Search

The present study examined the volatile profiles of

Ginseng (Panax ginseng C. A. Meyer) is a multifunctional medicinal herb used worldwide and is an economically important high-value crop in Korea. Here, we presented the mitochondrial genome of P. ginseng landrace ‘Jakyung’, which is one of the most common cultivars cultivated in Korean farms. The complete mitochondrial genome sequence was 464,661 bp in length and had a single circular form. The ginseng mitochondrial genome encoded 72 unique genes, including 45 protein-coding genes, 24 tRNA genes, and three rRNA genes. Nucleotide composition analysis revealed a GC content of 45.1%, with a slightly higher A + T bias (A, 27.1%; T, 27.8%; G, 22.5%; C, 22.6%). Phylogenetic analysis showed that P. ginseng was closely related to Daucus carota in the Apiales. This complete mitochondrial genome sequence of P. ginseng provides valuable genetic information for further studies of this important medicinal plant.

Panax ginseng C. A. Mey., a perennial herb belonging to the family Araliaceae, is a valuable medicinal plant with distinctive biological characteristics. However, comprehensive analyses of the mitochondrial genome (mitogenome) are lacking. In this study, we sequenced the complete mitogenome of ginseng based on long-read data from the Nanopore sequencing platform. The mitogenome was assembled into a &ldquo, master circle&rdquo, form of 464,705 bp and contained 72 unique genes. The genome had three large repeat regions, and 10.42% of the sequences were mitogenome sequences of plastid origin (MTPTs). In total, 278 variants (213 SNPs and 65 InDels) were discovered, most of which were identified in intergenic regions. The MTPT regions were mutational hotspots, harboring 74.5% of the variants. The ginseng mitogenome showed a higher mutation rate than that of the chloroplast genome, and this pattern is uncommon in plants. In addition, 10 Kompetitive allele-specific PCR (KASP) markers were developed from 10 SNPs, excluding those in MTPT regions. These markers accurately identified the genotypes of 59 Korean ginseng accessions and elucidated mitogenome diversity. These results provide insight into organellar genomes and genetic diversity in ginseng. Moreover, the complete mitogenome sequence and 10 KASP markers will be useful for ginseng research and breeding.

The development of molecular markers is one of the most useful methods for molecular breeding and marker-based molecular associated selections. Even though there is less information on the reference genome, molecular markers are indispensable tools for determination of genetic variation and identification of species with high levels of accuracy and reproducibility. The demand for molecular approaches for marker-based breeding and genetic discriminations in Panax species has greatly increased in recent times and has been successfully applied for various purposes. However, owing to the existence of diverse molecular techniques and differences in their principles and applications, there should be careful consideration while selecting appropriate marker types. In this review, we outline the recent status of different molecular marker applications in ginseng research and industrial fields. In addition, we discuss the basic principles, requirements, and advantages and disadvantages of the most widely used molecular markers, including restriction fragment length polymorphism, random amplified polymorphic DNA, sequence tag sites, simple sequence repeats, and single nucleotide polymorphisms.

Panax ginseng, commonly known as ginseng, is a well-known medicinal plant that has been used as traditional medicine in China and Korea. Research in the past few decades supports the pharmacological effects of ginseng. For example, ginseng roots (extracts) exhibit multiple medicinal effects, such as anticancer, antiaging, and protection against circulatory shock, in humans. In this review, we summarize the progress made so far in the ginseng proteomics, starting from sample preparation to establishments of proteomes and databases. Both gel-based (1-DE and 2-DE in combination with LC–MS/MS) and gel-free proteomics technologies have been applied on wide range of samples, collected during different growth and developmental stages and under normal or adverse stress conditions. In particular, comparative proteome analysis has been carried out to investigate the protein profiles of Oriental, American and Indian ginsengs using majorly root and leaf tissues. Moreover, identification of stress-responsive proteins was a key focus that led to the detection of some of the common proteins such as heat shock protein (HSP), ATPase, enolase, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and ribonuclease/ginseng major protein (GMP). Acquired proteomics-based knowledge has been very fruitful in providing better insight into the ginseng biology, opening a door for comparative and translation research of other important medicinal plants. However, due to the fact that proteins undergo various post-transcriptional and post-translational modifications, obtained proteomics data do not always complement the transcriptomics data perfectly; therefore, future efforts would require the utilization of an integrated/holistic molecular-genetic (or omics) approach to explore the biology of this golden plant.

