This experiment was carried out in order to collect the basic data on the standardization of the manufacturing process of Rehmannia glutinosa Libosch. var. purpurea Makino drying. By the drying methods of R. glutinosa, the content of water, inorganic components, reducing sugar, catalpol and benzo[α]pyrene were investigated. The water content was 15.6~17.2% when R. glutinosa was dried by cold-warm air moisture absorption drying method (CAMAD) at 60℃ during 6 days. Among of the inorganic components of R. glutinosa the K content was the most followed by P, Na, Ca and Mg. The reducing sugar content of R. glutinosa by the hot air drying method (HAD) was much more than that by the CAMAD. The catalpol content of R. glutinosa was not different by the drying temperature when it was dried by the CAMAD. The catalpol content of the large size tuber (about 50.0 g/unit) showed a tendency to increase from 60℃ until 70℃ drying temperature, but that of the small size tuber(about 4.0 g/unit) was decreased as being a trend as the drying temperature high when R. glutinosa was dried by the HAD, But the catalpol content R. glutinosa had a tendency to drop significantly at drying temperature above 80℃. The benzo[α]pyrene content was little detected when R. glutinosa was dried by both the SLD and the CAMAD, and the sampling by the HAD indicated within the scope of 5 μg/kg which was the scope to regulate by Korean food and drug administration. In conclusion, it seemed that an appropriate drying temperature of R. glutinosa by the CAMAD and the HAD was about 60℃ and about 70℃, respectively, when we consider the catalpol content and benzo[α]pyrene detection in the manufacturing process of drying R. glutinosa.

The objective of this study was to evaluate antioxidant activities of Rehmannia glutinosa (Raw Jihwang) by traditional method. The total phenol content of Rehmannia radix Preparata (the final cycle of Jihwang) was increased to 205%, compared with Rehmannia glutinosa. Antioxidant activities, determined by ferric-reducing antioxidant potential (FRAP), 2,2'-azinobis(3 ethybenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity, 1,1-diphenyl-2-picrydrazyl (DPPH) and hydroxyl radical scavenging activities, increased remarkably as the number of steaming-drying cycles increased. Especially, FRAP value increased 285%. Also, IC50 values for DPPH and hydroxyl radical scavenging activities of the final 9th-cycling product, decreased 48.4% and 76%, respectively, compared with those of Rehmannia glutinosa. Our result was suggested that antioxidant activities of Rehmannia radix Preparata improve according to the increasing number of steaming-drying cycles.

“고강”은 고품질･내병･다수성 지황 개발을 목표로 지황1호 실생 집단에서 우량 개체를 선발, 증식을 거친 후 2001～’02년 생산력 검정시험을 실시하였다. 그 결과 고품질이면서 병해에 강하고수량성이높아수원7호로계통명을부여한후, 2003～’05년까지 3년간 지역적응시험을 실시한 결과 우수성이 인정되어 2005년 12월 직무육성 품종 심의회를 거쳐 “고강”으로 명명하였는바 그 주요특성을 요약하면 다음과 같다. 1. 고강의 초형은 지면에서 약간 솟아오르는 형태를 나타내며, 생육중기 완전 전개된 잎중 가장 어린잎들의 색깔은 안토시아닌 색소를 거의 띠지 않아 대비품종과 구별된다. 2. 고강은 지황1호에 비하여 잎이 크고 많으며, 뿌리가 굵고 길다. 3. 고강은 병에 대한 저항성도 비교적 강하여 생산의 안정성이 높은 품종이다. 4. 고강은 2003～’05년까지 3년간 실시한 지역적응시험결과 수원 등 3지역에서 모두 증수되는 것으로 나타났고, 3지역의 10a당 평균 수량은 1,186kg로 지황1호 대비 13% 증수하였다. 5. 고강은 지황의 주요성분인 catalpol과 엑스함량이 지황1호에 비하여 많은 고품질 품종이다.

Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) are the most important angiogenic molecules associated with tumor-induced neovascularization. This study was carried out to investigate inhibitory effect of extracts from root of Rehmannia glutinosa LIBOSCHITZ (Rehmannia Radix and Rehmannia Radix Preparata) on endothelial cell proliferation. The methanol extracts from the medicinal herb were fractionated into n-hexane, ethyl acetate, n-butanol and aqueous fractions. Among the four fractions, the n-butanol fraction from R. Radix on exhibited highly effective inhibition (≈79% inhibition) on the binding of KDR/Flk-1-Fc to immobilized VEGF165 and then ethyl acetate fraction from R. Radix (≈45% inhibition) at the concentration of 100 μg/ml. The n-butanol fraction efficiently blocked the VEGF- and bFGF-induced HUVEC proliferation in a dose-dependent manner, but did not affect the growth of HT1080 human fibrosarcoma cells. The n-butanol fraction more efficiently blocked the binding of KDR/Flk-1-Fc to immobilized VEGF165 and VEGF- and bFGF-induced human umbilical vein endothelial cell proliferation than the fraction from R. Radix Preparata. Our results suggest that Rehmannia Radix may be used as a candidate for developing anti-angiogenic agent.

