The objectives of the current study were to evaluate the therapeutic effect of dioctahedral smectite (smectite) against calf diarrhea caused by pathogenic E. coli and/or Salmonella typhimurium. In this study, 20 calves (aged 2~3 months) with diarrhea were used for evaluation of the efficacy of smectite on calf diarrhea with 20% smectite suspension in PBS. Calves received 10 ml smectite suspension three times per day after feeding, and fecal samples were collected at the gate of treatment and on the first, second, third, fourth, and fifth day after administration. On the fifth day after treatment with smectite suspension, the diarrhea index showed a significant decrease in the treated group, compared to the control group (P<0.001). The number of pathogenic E. coli in feces of the treated group was significantly decreased, compared to each control group from the second day after treatment (P<0.001), and that of Salmonella typhimurium was significantly decreased from the first day after treatment (P<0.05). According to the results of the current study, 20% smectite suspension had a therapeutic effect on diarrhea caused by E. coli and/or Salmonella in calves.

This study was conducted in order to examine the safety of bee venom as an alternative for antibiotics using male ICR mice. Five-week-old male mice received a single intravenous injection of a dried honey bee venom at the concentration of 0.25 mg/kg (a clinical dose) or 0.5 mg/kg through the tail vein and various pathophysiological analyses were performed after three days. No significant differences in changes of body weight were observed between the saline-treated control group and the experimental groups. In the hematological analysis, none of the parameters were affected by bee venom. In blood biochemistry analysis, none of the markers were affected by administration of bee venom. Similarly, there were no significant effects on markers for liver, kidney, and skeletal muscle functions in all treated- groups. On macroscopic examination, no remarkable lesions were detected in these organs. Because there were no adverse effects of the bee venom in a single intravenous toxicity test for three days, it was concluded that bee venom could be a candidate for a safe natural antibiotic for use in the animal production industry.

Background : Ginseng seeds are one of short-lived seeds species which loose their viability easily in the condition of conventional storage. Cryopreservation using liquid nitrogen (LN) has been recommended as a alternative storage for this kind of germplasm short lived or dessiccation-sensitive. This study was performed to find out whether cryopreservation could assure initial viability not only for seeds with high germination rate but also for seeds with low germination rate (in aging process). Methods and Results : In this work, 3 cultivars of dehisced ginseng seeds were artificial aged in the condition of 40℃, 95% RH during 6 hours with 3 hours-interval. The germination rate of ginseng seeds was decreased to the range of 71 - 94% of initial viability by artificial aging treatment. After 24 hours of vapor-LN exposure on both of artificial aged and non treated seeds as a cryopreservation, germination rate for each cultivar was decreased with the range of 76 - 95% of initial viability. While the decreasing patterns of germination rate for each cultivar showed similar curves between before and after vapor-LN exposure, the aging effects could be slightly little by cryopreservation for 3 cultivars of ginseng seeds. Conclusion : From the above results, we may suggest that cryopreservation could be recommened for storage tool of dehisced ginseng seeds even with low viability also and expected to make slower seed aging process during preservation period through further study.

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 : This study examines the main growth characteristics, physiological stress, or diseases damage for 3 and 4-years-old ginseng in the new lines cultivated in Gyeonggi for development of new lines of Ginseng. Methods and Results : Eumseong 13, 14 and 15, which are two-years-old seedling in the new line ginseng were transplanted in 7 lines × 9 rows (70 plants)/1.62㎡ in 2015 along with Gumpoong and Yeonpoong as the control lines while Eumseong 16 and 17 as well as Chunpoong as the control line were transplanted in the same way in 2016. The ginseng shading was installed with the front post of 170 ㎝ high and the rear post of 100 ㎝ high using the rear line while as the sun blocking material, the aluminum shade plate was used. As for the new growth lines of 3 and 4-years ginseng in the new growth line, the aerial parts of the ginseng and underground part of ginseng were examined. In addition, the physiological stress such as the leaf discoloration, rusty root and root rot as well as the resistance against the disease were examined. It was found out that as for the aerial parts of 3-years-old ginseng in test line, Eumseong 16 and 17 were better in the growth than the control lines were while in 4 years old ginseng, Eumseong 13 and 14 were better in the growth than Gumpoong as the control line was. As for the root weight of the 4-years-old ginseng for the underground part, there was no significant difference compared with control line while in 3-years-old ginseng, Eumseong 16 and 17 were heavier. The leaf discoloration did not happen to both 3 and 4-years-old ginseng. As for the root rot, 3-years-old Eumseong 16 and 17 suffered less root rot in the same way as for the control line. 4-years-old Eumseong 14 and 15 had less root rot in the same way as for the control line. 3-years-old Eumseong 16 and 17 have less rusty root in the same way as the control line while as for 4-years-old ginseng, Eumseong 13, 14 and 15 showed a lot of rusty roots in the same way as the control line. Conclusion : Finally, considering the growth characteristics, physiological stress and root rusty of the aerial parts and underground parts of the ginseng, we selected Eumseong 16 and 17 as the excellent lines for 3-years-old ginseng.

