Background : Gentinae Macrophyllae Radix is one of the traditional medicines originated from the roots of multiple plants, Gentiana macrophylla Pall., Gentiana straminea Maxim., Gentiana crassicaulis Duthie ex Burkill and Gentiana dahurica Fisch., in Gentianaceae. Multi-origin traditional medicine usually has adulteration problem based on the morphological similarity and/or misunderstanding the species. Therefore, accurate and reliable identification criteria to ensure drug safety and quality is necessary. Methods and Results : We collected four original species of Gentinae Macrophyllae Radix from plantations and markets in China and Korea. DNA barcoding with four barcoding markers (Internal Transcribed Spacer (ITS), rbcL, trnL intron, trnL-F intergenic sapcer) was performed. Intra-specific variation was observed in ITS nucleotide sequence however, successfully distinct four original species based on the nucleotide discrepancy while trnL intron has no difference. trnL-F intergenic spacer has two transitions(T→C and A→G) sites only in G. crassicaulis and rbcL shows one transition(C→T) site in G. dahurica and G. macrophylla. Phylogenetic relationship analysis of the Gentinae Macrophyllae Radix revealed two major clades – clade I including three groups, G. macrophylla, G. straminea and G. dahurica, and clade II including G. crassicaulis. This aspects was shown more clear with multi-region combined analysis. Conclusion : DNA barcoding will be accurate and powerful criteria for the analysis the origin of Gentinae Macrophyllae Radix. However, single region analysis might be deficient such as trnL intron, rbcL and trnL-F intergenic spacer results in this research. Multi-region combined analysis based on the multiregional DNA barcode markers will be overcome the disadvantage and also increase the precision.

Background : Compositae is one of the largest plant families which has high probability of diversity and mutation. Though the various researches about the Compositae are ongoing, it is incomplete and need to be conducted the molecular genetic researches to back up the previous studies. This research was performed to identify the genetic divergence of Compositae plants based on the DNA barcoding for cpDNA-matK and rbcL regions. Methods and Results : For this studies, the genetic sequence analysis (SNP/InDel) and phylogenetic analysis were conducted by using Neighbor-Joining algorithm as targeting the nineteen specimens from 7 species which received a IT number along with NCBI Genbank database (http://ncbi.nlm.nih.gov) sequences. The result of matK sequence analysis, 68 SNP and 2 InDel regions (at nt 527-538bp and 695-706bp positions) were confirmed. Also 10 SNPs were found in rbcL region. The genetic divergence showed 0.000-0.059% in matK regions, and the mean was 0.024%. The highest distance were observed between Ligualria fischeri and the group composed with Aster tataricus and Solidago virgauria (2 and 3). The sequence divergence for rbcL regions showed 0.000-0.018%, and the mean was 0.005%. The highest sequence distance in rbcL region were observed between L. fischeri group and S. virgauria (HE574593). In result of phylogenetic analysis in matK region, the most species formed independent clade. A. tataricus in Aster genus and two samples of S. virguaria in Solidago genus were formed one same clade. S. virguaria(1) and A. spathulifolius(2) has been separated into independently for the plants belonging to same genus, respectively. A. spathulifolius showed differences with NCBI data. The rbcL formed one same clade except L. fischeri and Synurus deltoides. Conclusion : This study indicates that matK is more valuable than rbcL for the distinction among the species of Compositae. This results are expected to be used for the establishment of the classification system of Compositae as well as for the studies in the development of an authentication marker.

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 : Both Panax ginseng Meyer and Panax quinquefolius are obligate shade-loving plants whose natural habitats are broadleaved forests of Eastern Asia and North America. Panax species are easily damaged by photoinhibition when they are exposed to high temperatures or insufficient shade. In this study, a cytohistological study of the leaf structures of two of the most well-known Panax species was performed to better understand the physiological processes that limit photosynthesis. Leaves of ginseng plants grown in soil and hydroponic culture were sectioned for analysis. Methods and Results : Leaf structures of both Panax species were observed using a light microscope, scanning electron microscope, and transmission electron microscope. The mesostructure of both P. ginseng and P. quinquefolius frequently had one layer of noncylindrical palisade cells and three or four layers of spongy parenchymal cells. P. quinquefolius contained a similar number of stomata in the abaxial leaf surface but more tightly appressed enlarged grana stacks than P. ginseng. The adaxial surface of the epidermis in P. quinquefolius showed cuticle ridges with a pattern similar to that of P. ginseng. Conclusion : The anatomical leaf structure of both P. ginseng and P. quinquefolius shows that they are typical shade-loving sciophytes. Slight differences in chloroplast structure suggests that the two different species can be authenticated using transmission electron microscopy images, and light-resistant cultivar breeding can be performed via controlling photosynthesis efficiency.