This study investigated the root-associated bacterial endophytes of Panax ginseng at different ages by shotgun metagenomic analysis. After mapping metagenome data to the complete ginseng genome to identify unmapped sequences, we predicted the structure and functions of ginseng bacterial endophytes by metagenomic rapid annotation using subsystems technology analysis. While Proteobacteria and Actinobacteria were the predominant phyla in all samples (2–6-year-old roots), class Alphaproteobacteria was most abundant in 3-, 4-, and 5-year-old plants. We found that 3-year-old P. ginseng had a 0.66% unmapped rate against the whole ginseng genome and showed the greatest diversity of endophytic bacteria (α diversity = 299). Prediction of endophytic bacterial functions at different ages by SEED subsystem analysis revealed that siderophore and auxin-related traits—which are known to promote plant growth—were most highly represented in 3-year-old plants. This was supported by a gene frequency analysis of plant growth-promoting genes, including those responsible for solubilization of phosphate and nitrogen metabolism, using BLASTn. These results suggest that endophytic bacteria of the P. ginseng root affect plant growth. Furthermore, the isolation and purification of plant growth-promoting endophytes identified in this study could promote sustainable cultivation of ginseng in the future.

Background Licorice (Glycyrrhiza spp. L.) is used as a natural sweetener and medicinal herb in European and Asian countries. Molecular studies have been conducted to find differences between wild and cultivated species because most wild species are highly resistant to abiotic and biotic stresses compared with their cultivated species. However, few molecular markers have been developed for studying the genetic diversity and population structure of licorice species and to identify differences between cultivars. Thus, the present study aimed to develop a set of genomic simple sequence repeat (SSR) markers for molecular studies of these species. Methods In the present study, we developed polymorphic SSR markers based on whole-genomesequence data of Glycyrrhiza lepidota. Then, based on the sequence information, the polymorphic SSR markers were developed. The SSR markers were applied to 23 Glycyrrhiza individual plants. We also evaluated the phylogenetic relationships and interspecies transferability among samples. Results The genetic diversity analysis using these markers identified 2–23 alleles, and the major allele frequency, observed heterozygosity, genetic diversity, and polymorphism information content were 0.11–0.91, 0–0.90, 0.17–0.94, and 0.15–0.93, respectively. Interspecies transferability values were 93.5%, 91.6%, and 91.1% for G. echinata, G. glabra, and G. uralensis, respectively. Phylogenetic analysis clustered cultivated (group 1) and wild (group 2) species into three and two subgroups, respectively. The reported markers represent a valuable resource for the genetic characteri z ation of Glycyrrhiza spp. for theanalysis of its genetic variability, and as a tool for licorice transferability. This is the first intraspecific study in a collection of Glycyrrhiza spp. germplasm using SSR markers.

Bacillus cereus PgBE311, isolated from the root tissue of a 5-year-old Panax ginseng plant, showed activities against the fungal pathogens Cylindrocarpon destructans and Botrytis cinerea . Here, we report the genome sequence of B. cereus PgBE311.

Bacillus cereus PgBE311, isolated from the root tissue of a 5-year-old Panax ginseng plant, showed activities against the fungal pathogens Cylindrocarpon destructans and Botrytis cinerea. Here, we report the genome sequence of B. cereus PgBE311., Bacillus cereus PgBE311, isolated from the root tissue of a 5-year-old Panax ginseng plant, showed activities against the fungal pathogens Cylindrocarpon destructans and Botrytis cinerea. Here, we report the genome sequence of B. cereus PgBE311. The bacterium contains antibiotic-related gene clusters and has the potential to stimulate plant growth.

Chryseobacterium indologenes PgBE177, isolated from the root tissue of a 4-year-old Panax quinquefolius plant, showed antagonistic activity against Pseudomonas syringae pv. tomato DC3000, a bacterial pathogen., Chryseobacterium indologenes PgBE177, isolated from the root tissue of a 4-year-old Panax quinquefolius plant, showed antagonistic activity against Pseudomonas syringae pv. tomato DC3000, a bacterial pathogen. Here, we report the whole-genome sequence of C. indologenes PgBE177. The bacterium contains bacteriocin gene clusters and has the potential to stimulate plant growth.

Licorice (Glycyrrhiza glabra) is an important medicinal crop often used as health foods or medicine worldwide. The molecular genetics of licorice is under scarce owing to lack of molecular markers. Here, we have developed cleaved amplified polymorphic sequence (CAPS) and high-resolution melting (HRM) markers based on single nucleotide polymorphisms (SNP) by comparing the chloroplast genomes of two Glycyrrhiza species (G. glabra and G. lepidota). The CAPS and HRM markers were tested for diversity analysis with 24 Glycyrrhiza accessions. The restriction profiles generated with CAPS markers classified the accessions (2-4 genotypes) and melting curves (2-3) were obtained from the HRM markers. The number of alleles and major allele frequency were 2-6 and 0.31-0.92, respectively. The genetic distance and polymorphism information content values were 0.16-0.76 and 0.15-0.72, respectively. The phylogenetic relationships among the 24 accessions were estimated using a dendrogram, which classified them into four clades. Except clade III, the remaining three clades included the same species, confirming interspecies genetic correlation. These 18 CAPS and HRM markers might be helpful for genetic diversity assessment and rapid identification of licorice species.

