Due to modern trends with postponing child-bearing and getting worse living environment in women, an ovarian aging increased pregnancy failure and other complications with menopause or premature ovarian failure. Although several theories have been suggested such as mitochondrial malfunction, DNA damage/repair/methylation, caloric restriction, studies regarding ovarian aging-related molecular mechanisms for development of therapeutic methods are insufficient so far. Our objective is to determine molecular pathways of ovarian aging that result in pregnancy failure and other complications in women health to develop treatment strategies. This study is consisted of two parts: in Phase I stage, we analyzed distinct gene expression profile between young and aged mouse ovaries, and in Phase II stage several preferentially expressed genes in both ovaries were selected and analyzed their physiological functions and involved molecular networks related to ovarian aging for development of diagnostic markers and therapeutic methods. Ovaries from 10 week and 11 month-old FVB/NJ female mice with synchronized estrus cycle were collected for this study. A half of each ovary was used for RNA preparation and the other half for histological analysis. Using the Illumina HiSeq 2000 System, preferentially expressed genes were identified. Functional annotation database-based gene-set enrichment analyses and Pathway Studio® were employed to evaluate aging-related molecular networks. These findings were confirmed through qRT-PCR and immunohistochemistry. To validate RNA-Seq data, we examined expression patterns of marker genes (Amh, Bmp15 and Nobox) that were wellknown to be decreased in ovarian aging process. In young or aged ovary, preferentially expressed 876 genes were identified and extracellular matrix (ECM; p<0.001) and chromatin/nucleosome-related (p<0.001) protein-coded genes have the majority in these genes by GOTERM analysis. Amongthem, we selected several candidate genes and confirmed their expression profiles by qRT-PCR and immunohistochemistry followed by molecular network analysis. Regarding molecular interactions in these genes, PathwayStudio® was employed to predict aging-involved molecular networks in mouse ovary. Here we report a couple of candidate molecular networks and medicines (chemicals) for targeting these preferentially expressed genes/proteins. Further analyses are scheduled to produce transgenic animal models and with human ovarian tissues/cell lines.

Bitter melon (Momordica charantia, MC) has been used in traditional Korean medicine in treating diabetes. In addition, some reports were emerged, showing the antifertility activities of MC in mammals. We investigated the effects of ethanolic MC extract on the reproductive activity of golden hamsters whose spermatogenetic capacity is controlled by their photoperiods. The animals were divided into 4 groups: long photoperiod (LP) control, short photoperiod (SP) control, and LP animals treated with MC. The animals were orally ingested with low (0.03 g/kg) or high (0.15 g/kg) concentrations of the ethanolic extracts for 8 weeks on the daily basis. The control animals received the vehicle. The animals were then mated with age-matched females, experienced pregnancy. As results, the LP control animals showed active large testes but SP control animals displayed remarkably reduced testes. The animals treated with both concentrations of MC extracts demonstrated large testes, indicating fertile activity as animals in LP. LP control animals had litters as expected, but SP controls had no litters at all. MC extract showed the same results as LP animals in generating offsprings. These results suggest that the MC extract does not change the photoperiodic influence on reproductive activity of male golden hamsters.

Background : Codonopsis lanceolata is a perennial plant of Campanulaceae and mainly distributed in East Asia such as Korea, China, and Japan. C. lanceolata has a unique taste and aroma, and it is rich in minerals such as phosphorus and calcium, and vitamin B1 and B2, so our ancestors used the plant as medicinal herb and edible vegetable. However, systematic cultivation and development of varieties have not been achieved compared to demand or high added value. The genetic diversity and relationship analysis of the plants help to increase the efficiency of breeding through genetic variation. Methods and Results : Ten species of Codonopsis plants were used as materials and DNA was extracted from each 4 individuals per species and quantified at a concentration of 10 ng /㎕. The extracted DNA was pooled by species and PCR was performed using the EST-SSR marker developed based on C. lanceolata in the previous study. PCR amplification was carried out using a denaturation at 94℃ for 30 sec, annealing at 58℃ for 30 sec and extension at 72℃ for 30 sec, repeated for 35 total cycles. The PCR products were separated in a 4% agarose gell at 100 V for 40 min. Conclusion : In this study, C. lanceolata collections was determined among several Codonopsis species using these molecular marker. It is expected that the data of this study can be used as reference for genetic polymorphism analysis and related gene studies of Codonopsis species.

