and glycosides. The antioxidant activities of R-AG increased and changed in major compounds. In UVB exposed HDF cells, AGs did not affect cell viability and R-AG inhibited ROS more effectively than AG. Conclusion : From these results, R-AG can inhibit oxidative stress induced UVB in HDF cells.

Background : The isoflavonoids, triterpene and polysaccharides were know as major components in Astragalus membranaceus Bunge. Especially, isoflavonoids are classified as ononin, calycosin-glucoside of glycon binding and formononetin, calycosin of aglycone binding. In this study, we performed fermentation by a lot of microorganisms which were derived foods. Methods and Results : The A. membranaceus was extracted with 50% Ethanol and it was concentrated using rotary evaporator. In order to change in isoflavonoids, we fermented the A. membranaceus extracts with microorganisms which have β-glucosidase activity. we chose 20 microorganisms using esculin agar method and three strains with the highest activity of β -glucosidase were identified by pNP assay. The extracts of A. membranaceus were fermented during 3 days in sterilized distilled water, as 1 Brix and 3 Brix. The isoflavonoids of fermented extract, respectively days 0, 1, 2 and 3, were analyzed using High Pressure Liquid Chromatography (HPLC). In addition, the fermented A. membranaceus extracts was investigated about whitening effect using tyrosinase and radical scavenging activity by 1,1-diphenyl-2-picrylhydrazyl radical (DPPH). As a result of β-glucosidase activity by pNP assay, we selected two strains, Pediococcus pentosaceus (KACC 81017BP), Saccharomyces cerevisiae (KACC 83014BP). On the third day of fermentation of S. cerevisiae, isoflavone aglycone was the highest. The calycosin-glucoside and ononin in fermented A. membranaceus extract were reduced, calycosin and formononetin was increased compared to control. Also, we confirmed higher activity than control in tyrosinase inhibition rate on the third day of fermentation, as 78.38% and lower IC50 value of radical scavenging activity as 584.68 ㎍/㎖. Conclusion : From the above results, we may suggest that A. membranaceus aboveground parts might have useful as a safe material for functional food and pharmaceutics.

Background : Astragalus membranaceus belonging to the family of Leguminosae have been utilized as a traditional medicine. The aim of this study is to elucidate the basic information for breeding to superior Astragalus membranaceus cultivar. Methods and Results : Selection lines were developed by the medicinal crop breeding team of National Institute of Horticulture and Herbal Science (NIHHS), Rural Development Administration (RDA). The root of A. membranaceus lines were harvested in late October 2017. Calycosin-7-O-β-d-glucoside and calycosin were anlyzed by HPLC. The root yield of six selection lines were increased 8.9 - 74.8% compared with ‘Aseong’ (check variety). Also, the plant height of seven selection lines were shorter than ‘Aseong’. The calycosin–7-O-β -d-glucoside content was higher in six selection lines than ‘Aseong’. The calycosin content was higher in all selection lines except 1 line (1508 - 03) than that of ‘Aseong’. Conclusion : Three superior lines with short height and high yield were selected. These superior lines will conduct advanced yield trial to make varieties.

Astragalus membranaceus is a well known oriental medicinal herb. The polysaccharides of the aboveground parts (AMA) and the radix (AMR) of A. membranaceus are the most important functional constituents. Methods and Results: The aim of this study was to determine the effects of AMA and AMR on the oxidative damage induced in the skeletal muscle of rats subjected to exhaustive exercise. Sprague-Dawley rats were randomly divided into exercise and non-exercise groups; in the groups receiving the test compounds, AMA and AMR were administered orally for 30 days. Skeletal muscle samples were collected from each rat after running to exhaustion on a treadmill to determine the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) and the concentation of malondialdehyde (MDA). The antioxidant enzyme activities of SOD and GSH-Px of skeletal muscle of AMA- and AMR-treated groups were significantly higher than those of the control and commercial sports drink (SPD)-treated groups in exhaustive exercise rats. In addition, MDA concentrations in the skeletal muscle of the AMA- and AMR-treated groups were significantly lower than those of the control and SPD-treated groups. In the present study, the effects of AMA and AMR on exercise endurance capacity were also evaluated in mice subjected to a swimming exercise test. AMA and AMR supplementation prolonged the swimming time of mice and enhanced exercise endurance capacity. AMA and AMR possess the ability to retard and lower the production of blood lactate, and prevent the decrease of serum blood glucose. Conclusions: These results showed that, AMR and AMA exerted beneficial effect in mice, increasing the activity of the antioxidant systems and inhibiting oxidative stress induced by exhaustive exercise. The compounds improved exercise performance and showed anti-fatigue effects against exhaustive exercise.