Licorice (Glycyrrhiza glabra) is an important medicinal crop often used as health foods or medicine worldwide. The molecular genetics of licorice is under scarce owing to lack of molecular markers. Here, we have developed cleaved amplified polymorphic sequence (CAPS) and high-resolution melting (HRM) markers based on single nucleotide polymorphisms (SNP) by comparing the chloroplast genomes of two Glycyrrhiza species (G. glabra and G. lepidota). The CAPS and HRM markers were tested for diversity analysis with 24 Glycyrrhiza accessions. The restriction profiles generated with CAPS markers classified the accessions (2–4 genotypes) and melting curves (2–3) were obtained from the HRM markers. The number of alleles and major allele frequency were 2−6 and 0.31–0.92, respectively. The genetic distance and polymorphism information content values were 0.16–0.76 and 0.15–0.72, respectively. The phylogenetic relationships among the 24 accessions were estimated using a dendrogram, which classified them into four clades. Except clade III, the remaining three clades included the same species, confirming interspecies genetic correlation. These 18 CAPS and HRM markers might be helpful for genetic diversity assessment and rapid identification of licorice species.

Korean ginseng (Panax ginseng C.A. Meyer) has been widely used for medicinal purposes and contains potent plant secondary metabolites, including ginsenosides. To obtain transcriptomic data that offers a more comprehensive view of functional genomics in P. ginseng, we generated genome-wide transcriptome data from four different P. ginseng tissues using PacBio isoform sequencing (Iso-Seq) technology. A total of 135,317 assembled transcripts were generated with an average length of 3.2 kb and high assembly completeness. Of those unigenes, 67.5% were predicted to be complete full-length (FL) open reading frames (ORFs) and exhibited a high gene annotation rate. Furthermore, we successfully identified unique full-length genes involved in triterpenoid saponin synthesis and plant hormonal signaling pathways, including auxin and cytokinin. Studies on the functional genomics of P. ginseng seedlings have confirmed the rapid upregulation of negative feed-back loops by auxin and cytokinin signaling cues. The conserved evolutionary mechanisms in the auxin and cytokinin canonical signaling pathways of P. ginseng are more complex than those in Arabidopsis thaliana. Our analysis also revealed a more detailed view of transcriptome-wide alternative isoforms for 88 genes. Finally, transposable elements (TEs) were also identified, suggesting transcriptional activity of TEs in P. ginseng. In conclusion, our results suggest that long-read, full-length or partial-unigene data with high-quality assemblies are invaluable resources as transcriptomic references in P. ginseng and can be used for comparative analyses in closely related medicinal plants.

This study was performed to identify optimal harvesting time of ginseng seeds and to examine the effect of GA3 treatment for improvement of seed stratification rate. Ginseng seeds harvested from Land race, Chunpoong and Yun- poong cultivar in July 20 were tested for stratification rate. It was shown that stratification rates of land race, Yunpoong and Chunpoong cultivar were 94.1%, 93.1%, and 82.6%, respectively. Seeds of Chunpoong cultivar harvested 10-15 days later showed a comparable stratification rate to that of Land race, indicating that late harvest of Chunpoong seeds is beneficial for the increase of stratification rate. The higher stratification rate was found in mature seeds (92.3%) than immature seeds (37.8%), both of which were harvested in July 20. Stratification rate of mature seeds harvested in July 15 was 87.5%, dem- onstrating optimal harvesting time of ginseng seeds with higher stratification rate is after mid-July. An exponential growth of endosperms of ginseng seeds was observed from early June to mid-June and then slow growth was observed. There was no obvious growth of embryos from fertilization to mid-August. After the this time, embryos quickly grew until late October. Thus, appropriate stratification control is essential during the period (from early September to late October) in order to optimize embryo growth and development. While no increase of stratification rate was observed in seeds treated with 50 ppm of GA3, significant increases were observed in seeds treated with 100 ppm of GA3. At this concentration of GA3, the stratification rate of Land race, Chunpoong and Yunpoong cultivar was 95.0%, 95.3%, and 96.5%, respectively.

Ginseng Research Division, National Institute of Horticultural and Herbal Science, Rural Development AdministrationAbstract The geographical origin of commercial red ginseng concentrate was studied using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The ginsenoside content of domestic and Chinese red ginseng concentrates wasdetermined. Four types of suspected origin samples could be selected this technique. The LC-MS/MS data were statisticallyanalyzed on the basis of canonical function analysis and principal component analysis. Domestic and Chinese samplescould be discriminated via canonical function analysis using posterior probability. In addition, the mixture ratio (Koreanor Chinese origin) of the unknown origin specimen could be predicted based on the relationship between the mixingconcentration of red ginseng concentrates and principal component 1.Keywords: red ginseng concentrate, geographical origin, liquid chromatography-tandem mass spectrometry, discrimination

