Background : Cudrania tricuspidata Bureau is a widely used medicinal perennial woody plant. Obtaining information about the genetic diversity of plant populations is highly important for conservation and germplasm utilization. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific ecotypes of C. tricuspidata via amplification refractory mutation system (ARMS)-PCR analyses. We performed molecular authentication of twelve C. tricuspidata ecotypes from different regions using DNA sequences in the chloroplast TrnL-F intergenic region. Methods and Results : SNPs were identified based on the results of nucleotide sequence for the intergenic region of TrnL-TrnF gene (chloroplast). Molecular markers were designed for those SNPs with additional mutations on the second base from SNPs for amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). HRM pattern analyses were performed using the Mx3005P QPCR System (Agilent Technologies, CA, USA). Conclusion : We collected 12 individual lines of C. tricuspidata from various region in South Korea and China. Based on the nucleotide sequence in the trnL-trnF intergenic region of these lines, six SNPs and a deletion of 12 bps were identified and 12 individual lines were able to be grouped in one Korean ecotype and two different ecotypes of chinese lines, chinese line 1 and 2. The SNP markers developed in this study are useful for rapidly identifying these specific C. tricuspidata ecotypes collected from different regions.

The present study was carried out to identify traditional konwledges on Korean Campanulaceae plants and conduct a comprehensive review of them through analyzing phytochemistry and pharmacology of Korean Campanulaceae plants. According to the literature study, the ethnobotanical plants of Korean Campanulacae consisted of a total 18 taxa. Of them, 12 taxa including Platycodon grandiflorus, Adenophora triphylla var. japonica, Codonopsis lanceolata and others have been used as ethnomedicinal plants. These plants have been used for the treatment of 49 diseases such as cold, asthma and postnatal care. Phytochemical studies have identified the constituents present from Korean Campanulaceae (Adenophora, Codonopsis, Platycodon, Campanula and Asyneuma). A wide range of chemical compounds comprised 109 triterpenes, 8 sterols, 4 polyacetylenes, 21 alkaloids, 14 flavonoids, 14 phenolic acids, 11 phenolic glycosides, 8 phenylpropanoids and 22 other compounds. Pharmacological studies of these compounds have demonstrated immuno-stimulating, anti-inflammatory, anti-asthmatic, apophlegmatic and anti-allergic effects. They have also shown antioxidant, estrogenic, anti-diabetic, hepatoprotective, neuroprotective, antinociception and anti-tumor activities, as well as anti-obesity and cardiovascular effects. In light of traditional knowledge and phytochemical and pharmacological studies summarized, uses of Korean Campanulaceae based on traditional knowledge (for the treatment diseases and conditions of respiratory, pregnancy, childbirth, puerperium, genitourinary, circulatory, musculoskeletal and other systems) have been supported by phytochemical and pharmacological studies.

Leptalina unicolor is a hesperiid butterfly with a restricted distribution in Korea, Japan, China, and Russia. Recently, the number of individuals is rapidly decreased in their natural habitat. This skipper has been classified as an endangered species in the ‘Red Data Book’, and according to an IUCN report, is under threat of extinction. Therefore, the conservation and restoration of this species both locally and globally are urgently needed. A population of L. unicolor was found in an unknown habitat in Jaeyaksan, Miryang, Gyeongsangnam-do, Korea in 2011, and these individuals were used in the present study. Here, the life cycle, characteristics of each instar larva and breeding information were determined by breeding L. unicolor in the laboratory from 2012 to 2015. The results indicated that L. unicolor occurs twice a year and over-winter as a fifth instar larva. A springform female individual laid 17.25±5.52 eggs, and summer form laid 29.00±5.86 eggs; it takes 53.79±0.73 days for L. unicolor to develop from eggs to adults. After spawning, the eggs developed in 6.16±0.18 days, and larvae developed in 33.71±0.58 days; the pre-pupal stage to emergence required 14.22±0.31 days. Based on these results, we presented effective breeding information for the restoration and proliferation of the species. Several candidate plants species have been reported in the literature as a food source for L. unicolor, but we found that the preferred diet this butterfly was Miscanthus sinensis. The larvae could move easily when their preferred diet was planted in 80% the cage floor area Year-round breeding was achieved by placing overwintering individuals in low temperatures in autumn. To our knowledge, this is the first study to the biology of L. unicolor in Korea, and the results of this study could be used as preliminary information for the conservation and restoration of this species in its natural habitat.

