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 : The fungus Cylindrocarpon destructans (Zins) Scholten is the cause of root rot in many ginseng production areas. Root exudate composition and quantity vary in relation to plant nutritional status, but the impact of the differences on rhizosphere microbial communities is not known. Methods and Results : Five kinds of rotation crops, sudan grass soybean peanut sweet potato, perilla were grown for one year in ginseng garden harvested 6-year-old ginseng. The ratio of gram-negative bacteria, fungi, bacteria, total microbial biomass, aerobic/anaerobic microbes were increased by rotational crop cultivation, while the ratio of actinomycetes and the ratio of saturated to unsaturated fatty acids were decreased. The increase in the fungal density or the increase in the proportion of fungi to the bacteria tended to increase the incidence of root rot, but there was no significant difference. The yield of ginseng root showed a highly significant negative correlation with actinomycetes. The correlation between the soil chemical properties and the incidence of root rot was analyzed by cultivating 23 kinds of green manure crops for one year in field where cultivated ginseng continuously. The survival rate of ginseng showed a highly significant positive correlation with soil acidity and a highly significant negative correlation with nitrate nitrogen, and a significant negative correlation with soil salt concentration. Conclusion : Rotation crops improved soil microbial communities, lowered the rate of fungi and increased the proportion of bacteria, the survival rate of ginseng was significantly correlated with soil acidity, nitrate nitrogen and soil salinity.

Background: Some plants have harmful effects on fungi and bacteria as well as plants. Incorporating into soil as green manures are effective in reducing population densities of soil pathogens. Methods and Results: Twenty-three species of green manure crops were cultivated after the harvest of 6-year-old ginseng and then incorporated green manure into the soil at the flowering stage. The following year, the root rot ratio of 2-year-old ginseng and soil chemical properties were investigated. Sorghum sudanense, Helianthus annuus, and Helianthus tuberosus were relatively high in dry matter production. Without incorporating green manure into soil, NO3, EC (electric conductivity) and K were decreased by 95%, 79% and 65%, respectively. When green manure was incorporated to soil, P2O5 and NO3, were reduced by 41% and 25%, respectively. The survived root ratio of 2-year-old ginseng were significantly increased by 56.2%, 47.5%, and 47.3% in Sorghum sudanense, Ricinus communis and Helianthus tuberosus, respectively, In addition, there was a significant increase in Secale cereale, Chrysanthemum morifolium, Atractylodes macrocephala, and Smallanthus sonchifolius. The survived root ratio of ginseng showed a significant positive correlation with soil pH and a negative correlation with NO3, and EC. Conclusion: Cultivation of Chrysanthemum family mainly using rhizome and root as green manure was effective for root rot disease of ginseng.

Sexual dimorphism is the most conspicuous difference between the sexes. This study examines possible sexual dimorphism and the relative growth patterns of morphometric characteristics in the marine medaka, Oryzias dancena for their potential to help differentiate between males and females of this species. The von Bertalanffy growth parameters estimated by a non-linear regression method were L∞=30.2 mm, K=3.22/year, and τ0=-0.05. All 18 characteristics measured showed a difference between males and females from 70 days after hatching. Each of these characteristics were significantly different between sexes (ANCOVA, P<0.05), and the ratio of standard length between sexes showed that males were larger than females for all five morphometric measurements. Fin length measurements were taken for 21 distances of anal fin and 7 distances of dorsal fin between landmarks. There were all differences for all dorsal fin rays between the males and the females and there is significant difference in 70 days after their hatch when the sexual dimorphism is presented. The significant difference (P<0.05) in fin ray for male and female was more greatly seen as they grow. Male marine medaka showed more rapid growth than females, with longer length, dorsal fins and anal fins. Differences in these characteristics will be useful during experiments when it is necessary to differentiate between sexes of marine medaka.

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 : The minor saponins produced by the hydrolysis of a major saponins sugar. The minor saponins has high absorption and efficacy compared to major saponin. The acid treatment, heat treatment and fermentation with minor saponin research has been actively conducted. This study was performed in order to investigate the bioconversion of ginsenoside Rg5 of fermented wild ginseng adventitious roots by using lactic acid bacteria. Methods and Results : 20g adventitious roots of ginseng was added to water (10-fold v/w). 10% (v/v) of lactic acid bacteria (Pediococcus pentosaceus HLJG0702[KACC 81017BP]) were inoculated with wild ginseng adventitious roots. For the fermentation process the inoculated samples were transferred to culture room for 1, 3 and 5 days. The fermented samples were dried at room temperature and extracted with 70% ethanol. Extract was concentrated completely at 50 ℃ and Rg5 was analysed by using HPLC. Results showed no significant difference the dry weight of non-fermented and fermented wild ginseng adventitious roots. During the fermentation process, the pH changed from 5.7 to 4.2. HPLC analysis showed higher ginsenoside Rg5 (39.588 mg/g) at 3 days. Conclusion : The fermentation of ginseng root can increase the Rg5 contents and minor saponin composition. This process may be used to enhance the minor saponin thereby increasing in fermented property of wild ginseng adventitious roots.