Background : The major components in Astragalus membranaceus are isoflavonoids, triterpene and polysaccharides. Also, isoflavonoids was composed to aglycon and glucoside derivatives. In this study, we performed fermentation process (enzyme treatment) and steam processing (high temperature and pressure) to increase aglycon such as formononetin and calycosin. Methods and Results : The steam processing was performed at a lot of conditions, such as temperature (80, 100, 120℃) and time (30, 60 and 120 min). Fermentation processing carried out by A. membranaceus extract fermented with microorganisms which have high β -glucosidase activity (selection by esculin agar method) for detached glucose from isoflavonoids, converted to algycon. The isoflavonoids were analyzed using high pressure liquid chromatography (HPLC) in fermentation product of A. membranaceu extract by treated with steam processing at various conditions. As a result of β-glucosidase activity by pNP assay, we selected three strains, Pediococcus pentosaceus, Weissella cibaria and Saccharomyces cerevisiae. Isoflavone aglycon was the highest in fermentation product by S. cerevisiae for 3 days, but change of isoflavonoids was not observed in steam processing. The calycosin-glucoside and ononin were reduced in fermentation product of A. membranaceus extract, whereas calycosin and formononetin was increased. Conclusion : This results showed that isoflavone glycoside converted to isoflavone aglycon in A. membranaceus by fermentation process, it seems to be available for industrial use.

Background: This study were to investigate the effect of Pediococcus pentosaceus fermented Radix astragali (AMRP) and non-fermented products (AMRNP) on collagen synthesis in the cultures of human dermal fibroblasts, and their inhibitory effects on the matrix-degrading enzymes (collagenase, elastase, and gelatinase). Methods and Results: Both AMRP and AMRNP significantly improved cell growth and proliferation of HDF cells. However, the enzyme-linked immunosorbent assay and Western blot analysis demonstrated that AMRP, but not AMRNP, significantly and dose-dependently stimulated the biosynthesis of type I procollagen in both aged (74 y) and young (21 y) HDF cells. Real-time reverse transcription-polymerase chain reaction revealed that expression of type I, type III procollagen and transforming growth factor β1 (TGF-β1) mRNA was significantly stronger in AMRP-treated HDF cells than that of AMRNP-treated and un-treated HDF cells. The AMRP revealed an increase in astragaloside Ⅳ only depending on increase in fermentation period, because other astragalside converted to astragaloside Ⅳ, which it detached acyl group by fermentation processing of Pediococcus pentosaceus. Conclusion: The results also suggested that AMRP could stimulate the collagen biosynthesis in human dermal fibroblasts, which is, associated with the regulation of procollagen biosynthesis resulting from AMRP-induced TGF-β1 expression and the mitogenic activity in HDF cells, and therefore, is expected to reduce the age-dependent loss of extracellular matrix proteins.

Background: Polysaccharides are the most important functional constituent in Astragalus membranaceus. The purpose of the present study was to evaluate the effect of polysaccharides isolated from the aboveground parts of A. membranaceus (AMA) and polysaccharides isolated from the roots of A. membranaceus (AMR) immune function by modulated cytotoxic T cell and Th1- and Th2- related cytokines kinetics. Methods and Results: Sprague-Dawley rats were randomly divided into exhaustive exercise case groups and non-exercise case, AMA and AMR samples were administered orally for 30 days (500 ㎎/㎏/day and 10 ㎎/㎏/day, respectively) and were compared to those rats in the groups fed commercial sports drink (SPD) and vehicle. Both exhaustive exercise groups and non-exercise groups had a lower ratio of CD4+ and CD8+ cells in the spleens of the rat fed AMA and AMR compared to those in the rats fed SPD and vehicle group. These results suggested that AMA and AMR promote an increase in the proportion of cytotoxic T cells. The IL-4- producing T lymphocytes decreased significantly in the AMR (10 ㎎/㎏/day) group compared to SPD and vehicle, whereas the AMA group increased the IL-4 concentration more than the SPD and vehicle in exhaustive exercise group. However, the populations of IFN-γ-producing T lymphocytes of AMR and AMA increased. AMA decreased the concentration of IFN-γ to inhibit the Th1 response and thereby increased the concentration of IL-4 to induce a Th2 response that was related to humoral immunity in the non-exercise group. Conclusions: These results showed that, in addition to Th1/Th2 regulation, AMR and AMA played an important immuno-modulatory role after exhaustive exercise-induced Th1/Th2 lymphocyte imbalance, which might be correlated with cytokine producing immunoregulatory cells.