Agrobacterium rhizogenes ATCC15834에 의해 형질전환된 지황 (Rehmannia glutinosa)의 모상근으로부터 유식물체를 재분화 시키기 위한 최적 조건을 탐색하였다. 모상근 절편으로부터 유식물체의 재분화는 0.5 mg/l BA가 첨가된 SH배지 (3% sucrose, pH 5.8)에서 배양 5주 후 이루어졌다. 이와 같은 유식물체는 식물생장조절물질이 포함되지 않은 기본배지로 옮겼을 때 발근이 이루어졌다. 형질전환된 모상근 유래 식물체는 잎으로부터 직접 재분화된 식물체와 비교했을 때 잎이 오므라드는 경향을 보였고 줄기는 절간이 짧았으며, 뿌리는 가늘고 수량이 많은 특징을 보였다. 생체량 비교에서 형질전환식물체는 식물 하나의 무게가 1.28 g (생중량)으로 대조구인 잎 유래 재분화식물체 (0.53 g)에 비해 높았으며, 지표물질인 catalpol의 함량에 있어서도 차이를 보여주었다.

Superoxide dismutases (SOD) are metalloenzymes that convert O2- to H2O2. Rehmannia glutinosa is highly tolerant to paraquat-induced oxidative stress. The primary objective of this study was to characterize regulation of SOD gene expression in R. glutinosa in response to oxidative stresses and hormones. A full-length putative SOD clone (RgCu-ZnSOD1) was isolated from the leaf cDNA library of R. glutinosa using an expressed sequence tag clone as a probe. RgCu-ZnSOD1 cDNA is 777 bp in length and contains an open reading frame for a polypeptide consisted of 152 amino acid residues. The deduced amino acid sequence of the clone shows highest sequence similarity to the cytosolic Cu-ZnSODs. The two to three major bands with several minor ones on the Southern blots indicate that RgCu-ZnSOD1 is a member of a small multi-gene family. RgCuZnSOD1 mRNA was constitutively expressed in the leaf, flower and root. The expression of RgCu-ZnSOD1 mRNA was increased about 20% by wounding and paraquat, but decreased over 50% by ethylene and GA3. This result indicates that the RgCu-ZnSOD1 expression is regulated differentially by different stresses and phytohormones at the transcription level. The RgCu-ZnSOD1 sequence and information on its regulation will be useful in investigating the role of SOD in the paraquat tolerance of R. glutinosa.

Ascorbate peroxidase (APX) plays a crucial role in the detoxification of hydrogen peroxide. APX activity is maintained significantly higher in the paraquat­treated leaves of the paraquat-tolerant Rehmannia glutinos. This study was conducted to understand structural and regulatory characteristics of APX gene in R. glutinosa. A putative APX cDNA clone (RgAPX1) was isolated from a leaf cDNA library using a partially sequenced expressed sequence tag clone. RgAPX1 is consisted of 1148 bp nucleotides and contains an open reading frame encoding a 250 amino acid-long polypeptide. Deduced RgAPX1 amino acid sequence shares higher sequence similarity to cytosolic APXs. RgAPX1. expression was higher in the leaf than in the flower and root. Southern blot result indicates the presence of one or two RgAPX1-related genes in R. glutinosa genome. RgAPX1 transcription was affected differentially by various stresses and phytohormone. The results indicate that RgAPXl is constitutively expressed in most tissues and its expression is modulated for the immediate and efficient detoxification of H2O2 under normal and stress conditions.

Resveratrol, which is both a phytoalexin with antifungal activity and a phytochemical associated with reduced cancer risk and reduced cardiovascular disease, is synthesized in a limited number of plant species including peanut. Resveratrol synthesis is catalyzed by the enzyme stilbene synthase including resveratrol synthase (RS). Resveratrol synthase gene (RS3) obtained from peanut, Arachis hypogaea, Fabaceae has been transferred into chinese foxglove, Rehmannia glutinosa by using Agrobacterium mediated transformation. RS t-DNA introduced to chinese foxglove (R. glutinosa L) by transformation and its reaction product, resveratrol-3-O-β-D-glucoside was isolated and characterized using HPLC. Also its biological effects was tested in inhibition of the lipid peroxidation of mouse LDL by glycosylated stilbenes derivatives obtained from transgenic plants. Resveratrol-3-O-β-D-glucoside isolated from transgenic R. glutinosa L. showed antimicrobial activity of the growth inhibition zone against Escherichia coli and Salmonella typhimurium. Therefore, this compound can be contributed to be useful as a phytoalexin for plant health as well as a phytochemical for human health.