Background : For cultivation of varieties of ginseng, the pure line selection method, which is to select the best among those cultivated in farms for pedigree breeding, replicated yield trials and regional adaptation trials before registering as a new variety, is widely used. Although there are 25 registered varieties of ginseng in Korea, the quality of ginseng is declining together with the amount of harvest being decreased by 15 - 20% due to the heat injuries and diseases from the warming & abnormal climate. Thus, the needs for development of disaster-resistant varieties with better chances of surviving through high temperature, salts and disease are increasing. Therefore, this study is to cultivate disaster-resistant varieties among those selected for their disaster tolerance and salt tolerance through regional adaptation trials. Methods and Results : As a result of examining the growth characteristics of the selected 2-6-years-old varieties used in the study, among the 6-years-old crops, Eumseong 9 and Eumseong 10 showed superior growth in aboveground parts, Eumseong 10 showed superior growth in underground parts, among the 5-years-old crops, Eumseong 15 showed superior growth in both above and below aerial parts, and among the 4-years-old crops, Eumseong 9 and Eumseong 16 showed superior growth while the growth in the underground parts were satisfactory in the order of Eumseong 16 > Eumseong 10 > Eumseong 12 > Eumseong 9. Among the 3-years-old crops, the most superior growth in both above and below aerial parts was observed in Goryeo 2 with the weight of the underground part, in the 2-years-old crops, Eumseong 26 showed the most superior growth in both above-ground and underground parts. Conclusion : Based on the above results, Eumseong 10 among the 6-year-old crops, Eumseong 15 among the 5-years-old crops, Eumseong 9 & Eumseong 16 among the 4-years-old crops, Goryeo 2 among the 3-years-old crops and Eumseon 26 among the 2-years-old crops showed the most superior growth among the selected varieties. The growth characteristics of both above and below aerial parts in each year will continuously be monitored.

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: In Korea, 6-year-old ginseng root is economically more important than 4 or 5-year-old roots. In general, the root age is determined by counting the number of stem vestiges. However, this method does not accurately estimate ginseng root age. Methods and Results: In this study, the stem vestige counting method was used to survey a total of 18,395 fresh ginsengs cultured in 2014, and 2015, to determine the accuracy of this method. The proportion of 6-year-old roots, with more than four stem vestiges, was 46.1% in 2014. For the cultivar Chunpoong cultivated in Eumseong and Goesan countries in 2015, the proportion of more than four stem vestiges was 55.9%, and 43.5%, respectively. The proportion of more than four stem vestiges for the Gumpoong cultivated in Eumseong and Yangpyeong countries was 67.0%, and 35.1%, respectively, whereas that for the cultivar Yunpoong was 36.0% and 61.0%, respectively. Moreover, it was confirmed that differences in the levels of Rg1 will enable root age determination. Conclusions: Root age determination by the stem vestige test was found to differ depending on the environmental and cultivation conditions. To determine the age of ginseng roots, a comprehensive method, such as counting stem vestiges and evaluating differences in ginsenoside levels, should be applied.

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 geographical origin of Panax ginseng Meyer, a valuable medicinal plant, is important to both ginseng producers and consumers in the context of economic profit and human health benefits. We therefore aimed to discriminate between the cultivation regions of ginseng using the stable isotope ratios of C, N, O, and S, which are abundant bio-elements in living organisms. Methods and Results : The C, N, O, and S stable isotope ratios were measured by isotope ratio mass spectrometer, and then these isotope ratios profiling was statistically analyzed with chemometrics. The various isotope ratios found in Panax ginseng roots were significantly influenced by region, cultivar, and the interactions between these two factors (P ≤ 0.0002). In particular, δ18O was lower in ginseng roots grown at high altitudes (r = −0.47), while δ34S was higher in ginseng roots grown close to coastal areas (r = −0.48). Chemometric results provided discrimination between the majorities of different cultivation regions. Conclusion : Our case study extends the understanding about the variation of C, N, O, and S stable isotope ratios in ginseng root depending on cultivation region. Hence, the analysis of stable isotope ratios is a suitable tool for discrimination between the regional origins of ginseng samples from Korea, with potential application to other countries.

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.