This study analyzed the changes in physicochemical components of Astragalus membranaceus (AM) fermented with Phellinus linteus. Moisture content, pH, total acidity, total reducing sugar content, extraction yield, free sugar content, free amino acid and isoflavonoid (calycosin, formononetin) were investigated. The moisture content was increased during fermentation with Phellinus linteus. The pH level increased while the total acidity significantly decreased during fermentation. The reducing sugar content were in the range of 0.32~0.61%. The extraction yield using water was higher than that using 80% ethanol. The major free sugars were identified as glucose, fructose, sucrose and the content of free sugars decreased through fermentation. However, the glucose and sucrose contents of the water extracts were increased. In addition, the free amino acid increased significantly during fermentation. Finally, calycosin and formononetins contents in water extracts of after 30 days of AM fermentaion with Phellinus linteus were (3.91 mg/100 g) and (1.38 mg/100 g), respectively. These results suggest that fermentation with Phellinus linteus could be used to increase the bioactivity of AM. The mycelium-fermented AM could be a valuable source of functional material and edible resource for industry.

Background: In recent years, adjuvants have received increasing attention owing to the development of purified subunit and synthetic vaccines which are poor immunogens and require additional adjuvants to evoke an immune response. Therefore, immunologic adjuvants have been developed and tested. Plant polysaccharides have been recognized as effective biological response modifiers with low toxicity.Methods and Results: In this study, the polysaccharide from the aboveground part of Astragalus membranaceus Bunge containing immunomodulating arabino-3,6-galactan was evaluated for its hemolytic activity and adjuvant potential in the specific cellular and humoral immune responses to ovalbumin. The polysaccharide from the aboveground part of Astragalus membranaceus Bunge was co-immunized with the purified Vi capsular polysaccharide of Salmonella typhi vaccine in mice. The polysaccharide from the aboveground part of Astragalus membranaceus Bunge did not induce any hemolytic activity or side effects at doses up to 500㎍/㎖. The concanavalin A-, lipopolysaccharide-, and ovalbumin-induced splenocyte proliferation and serum ovalbumin-specific IgG, IgG1 and IgG2b antibody titers in immunized mice were significantly enhanced by AMA. Pharmacological data revealed that the polysaccharide from the aboveground part of Astragalus membranaceus Bunge increased antigen-specific antibody levels in immunized mice. The polysaccharide from the aboveground part of Astragalus membranaceus Bunge-adjuvanted purified Vi capsular polysaccharide of Salmonella typhi vaccine improved the proliferation of splenocytes and macrophages as well as stimulated cytokine production.Conclusions: These results suggest that the polysaccharide from the aboveground part of Astragalus membranaceus Bunge-adjuvanted vaccines enhanced humoral and cellular immunity and that the polysaccharide from the aboveground part of Astragalus membranaceus Bunge is a safe and efficacious adjuvant candidate suitable for use in prophylactic and therapeutic vaccines.