Rehmannia glutinosa shows a high level of resistance to the non-selective herbicide paraquat. To characterize the antioxidant enzyme system of R. glutinosa, we comparatively examined the responses of antioxidant enzymes to UV, wounding and a general elicitor yeast extract in R. glutinosa and soybean. The levels of enzyme activities of the two plant species were drastically different between those per fresh weight (general activity) and per protein (specific activity) bases. The general activities of superoxide dismutase (SOD), peroxidase (POX), catalase (CAT), and glutathione reductase (GR) were lower, but that of ascorbate peroxidase (APX) was higher in R. glutinosa than in soybean. The specific activities of the enzymes, however, were about two- to seven-fold higher in R. glutinosa than in soybean, except that of CAT, which was about 12-fold higher in soybean. The general and specific enzyme activities of R. glutinosa relative to those of soybean showed a consistent increase in responses to the stresses only in SOD. The specific activities of SOD and APX were higher in R. glutinosa in all stress treatments. The results might suggest a relatively higher contribution of SOD and APX to the stress tolerance.

지황의 안정적인 저장으로 종근 비용을 절감하고자 재래종과 생육특성이 상이한 지황 1호를 공시하여 저장 후 120일 동안의 저장방법에 따른 종근 특성을 분석한 결과는 다음과 같다. 지황의 저장방법별 온도변화는 움저장에서 7.2∼13.8로 가장 적었고 스치로폼상자내 흙을 충진한 저장에서 높았으며, 습도는 움저장에서 98.2%로 가장 높았다. 당도와 중량 감모율 그리고 부패율은 저장기간이 경과할수록 증가하는 경향을 보였으며, 왕겨를 충진한 망사와 스치

The objectives of this study were to establish the genetic transformation system of stilbene synthase in Rehmannia glutinosa. Resveratrol, which is both a phytoalexin with antifungal activity and a phytochemical associated with reduced cancer risk and reduced cardiovascular disease, is synthesized in a limited number of plant species including peanut. Resveratrol synthesis is catalyzed by the enzyme stilbene synthase including resveratrol synthase (RS). Stilbene synthase gene (RS3) obtained from peanut, Arachis hypogaea, Fabaceae has been transferred into chinese foxglove, Rehmannia glutinosa by using Agrobacterium mediated transformation. PCR analysis with RS3 primer confirmed that the targeted gene was introduced into the plant genome, 904 bp in size. Further analyses of identification of transformation using developed other molecular techniques and transgenic plants that RS t-DNA introduced to chinese foxglove (R. glutinosa L) and its reaction product, stilbene such as resveratrol will be isolate and characterize using NMR, MS, and HPLC.

Using Agrobacterium-me야ated transformation method the auxin-regulated cotton GST (Gh-5) constructs were used to transform Rehmannia glutinosa L. The PCR analysis was conducted to verify transgenicity. Based on the PCR analysis, there was verified that the 988 bp DNA band had showed in transgenic plant genomes in PCR anaJysis using Gh5-1 and Gh5-2 primers. The effects of cocultivation with Agrobacterium tumefaciens, regeneration and selection conditions on the transformation efficiency of Chinese foxglove (Rehmannia glutinosa L.) were investigated. Factors such as cocultivation period, use of acetosyringone, postcultivation in darkness, and different kanamycin concentrations for selection were assessed. In vitro regeneration, the number of leaves, shoot lengths and numbers on MS medium were superior to on B5 and WPM medium, and the shoot formation rate was highest level of 95% in cultured base part containing leaf stalk. Addition of acetosyringone at concentration of 200μM to cocultivation medium and 3-day of cocultivation improved transformation frequencies. Exposure of explants to darkness for 4 weeks on selection medium resulted in further increased the regeneration frequency of transgenic shoots. In PCR analysis, the amplified fragments of Gh5 gene were detected (988 bp), and GST-expressing transgenic R. glutinosa L. plants had approximately three-fold higher activity in leaf extracts compared with control plant.

전라북도의 진안, 장수, 정읍 등의 3개 지역에서 재배하고 있는 지황에 바이러스 병징이 관찰되어 ELISA 방법으로 TMV의 감염 여부를 확인한 결과 3개 지역 모두에서 ELISA 양성 반응을 나타내었다. N. glutinosa를 이용, 순수분리 및 증식한 이병주의 바이러스 입자를 관찰한 결과, 폭 18nm, 길이 300nm의 막대 모양의 바이러스가 관찰되었다. 또한 epoxy수지를 통해 이병조직의 세포질내에서 많은 바이러스 입자가 군집 또는 산재되어 있음이 관찰되었다. TMV의 병징은 즙액 접종 4주 후부터 담배 잎의 본엽에 부분적으로 반점이 보이다가 4­6주 후에는 황화 현상과 괴사 현상이 나타났다. 이병주로부터 total RNA를 분리하여 RT­PCR을 수행한 결과 531bp DNA 증폭 산물을 얻었으며 genetyx win program을 이용하여 외피단백질의 염기수준에서 비교 분석한 결과, TMV­T, V, To 세 계통에서 각각 94.83％로 가장 높게 나타났고, TMV­P계통에서 67.35％로 가장 낮게 나타났다. 아미노산 수준에서는 TMV­T, V, To 세 계통에서 각각 97.65％로 가장 높게 나타났고, TMV­P계통에서 72.22％로 가장 낮게 나타났다. 기타 계통들간에는 TMV­152, F 계통에서 94.05％, TMV­C 계통에서 68.63％ 유사도를 나타냈다.