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 : Wild-cultivated ginseng (WCG) prices are very different according to root ages. Generally, two methods are used to discriminate the root ages of Panax ginseng C.A. Meyer. The first method is the yearly determination method by the ring dyeing method, and the second method is the confirmation the number of stem vestiges in the rhizome. In this study, we analyzed the agronomic and growth characteristics of the WCG cultivated in Korea. In this study, to determine the appropriate root ages discrimination method for the determination of the root ages of WCG, the root ages of WCG and cultivated ginseng was examined. Methods and Results : We examined the cultivated ginseng (CG) and WCG that was collected and sold by regional groups at the Korean market. WCG does not form annual rings, which are clear and regular in wild ginseng. Therefore, it is impossible to identify the age of WCG by using the annual growth rings staining method. However, the age can be estimated by determining the number of stem vestiges in the rhizome. Conclusion : From the results of the Study on identification of root age for quality evealuation in WCG in Korea. Appropriate root ages discrimination method of WCG was confirmation the number of stem vestiges in the rhizome.

Background : Panax ginseng C.A. Meyer is a representative medicinal plants and it has been used in traditional medicine because the plant have many effective component such as saponins. To obtain a wild-cultivated ginseng (WCG) which is similar to wild ginseng (WG), this method usually performed in a mountain using seeds or seedlings of cultivated ginseng (CG) and WG. WCG is very expensive because it is difficult to cultivate. However, systematic cultivation method have not been developed compared to their high added value. Furthermore, very high price of WCG caused the problem that Panax notoginseng or Panax quinquefolium are sold as WCG in Korean market. This is concerned as a serious problem to consumers. In this study, we analyzed the agronomic and growth characteristics of the WCG cultivated in Korea. Methods and Results : We examined the WCG that was collected and sold by regional groups at the Korean market. The root age, growth conditions, and quality level of the cultivated WCG were confirmed. WCG samples were collected from five areas in Korea (Bucheon, Cheongju, Hoengseong, Judeok and Ulsan). The main root diameter, root shape index, rhizome length, and root weight showed high level of variation and they did not form annual rings. Conclusion : Agronomic and growth characteristics of WCG showed high variations according to cultivating regions.

Background : The cultivation of wild greens in a forest farming system is an attractive alternative to wild harvesting, due to its much lower production cost compared with conventional cultivation, and its provision of a second income to the landowner. Yet little is known about the conditions that would maximize the growth and antioxidant activities of wild greens in a forest farming system. Thus, this study was conducted to evaluate the optimal conditions that would maximize antioxidant activities of Ligularia fischeri (Ledeb.) Turcz., being cultivated in three different cultivation systems in Korea. Methods and Results : After the fibrous roots of L. fischeri were planted in three different cultivation systems, this study was conducted to assess the effect on health-related properties such as total phenolic contents, flavonoids, DPPH (1,1-diphenyl-2-picrylhydrasyl) free radical scavenging activities and reducing power. From these harvests in different sites, extracts were prepared using methanol. Total phenolic content in forest farming system (1.061 ㎎·GAE/㎖) was higher than that in other products harvested in conventional and greenhouse system (0.666㎎·GAE/㎖). Also, flavonoid content was higher in forest farming system (0.124 ㎎·QE/ ㎖), compared to conventional and greenhouse system (0.084 ㎎·QE/㎖). Conclusions : Antioxidant activity and cultivation system seem to correlate with total polyphenol and flavonoid contents.

Background : Platycodon grandiflorum has been used as food material and a traditional medicine in Korea. In order to develop an efficient in vitro micropropagation technique for a rare plant species and conservation for inbred line of plant breeding. Methods and Results : Plant regeneration via organogenesis and somatic embryogenesis was investigated in Platycodon grandiflorum. Leaf, stem, root tissues of 7-day-old seedlings were cultured on 1/2MS medium containing various concentration (0, 0.5, 1 and 2 ㎎/L) of IBA, BA and NAA. The results showed that 1/2MS medium supplemented with BA+NAA 2.0 ㎎/L yielded the highest callus formation ratio of 73.5%. When various tissues (leaf, stem, root) were tested on 1/2MS medium containing BA 2.0 ㎎/L+ NAA 2.0 ㎎/L for somatic embryogenesis, the optimum tissue for embryogenic shoot induction was stem tissue. Callus were cultured on MS medium containing various concentration (0, 0.5 and 1 ㎎/L) of BA and NAA. The best rooting rate was achieved by three different medium (B5, 1/2MS and MS) and 1/2MS medium cultured the highest rooting ratio (82%). Conclusion : This study suggested that above micropropagation techniques can be used for rapid multiplication as well as in vitro or in vivo conservation of the Platycodon species.

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 : In the herbal medicine market, Angelica gigas, Angelica sinensis, and Angelica acutiloba are all called "Danggui" and used confusingly. We aimed to assess the genetic diversity and relationships among 14 Angelica species collected from different global seed companies. Toward this aim we developed DNA markers to differentiate the Angelica species. Methods and Results : A total of 14 Angelica species, A. gigas, A. acutiloba, A. sinensis, A. pachycarpa, A. hendersonii, A. arguta, A. keiskei, A. atropurpurea, A. dahurica, A. genuflexa, A. tenuissima, A. archangelica, A. taiwaniana, and A. hispanica were collected. The genetic diversity of all 14 species was analyzed by using five chloroplast DNA-based simple sequence repeat (SSR) markers and employing the DNA fragment analysis method. Each primer amplified 3 - 12 bands, with an average of 6.6 bands. Based on the genetic diversity analysis, these species were classified into specific species groups. The cluster dendrogram showed that the similarity coefficients ranged from 0.77 to 1.00. Conclusions : These findings could be used for further research on cultivar development by using molecular breeding techniques and for conservation of the genetic diversity of Angelica species. The analysis of polymorphic SSRs could provide an important experimental tool for examining a range of issues in plant genetics.