*Ginseng Research Division, Department of Herbal Crop Research, NIHHS, RDA, Eumseong 369-873, Korea.**Department of Plant Bioscience, Pusan National University, Miryang 627-706, Korea.***Department of Plant Resources, Chungbuk National University, Cheongju 361-763, Korea.****National Agrobiodiversity Center, National Academy of Agricultural Science, RDA, Suwon 441-853, Korea.ABSTRACT : The transcriptomes of four ginseng accessions such as Cheonryang (Korean ginseng cultivar), Yunpoong(Korean ginseng cultivar), G03080 (breeding line of Korean ginseng), and P. quinquefolius (American ginseng) was charac-terized. As a result of sequencing, total lengths of the reads in each sample were 156.42 Mb (Cheonryang cultivar), 161.95Mb (Yunpoong cultivar), 165.07 Mb (G03080 breeding line), and 166.48 Mb (P. quinquefolius). Using a BLAST searchagainst the Phytozome databases with an arbitrary expectation value of 1E-10, over 20,000 unigenes were functionally anno-tated and classified using DAVID software, and were found in response to external stress in the G03080 breeding line, as wellas in the Cheonryang cultivar, which was associated with the ion binding term. Finally, unigenes related to transmembranetransporter activity were observed in Cheonryang and P. quinquefolius , which involves controlling osmotic pressure and tur-gor pressure within the cell. The expression patterns were analyzed to identify dehydrin family genes that were abundantlydetected in the Cheonryang cultivar and the G03080 breeding line. In addition, the Yunpoong cultivar and P. quinquefoliusaccession had higher expression of heat shock proteins expressed in Ricinus communis. These results will be a valuableresource for understanding the structure and function of the ginseng transcriptomes.Key Words : Panax ginseng, P. quinquefolius, Transcriptomes, Next Generation Sequencing

*Department of Plant Bioscience, Pusan National University, Miryang 627-706, Korea.**Ginseng Research Division, Department of Herbal Crop Research, NIHHS, RDA, Eumseong 369-873, Korea.ABSTRACT : Salt stress is one of the major abiotic stresses affecting the yield of ginseng (Panax ginseng C. A. Meyer). Theobjective of this study was to identify bio-marker, which is early responsive in salt stress in ginseng, using proteomicsapproach. Ginseng plants were exposed to 5 ds/m salt concentration and samples were harvested at 0, 6, 12 and 18 hoursafter exposure. Total proteins were extracted from ginseng leaves treated with salt stress using Mg/NP-40 buffer and wereseparated on high resolution 2-DE. Approximately 1003±240 (0 h), 992±166 (6 h), 1051±51 (12 h) and 990±160 (18 h)spots were detected in colloidal CBB stained 2D maps. Among these, 8 spots were differentially expressed and were identi-fied by using MALDI-TOF/TOF MS or/and LC-MS/MS. Ethylene response sensor-1 (spot GL 1), nucleotide binding protein(spot GL 2), carbonic anhydrase-1 (spot GL 3), thylakoid lumenal 17.9 kDa protein (spot GL 4) and Chlorophyll a/b bindingprotein (spot GL 5, GL 6) were up-regulated at the 12 and 18 hour, while RuBisCO activase B (spot GL 7) and DNA helicase(spot GL 8) were down-regulated. Thus, we suggest that these proteins might participate in the early response to salt stressin ginseng leaves.Key Words : Ginseng, Protein Biomarker, Proteomics, Salt Stress, 2-DE

Korean ginseng (Panax ginseng) and American ginseng (Panax quinquefolius) are widely used medicinal plants with similar morphology but different medicinal efficacy. Roots, flowers, and processed products of Korean and American ginseng can be difficult to differentiate from each other, leading to illegal trade in which one species is sold as the other. This study was carried out to develop convenient and reliable chloroplast genome-derived DNA markers for authentication of Korean and American ginseng in commercial processed products. One codominant marker could reproducibly identify both species and intentional mixtures of the two species. We further developed a set of species-unique dominant DNA markers. Each species-specific dominant marker could detect 1% cross contamination with other species by low resolution agarose gel electrophoresis or quantitative polymerase chain reaction. Both markers were successfully applied to evaluate the original species from various processed ginseng products purchased from markets in Korea and China. We believe that high-throughput application of this marker system will eradicate illegal trade and promote confident marketing for both species to increase the value of Korean as well as American ginseng in Korea and worldwide.