Background : Angelica dahurica is the dried root of Angelica dahurica Bentham et Hooker f., a perennial plant belonging to the Umbelliferae family. Since its peak at 63 hectares in 2000, the cultivated area of Angelica dahurica dropped to its lowest level at 14 hectares in 2014 although there had been a short bounce back to 59 hectares in 2007. The decrease in the cultivated area has created a vicious cycle where even a small change in production amounts causes Angelica dahurica prices to fluctuate, leading farmers to become reluctant to cultivate the species. Therefore, this study aims to provide growers with new information on Angelica dahurica cultivation technology development, based on surveys and analyses of the difficulties and needs the farmers have. Methods and Results : Setting the nationwide Angelica dahurica farming households as the population, this study looked into 11 households of growers from main producing areas including Yeongju, Yeongcheon, and Bonghwa. Researchers carried out the survey by visiting the farmers and conducting face-to-face interviews using a prepared questionnaire. The survey was conducted for 2 months (from July to August 2016) to identify difficulties and specific characteristics of farming Angelica dahurica. SAS 9.2 was used for the statistical analysis. Conclusion : The surveyed types of difficulties identified were varieties, seeding, fertilization, pests, water management, weeding, soil, and drying. The results indicate that seeding (2.2) is the most serious difficulty, and water management (2.5), weeding (3.1), fertilization (3.7), pests (3.9) and others followed. Regarding specific technical problems, browning roots in the process of drying (61.5%) was the biggest concern along with germination after seeding (23.1%) and water management in the fields (15.4%). In response to the question about their willingness to expand farming in the future, no households showed intentions of expansion, while 36.4% reported intentions to reduce and 63.6% to maintain their current Angelica dahurica cultivation. The causes of weakening supply include low prices (54.5%), unreasonable pricing (36.4%), and difficulties in selling due to lack of retailers. Therefore, it seems necessary for the government to support sustainable Angelica dahurica farming by taking measures such as contract farming with herbal medicine circles, a minimum production cost guarantee system, or joint purchasing by the Agricultural Cooperative Association.

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 : In previous studies, adlay seeds showed a prevalence of diversified fungal flora with the predominant fungal genera being Fusarium (45.6%) and In vitro test showed that fungal toxins like Fumonisin and Zearalenone were produced by Fusarium fujikuroi, F. asiaticum and F. graminearum. Because of this we performed experiments to selecting disinfective chemicals for controlling the Fusarium contamination in the adlay seed. Methods and Results : We carried out the chemical efficacy test such as seed disinfectants selection test appling before planting and pesticides selection test using in the earing season. In the present study, eleven different commercially available seed disinfectant were applied to the adlay seeds. Among 11 seed disinfectants, Hexaconazole+Prochloraz emulsifiable concentrate (EC) and Prochloraz emulsifiable concentrate (EC) had control value of 80% or above against Fusarium species tested. In the pesticides selection test, seven different commercially available pesticides for Fusarium blight (Scab) control were applied and we observed that Metconazole suspension concentrate(SC) strongly inhibited the mycelial growth of 10 Fusarium species all. Conclusion : From the above results, we selected Hexaconazole+Prochloraz EC and Prochloraz EC as a seed disinfectants and Metconazole SC as a pesticide using in the earing season for Scab control.