The genus Tilia is characterized by linear form bracts of which the lower part is attached to the peduncle of a cyme. This character is distinguished from the others genus of Malvaceae. The purpose of this study is verifying the phylogenetic relationship of genus Tilia. Phylogenetic analyses were conducted to evaluate relationships of 10 taxa of Tilia in Korea and Japan including one outgroup (Gossypium hirsutum). The molecular phylogenetic analyses were conducted with sequences based on ITS, trnL-F and rpl32-trnL region. The combined data result of ITS, trnL-F and rpl32-trnL was formed by 6 clades. T. kiusiana situated as the most basal clade. T. amurensis, T. taquetii and T. rufa are composed a clade. T. koreana, T. insularis and T. japonica was formed independent clade. T. insularis has the closest relationship with T. japonica. T. miqueliana, T. mandshurica, and T. megaphylla are composed a clade and showed a sister relationship than other species.

The purpose of this study was to determine the optimum re-extraction conditions of ethanol from catechin in Korean green tea (Camellia sinensis L.) using response surface methodology (RSM). The experiments were carried out according to a five level and two variable central composite design (CCD). The two independent variables were solvent ratio to sample content (1, 4, 7, 10, 13 mL/g) and extraction temperature (-20, -10, 0, 10, 20℃) on the dependent variables including yield, epigallocatechin (EGC), epicathchin (EC), epigallocatechingallate (EGCG), epicatechingallate (ECG), total catechin and caffeine. ANOVA results showed that Coefficients of determination (R2) of estimated models for dependent variables were ranged from 0.9054~0.9778, while R2 of caffeine were estimated 0.8770. The optimum ranges for the maximized extraction including yield, EGC, EC, EGCG, ECG, caffeine and total catechin were 4.5~7.5 mL/g in ratio of ethanol to sample and -8~8℃ in extraction temperature. The actual values of yield, EGC, EC, EGCG, ECG, caffeine and total catechin, respectively, at the optimized conditions were 35.02%, 13.31%, 3.978%, 19.11%, 4.29%, 5.30% and 40.68%

Background : Korean Indigenous Hylotelephium erythrostictum is widely distributed in South Korea and is used in Korean traditional medicine. In this study, the phylogenetic analysis of Korean native Hylotelephium erythrostictum and related plants on Internal Transcribed Spacer (ITS) sequences were investigated to distinguish its origin. Methods and Results : The phylogenetic analysis of 6 species of Hylotelephium were investigated by ITS. The dendrogram was constructed by UPGMA(Unweighted Pair Group Method with Arithmetic Mean) clustering algorithm based on genetic similarity of ITS. In the ITS sequence analysis, the size of total was varied from 676 to 779 bp. The size of ITS 1 was rated at 287bp, while ITS 2 was rated at 123bp. The G+C content of ITS region was ranged from 60 to 66%. In the ITS tree, six species of Hylotelephium were monophyletic, and H. viviparum was the first branching within the clade. Conclusion : H. spectabile formed a clade with H. erythrostictum, while H. verticillatum formed with H. viviparum.