Background : This experiment was conducted to select GAP applying seed disinfectants in Astragalus membranaceus and Platycodon grandiflorum. Methods and Results : We carried out the chemical efficacy and injury test. For the efficacy test, we investigated fungal detection rate by seed disinfectants and for the chemical injury, we investigated germination rate and emergence rate by seed disinfectants in reference amount and fold amount. These experiments carried out two times. The results obtained are as follows. In the experiment for seed disinfectants selection of Astragalus membranaceus, all tested chemicals such as Tebuconazole emulsifiable concentrate(EC), Thiophanate-methyl + Triflumizole wettable powder(WP), Prochloraz copper chloride complex+Tebuconazole suspension concentrate(SC), Prochloraz emulsifiable concentrate(EC), Fludioxonil wetting liquid(WL) and Hexaconazol+Prochloraz emulsifiable concentrate(EC) had control value of 80% or above against seed contaminated fungi. However two chemicals such as Tebuconazole EC and Prochloraz copper chloride complex+Tebuconazole SC and two chemicals such as Prochloraz EC and Hexaconazol+Prochloraz EC exhibited chemical injury significantly in reference amount and in fold amount respectively, compared to non treated control. In the case of seed disinfectants selection of Platycodon grandiflorum, Prochloraz copper chloride complex+Tebuconazole SC, Prochloraz EC and Hexaconazol+Prochloraz EC had control value of above 80% against seed contaminated fungi except Thiophanate-methyl+Triflumizole WP and Fludioxonil WL. However Hexaconazol+Prochloraz EC and Prochloraz copper chloride complex+Tebuconazole SC exhibited chemical injury significantly in reference amount and in fold amount respectively, compared to non treated control. Conclusion : From the above results, we finally selected three items of seed disinfectants including Thiophanate-methyl+Triflumizole WP and Fludioxonil WL in Astragalus membranaceus and Prochloraz emulsion in Platycodon grandiflorum.

Background : This study, the fraction for testing the efficacy of the Astragalus extract was concentrated active ingredient. The concentrated fraction was applied to a cosmetic material that Astragalus testing results confirmed that the improved efficacy. Methods and Results : The fractions were performed using an n-butanol solvent for increasing the efficacy of the Astragalus extract, by using the material fractions collected to compare and ultimately an increase in whitening and wrinkle efficacy. The solvent to be used in the fractions was used for the n-butanol good dissolution to the effective substance(Astragaloside, Isoflavonoid). It increased approximately 6.5 times the sample extract and the n-butanol fraction of the Astragalus as a result Astragaloside 15 ppm, 97 ppm respectively analyzed by HPLC equipment, isoflavonoid content was confirmed by an increase in the content of the fractions increased 4.5 times to 280 ppm, 1,260 ppm. Tyrosinase inhibitory effect, respectively IC50 5.70 mg/mL, IC50 1.02 mg/mL to, Collagenase producing ability is IC50 4.88 mg/mL, IC50 0.93 mg/mL with n-butanol fraction was good whitening, anti-wrinkle efficacy than the extract. Sensory evaluation was conducted in the same amount of sample, using a purified Astragalus cosmetics received high marks. Stability evaluation(MTT assay) was checked for more than 100% cell viability at the concentration 2,000 ppm. Conclusion : n-butanol fraction of Astragalus was subjected to a component analysis and In vitro test, it was confirmed an increase active ingredient content. The results of sensory evaluation and stability evaluation, it was confirmed been made to improve qualities as a cosmetic materials.

Background : ROS produced by oxidative stress damaged endothelial cells, and cause a variety of vascular complications. In diabetic hyperglycemia state, ROS increase. The polyol pathway occur in diabetic complications, the excess glucose is absorbed into the polyol pathway when aldose reductase increased, NADPH changes it to sorbitol. Glutathione (GSH) removes ROS. GSH level is reduced by glutathione reductase, using NADPH as an electron donor. Activation of the polyol pathway decrease NADPH, and GSH also reduced. As a result, ROS is increased. In diabetic hyperglycemia state, Glycolysis increases. Effects of increased glycolysis, protein kinase C (PKC) is increased. NAD(P)H oxidase, stimulated by PKC-dependent pathway, increases ROS in the cell. In this study, we measured the ROS scavenging activity of 5 natural products (Lycii fructus, Astragalus membranaceus, Cassia Tora, Polygonatum odoratum, Rubus Coreanus), to confirm the efficacy as diabetic antioxidants. Methods and Results : We extracted 5 natural product by distilled water and ethanol. DPPH radical scavenging activity was significantly higher in Lycii fructus, Rubus coreanus. ABTS radical scavenging activity was better Rubus coreanus, Lycii fructus, Cassia Tora. In addition to, Rubus coreanus, Cassia Tora, Lycii fructus was comparatively higher reducing power activity than other natural products. And total phenolic and flavonoid contents were much higher in Rubus coreanus compared with other extracts. Conclusion : These results suggest that Lycii fructus, Rubus coreanus can be applied as diabetic antioxidant that prevent vascular complications caused by ROS.