Background : Ginseng rain cover farming is expanding around Jinan county and Jangsu country of North Jeolla Province. Some farmers doing ginseng rain cover farming have suffered from difficulties due to hot weather damages. However, it is a situation that the study on mitigation techniques for high temperature damage do not exist with ginseng rain cover farming. Methods and Results : The test covering work was firstly done on April 28th for heat block film+90% black light blocking net, blue double sided film, and PE film+75% black light blocking net and when it comes to second treatment, 30% and 40% shading were implemented for heat block film group and blue double sided film group respectively and 75% black light blocking net was installed on PE film+75% shading group. When it comes to micro-climate measurement in rain cover facility, temperature, humidity and light intensity were measured during the growing period of ginseng. The results are as follows. Regarding the light transmittance (per PAR, 10 am in clear day) in facility with 1st covering, light block film covered group (LBF), blue double sided film group (BDF) and PE film group have 12.9±1.8%, has 11.6±1.0% and 27.1±1.1% respectively and after 2nd covering, in LBF groups, 30% blocking, 40% blcoking and no blocking have 10.6±1.3%, 8.2±0.9% and 12.9±1.8% and in BDF groups, 30% blocking, 40% blcoking and no blocking have 9.4±0.8%, 7.9±0.7% and 11.6±1.0 respectively and PE film group has 10.6±0.7%. Relative humidity also showed the same trend as temperature. The average monthly amount of light and maximum light intensity were lower in 30% and 40% light blocking groups of LBF and BDF and a little higher in no light blocking group compared to PE film group. The degree of high temperature damage was 1 in no LBF of BDF, but no LBF of LBF was so bad like 3. However, there was no high temperature damage in the test groups of blocking films or BDF with 30% and 40% light blocked light screens. Regarding root weight, all secondly treated groups of LBF group and BDF group were lighter compared to 4.36g of PE film group and especially, prism sheetof no light blocking group has 2.5g and BDF of of no light blocking group has 3.21g. 30 % and 40% light blocking groups of LBF group and BDF group were light with 3.20~4.07g. Conclusion : Regarding the analysis result on micro-climate in facility with different cover materials for 2 years old ginseng in ginseng rain cover farming of Gyeonggi Province, the covering method suitable for high temperature damage mitigation in ginseng rain cover farming was that 1st covering was done by PE film+75% black light blocking net and then 75% black light blocking net is additionally covered at a time when outside temperatures reaches 30℃.

Background : Ginseng (Panax ginseng C.A. Meyer) is one of the most important medicinal plants in Korea, but its yields are often reduced by a variety of root pathogens. The root rot of ginseng is a destructive soil-borne disease caused by Cylindrocarpon destructans (teleomorph: Ilyonectria radicicola). To monitor contamination with C. destructans in ginseng harvested in 2015 were sampled from 57 different growing fields. The spore number of C. destructans was quantified by use of a specific primers and selective media (radicicol) in soils of ginseng fields. Methods and Results : The ginseng samples were surface-sterilized and placed on potato dextrose agar plates for 7 day incubation at 20℃. Emerging fungal colonies were counted primarily based on colony and conidia morphology. Further species level identification was confirmed by ITS rDNA sequencing. For quantification of the soil-borne C. destructans, the genomic DNA was extracted from the soil using a NucleoSpin soil kit (MN, Germany). Density of C. destructans was determined by species specific real time PCR (qPCR). The qPCR was completed by running a melting curve analysis. Conclusion : The C. destructans associated with root rot disease of ginseng were detected in more than 60% in pyeongtaek-1, pochenon-1, jecheon-1, chungju-1 and jinan-4. As results of the study, the correlation between pathogen density and identification clearly clarified in the soil.

Background : Ginseng (Panax ginseng C. A. Meyer) is a precious herb plant belonging to Araliaceae family especially in Asia and it has been cultivated more than a thousand years as a traditional medicine. Due to their pharmacological efficacy, old ginseng plants are traded high price, however, there are no crucial criteria to determine the ginseng age. To prevent illegal transactions, we assessed the telomere of ginseng roots based on modifications of the assays reported previously. Methods and Results : It is known that telomere length of ginseng root is shorter upon organismal aging. In this study, to support the determination of ginseng age, we modified and investigated methods through telomere analysis. Firstly, we examined the southern blot analysis whether telomere length depends on ginseng age. Based on previous study, we measured telomerase activity that is correlate with age. Furthermore, telomeric DNA was quantified by fluorescence in situ hybridization (FISH) to corroborate telomere shortening. The older ginseng root was shown short telomere length to compare with younger ginseng roots. Also enzyme activity of telomerase and amount of telomeric DNA represented decrease patterns upon age. Conclusion : Taken together, it is help to determine the age of ginseng through various methods using telomere because the results shown to positive correlation between telomeric characteristics and age for ginseng.

