RAPD 마커를 이용하여 인삼 품종 및 육성계통의 유전적 다양성 및 유연관계를 분석한 결과는 다음과 같다. 1. 총 130개의 primer 중 polymorphism을 나타내는 70개의primer를 선발하였고, 그 중 재현성이 있으면서 polymorphism이 높은 25개의 primer를 선발하였다. 증폭된 DNA 단편의수는 189개이고, PCR 산물은 100 ~ 2,800 bp 범위로 증폭되었다. 2. 각 primer에 의해 증폭된 DNA 단편의 수는 3개 ~ 17개로 다양하였으며, primer 한 개당 평균 7.6개의 DNA 단편이증폭되었다. OPD19 primer를 이용한 유전분석 결과, 총 5개의 유전양상이 나타났는데, 약 500 ~ 1,300 bp의 증폭산물에서품종 및 계통 간 유전적 다형성을 나타냈다. 3. 선발된 primer별 대립인자는 최소 1.33에서 최대 2.00의 범위였고, 평균 1.709이었다. primer별 유전적 다양성은 OPD15가 가장 높았고, OPF2가 가장 낮은 값을 나타내었다. 본 연구에서 분석에 이용된 25개의 RAPD primer 중에서 D15, D19, B5, A19등은 인삼 품종과 계통에서 비교적 높은 수의 대립단편과 높은 유전적 다양성 값을 나타내는 primer였다. 4. 유사도 계수 0.98을 기준으로 24개의 품종 및 계통을 대상으로 군집분석을 수행한 결과, 미국에서 수집 육성된 G04116과 국내 품종인 천풍, 연풍 그리고 국내 육성 계통인 G04009, G04026, G04069, G04084는 그룹을 형성하지 않았고, 17개의 품종 및 계통은 2그룹으로 분류되었다. I 그룹에는 고풍, 금풍과 12계통(85%), II 그룹에 3계통(15%)이 포함되었다.

Background : Korean ginseng require 3 - 4 years to produce mature seeds from their mother plants. Therfore, it takes over 20 years to genetic fixation by artificial crossing of 8 generation. Anther culture is a useful method for obtaining homozygotes in only one generation. However, there is not much research on ginseng yet. In this study, we investigated the callus induction of anther depending on the type and concentration of the plant growth regulators in Panax ginseng C. A. Meyer. Methods and Results : Flower buds of P. ginseng were cold pretreated at 2 days before the anthers were plated on the induction medium. The flower buds were immersed in 70% ethanol for 30 sec min, washed two times with sterile distilled water, surface-sterilized in 2% sodium hypochlorite solution for 20 min, then rinsed five times with sterile distilled water. The anthers were placed on Petri dishes containting fifteen different concentrations and combinations of 2,4-D, NAA, BAP and KT. Callus induction was significantly influenced by the type and combination of plant growth regulators. The highest callus induction rate was observed in GR5 medium at 79.2%. The 2,4-D mediums had significantly higher callus induction than the NAA medium, and 2,4-D 1 ㎎/ℓ have a higher callus induction rate than the other concentrations. The increase of callus induction rate was not observed by the addition of cytokinin, but the callus induction rate was gradually decreased as the BAP concentration was increased. There was no difference in callus induction rate between BAP and KT. Conclusion : The important factor for inducing callus of ginseng anther was the addition of 2,4-D, and no effect of cytokinin addition could be found.

Background : Mutagenesis is one of the most powerful method for genetic variation, however there is little research history in P. ginseng. The seeds and vegetative tissues are materials commonly used for mutagenesis, but there is a problem that chimeras known as sectoral differences can occur and unintentional poor selection can occur owing to the influence of the growth environment. The combination of in vitro culture and mutagenesis can eliminate the effects of the environmental factors. Methods and Results : Gamma irradiation was carried out using a 60Co irradiator (3000Ci, Nordion Inc., Ottawa, Canada) of the Korea Atomic Energy Research Institute, Jeongeup, Korea. The explants containing the somatic embryos were treated with 20, 40, 60, 80, and 100 Gy. Gamma-rays affected the somatic embryos produced in vitro depending on the dose. The survival rate of somatic embryo was significantly influenced by gamma irradiation. Also, the development of the embryo was changed by gamma irradiation depending on each dose. When somatic embryos were transferred to the medium supplemented with GA3, they started to germinate but were affected significantly by gamma irradiation dose. When germinated shoots were transferred to 1/2 SH medium to induce rooting, as the concentration of gamma ray treatment increased, the rooting gradually decreased. Conclusion : Considering the overall survival rate, germination rate, embryo development, and rooting rate, the optimal dose of gamma rays to be used with somatic embryo is estimated to be between 60 Gy and 80 Gy.