*Department of Herbal Crop Research, NIHHS, RDA, Eumseong 369-873, Korea.**National Agrobiodiversity Center, NAAS, RDA, Suwon 441-707, Korea.***Experiment Research Institute of National Agricultural Products Quality Management Service, MIFAFF, Seoul 150-043, Korea.****Department of Food Science and Technology, Seoul Women’s University, Seoul 139-774, Korea.*****Department of Crop Science and Biotechnology, Dankook University, Cheonan 330-714, Korea.ABSTRACT : The development of random amplified polymorphic DNA (RAPD) and expressed sequence tag-derivedsimple sequence repeats (EST-SSRs) provided a useful tool for investigating Korean ginseng genetic diversity. In this study,18 polymorphic markers (7 RAPD and 11 EST-SSR) selected to assess the genetic diversity in 31 ginseng accessions (11Korean ginseng cultivars and 20 breeding lines). In RAPD analysis, a total of 53 unique polymorphic bands were obtainedfrom ginseng accessions and number of amplicons ranged from 4 to 11 with a mean of 7.5 bands. Pair-wise genetic similaritycoefficient (Nei) among all pairs of ginseng accessions varied from 0.01 to 0.32, with a mean of 0.11. On the basis of theresulting data, the 31 ginseng accessions were grouped into six clusters. As a result of EST-SSR analysis, 11 EST-SSR mark-ers detected polymorphisms among the 31 ginseng accessions and revealed 49 alleles with a mean of 4.45 alleles per primer.The polymorphism information content (PIC) value ranged from 0.06 to 0.31, with an average of 0.198. The 31 ginsengaccessions were classified into five groups by cluster analysis based on Nei’s genetic distances. Consequently, the results ofginseng-specific RAPD and EST-SSR markers may prove useful for the evaluation of genetic diversity and discrimination ofKorean ginseng cultivars and breeding lines.Key Words : Korean Ginseng, Cultivar, Breeding Line, Genetic Characteristics, DNA Marker, RAPD, EST-SSR

The geographical origin of red ginseng extract (RGE) was studied using a mass spectrometry based electronic nose. Imported RGE and domestic RGE were diluted to 12 o Bx. The treated RGE was analyzed, and discriminant function analysis (DFA) was used for discriminating of geographical origins. The DFA plots indicated a significant separation of imported RGE and domestic RGE. The F-value of discriminant function first score (DF1) was much higher than that of discriminant function second score (DF2), indicating that discrimination was mainly affected by DF1. Based on DF1, the concentration of domestic RGE to imported RGE shifted to the left side of DFA plot, and the mixing ratio highly correlated to DF1 value. Unknown sample (#2) was closely located to the sample of mixed imported : domestic (6:4) RGE. In the bar graph, the DF1 value correlated to the mixing ratio. Unknown samples (#2) were thought to be mixed with the imported RGE. This technique could be used to efficiently differentiate the geographical origin of RGE.

Korean ginseng (P. ginseng C. A. Meyer) is one of the most important medicinal plant in the world. Under- standing genetic variability among the assortment of Korean ginseng is important for breeding. The aim of this study was to molecularly characterize Korean ginseng cultivar and breeding lines through the use of eight previously reported STS markers (MFGp183, MFGp130, MFGp110, UFGp74, UFGp163, MFGp108, MFGp81 and UFGp156). All STS markers produced interpretable electropherograms from 31 accessions consisting of 11 Korean ginseng cultivars and 20 breeding lines. When eight STS markers were combined, we identified to total 19 genetic patterns; in particular, nine cultivars (Chun- poong, Yunpoong, Gopoong, Gumpoong, Sunpoong, Sunone, Cheongseon, Sunhyang, Cheonryang) and 5 breeding lines (G08012, G04079, G04075, G08036, G04110) in ginseng samples can be discriminated from the others. Together with other available markers, these STS markers will contribute to the management of ginseng genetic resources and the protection of breeders' rights.

Department of Herbal Crop Research, NIHHS, RDA, Eumseong 369-873, Korea.ABSTRACT: This study was carried out to investigate the difference of the content of soil chemical components and growthcharacteristics in five years old ginseng affected by application of manure in paddy-converted field. As all livestock manureregardless of kinds increased along with the whole soil chemical component, including the pH and EC in 2008. Change in theEC of control plot was slightly increased but not exceeded 1 ds/m over the years. However, the changes in the EC of livestockmanure regardless of kinds and amounts were highly increased and irregularly exceeded 1.5 ds/m in 2012. The 5 years oldginseng root fresh weight, treatment of fertilizing pig manure compost 4 ton per 10 areas (PMC 4t on/10a) and fowl manurecompost 4 ton per 10 areas (FMC 4 ton/10a), were superior to the others. But there were no difference between PMC 4 ton/10a, FMC 4 ton/10a and control. The standing crop rate 39.6%, treatment of fertilizing cattle manure compost 4 ton per10 areas (CMC 4 ton/10a), was best in all livestock manure. However that was relatively lower than control. Physiologicaldisorder occurrence rates of livestock manure related with leaf and root of ginseng were also higher than that of control. Ifexcessively using non-decomposed livestock manure, It would be caused physiological disorder in many ways. It is a bigproblem to be producing the quality ginseng. More research is needed to find out the economic and effective fertilizer.Key Words : Korean Ginseng, Paddy-Converted Field, Livestock Manure, Physiological Disorder