Background : This study was performed to investigate by antioxidant activity, total phenolic contents, total flavonoid contents, and effective component of Astragalus membranaceus treated with different artificial light Sources (fluorescent lamp, red, blue, green, white, LEP). Methods and Results : We investigated the effects of various artificial light sources on the DPPH radical activity, total phenol and flavonoid contents, tyrosinase activity and main flavonoid compounds contents (formononetin and calycosin) and other biological activities in A. membranaceus. Antioxidant activities were 53.6% as the highest level of activity under LEP light. Growth under LEP light also produced the highest total phenolic and flavonoid contents of 36.05 and 5.94 mg/ml, respectively. Extracts from plants grown under LEP light caused the highest inhibition of tyrosinase activity with inhibition of 35.37, 61.87, and 65.49%, respectively, for extract concentrations of 100 μg/ml, 500 μg/ml, and 1000 μg/ml compared with other artificial light treatments. Conclusion : Little information is available on the influence of LED and LEP light sources on antioxidant production or other biological activities in A. membranaceus. Our goal in this study was to determine the effects of LED and LEP artificial light sources on the production of new functional compounds in A. membranaceus.

Background : Astragalus membranaceus is one of the most widely used traditional medicinal herbs in Korea. Studies on the genomic of A. membranaceus resources have not been carried out so far. The present study was carried out to discriminate A. membranaceus based on genetic diversity using genomic simple sequence repeat (SSR) markers. Methods and Results : We collected 5 A. membranaceus lines: Asung, Poongsung, Am-Jecheon, Am-Sancheong, and Am-China. One hundred mg of fresh leaves were used for genomic DNA extraction using the DNeasy plant DNA isolation kit (Qiagen GmbH, Hilden, Germany). 450,449 contigs were searched for 147,766 SSR candidate loci in this study using the MicroSAtellite identification tool (MISA). We selected 949 A. membranaceus genomic SSR markers that were showed variation for the five collections in silico screening with CLC genomics workbench program. The genetic diversity of all A. membranaceus resources was analyzed using 17 SSR markers employing the DNA fragment analysis method. Based on the genetic diversity analysis, these lines were classified into four distinct groups. Conclusion : These findings could be used for further research on cultivar development using molecular breeding techniques and for conservation of the genetic diversity of A. membranaceus. Furthermore, the markers could be used for marker-assisted selection for crop breeding.

This study analyzed the physicochemical characteristics of Astragalus membranaceus (AM) fermented with seven different mushroom mycelia. Physicochemical characteristics, such as contents of moisture, pH, total reducing sugars, free sugar, and isoflavonoid, were investigated. The moisture content was increased in most of the samples. The pH values of AM fermented with Phellinus linteus and Flammulina velutipes were increased, while the pH of other samples were similar to that of non-fermented AM. The reducing sugar content was in the range of 211.69~391.74 mg/100 g. The extraction yield using water was higher than that when extracted with 80% ethanol. The free sugar content was increased through fermentation with mushroom mycelia. However, the glucose contents of the 80% ethanol and water extracts were decreased. Finally, the calycosin and formononetin contents in 80% ethanol and water extracts of AM fermented with Phellinus linteus were 2,549.24 mg/g, and 827.66 mg/g for calycosin, and 1,366.69 mg/g and 221.28 mg/g for formononetin, respectively. These results suggest that fermentation with mushroom mycelia could be used to increase the bioactivity of AM. The mycelium-fermented AM might be a valuable source of functional material and edible resource for industry.