Background : Panax ginseng C. A. Meyer is an important medicinal herb and their major pharmacologically active components are ginsenosides. Many studies have reported that various ginsenosides are effective in the treatment of human diseases such as cancer, cardiovascular and alzheimer’s disease. However, it is difficult to create new desirable variation such as increased ginsenosides. Since the frequency of spontaneous mutations is rare naturally and difficult to apply to plant breeding, artificial mutation inductions are necessary to obtain noble genetic resources having desirable traits. Methods and Results : Gamma irradiation was carried out using a 60Co irradiator (3000Ci, Nordion Inc., Ottawa, Canada) of the Korea Atomic Energy Research Institute, Jeongeup, Korea. One-year-old ginsengs were treated at 20, 40, 60, 80, and 100 Gy. The emergence rate was significantly decreased with increasing gamma doses. When irradiated at more than 80 Gy, it did not emerge at all. Survival rate was also significantly influenced by gamma-ray dose. The maximum survival rate were recorded in control. The survival rate of 20 Gy was 22.0% and only 1.0% survived when 40 Gy dose was treated. There was no survival when irradiated at more than 60 Gy. Gamma irradiation also affected the phenotype of emerged plants. The growth of plants derived from irradiated one-year-old ginsengs was decreased and the leaves became variegated or curled. Also, rusty roots resulting from physiological disorders were observed and the rhizomes, which is a kind of dormant bud, were destroyed completely. Conclusion : Based on these results, we estimated the optimal dose for mutation induction by gamma-ray treatment of one-year-old ginseng is to be less than 20 Gy.

Background : The conventional ginseng breeding is time-consuming and labor-intensive. Furthermore, it is difficult to create new desirable variation such as resistant to various stresses and increased ginsenosides. Therefore, creating variants remains a serious challenge of ginseng breeding. Methods and Results : Gamma irradiation was carried out using a 60Co irradiator (3000Ci, Nordion Inc., Ottawa, Canada) of the Korea Atomic Energy Research Institute, Jeongeup, Korea. The dehiscent seeds were treated 20, 40, 60, 80, 100, 120, 150, 200 and 400 Gy. The seed germination was greatly influenced by gamma-ray treatment. It was found that the germination rate decreased significantly when treated at 60 Gy or more. The maximum survival rate was achieved at 20 Gy but there was no significant difference from control. Following exposure to 40 Gy or more, survival rate was declined compared to the control. Gamma irradiation affected not only the germination and survival rate but also the growth of plants germinated from seeds. As gamma irradiation dose increased, the size of the plant decreased sharply and it also had a negative effect on root development. Conclusion : Considering the extent of reduction in the germination and survival rates, as well as growth reduction, the optimal dose of gamma-ray for inducing mutation in ginseng dehiscent seeds was estimated at approximately 40 Gy, because the germination rate observed at 40 Gy was similar to that of the observed in the control, while the survival rate was 50% of the control.