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 : 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 : Ginseng root rot caused by soil-borne pathogens, Cylindrocarpon destructans and Fusarium solani, is a major factors of replanting failure in ginseng cultivation. Some of the phenolic compounds detected in the soil of commercially cultivated American ginseng could inhibit the seed germination and seedling growth of American ginseng. Our study is to investigate the causes of replanting failure of ginseng by overhead flooding treatment and soil incorporation of ginseng fine root in soil infected with root rot pathogens. Methods and Results : To make soil occurring continuous cropping injury, 2-year-old ginseng infected with Cylindrocarpon destructans replanted in soil cultivated ginseng for 5 years. Treatment are as follows: 1) control, 2) water of 2ℓ was irrigate infected soil of 20ℓ, 3) ginseng fine root of 20g was mixed with infected soil of 20ℓ. Soil pH was increased, while other inorganic components were significantly reduced by overhead flooding treatment. Soil incorporation of ginseng fine root decreased soil pH, but increased EC, NO3, P2O5 and K, meanwhile, did not affected changes in organic matter, calcium, magnesium, sodium. Irrigation treatment in soil occurring replanted failure promoted distinctly above-ground growth of ginseng, and inhibited the occurrence of root rot because inorganic nutrient like NO3, P2O5 and K were decreased to optimal levels, and the density of soil pathogens could be reduced. Growth of ginseng was not inhibited, while root rot was promoted by soil incorporation of ginseng fine root. Conclusion : Irrigation treatment was effective in promoting the growth of ginseng and inhibited root rot distinctly. Ginseng fine root remaining in the soil after ginseng harvest did not affect the above-ground growth of ginseng, while promoted the occurrence of root rot.

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.

Background : This study was carried out to understand the effect of seedling weight (SW) on growth and flowering in Panax ginseng. Methods and Results : The testing materials were Chunpoong (CP), Yunpoong (YP) and Jakyeongjong (JK). The increase of seedling (1yr) weight led to an increase in ratio of flowering plant and in number of flower per plant. The seed setting rate of two year-old plant (CP, YP, JK) increased with increase of SW at the planting time (PT) and number of flower per plant of three year-old plant (CP, YP) increased also. In the two year-old plant (JK), the ratio of three leaves per plant was 8.8, 19.6, 31.0, 42.0, 44.7 and 58.2%, respectively, in the SW of >0.6, 0.6~0.8, 0.8~1.0, 1.02~1.2, 1.2~1.4 and 1.4g<. The growth of ginseng plant was good with increase of SW at the PT. Conclusion : There was a highly positive correlation between seedling weight and flowering characteristics.

Background : Due to immature development of embryo in ripened berries, dehiscence process is required for the proper germination of ginseng seeds. Such process involves the preparation of the container with alternating layers of seed and moist sand. In order to make sand fully moist, water sprayer has been usually used by farmers, which is labor intensive, time consuming and causing uneven sand moisture. Methods and Results : In this study, we investigated the effects of different stratification methods on dehiscence ratio of ginseng seeds. Ginseng seeds were stratified for 90 days in a total of 12 different treatments and the dehiscence ratios were compared; drainage methods, drainage time, the ratio between ginseng seeds and sand, and etc. Seed stratification process was performed according to the guideline of ginseng GAP. One thousand ginseng seeds were used for each treatment. It was found that the average of dehiscence of the 12 treatments was 84.6 %. The highest dehiscence ratio (90.3 %) was observed in the seeds that were treated with water soaking, immediately followed by drainage. Higher ratio was also observed in the seeds that were soaked for 60 min, followed by drainage. Therefore, our findings indicate that ginseng seeds soaked in water less than 60 min could dehisce more efficiently than traditional method. Conclusion : Our study demonstrated that ginseng seeds that are subjected to water soaking and then drainage showed better ration of dehiscence. This method will eventually decrease the time and labor used for seed stratification.