This study describes the identification of Panax species using mitochondrial consensus primers. Initially, a total of thirty primers were tested in ten Korean ginseng cultivars and two foreign Panax species, P. quinquefolius and P. notoginseng. In the polymerase chain reaction (PCR) amplification results, three primers (cox1, nad1/2-3 and nad2/1-2) generated co-dominant polymorphic banding patterns discriminating Korean ginseng cultivars from P. quinquefolius and P. notoginseng. However, these primers could not generated polymorphisms among the Korean ginseng cultivars, and simply represented species-specific polymorphisms for P. quinquefolius and P. notoginseng. Primers PQ91 and PN418 were designed from the consensus sequence of nad1/2-3 region. Two banding patterns (A or B) were detected in PQ91. Korean ginseng cultivars and P. notoginseng shared the same banding pattern (A type) and P. quinquefolius was identified another banding pattern (B type). In the case of PN418, two banding patterns (A or B) were detected in the Korean ginseng cultivars and two foreign Panax species. Korean ginseng cultivars and P. quinquefolius shared the same banding pattern (A type) and P. notoginseng was identified another banding pattern (B type). The combination banding patterns of three Panax species, Korean ginseng cultivars (Panax ginseng C. A. Mey.), P. quinquefolius and P. notoginseng, was identified as `AA`, `BA` and `AB`, respectively. Consequently, PQ91 and PN418 primer sets can be used to distinguish among Panax species.

We established that regenerated plants of Panax ginseng were directly transplanted into the soil without the acclimatization process. When regenerated plants were treated at the concentration of 0.4 mg l -1 for 9 hr before transplantation into the soil, our results showed that the survival rate of them was the highest. The optimal size of the root was above 4 cm in length and 0.4 g by weight before transplantation into the soil. The highest survival rate increased to 59.6%. After 4 years of cultivation in the fields, our results sh owed that there were no significant morphological differences between regenerated plants and wild-type ginseng. Also, we obtained healthy seeds from regenerated plants. This study provides the basis for a relatively rapid multiplication of selected genotypes and will allow a much more rapid evaluation of the germplasm of the unexplored and unstudied Korean ginseng. Key words - Acclimatization, Gibberellic acid, Ginseng, Tissue culture * 교신저자(E-mail): choije@cnu.ac.kr

Panax ginseng roots are well known for their medicinal properties and have been used in Korean and Chinese traditional medicines for 1000s of years. However, the medicinal value of P. ginseng fruits remain poorly characterized. In this study, we used an integrated biochemical, proteomics, and metabolomics approach to look into the medicinal properties of ginseng fruits. DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS [2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid)] assays showed higher antioxidant activities in ginseng fruits than leaves or roots. Two-dimensional gel electrophoresis (2-DE) profiling of ginseng fruit proteins (cv. Cheongsun) showed more than 400 spots wherein a total of 81 protein spots were identified by mass spectrometry using NCBInr, UniRef, and an in-house developed RNAseq (59,251 protein sequences)-based databases. Gene ontology analysis showed that most of the identified proteins were related to the hydrolase (18%), oxidoreductase (16%), and ATP binding (15%) activities. Further, a comparative proteome analysis of four cultivars of ginseng fruits (cvs. Yunpoong, Gumpoong, Chunpoong, and Cheongsun) led to the identification of 22 differentially modulated protein spots. Using gas chromatography-time of flight mass spectrometry (GC-TOF MS), 66 metabolites including amino acids, sugars, organic acids, phenolic acids, phytosterols, tocopherols, and policosanols were identified and quantified. Some of these are well known medicinal compounds and were not previously identified in ginseng. Interestingly, the concentration of almost all metabolites was higher in the Chunpoong and Gumpoong cultivars. Parallel comparison of the four cultivars also revealed higher amounts of the medicinal metabolites in Chunpoong and Gumpoong cultivars. Taken together, our results demonstrate that ginseng fruits are a rich source of medicinal compounds with potential beneficial health effects.

Consumers are increasingly concerned about the origin of foods, so the geographical origin of foods has been a major topic of debate and extensive research. Various instrumental methods (e.g. high performance liquid chromatography (HPLC), gas chromatography (GC), capillary electrophoresis (CE), electronic nose, near-infrared spectroscopy (NIRS), nuclear magnetic resonance spectroscopy (NMR), DNA analysis, multi-isotope analysis) in conjunction with statistical analysis, were developed and applied in attempt to provide reliable answers to their geographical origin. This study reviews current developments in the application of various methods for a clear geographical origin of foods. The limitation of discrimination analysis for geographical origin was also discussed.