Background : Ixeris dentata is the perennial herbaceous plant in the Ixeris genera within the Compositae family. The whole plant has been used traditionally as herbal medicine. It is widely distributed in South Korea and the genetic difference among the plants harvested from different regions may differ due to the disparity in cultivation climate. Therefore, this research was performed to discriminate the I. dentata that are collected from four locations in South Korea based on sequence analysis of nrDNA-ITS region. Methods and Results : Genomic DNA was extracted from I. dentata obtained from Goesan, Dangjin, Yangpyeong, and Chuncheon. I. stonolifera was included for a comparison of genetic distance with I. dentata. PCR amplification was performed by using an universal barcode nrDNA-ITS primer for DNA barcoding. After sequencing, the data was aligned using ClustalW multiple alignment tool in BioEdit version 7.2.5 software. SNP and phylogenetic analysis were conducted with the MEGA7 program. Phylogenetic analysis was presented with Neighbor-joining method using the K2P model. Statistical significance are evaluated using bootstrap (1,000 replicates). PCR products were amplified with about 800bp length for the ITS1-4 sequences of all samples. Nineteen SNPs were detected within a 578bp fragment of the aligned sequences. The mean GC content was 51.88% for ITS1-4 sequences of them. The interspecific genetic distance between I. dentata and I. stonolifera was 0.029%. The highest region-specific distance was confirmed to 0.010% between the plants from Dangjin with Goesan and Chuncheon group. Meanwhile, the mean intraspecific distance among the plants from Yangpyeong was 0.002%. Conclusion : In the phylogenetic analysis, the plants from Goesan and Chuncheon were placed within the same clade, while the plants from Dangjin formed independent clade. On the other hand, the plants from Yangpyeong were not grouped into one clade followed by the intraspecies variation. In conclusion, the data from the research will be useful for the regional identification of Dangjin, even if it is required to perform some additional researches.

In this study, the phylogenetic relationship of Korean Hydrangea was evaluated by using sequenced three chloroplast regions and ITS region, including the 7 taxa. The result of phylogenetic analysis indicated that Korean Hydrangea, 7 taxa formed the monophyletic group. This analysis also revealed that subsect. Macrophyllae of Korea was separated into two groups; H. serrata f. acuminate and H. macrophylla group. The H. serrata f. acuminta group was included with H. serrata f. buergeri and H. serrata f. fertilis. These three species form a monophyletic clade, with no significant differences between their nucleotide sequences. The H. serrata f. acuminta group showed a monophyletic group with H. serrata f. buergeri and H. serrata f. fertilis and there is significant differences between their nucleotide sequences. H. macrophylla group was an independent clade distinguished by H. serrate f. acuminate group. Subsect. Petalanthe, Heteromallae and Calyptranthae form a monophyletic group. H. petiolaris which is located in Subsect. Calyptranthae was separated into two subgroups; First subgroup: Jeju island (except for Mt. Halla) and Second subgroup: Ulleung island and Japan. Additional studies of two subgroups of H. petiolaris should be conducted a geographical study and add more samples.

Background : With the increasing demand of the mistletoe in larger quantities for cancer therapy, it has been depleted from its natural habitat in the Far East countries including Korea because of overharvesting for high-value products (e.g., lectins and viscotoxins). The rapid multiplication of mistletoe by tissue culture can help this problem and provide the benefits in the phamaceutical industry. Methods and Results : Mistletoe plants growing on oak trees were collected and their leaf and stem segments were inoculated on MS basal medium supplemented with various concentrations of growth regulators. Calli were induced only from stem explants on MS medium containing 2,4-D and transferred onto MS medium supplemented with various concentrations of 2,4-D, NAA and BAP, respectively, for their propagation. The best callus multiplication rate of more than 15 folds (759 mg) was obtained in treatment of 2,4-D (4 mg/L) that produced yellowish-green, white and friable callus on this medium. To compare biochemical characterization, lectins were partially purified from natural mistletoe plants (nML) and in vitro cultured mistletoe calli (cML), respectively. The former was purified by lactose-agarose affinity chromatography and the latter was done by ammonium sulfate precipitation followed Sephadex G-25 chromatography. Both nML and cML showed similar molecular weight on SDS-PAGE and Western blot analyses. In addition, they showed similar carbohydrate-binding specificities and hemagglutination activities. Conclusion : From the above results, we may suggest that nML and cML showed the similarity in biochemical characters.