Background : This study was conducted to determine the impact of temperature elevated and the effect of transplanting times based on climate change scenario on growth of 2-year-old korean ginseng (Panax ginseng C. A. Meyer.) in temperature gradient chambers (TGC). Methods and Results : As a plant materials, ‘Yunpoong’ was cultivated in TGC at ambient temperature(Amb), Amb+2℃, Amb+4℃ and Amb+6℃ respectively. Ginseng was also transplanted on March 29, April 12 and 26 respectively. Investigation on characteristic of aerial parts were carried out on 28, 56, 84 and 112 days after transplanting and characteristic of roots were conducted on October 19. As transplanting time was faster and temperature was higher, the growth of aerial parts were increased. Compared with those of ginseng transplanted on March 29 with Amb, the root weight which tend to decrease depending on late transplanting time and high temperature decreased about 11.1%, 35.4% and 42.4% in Amb+ 2℃, Amb+4℃ and Amb+6℃ respectively. Ginseng transplanted on April 12 and 26 decreased about 20.9%, 33.9% respectively. Conclusion : Consequently, the more transplanting time extend, the more quantity increased in all temperature treatment. So, it is possible to increase in quantity to advance transplanting time although high temperature will be caused by the climate change.

Background : When ginseng seeds were gathered, the seeds were unripe. To grow immature embryo definitely, special treatment called dehiscence must be performed. Even though dehiscence is completed, most ginseng seeds are on enforced dormancy. The breaking seed dormancy is generally achieved using cold treatment. Also it is reported that gibberellin treatment can replace the treatment. It is very time consuming process in order to develop new ginseng cultivar because ginseng flowers after 3 years of growth. To shorten the ginseng breeding period, it is necessary to establish fast generation progress. Therefore, this study examined the possibility of breaking seed dormancy of ginseng using GA3 treatment and alternating temperature. Methods and Results : Seeds were obtained from local variety fruit which is not inbred. Gibberellin of 100 ppm was treated at seeds for 24 hours. Fixed cold condition was treated on both –2℃ and 2℃. Alternating cold condition was treated on 2℃ and then –2℃, finally 2℃. Fixed and alternating temperature was continued for 15, 30, 45, 60, 90 days that 15 days of alternating temperature is first 2℃ for 5days and then -2℃ for 5days, finally 2℃ for 5days. The other treatment periods such as 30, 45, 60, 90 days mean 10, 15, 20, 30 days respectively. Each of 48 seeds were sowed on tray in greenhouse at 3 replication. Experimental plot was completely randomized. Conclusion : Seeds untreated with GA3 were germinated little and there is no difference between 2℃ and –2℃. Alternating temperature until 60days made no difference with fixed temperature but germination rate increased up to 70.8% when seeds were treated for 90days. Germination of seeds treated with GA3 is much higher than untreated seeds especially combined with alternating temperature.

Background : Correct identification of Panax species is important to ensure food quality, safety, authenticity and health for consumers. This paper describes a high resolution melting (HRM) analysis based method using internal transcribed spacer (ITS) and 5.8S ribosomal DNA barcoding regions as target (Bar-HRM) to obtain barcoding information for the major Panax species and to identify the origin of ginseng plant. Methods and Results : A PCR-based approach, Bar-HRM was developed to discriminate among Panax species. In this study, the ITS1, ITS2, and 5.8S rDNA genes were targeted for testing, since these have been identified as suitable genes for use in the identification of Panax species. The HRM analysis generated cluster patterns that were specific and sensitive enough to detect small sequence differences among the tested Panax species. Conclusion : The results of this study show that the HRM curve analysis of the ITS regions and 5.8S rDNA sequences is a simple, quick, and reproducible method. It can simultaneously identify three Panax species and screen for variants. Thus, ITS1HRM and 5.8SHRM primer sets can be used to distinguish among Panax species.

The influence of triploidization on cell and nucleus size characteristics of the same tissues of erythrocyte, retina, kidney, hepatocyte and midgut epithelium in marine medaka, Oryzias dancena has been determined histologically. Induced triploid fish are produced by cold shock treatments. Likewise, the size of horizontal cell nucleus in inner nuclear layer of retina, ganglion cell nucleus in ganglion cell layer of retina, proximal tubule cell of kidney, hepatocytes and nuclear height of midgut epithelium all appear to be significantly larger than diploid (P<0.05). On the other hand, retina thickness is larger in diploid than induced triploid (P<0.05). Induced triploid shows low density of cell number. Results of this study suggest that same characteristics in the induced triploid exhibiting larger cells and nucleus sizes with fewer number of cells than the diploid can be useful criteria for the distinction between diploid and induced triploid, and also the ploidy level in marine medaka.