Background : Ginseng is a perennial, and damage occurs due to various diseases and pests. As a result, yield decreases and quality deteriorates. In particular, it is impossible to be repeatedly cultivated due to root rot, soil-borne disease. For this, in order to control root rot and repeatedly cultivate ginseng by using soil fumigation from virgin soil, this experiment was conducted. Methods and Results : This experiment was performed in ginseng farm field, Geumsan-gun, Chungnam province. And total area was 3ha. And the experiment was conducted in soil where apple trees had been grown. Apple trees were rooted up in 2006. And soil was fumigated in May 2007. With regard to soil fumigant, soil was treated with Basamid of 40kg/10a. After soil fumigation, 5 ton/10a of cattle manure fermented for 1 year was put in the soil in June, and then the soil was cared for by being plowed and rotary-tilled as occasion. Ginseng seeds were sowed in the soil cared for in October 2007. And 4-year-old ginseng was harvested in 2011. The soil from which ginseng was harvested was fumigated by the same method in May 2012, and then ginseng seeds were sowed in 1.5ha in October 2012 and in 1.5ha in October 2013. In October 2013, 3-year-old and 4-year-old ginseng was harvested. And ginseng growth characteristics and root rot incidence was examined. It was shown that 4-year-old ginseng yield after the fumigation of virgin soil was nearly 2 times as high as that of conventional virgin soil cultivation. And root rot didn't occur. After that time, the soil was fumigated again. And when ginseng was cultivated, 3-year-old and 4-year-old ginseng yield was 650kg and 960kg per 10a. And it was shown that root rot incidence was 1.3% and 15.3% in 3-year-old and 4-year-old ginseng respectively. Conclusion : This study showed the results where ginseng can be cultivated repeatedly if soil was fumigated again after ginseng was harvested following the soil fumigation from virgin soil so as to control the ginseng root rot and cultivate ginseng repeatedly.

Background : Ixeris genus has been used in traditional medicines as stomachics, sedatives, and diuretics. Ixeris dentata var albiflora is a kind of perennial herbaceous plant and one of the plants of the genus Ixeris (Asteraceae). It is well-known for edible wild vegetable in Korea, China, Japan, and Mongolia. Specially, Korean has its root and young leaf with appetizing vegetable due to bitter taste. Methods and Results : We isolated 8 genes that are involved in carotenoid biosynthesis using the Illumina/Solexa HiSeq2000 platform. In this study, a full-length cDNA clone encoding phytoene synthase (IdPSY), phytoene desaturase (IdPDS), ξ-carotene desaturase (IdZDS), lycopene β-cyclase (IdLCYB), and zeaxanthin epoxidase (IdZEP) and partial-length cDNA clones encoding lycopene ε-cyclase (IdLCYE), ε-ring carotene hydroxylase (IdCHXE), and β-ring carotene hydroxylase (IdCHXB2) were identified in I. dentata. The theoretical molecular weight (MW) and isoelectric point values of 8 genes were investigated. Sequence analyses revealed that these proteins shared high identity and conserved domains with their orthologous genes. IdPSY, IdPDS, IdZDS, IdLCYB, IdCHXB2, and IdZEP were constitutively expressed in the roots, stems, leaves, and flowers of I. dentata. Conclusion : Our study on the biosynthesis of carotenoids in I. dentata will provide basic data for elucidating the contribution of carotenoids to the considerable medicinal properties of I. dentata.

Background : Ginseng widely cultivated as a major medicinal herb in Korea, is economically important crop for farmer. Ginseng root disease caused by soil borne pathogens is main factors restricting the quantity and quality of ginseng. The disease can result in harvest loss of up to 20~70% and limits the replanting of ginseng under same field for long time. The traditional control method of agrochemical use is not recommend to control soil borne disease because of difficulty in use and unstable effect. The objective of this study was to evaluate the efficacy of several antagonistic microbes for developing biological control method of ginseng root rot. Methods and Results : To select biocontrol agents against ginseng soil borne disease, several bacteria were isolated from ginseng root and rhizosphere soil evaluated in vitro screening of antifungal bacterial against ginseng root pathogens. Two antagonistic bacteria, ES17 and CJ4, showed the strongest inhibition effect against ginseng root pathogen. In the pot experiment under greenhouse conditions, ginseng seedling dipped in bacterial suspension at inoculum density of 106 cfu/ml for 1 hour were planted in pot containing inoculum. Control effect was examined depend on disease severity index at 30 days after inoculation. Ginseng root treated with CJ4 and ES17 isolate reduced root rot disease development on the ginseng root with degrees of control efficacy of 85% and 70%, respectively. Conclusion : Two biocontrol agent, Burkholderia ambifaria CJ4 and Paenibacillus strain ES17, had strong antifungal efficacy against ginseng soil borne pathogens. These results obtained from in vitro test and pot experiment suggest the potential applicability of the biocontrol agent to control ginseng root rot caused by various soil borne pathogens.