본 연구에서는 기호성을 향상시킬 수 있는 고부가가치 간장을 개발하고자 황기 첨가량에 따른 간장의 이화학적 특성 및 관능평가를 실시하였다. 식염변화에서는 황기 10% 첨가 간장에서 약간 감소하는 경향을 보였다. pH 변화는 실험군 모두 숙성기간에 따른 변화는 뚜렷하지 않았으며, 가용성 고형분 함량은 황기 10% 첨가 간장를 제외하고 모두 2개월까지 급격히 증가되었으며 그 이후 일정한 양상 을 보였다. 그 반면 황기 10% 첨가 간장는 숙성 초기에 증가되었지만 무첨가 및 황기 5% 첨가 간장 보다 낮은 경향을 보였다. 갈색도 변화는 숙성이 진행될수록 증가되 는 경향을 보였으며 조단백질 함량은 무첨가 및 황기 5% 첨가 간장은 숙성 기간이 지남에 따라 증가되는 경향을 보였지만 황기 10% 첨가구는 숙성 기간에 따른 차이를 보지 않았다. 유리 아미노산 측정결과 황기 5% 첨가 간장의 6개월 때가 가장 높은 함량을 나타내었다. 관능평가에서는 황기 5% 첨가 간장의 숙성기간이 증가됨에 따라 기호도가 점차 향상되었으며 이러한 결과를 미루어 보아 황기 5% 첨가 간장이 황기 무첨가 간장에 비해 염도가 낮고 아미노 태질소가 높으며 기호도는 6개월 때가 가장 우수하였으며, 풍미가 좋은 간장으로의 활용 가능성이 있음을 제시하였 다.

Background : A series of studies were conducted to optimize adventitious root induction in vitro from explants of Astragalus membranaceus using various nutrient media supplemented with plant hormones. Methods and Results : Levels of active components were analyzed from adventitious roots induced under different media conditions. Among the different media conditions, Murashige and Skoog medium supplemented with 1.0㎎• ℓ −1 indole-3-butyric acid resulted in the greatest adventitious root induction rate. The amount of the major active component of the adventitious roots of Ama1, calycosin-7-O-β-D-glucopyranoside was higher than that of other adventitious root samples. Conclusions : These results suggest that the adventitious roots of A. membranaceus could be used for the commercial production of medicines.

황기는 콩과에 속하는 다년생 초본으로 단너삼이라고도 불리며 한국을 비롯한 중국, 몽고 등 아시아 지역에서 자생한다. 한국에서는 한약재와 식품을 주 목적으로 재배하며 주 이용부위인 뿌리는 독성이 없어 안정하면서도 다양한 약리효능 때문에 소비가 증대되고 있다. 하지만 황기는 뿌리작물로써 연작장해가 심하고 재배 시 노동력과 비용이 많이 소모되기 때문에 이러한 문제점을 극복하기 위해 기내배양 조건을 구축하고자 하였다. 실험재료는 황기품종인 아성과 풍성을 이용하였다. 각 종자를 기내에서 발아시키기 위해 1% NaOCl에 5분동안 침지하고 다시 70% Et-OH에 3분간 침지하여 표면살균 후 멸균수에서 3회 세척하였다. 종자발아를 유도하기 위해 30g/L sucrose가 함유된1/2MS 배지를 25℃ 인큐베이터에서 16시간의 광조건 3,000룩스(lux) 광량으로 배양하였다. 부정근을 유도하기 위하여 발아된 유식물의 잎, 줄기, 뿌리 절편을 0.5 × 0.5 mm 로 절취하여 3～5 mg/L 3-indolybutyric acid(IBA)가 첨가된 1/2MS 고체배지를 사용하였으며 25℃, 암조건의 인큐베이터에서 3주간 배양하였다. 부정근의 증식은 고체배양 할 때에는 1～5 mg/L IBA가 첨가된 MS 배지에 캘러스를 치상하여 25℃, 암조건의 인큐베이터에서 3주간 배양하였고 액체배양은 0.5 와 1.0mg/L IBA가 첨가된 MS 배지에 부정근 생체를 0.2g으로 정량하여 25℃, 120rpm, 암조건의 진탕배양기에서 배양하였다. 그 결과 캘러스의 유도는 아성뿌리절편에서 IBA 3 mg/L, 풍성뿌리절편에서 IBA 4 mg/L 일 때 가장 높은 유도율을 보였다. 캘러스의 유도율이 가장 우수한 조건에서 얻어진 부정근을 이용하여 고체배양을 실시하였으며, 아성은 IBA 3 mg/L, 풍성은 IBA 5 mg/L에서 높은 증식률을 나타냈다. 액체배지의 증식은 MS액체배지에 IBA 0.5와 1.0mg/L 농도로 수행하였다. 그 결과 IBA 1.0mg/L의 MS배지에서 대조군에 비해 약 2배의 부정근 생산량을 보였다. 따라서 본 연구에서 얻어진 결과를 바탕으로 황기의 유효성분 대량생산을 위한 기내배양 시스템을 구축할 수 있을 것이라 사료된다.