Background : Recently, the shortage of first planted ginseng field has increased direct seeding cultivation in paddy. Therefore, the demand for the development of ginseng varieties suitable for direct seeding cultivation in paddy has increased. So the aim of this study was selection ginseng varieties and germplasms suitable for direct seeding cultivation in paddy. Methods and Results : We used 13 varieties and 191 lines of Korean ginseng (Panax ginseng C. A. Meyer) in this study. Ginseng Seeds were harvested at the end of July 2013, sown in mid-November, and were grown for four years in paddy. In 2017, the survey was conducted on 27 items of growth characteristics in accordance with the standards for survey of ginseng variety, and the degree of diseases and physiological disorders. Among ginseng varieties, the growth of Gopoong, Sunwon and Gwumjin were excellent. However, the incidence of physiological disorder was low in Gumpoong and Geumsun. Among the genetic sources, the growth of 7 ginseng lines, such as G04092, G04098, M05015, G07053, G10069 and G10093, were excellent. However, the incidence of physiological disorder was low in 10 lines of ginseng, such as G04020, G04061, G04081, G04092, G05054, G07052, G07053, G10069 and G10093. Conclusion : This result suggests that 5 varieties and 10 lines of ginseng showed outstanding growths in paddy. In the future, we will review the characteristics of root and the content of ginsenosides of ginseng variety and elite lines.

Background : One generation of ginseng is four years, so it takes a long time to breed. So far, a total of 18 ginseng varieties have been developed under pure line selection. However the pure line selection can reach the limit, because ginseng’s gene pool is very small. A solution to this is the need for cross breeding. Therefore, this study carries out cross breeding for the introduction of excellent character and expansion of variation and check the F1 generation. Methods and Results : As plant materials, Chunpoong, Yunpoong, Gumpoong, Chungsun and Cheonryang were used in this study. In order to cultivate the F1 of ginseng, we cross-bred ginseng varieties from mid-May to late May 2013 and harvested seeds from late July. Ginseng seeds were sown on November 15 and cultivated in 2014 for one year in the field and then transplanted into the main field in 2015. In 2017, the survey was conducted on 27 items of growth characteristics in accordance with the standards for survey of ginseng variety, such as length of leaf, length of stem and width leaf, and the degree of diseases and physiological disorders. In a combination of cross-breeding in domestic ginseng varieties, the germination time and growth characteristics of F1 were not much different from the parent plants, but the incidence of disease was reduced. Conclusion : According to this study, there was a tendency for expressing dominant figures in the crossbreeding between ginseng varieties. In the future, we will review the characteristics of root and the content of ginsenosides of F1.

Background : Recently, there is an urgent demand for development of new varieties with enhanced resistance to various biotic/abiotic stresses. One generation of ginseng is four years, so it takes a long time to breed. To increase the efficiency of the ginseng breeding and save time and effort, it is necessary to build a ginseng core collection. This study examined the major growth characteristics of genetic sources for the establishment of core collection. Methods and Results : As plant materials, 187 ginseng lines collected in 2003 were used in this study. Ginseng Seeds were harvested at the end of July 2014, sown in mid-November, and cultivated in 2015 for one year in the field and then transplanted into the main field in 2016. All phenotypes including agronomic characteristics, seed yield, and physiological response to biotic/abiotic stresses were investigated according to the ginseng GAP and UPOV guidelines. The stem colors of the collected ginseng germplasm were classified as the five classes; light green, green, light purple, purple and dark purple, but most of them were purple or dark purple. The locations of distribution of anthocyanin coloration in stem were classified into the four classes; proximal end, proximal end and petiole part, the entire stem and the stem with purple not revealed, but most of them were petiole part or the entire stem. The shapes of leaflet were classified the four classes; the long elliptic, elliptic, slender type, and spatulate type. Most of surveyed lines were elliptic type. When the leaflet of ginseng was laterally cut, the shape was classified into the three types; concave type, plane type and convex type. Most of surveyed lines were red berry color. Conclusion : The genetic pool of ginseng is known to be narrow. The results of this study showed similar characteristics among the ginseng fields collected. In the future, we will carry out the survey about quantitative characteristics and correlations of ginseng genetic resources for establishment of Korean ginseng core collection.