Background : Codonopsis lanceolata is a perennial plant of Campanulaceae with characteristic flavor and aroma and this plant has saponin, flavonoid, and inulin, which are reported to have physiological activity and antioxidant activity. In contrast, breeding or study of C. lanceolata varieties had not been done for a long time. Genetic polymorphism and phylogenetic relationship analysis of the plants by region of the crops can help the collection of genetic backgroud data for variety development. Methods and Results : In this study, we collected 26 C. lanceolata lines (95 individual plants) from 26 regions in Korea. We genotyped the collected lines using SSR markers developed in the previous study and analyzed the population structure based on the results. Population structures were analyzed using model-based STRUCTURE software (version 2.3.4) using the following parameters: Number of clusters (K) set = 1 to 12; Number of Iterations = 5; Length of Burning Period = 100,000; Number of MCMC (Markov Chain Monte Carlo) Reps after Burnin = 100,000. As a result, Of the 26 collections, were genetically grouped into 6 or 7 groups. Conclusion : The 26 C. lanceolata collections (95 individual plants) were genetically grouped but not grouped by collected regions. These results suggest that C. lanceolata has diverse genetic backgrounds and this data could be used as a basis for genetic polymorphism analysis of Codonopsis species.

Background : Licorice has been used as a source of medicine and a food material in East-Asia. Recently, demand for licorice increased in market due to a growing interest in health. Thus we conducted breeding research to solve the problems associated with domestically cultivated licorice such as low productivity and low glycyrrhizin content. Methods and Results : We crossed European licorice (G. glabra L.; female parent) and Chinese licorice (G. uralensis Fisch; male parent) in the greenhouse in May 2007. In September 2007, crossed and germinated seeds were retrieved and sown in the greenhouse. In June 2008, stolons were separated from the F1 licorice seedlings and cultivated, resulting in 32 clonal lines of interspecific hybrids. Among them we selected good lines and then conducted the replicated yield trials (RYT) in 2012-2013 and local adaptability test (LAT) in 2014-2015. The results, GLYES9 showed that was elect of stem, oblong of leaf shape, red-brown of root color. Glycyrrhizin conten of GLYES9 (3.0%) was higher than G. uralensis (1.9%) at four regions from 2014 to 2015. GLYES9 was less than 10% in the desease of brown spot (G. uralensis was more than 30%). The root yield of GLYES9 was 4.31 ton per hectare, which was increased 193% compared with a check variety of G. uralensis. Therfore, we named GLYES9 as new cultivar ‘Dagam’. Conclusion : Depending on the above results, we have developed a new licorice cultivar ‘Dagam’ by the medicinal crop breeding team of National Institute of Horticulture and Herbal Science, RDA, in 2015. It showed brown spot disease resistant, high-glycyrrhizn content and high-yielding than colleted Glycyrrhiza spp.