Background : Most of cases in korea, distributed herbal medications are storaged in room temperature and it causes many insect pests during storage. Despite the fact, there is little research about storage conditions. Temperature and humidity in storehouse are highly related with pests’ overwintering, generation length and the number of occurrence in summer. Consequently, in this research, major pests’ overwintering was studied by reviewing facility condition, temperature and humidity in major regional storehouses. The results of this would help to develop how to manage pest occurrence in distributed herbal medications. Methods and Results : For this study, storage facilities was investigated at different regions including Gyeongdong Market in Seoul, the pharmaceutical company in Daegu and farmhouse in Youngcheon by monitering temperature, humidity, ventilator, air conditioning and heating on ground and underground places from June 2014 to July 2016. including Upon investigation, most of building using for storage had simple ventilators. Whereas in case of farmhouse, it is vulnerable to maintaining quality. Because most farmhouse had temporary buildings which are difficult to control temperature and humidity. In addition, among the other surveyed warehouses, Basement in the Gyeongdong Market had most favorable condition for pest overwintering. The lowest temperature of it was seldom below zero maintaining level of about 1.5℃ and average temperature was about 10℃ in underground warehouse during winter. And the ground warehouse of Daegu had similar patterns in temperature and humidity with that of Seoul in that being over 0℃ even in the winter. In Youngcheon, the temperature of farmhouse was below zero degree and the daily range of temperature and humidity was wide. It caused difficulty of overwintering of insect pests but possible to inhabit. Conclusion : According to a study by Han-Mo, Koo(1997), it was impossible to overwinter for Lasioderma serricorne F. which is dominant medicinal plants’ pest in north of the Chungcheong region’s warehouse due to low temperature. But in case of investigated warehouses, it was possible to overwinter because of the high temperature in office building or basement storage area. In order to make prediction of pest occurrence and comprehensive measures, it is urgent to take more investigation by regional groups and types of storage.

Background : Recently, with the growth of traditional medicine industry, management of the quality of herbal medications as a raw material for functional food and medicine becomes more important. One of the most difficult problems of quality control of medicinal plants is management in storage insects. The problem of storage insects is a matter that must be resolved in order to safely obtain raw materials in herbal sector. But the basic prevention measures as well as grasp of the exact situation in the field are not yet. In this study, we investigated to see the problems and solutions through a review of storage pest management system of medicinal plants and the relevant provisions in herbal sector. Methods and Results : For this study, we examined the Status of domestic distribution, storage pest control techniques, news, domestic and import regulations. Storage pests that occur in herbal medications have dropped commercial value and it leads to consumer mistrust. A report said that consumers who had found at least once a pest and microbial contamination when they buy a herbal medications is more than 50% (2004, Ministry of Food and Drug Safety). In the past, those who want to solve these problems had tried through the sulfur fumigation. However, due to concerns about the risk to humans nowadays it was banned in many countries. According to the latest editions of Korean Pharmacopoeia (KP), it is allowed to use a suitable fumigant. but the domestic and imported medicinal plants during distribution can not be used it. Because currently registered fumigant (aluminium phosphide) is allowed only imported medicinal plants in the quarantine process. Chemical fumigation is inappropriate for use in foods and pharmaceuticals so the introduction of eco-friendly control methods is urgent. Eco-friendly methods of pest control that is currently being developed include a cold storage, packaging, high temperature and frozen method etc. Conclusion : These methods should be considered potential cost and Feasibility and it should meet the condition that stated “"Do not change the therapeutic effects of the herbal medications.” in the KP. Also supporting policy for expanding low temperature storage facilities in distribution area will raise possibility of success.