Background : Endophytes are generally regarded as beneficial microorganisms that live in plant tissues without disease symptoms. The endophytic species may differ depending on the plant age, the sampled time, the plant genotype, and the tissue. Although numerous endophytes have been identified from various plants, little is known about the endophytic bacteria of Panax quinquefolius, a useful herb plant. Therefore, the aim of this study is to investigate diversity and distribution of endophytic bacteria from 2-years-old to 6-years-old P. quinquefolius and to evaluate antimicrobial activity of isolates. Methods and Results : Initially, 2-years-old to 6-years-old plants were collected and sterilized with 70% ethanol and sodium hypochlorite, subsequently we prepared individual suspensions which were mixed with sample and sterile distilled water. A total of 88 single colonies were obtained from the LB agar plates spreading suspension. Using 16S rDNA sequencing, the taxonomic status of the isolates were determined consequently 42 species were identified. The 42 species were classified into 4 phylum; Proteobacteria (64%), Firmicute (27%), Actinobacteria (8%), and Bacteroidetes (1%). Based on age, the isolates of 5-years-old plant showed highest diversity. Moreover, Actinomycetales, Bacillales, and Pseudomonadales were equally dispersed as predominant orders in 5-years-old plant. The antagonistic activities of isolates against a phytopathogen, Pseudomonas syringae pv. tomato DC3000 : GFP (Pst : GFP) were performed using dual culture assays. We measured antibacterial activity by quantifying fluorescence of Pst:GFP which representing pathogen growth. As a result, 36 isolates inhibited growth of Pst:GFP. Interestingly, all species belonging Pseudomonas in this study showed strong antibacterial activity against Pst : GFP. Conclusion : These results improve our understanding of the structure and diversity of the endophytic bacteria of P. quinquefolius. Furthermore, we suggest that bacterial endophytes with antimicrobial activity might have useful as materials for biocontrol agents.

Background : Recently, there is a urgent demand for development of new varieties with enhanced resistance to various biotic/abiotic stresses. In order to develop ginseng varieties with such traits, systematic breeding program and comprehensive field studies are prerequisite. Methods and Results : ‘Jinwon' seeds were collected in 2003 from the farmer field of Pocheon. Physiological investigation and propagation were conducted from 2008 to 2010. It was given the name 'Eumseong No. 11 through the observed yield trial from 2011 to 2013 and local adaptability was carried out from 2014 to 2017. All phenotypes including agronomic characteristics, seed yield, and physiological response to biotic/abiotic stresses were investigated according to the ginseng GAP and UPOV guidelines. Red leaf and red berry at maturing stage were observed. The time of emergence, flowering and berry maturity of the ‘Jinwon’ were faster than those of ‘Chunpoong’. Stem length of ‘Jinwon’ was longer than that of ‘Chunpoong’, whereas stem diameter was thicker than that of ‘Chunpoong’. Main root length was similar but main root diameter is thicker than that of ‘Chunpoong’. The yield performance of this variety was 576 ㎏/10 a in local adaptability test for two years, which is 9% higher than that of ‘Chunpoong’. ‘Jinwon’ showed strong resistance at high temperatures. Conclusion : Our study demonstrated that ‘Jinwon’ is an ideal variety with heavier root weight and enhanced stress resistance and contribute will enhance biotic/abiotic stress resistance and increase the farmers' income.

Background : This study aimed to investigate the effect of elevated temperature by climate change on growth and ginsenoside contents of Korean ginseng, which requires low temperature during growth period. Methods and Results : Yunpoong was transplanted in temperature gradient chamber (TGC) for high temperature treatment and then growth characteristics and ginsenoside contents were investigated by growth period. Stem length, petiole length, leaf length and leaf width were increased as the temperature was higher than Ambient Temperature (Amb), but stalk diameter was decreased and small petiole length, number of petiole and leaf were no significant difference. Root growth, whereas, including length diameter and weight were decreased as the temperature was higher than Amb. Compared with Amb, root length, diameter and weight were decreased by 19.6%, 22.4% and 41.5% at Amb+6, respectively. The higher the temperatures are associated with the more ginsenoside contents. Compared to Amb, total ginsenoside contents were increased by 68% and 78% at Amb+4℃ and Amb+6℃, respectively. Almost all contents of individual ginsenoside, with the exception of Rg1, made a significant difference. Conclusion : When temperatures rises during growth period, the growth of ginseng was decreased but, ginsenoside contents were increased.