Background : Lectins were individually isolated from natural Korean mistletoe (nML) and its in vitro cultured calli (cML). Both of the lectins showed the difference in bioactivities such as cytotoxicity and cytokine induction. Methods and Results : Target cells (1 x 104 cells/well) were seeded independently into each well of a 96-well culture plate and incubated with different concentrations of each lectin. Survivability of target cells was determined by CCK-8 kit (Sigma, USA) according to manufacturer’s directions. The nML showed 46, 34 and 5.5 times stronger than cML in cytotoxicity (IC50) to human melanoma cell line (SK-MEL-28), human carcinoma cell line (NCI-H1650) and murine macrophage (RAW 264.7), respectively. In addition, respective lectins directly stimulated macrophage RAW 264.7 but they showed the difference in enhanced productivity of some inflammatory cytokines. Compared with cML, the nML induced both TNF-α and IL-1β at its low concentration. Administration of two kinds of lectins (10-1000 μ g/kg body weight) to ICR mouse did not show any significant changes on the level of alanine transaminase (ALT), glutamate-oxaloacetic transaminase (GOT) and blood urea nitrogen (BUN) in sera. Conclusion : From the above results, we may suggest that nML and cML showed differences in cytotoxic effects and cytokine production due to the difference in amounts from sources.

Background : Dibenzocyclooctadiene lignans are secondary metabolites present abundantly in the fruits belonging to the genus Schisandra. According to previous studies, Schisandra lignans exhibit anti-inflammatory, anti-cancer and anti-diabetic properties, as well as an inhibitory effect on platelet aggregation. Therefore, establishing the Korean “Omija” (Schisandra chinensis) as a lignanrich source, in addition to identifying and quantifying the lignans, is extremely valuable. Methods and Results : Dibenzocyclooctadiene lignans were analyzed with liquid chromatography using diode array detection/ mass spectrometry, from methanol extracts subsequently identified by a constructed chemical library of 50 lignans. A total of 27 components of lignan including gomisin S were identified, of which schisandrin, gomisin A, gomisin N, deoxyschisandrin, γ- schisandrin, and schisandrin C were identified as the major components in the Korean Omija, Schisandra chinensis. These compounds were divided into two groups, S-biphenyl and R-biphenyl based on the configurations of the stereoisomers structures with contents of 661.7 and 1350.1㎎ per 100 g dry weight, respectively. The total lignan content averaged 2011.4㎎ per 100 g dry weight, of which schisandrin and gomisin N comprised the majority (771.8 and 420.5㎎ per 100 g dry weight respectively). Conclusions : Lignans which are present in high quantities in the ripe fruit of Schisandra chinensis are important functional compounds that play a major role in the prevention and treatment of human diseases.

Polygonatum is a genus placed in the family Liliaceae, distributed throughout the Northern Hemisphere and 16 of the species are grown naturally in Korea. In oriental medicine, the rhizomes of Polygonatum have been used as two different medicines, Okjuk (Polygonati odorati Rhizoma) and Hwangjeong (Polygonati Rhizoma). However, it is difficult to identify the morphological and chemical differences between the medicinal groups and thus easy to confuse the one with the other. Therefore, a clear classification standard needs to be established so as to be able to discriminate between them. In the study, the morphological characteristics of the plants, Polygonatum spp., were examined. Then, the differences in SNPs among the DNA sequences of 7 of the Polygonatum spp. and 1 of the Disporum spp. were analyzed by DNA barcoding with rpoC1, rpoB2, matK, and psbA-trnH of the cpDNA region. In the results, three regions, rpoC1, rpoB2, and matK were useful for discriminating the species, P. stenophyllum and P. sibiricum. Furthermore, it was possible to discriminate the individual germplasm within the species by using the combination of the results obtained from rpoB2, rpoC1, and matK.