Background : Korean mountain ginseng (Panax ginseng C.A. Meyer) are difficult to industrially apply because of its scarcity and high cost. Advances in plant biotechnology have made it possible to produce mountain ginseng on a large scale using adventitious root cultures in bio-reactors. This study was conducted to develop a cosmetic emulsion using ginsenoside and physiological activity - enhanced raw materials by fermentation process. Methods and Results : Wild ginseng adventitious roots were fermented with Pediococcus pentosaceus HLJG 0702 (KACC 81017BP). ginsenoside contents was analysed by using HPLC. Antioxidant activity was measured by DPPH and ABTS radical scavenging activity and whitening effect was measured by tyrosinase inhibitory activity. After microfluidizer processing was performed to prepare emulsions with homogenized particles, particle size and distribution were measured through a transmission electron microscop e(TEM). Particle stability compares pH, viscosity, light and zeta potential. When fermented with Pediococcus pentosaceus HLJG 0702, the highest change rates of Rg3, Rk1 and Rg5 were shown and the antioxidant activity was increased. The whitening effect was 73.2 ± 0.9% when treated at 100 ㎍/㎖, 1.5 times higher than the control. The optimum particle size and distribution were shown to be 418.0 ± 14.9 ㎚ for 6 times treatment with 0 - 10 times microfluidizer treatment. Stability was about 3% in pH, viscosity and light test. the zeta potential was found to be homogeneous at –33.33 mV. Conclusion : Pediococcus pentosaceus HLJG 0702 Fermented Wild ginseng adventitious roots were found to have effective ingredients and improved physiological activity. We have also developed emulsions that exhibit optimal particle size and distribution

Background : Adenophora triphylla var. japonica (Regel) H. Hara shows vegetative growth by radical leaf until 1 year after sowing and shows reproductive growth during the second year and there is a characteristic of bolting by turning into cauline leaf. In addition, the phenotypes of plants varies even though they are belonging to the same species. For this reason, there is a limit for the classification of the species by the method of visual examination. Methods and Results : Simple sequence repeat (SSR) markers were developed based on the genomic sequence of A. triphylla using next generation sequencing to prepare the basis of molecular breeding and analyze the genetic diversity. Ninety-five primer sets including tri-, tetra- and penta-nucleotide motif types were randomly selected and they were applied to mixed genomic DNA and finally 39 primer sets showing from two to four bands were selected and used for genetic relationship analysis. Conclusions : Using the next generation sequencing, 39 polymorphic SSR markers were developed.

A monitoring system for a field magnetometer was configured with assistance of a Raspberry Pi as a data logger. The suggested geomagnetic system uses a semi-real-time data transmission module. The system consists of two parts: a field-observation part and a data-center part. The field-observation part comprises a Raspberry Pi, magnetometer, LTE router, and power source, while the data center part takes samples at the site. The collected magnetometer data are then sent to the data center through the LTE router. The newly designed monitoring system was deployed and checked in Jeju-do island, and found to operate stably. The suggested system is promising in that it is simple and cost saving, providing at least physical insight and knowledge on the complex natural phenomena.

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 : 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 : Codonopsis lanceolata is a flowering perennial climber. The roots are used as medicinal materials or vegetables. C. lanceolata is distributed in India and East Asia such as China, Japan as well as Korea. Recently, demand for C. lanceolata is increasing as a healthy food. In South Korea, this plant is widely cultivated in Gangwon-do province. Although, C. lanceolata is one of the most important medicinal plants in Korea, an elite, inbred line or a variety has not been developed yet. Simple sequence repeat (SSR) marker is a powerful tool for analysis of genetic relationships. In addition, it is a useful tool for studying the non-reference plant genome, due to its even distribution throughout the genome, as well as its high polymorphism between individuals. Methods and Results : We constructed microsatellite-enrichment libraries using C. lanceolata genomic DNA, and obtained a total of 226 non-redundant contig sequences. Routine PCR was performed using gDNA as templates for the polymorphic markers screening. Finally, total 15 polymorphic SSR markers based on C. lanceolata genomic sequences were successfully developed. These markers were applied to 53 C. lanceolata collected from Korea. 103 alleles of the 15 SSR markers ranged from 3 to 19 alleles at each locus, with an average of 6.87. The average of observed heterozygosity and genetic diversity were 0.42 and 0.62, respectively. The average of polymorphism information content (PIC) value was 0.57. The genetic distance value ranged from 0.73 to 0.93, and there was no observed distinct group according to the collecting areas. Conclusion : We developed 15 novel SSR markers from C. lanceolata genomic sequences for further genetic studies. Also, we concluded that the lineage of C. lanceolata collected in Korea has not been established systematically.