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 : This study was carried out to distribute superior cultivars to farmers by analyzing growth characteristics of ginseng and effectively set the target for breeding by investigating annual agronomic characteristics of ginseng. Methods and Results : As a plant materials, Chunpoong (CP), Yunpoong (YP), Gopoong(GP), Gumpoong (GU) and Sunun (SU) were transplanted in Eumseong (ES), Geumsan (GS) and Yeoncheon (YC) on March 2013. Cultivation and management were conducted on the basis of the standard guidelines. According to stem and petiole by region, YC represent the largest stem diameter, leaf length and petiole length of cultivars, followed by those in ES and those in GS. Leaf growth of cultivars was overall the best in YC. Although CP had long and narrow leaves in all regions, YP and SU had short and narrow leaves. Root weight of all cultivars was the best in YC, but there was no significant differences by them. Growth of CP, YP and GP was generally good in ES. Growth of cultivars was relatively even in GS, but it was poor compared to cultivars which were cultivated in ES and YC. Conclusion : There were considerable differences in root growth of cultivars. However, this results suggest that CP of 4-year-old is produced stably in some regions because it had a small coefficient of variation.

Background : Panax ginseng C. A. Meyer is a slow-growing perennial herb that is cultivated in shading condition. Climate change occur around the world that make a lot of problem such as damage of high temperature, drought, salinity and disease. The problems lower the ginseng productivity that cause income reduction of farmers. To achieve stable ginseng production, development of elite varieities resistant to abiotic and biotic stresses is consistently required. It is very time consuming process in order to develop new ginseng varieties because ginseng flowers after 3 years of growth. So, early selection system of elite line must be established. This study was conducted to develope efficient ginseng breeding techniques for early identification of heat or salinity resistance. Methods and Results : Ginseng petioles was soaked in mixed salts solution consisting of KNO3, KH2PO4, MgSO4․H2O in order to test resistant or susceptible salinity. The degree of resistance was quantified according to damage size. Also, ginseng lines transplanted in pot were treated 46℃ for 1 hour and then chlorophyll fluorescence reaction were measured in order to test resistant or susceptible high-temperature. The measured values such as Fm/Fo, Fv/Fm, Rfd were differentiated between resistant and susceptible line. Conclusion : Several lines showed that they are resistance to high temperature or salinity. The selected lines will be utilized for parents to develop new varieties.

Background : The P. ginseng breeding line G07006, was selected for salt tolerance through salinity screening of mature leaves at the NIHHS of the RDA in 2014-2016. However, it is difficult to maintain a genetically stable breeding line of cross-pollinating crop in the field. Therefore molecular marker required to identify and maintain breeding line G07006. Methods and Results : DNA was extracted following the CTAB DNA extraction protocol (Doyle and Doyle, 1987) with modifications. A pair-end (PE) library was constructed and sequenced using an Illumina MiSeq platform by Lab Genomics, Inc. (Seongnam, Korea). Approximately 4.0 Gb of sequencing data were obtained, and de novo assembled by a CLC genome assembler(v. beta 4.6, CLC Inc., Rarhus, Denmark). The complete chloroplast(CP) genome size is 156,356 bp, including two inverted repeats (IRs) of 52,060 bp, separated by the large single-copy (LSC 86,174 bp) and small single-copy (SSC 18,122 bp) regions. This CP genome encodes 114 unigenes (80 protein-coding genes, four rRNA genes, and 30 tRNA genes), in which 18 are duplicated in the IR regions. Conclusion : This complete chloroplast DNA sequence will provide conducive to discriminate line G070006 (salt-tolerant) and further enhancing genetic improvement program of this important medical plant.