This study was conducted to observe the influences of Crataegus pinnatifida BUNGE extract on lipid concentration in hypercholesterolemic rats (Sprague Dawley, male) fed the experimental diets for 5 weeks. Body weight gain was remarkably lower in the Crataegus pinnatifida BUNGE extract intake group than in the hypercholesterolemic rats. Total cholesterol concentration was remarkably lower in the extract of Crataegus pinnatifida BUNGE supplement in hypercholesterolemic rats than in the hypercholesterolemic model. Whereas, concentrations of HDL-cholesterol, ratio of HDL-cholesterol concentration to total cholesterol were remarkably higher in the extract of Crataegus pinnatifida BUNGE supplement in hypercholesterolemic rats than in the hypercholesterolemic model. Atherosclerotic index (AI), cardiac risk factor (CRF) were remarkably lower in the extract of Crataegus pinnatifida BUNGE supplement in hypercholesterolemic rats than in the hypercholesterolemic model. From the above research, Crataegus pinnatifida BUNGE extract was effective on the lipid concentrations in sera of dietary hypercholesterolemic rats.

본 연구에서는 한국산 봄배추와 일본산 봄배추의 품질특 성과 기능성을 살펴보고, 이 배추를 이용하여 김치를 제조 한 후 5℃에서 4주 동안 발효하여 이화학적 특성 및in vitro 건강 기능성의 차이를 살펴보았다. 품질특성으로 수분함량, pH, 산도, 총균수, Lactobacillus sp.의 수, Leuconostoc sp.의 수, 조직감, 관능평가를 실시하였으며 적숙기에서의 항산화, in vitro 항암 기능성 실험을 하였다. 일본산 배추는 97.1%의 수분을 함유하였고 한국산 배추는 92.4%의 수분 을 함유한 것으로 나타나 일본산배추가 한국산 배추에 비해 높은 수분을 함유하고 있었다. 생배추와 절임배추에서 한 국산 배추가 일본산 배추보다 탄력성이 높았고 4주간 숙성 한 김치의 탄력성은 한국산 김치가 53.5%로, 일본산 김치 (41.4%)보다 월등히 높은 것을 확인할 수 있었다. 한국산 배추로 제조하여 발효한 김치는 총균수는 낮으면서 젖산균 인 Lactobacillus sp.와 Leuconostoc sp.의 수는 높게 나타났 다. 관능평가에서 한국산 배추로 제조한 김치의 관능적 만 족도가 높게 평가되었다. 발효 3주차의 김치를 이용하여 DPPH radical 소거능을 측정한 결과 한국산 배추로 제조한 김치는 83.2%, 일본산 배추로 제조한 김치는 46.1%로 한국 산 배추로 제조한 김치에서 약 2배 정도 높은 DPPH radical 소거효과를 나타냈다. AGS 인체 위암세포에 김치의 메탄 올 추출물을 1 mg/mL의 농도로 처리했을 때 한국산 배추로 제조한 김치는 45%, 일본산 배추로 제조한 김치는 26%의 암세포 성장 저해율을 나타냈고, 2 mg/mL의 농도에서는 각각 97%, 74%의 암세포 성장 저해율을 나타내어 한국산 배추로 제조한 김치에서 저해 효과가 높은 것으로 나타났 다. 결국 한국산 배추의 품질이 우수하고 이로 제조한 김치 가 일본산 배추로 제조한 김치보다 품질이 우수하고 더 높은 항산화 및 항암기능성(AGS 위암세포)을 나타내었다.

This study was conducted to obtain the basic data for using the Artemisia anuua as a new economic crop,thus Artemisia anuua was investigated their cultivation characteristics, yield, and variation of artemisinin contents by plantingdensity and harvesting times. Seed characteristics of A. anuua have observed micro-size, and their germination optimumtemperature was at 15 to 20 celsius degree. Planting density on the yield of A. anuua was increased high density better thanlow density. The highest yield was planted in the space of 30×10㎝. Moreover, optimum harvesting time of A. anuua wasinvestigated in early september and a periods of most highly detected artemisinin was time of before and after bloomingof A. anuua.Key Words : Art