Background : Platycodon grandiflorum is a perennial plant and a member of Campanulaceae family. Since ancient times, they have been using P. grandiflorum as an important medicinal plant in Korea. Platycodin D is the most abundant saponin derived from P. grandiflorum and pharmacologically active component. Cytochrome P450s (CYPs) are important enzymes in the saponin biosynthesis. CYP is, in general, the terminal oxidase enzymes and essential roles in saponin biosynthesis pathway by hydroxylation or oxidaition of triterpene skeletons. Methods and Results : We tried to identify CYP genes related to saponin biosynthesis of P. grandiflorum through RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obtained 122,663 contigs and found 191 putative CYP genes. Familes of CYP716, CYP708, CYP93 and CYP51 were selected as putative saponin biosynthesis related gene families using phylogenetic relationship analysis. Conclusion : The results in this study could help to find the CYPs related to saponin biosynthesis pathway of P. grandiflorum.

Background : Medicinal crop has been used in the traditional Asian medicinal methods. From ancient times, various kinds of medicinal crop are being cultivated in East Aisa including Korea, China and Japan. In Korea, they used a variety of medicinal plants in folk medicine and oriental medicine since ancient times. Molecular markers can be widely used in a variety of settings such as effective-loci estimation, genetic-diversity characterization, allelic-effect studies, gene-flow studies, quantitative-trait locus (QTL) mapping, and evolutionary studies. The genetic analyses of crops require large numbers of useful molecular markers for genetic or QTL identification, comparative mapping and breeding. Studying the genetic diversity and genetic relationship of crops can assist breeders. Crop genetics within a breeding program enable the economic and effective cultivar development. We tried to develop a variety of molecular markers from Angelica gigas genomic sequences for genetic studies and breeding. Methods and Results : A. gigas resources cultivated in Republic of Korea were collected. Fresh leaves were ground with liquid nitrogen and gDNA was extracted using a DNeasy Plant Mini kit (Qiagen, Valencia, CA, USA). We sequenced the whole genomes of five A. gigas accessions using Illumina HiSeq 2500 platform and identified genomic Simple Sequence Repeat (SSR) and InDel markers. DNA amplification was conducted using the PCR system (Bio-Rad T-100 Thermal Cycler). PCR products were separated by capillary electrophoresis on the ABI 3730 DNA analyzer (Applied Biosystems) and Fragment analyzer automated CE system (Advanced Analytical Technologies, Ankeny, IA, USA). Conclusion : We developed novel SSR and InDel markers from A. gigas genomic sequences for further genetic studies and breeding.

Background : Angelica gigas, also called Dang Gui or Korean Angelica, is a major medicinal herb used in Asian countries such as Korea, Japan and China. A. gigas has many active constituents such as dercursin, decursinol angelate, nodakenetin, nodakenin, β-sisterol or α -pinene. But, there is no research on the gexpression of the genes related to saponin biosynthesis from A. gigas. In this study, we compared the expression of saponin biosynthesis related genes from various organs of A. gigas. Methods and Results : The reads of Angelica gigas mRNAs were produced using Illumina Hiseq 2000, and the reads were assembled to produce 113,597 contigs using CLC　Genomic Workbench. To select the saponin biosynthesis genes, assembled contigs were subjected to BLAST analysis at NCBI site. RNAs were extracted from five tissues, roots, stems, flowers, old leaves and young leaves of A. gigas. We produced total of 16 gene specific primers and used for RT-PCR. PCR conditions composed pre-denaturation at 95℃ for 3min, then 35 cycles of 95℃ for 30 sec, 57℃ for 30sec and 72℃ for 1min, and a final extension at 72℃ for 5min. Electrophoresis performed at 100 V, 30 min using 1.2% gel. Our experiment shows that A. gigas has several genes related to saponin biosynthesis and the genes were expressed from variety of organs. Conclusion : From the above results, we may suggest A. gigas genes related to the biosynthesis of saponins.