Background: Developing new ginseng cultivars is a significant time-consuming process owing to the three years of growth required for ginseng to flower. To shorten the ginseng breeding process, it is necessary to establish rapid progression through each generation. In this study, we examined it was possible to rapidly break ginseng seed dormancy using gibberellic acid (GA3) treatment and alternating temperature. Methods and Results: Seeds were obtained from local variety. Seeds were treated with either GA3 at a concentration of 100 ㎎/ℓ, constant temperature (−2℃ and 2℃), alternating temperature (2℃ followed by −2℃, followed by 2℃) or a combination GA3 and temperature treatment. Following experimental treatment, seeds were sown into trays and placed in a greenhouse. Low germination rates were observed in seeds that did not receive GA3 treatment, which were similar following 2℃ and −2℃ constant temperature treatment. Germination rates increased in proportion to GA3 and more so when combined with alternating temperature treatment. In additon, stem and leaf lengths of the resulting ginseng plants were increased following GA3 treatment, although no synergistic effect was observed with alternating temperature treatment. Conclusions: These results suggest that a combination GA3 and alternating temperature treatment enhances ginseng seed germination, which can contribute to shortening the time required to progress through a single ginseng generation for breeding.

Background : Recently, there is a urgent demand for development of new varieties with enhanced resistance to various biotic/abiotic stresses. In order to develop ginseng varieties with such traits, systematic breeding program and comprehensive field studies are prerequisite. Methods and Results : ‘Cheonmyeong' seeds were collected in 2008 from the farmer field of Buyeo. Physiological investigation and propagation were conducted from 2009 to 2011. It was given the name 'Eumseong No. 8 through the observed yield trial from 2012 to 2013 and local adaptability was carried out from 2014 to 2015. All phenotypes including agronomic characteristics, seed yield, and physiological response to biotic/abiotic stresses were investigated according to the ginseng GAP and UPOV guidelines. Yellow-red leaf and pink berry at maturing stage were observed. The time of emergence, flowering and berry maturity of the ‘Cheonmyeong’ were faster than those of ‘Chunpoong’. Stem length of ‘Cheonmyeong’ was shorter than that of ‘Chunpoong’, whereas stem diameter was thicker than that of ‘Chunpoong’. Main root length was shorter but main root diameter is thicker than that of ‘Chunpoong’. Number of seeds of ‘Cheonmyeong’ was fewer than that of ‘Chunpoong’ but 1,000-seeds weight and stratification rate were higher than those of ‘Chunpoong’. The yield performance of this variety was 661 kg/10 a in local adaptability test for two years, which is 22% higher than that of ‘Chunpoong’. ‘Cheonmyeong’ showed strong resistance to phytophthora blight, mulberry mealybug and nematode and moderate resistance to alternaria blight. ‘Cheonmyeong’ did almost not occur yellow spot of aerial part and rusty skin of root, show moderate resistance at high temperatures. Conclusion : Our study demonstrated that ‘Cheonmyeong’ is an ideal variety with heavier root weight and enhanced stress resistance and contribute will enhance biotic/abiotic stress resistance and increase the farmers' income.

Background : Water uptake and flow across cellular membranes is a fundamental requirement for plant growth and development, and plant water status is important not only for plant growth under favorable conditions but also for ability of a plant to tolerate adverse environmental conditions. Thus identification of plasma membrane water channel genes (aquaporins) in ginseng provides extensive information for functional studies and the development of markers for salinity stress tolerance. Methods and Results : For salinity treatment, the plants were grown for 4 weeks in culture medium gelled with 0.8% Phytoagar, and the old media were replaced with the fresh medium containing NaCl at 0, 50, 100, 200 and 400 mM, respectively. The samples for stress treated and non-stressed plants were collected from 6h to 72h, and frozen immediately into liquid nitrogen. According to the sequence information from the assembled transcripts, four primer pairs were designed from the aquaporin gene regions. In order to determine the pattern of aquaporins expression in ginseng seedlings to salinity stress, we conducted semi-quantitative RT-PCR. Conclusion : A tonoplast intrinsic protein 1 (TIP1)-type aquaporin is not only believed to be essential for plant life, but also to be beneficial for growth under salinity stress. Therefore, a deeper understanding of aquaporin genes in ginseng will be essential for crop improvement, which could help us to understand the molecular genetic basis for the ginseng genetic improvement and also provide the functional genetic resources for selective breeding and transgenic research.