This study was carried out to research microorganisms having the antifungal activity against ginseng Alternaria blight pathogen Alternaria panax and ginseng anthracnose pathogen Colletotrichum gloeosporioides. Eleven Bacillus strains. were isolated from Korean traditional soybean paste and Kimchi. Among the 11 isolates, DJ5, KC1, KC2 and KC4 showing antagonistic activity on the mycelial growth of A. panax and C. gloeosporioides in pairing culture were finally selected as the antagonistic microorganisms. Based on 16s rRNA sequence and phylogenetic tree analysis, they were identified as Bacillus spp.. The selected microorganisms were investigated antagonistic activity by measured leaf-segment colonization in pot test. When Bacillus sp. were injected after A. panax treatment, KC1, KC2 and KC4 showed similar effect to chemical pesticides treated control. To measure preventive effect of Bacillus sp, antagonistic microorganisms were injected and C. gloeosporioides was treated in pot. When measuring the effectiveness for the prevention of Anthracnose, All Bacillus spp. showed approximately 83~90 % degree of superior preventive effect. In general, The four Bacillus spp. isolated from Korean traditional fermented foods showed therapeutic effect of Alternaria blight and preventive effect of Anthracnose.

These studies were conducted to provide basic information on Korean ginseng cultivars and breeding lines (Panax ginseng C. A. Mey.) and to identify the variations that can be utilized in ginseng breeding programs. The agronomic characteristics was used to clarify the genetic relationships among Korean ginseng cultivars and breeding lines and to classify them into distinct genetic groups. Angle of petiole and number of fibrous root showed a wide variation from and 0~5, respectively. The average plant length was 54.2cm with a range of 37.9~64.8cm and the average stem diameter was 5.6mm with a range of 4.0~7.5mm. The average stem length was 31.9cm with a range of 21.8~37.9cm and the average root weight was 38.1 g with a range of 23.0~52.0 g. The 24 Korean ginseng cultivars and breeding lines were classified into 4 groups based on agronomic characteristics using the complete linkage cluster analysis. The I, II, III and IV groups included the 60.8%, 7.4%, 13.1% and 8.7% of the cultivars and breeding lines, respectively. The breeding lines in group I could be characterized as the group with the highest growth characters and yield components, such as plant length, stem diameter and root weight. The root weight, the yield component, had highly significant positive correlations with stem diameter, plant length and stem length.

In order to develop a novel system for the discrimination of five ginseng cultivars (Panax ginseng Meyer), single nucleotide polymorphism (SNP) genotyping assays with real-time polymerase chain reaction were conducted. Nucleotide substitution in gDNA library clones of P. ginseng cv. Yunpoong was targeted for the SNP genotyping assay. From these SNP sites, a set of modified SNP specific fluorescence probes (PGP74, PGP110, and PGP130) and novel primer sets have been developed to distinguish among five ginseng cultivars. The combination of the SNP type of the five cultivars, Chungpoong, Yunpoong, Gopoong, Kumpoong, and Sunpoong, was identified as 'ATA', 'GCC', 'GTA', 'GCA', and 'ACC', respectively. This study represents the first report of the identification of ginseng cultivars by fluorescence probes. An SNP genotyping assay using fluorescence probes could prove useful for the identification of ginseng cultivars and ginseng seed management systems and guarantee the purity of ginseng seed.

In this study, simple sequence repeat (SSR) analyses were utilized for evaluation of genetic diversity and dis- crimination of 17 accessions. Five cultivars, which were developed from Korea, and 12 foreign accessions, which were col- lected from China, Japan, Russia and USA, were evaluated by nine markers out of 22 SSR markers. A total of 39 alleles were detected, ranging from 2 to 8, with an average of 4.3 alleles per locus. The expected heterozygosity and PIC values were 0.627 and 0.553, with a range from 0.21 (GB-PG-078) to 0.76 (GB-PG-142) and from 0.19 (GB-PG-078) to 0.70 (GB-PG- 142), respectively. Four makers out of nine SSR markers, GB-PG-026, GB-PG-043, GB-PG-142 and GB-PG-177, were selected as key factors for discrimination of Korean ginseng cultivars and foreign accessions. All of Korean ginseng cultivars and foreign accessions were individually by the combination of four SSR markers. Consequently, the SSR markers devel- oped in this study may prove useful for the evaluation of genetic diversity and discrimination of Korean ginseng cultivars and foreign accessions.

The principal objective of this study was to develop a discrimination method using SSR markers in Korean ginseng cultivars. Five cultivars--Chunpoong, Yunpoong, Gopoong, Sunpoong, and Kumpoong--were evaluated by nine markers out of 22 SSR markers. A total of 23 alleles were detected, ranging from 1 to 4, with an average of 2.6 alleles per locus, and an averages of gene diversity (GD) of 0.480. Nine markers were tested in order to distinguish among five Korean ginseng cultivars. Two markers out of nine SSR markers, GB-PG-065 and GB-PG-142, were selected as key markers for discrimination among Korean ginseng cultivars. Two genotypes were detected in GB-PG-065. Chunpoong and Kumpoong shared the same allele type, and Yunpoong, Gopoong, and Sunpoong shared another identical allele type. In the case of GB-PG-142, a specific allele type differentiated from those of other four cultivars was observed only in Sunpoong cultivar. Consequently, the SSR markers developed in this study may prove useful for the identification of Korean ginseng cultivars and the development of ginseng seed management systems, as well as tests to guarantee the purity of ginseng seeds.