Background : Codonopsis is a flowering plants belong to the family Campanulaceae, and has many kinds of medicinal properties. As currently recognized, two other groups, Campanumoea and Leptocodon, are included in the Codonopsis. The enlarged genus Codonopsis is distributed in Eastern, Southern, Central, and Southeastern Asia. C. lanceolata, C. clematidea and C. pilosula has many kinds of medicinal properties and this plants are used as medicinal and edible plants. C. ovata and C. mollis are distributed in Pakistan Kashmir and Himalaya mountains at an altitude of about 3,000 m, and flowers bloom in July to August. Methods and Results : In this study, we analyzed the genetic diversity of 5 Codonopsis species using 8 SSR markers base on C. lancelolata genomic sequences. Samples were obtained from fresh leaves of 5 plants from each species and genomic DNA was extracted using CTAB method. PCR was performed in total 20μl reaction volume containing 20 ng of DNA template and 5 pmole of primers. PCR conditions composed pre-denaturation at 95℃ for 5 min, then 35 cycles of 95°C for 30 sec, 60°C for 30 sec and 72°C for 30 sec, and a final extension at 72℃ for 30 min. The amplified band sizes ranged from 74 to 301 bp and clearly showed single or doble bands in eletrophoresis. From the phylogenetic analysis, C. lanceolata was grouped together, but the others were not grouped together according to the species. Conclusion : We concluded that C. lanceolata cultivated in Korea is different from the other species, and the eight SSR markers used in this study are able to distinguish C. lanceolata from the other species.

Background : Platycodon grandiflorum is a perennial plant and a member of Camanulaceae family. Since ancient times, they have been using P. grandiflorum as an important medicinal plant in Korea. Platycodin D is the most abundant saponin derived from P. grandiflorum and pharmacologically active component. UDP-glycosyltransferases (UGTs) are important enzymes in the saponin biosynthesis. UGT is a glycosyltransferase and act on the final step of the secondary metabolite biosynthesis. Methods and Results : We tried to identify UGT genes related to saponin biosynthesis of P. grandiflorum through RNA-seq analysis. The sequencing was performed using Illumina Hi-Seq platform after cDNA library preparation. The produced reads were assembled using CLC Genomics Workbench software (CLC Bio, Inc.). We obtained 122,663 contigs and found 137 putative UGT genes. Familes of UGT71, UGT73, and UGT74 were selected as putative saponin biosynthesis related gene families using phylogenetic relationship analysis. qPCR condition about UGT73 is preheating 94℃ 180 sec, denaturation 94℃ 60 sec, annealing 53℃ 60 sec, extension 72℃ 90 sec, final extension 72℃ 600 sec, 45 cycles repeated. Conclusion : The results in this study could help to find the UGTs related to saponin biosynthesis pathway of P. grandiflorum.

Background : Codonopsis lanceolata is used as a natural medicine or vegetables. It originates in East Asia such as Korea, Japan and China. C. lanceolata roots contain various chemical compounds including saponins like Panax ginseng. Although C. lanceolata are cultivated in different regions of South Korea, no variety has been developed. Therefore, it is necessary to develop discriminating methods such as molecular markers in C. lanceolata species. Methods and Results : To find simple sequence repeat (SSR) markers, we sequenced C. lanceolata genomic DNA using Illumina HiSeq 2000 System. A total of 250,455 putative SSR loci were obtained, and 26,334 non-redundant primers were designed to amplify these SSRs. Di-nucleotied repeats were the most abundant SSR reapeats, accounting for 89.53% (23,578) of primer designed SSRs. Tri-nucleotide, tetra-nucleotide and penta-nucleotide accounted for 8.44% (2,223), 1.3%, (348) 0.2% (55), respectively. Tri-, tetra-, and penta-nucleotide (total of 2,626 SSRs) were investigated in silico to identify polymorphism between individuals. Consequently, 573 SSRs showed polymorphism. Forty genomic SSR markers were tested in 16 C. lanceolata plants for determination of PCR amplification and polymorphism. From these primers, 27 (67.5%) amplified products and the average polymorphism information content (PIC) value was 0.52. Conclusion : We development 27 SSR markers from C. lanceolata using NGS, and it could be used for breeding of new varieties in the future.