Recently, new ginseng cultivars having superior agricultural traits have been developed in Korea. For newly developed plant cultivars, the identifi cation of distinctiveness is very important factors not only in plant cultivar management but also in breeding programs. Thus, eighty-fi ve inter simple sequence repeat (ISSR) primers were applied to detect polymorphisms among six major Korean ginseng cultivars and two foreign ginsengs. A total of 197 polymorphic bands with an average 5.8 polymorphic bands and 2.9 banding patterns per assay unit across six Korean ginseng cultivars and foreign ginsengs from 236 amplifi ed ISSR loci with an average 6.9 loci per assay unit were generated by 34 out of 85 ISSR primers. Three species of Panax ginseng including the Korean ginseng cultivars, P. quinquefolius, and P. notoginseng, could be readily discriminated using most tested primers. UBC-821, UBC-868, and UBC-878 generated polymorphic bands among the six Korean ginseng cultivars, and could distinguish them from foreign ginsengs. Sequence characterized amplifi ed region (SCAR) marker system was introduced in order to increase the reproducibility of the polymorphism. One SCAR marker, PgI821C650, was successfully converted from the randomly amplifi ed polymorphism by UBC-821. It showed the expected dominant polymorphism among ginseng samples. In addition, the specifi c polymorphism for Sunwon was generated by treating Taq I restriction enzyme to polymerase chain reaction products of PgI821C650. These results will serve as useful DNA markers for identifi cation of Korean ginseng, especially Sunwon cultivar, seed management, and molecular breeding program supplemented with marker-assisted selection.

This study describes an efficient approach for developing sequence tagged sites (STS) for Panax ginseng C.A. MEYER, and their applications for line discrimination. By using the methylation filtering (MF) technique, a genomic library was constructed, in which clone inserts were derived from the hypomethylated regions of ginseng genome. A methylation unfiltered genomic library was also constructed and the clone inserts were compared to those from the MF library in terms of sequence characteristics. Sequence analysis revealed that MF efficiently enriched the protein coding region of P. ginseng, for which the repetitive DNA appeared to be as little as 2.5 fold lower than clones in the unfiltered library, and also indicated that the P. ginseng genome may contain a large fraction of methylated repetitive DNA elements. A total of 99 and 100 highly stringent STS primer sets were designed from the filtered and unfiltered library, respectively. Amplification products were tested for latent polymorphism across six cultivars of P. ginseng and other 2 Panax species using six endonucleases recognizing four-bases. STS primer sets described here will be useful for marker-assisted selection, genome mapping and line discrimination of P. ginseng or its cultivars from other Panax species.

－ The root of Astragalus membranaceus is a traditional folk medicine that has been used for many therapeutic purposes in Asia. It reportedly acts as an immunostimulant, tonic, hepatoprotective, diuretic, antidiabetic, analgesic, expectorant, sedative, and anticancer drug. In this study, metabolomic profiling was applied to the roots of A. membranaceus of different ages using NMR coupled with two multivariate statistical analysis methods: such as principal components analysis (PCA) and canonical discriminant analysis (CDA). This allowed various metabolites to be assigned in NMR spectra, including γ-aminobutyric acid (GABA), aspartic acid, succinic acid, glutamic acid, glutamine, N-acetyl aspartic acid, acetic acid, arginine, alanine, threonine, lactic acid, and valine. The score plot from PCA and also CDA allowed a clear separation between samples according to age. Keywords: Fingerprinting analysis, Astragalus membranaceus , 1 H NMR spectroscopy, Principal component analysis, Canonical discriminant analysis

The present study describes the molecular authentication of 21 Korean Artemisia species using PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) technique based on the trnL-F sequences in chloroplast DNA. Five different banding patterns were generated from 21 Artemisia species using HinfI restriction enzyme. A. apiacea, A. keiskeana and A. sieversiana have specific banding patterns. The remaining 18 species had shared two banding patterns. Phylogenetic analysis based on trnL-F sequence variations showed results similar to PCR-RFLP banding patterns. It suggested that the trnL-F region does not have sufficient variations to identify the 21 Artemisia species. However, the specific banding patterns for A. apiacea, A. keiskeana and A. sieversiana can be utilized as a DNA marker for discriminating them from other Artemisia species. These markers will be also useful for developing A. apiacea, A. keiskeana and A. sieversiana into new medicine and food based on their efficacy.