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: In the herbal medicine market, Angelica gigas, Angelica sinensis, and Angelica acutiloba are all called "Danggui" and used confusingly. We aimed to assess the genetic diversity and relationships among 14 Angelica species collected from different global seed companies. Toward this aim we developed DNA markers to differentiate the Angelica species. Methods and Results: A total of 14 Angelica species, A. gigas, A. acutiloba, A. sinensis, A. pachycarpa, A. hendersonii, A. arguta, A. keiskei, A. atropurpurea, A. dahurica, A. genuflexa, A. tenuissima, A. archangelica, A. taiwaniana, and A. hispanica were collected. The genetic diversity of all 14 species was analyzed by using five chloroplast DNA-based simple sequence repeat (SSR) markers and employing the DNA fragment analysis method. Each primer amplified 3 - 12 bands, with an average of 6.6 bands. Based on the genetic diversity analysis, these species were classified into specific species groups. The cluster dendrogram showed that the similarity coefficients ranged from 0.77 to 1.00. Conclusions: These findings could be used for further research on cultivar development by using molecular breeding techniques and for conservation of the genetic diversity of Angelica species. The analysis of polymorphic SSRs could provide an important experimental tool for examining a range of issues in plant genetics.

Background : Wild-cultivated P. ginseng (WCG) is a specific ginseng in Korea which depends on artificial forest growth method. To obtain a WCG which is similar to wild ginseng (WG), this method usually performed in a mountain using seeds or seedlings of cultivated ginseng (CG) and WG. Recently, very high price of WCG caused the problem that Panax notoginseng or Panax quinquefolium are sold as WCG in Korean market. This is concerned as a serious problem to consumers. In this study, we tried to develop a method to discriminate WCG, CG or WG using simple sequence repeat (SSR) markers and phylogenetic analysis. Methods and Results : WCG samples (3, 5, or 6-years old) were collected in Hoengseong, Gangwondo. DNA extraction was performed using CTAB method. SSR markers were collected from the published papers. After test PCR using the markers, one of the primer pair was labeled with fluorescence dye (FAM, NED, PET, or VIC) and Gene Scan analysis were performed. NTsys-PC program was used for the phylogenetic analysis of the data. Eight SSR markers were collected from the published literature and used for the analysis. From the 8 tested SSR markers, 7 SSR markers showed polymorphism between varieties. GenScan analysis were performed using the selected SSR markers to analyze the phylogenetic relationship of WCG. Conclusion : Phylogenetic analysis showed the relationship between WCG and P. ginseng cultivars and the seven SSR markers used in this study are able to distinguish Wild-cultivated P. ginseng.

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 : ‘baek-chul’(White atractylodes rhizome) widely used in traditional herbal remedies in Asia. A. Japonica and A. Macrocephala are used as ‘baek-chul’ in Japanese Pharmacopoeia but only A. Macrocephala is used as ‘baek-chul’ in Chinese Pharmacopoeia. Based on morphologic observation A. japonica has small infloresence diameter, white flowers and gynodioecism, whereas A. macrocephala has large inflorescence diameter, red flowers ,monoecism and developed rhizomes. but The distinction of these isn't easy. SSRs are very useful molecular markers for species identification. In this study, genetic diversity and identification between A. Japonica and A. Macrocephala were confirmed by SSR marker. Methods and Results : DNAs were extracted from leaf tissue of A. Japonica, A. Macrocephala and A. Japonica × A. Macrocephala (Breeding varieties, ‘Dachul’) using DNeasy plant Mini Kit (Qiagen, Hilen, Germany). these plants cultivated from RDA(Eumseong) and used for PCR amplification. The relative concentration of the extracted DNA was estimated Nano Drop ND-1000 (NanoDrop Technologies, Wilmington, De, USA) And final DNA concentration was adjusted to 5.5ng/μL. In this study 8 primer pairs were tested on 4 A. Japonica, 4 A. Macrocephala, 2 ‘Dachul’. These primers showed high polymorphism among and within four populations of A. macrocephala.(Zheng et al.). We detected interspecific and intraspecific SSR polymorphism by 3 primer pairs. Conclusion : The results showed that these markers were found to be useful for diversity analysis as they distinguished among Atractylodes spp. and also A.Macrocephala. This work is intended to serve as the basis for the breeding of new varieties in white atractylodes rhizome.

Background : Astragalus membranaceus is one of the most widely used traditional medicinal herbs in Korea. Studies on the genomic of A. membranaceus resources have not been carried out so far. The present study was carried out to discriminate A. membranaceus based on genetic diversity using genomic simple sequence repeat (SSR) markers. Methods and Results : We collected 5 A. membranaceus lines: Asung, Poongsung, Am-Jecheon, Am-Sancheong, and Am-China. One hundred mg of fresh leaves were used for genomic DNA extraction using the DNeasy plant DNA isolation kit (Qiagen GmbH, Hilden, Germany). 450,449 contigs were searched for 147,766 SSR candidate loci in this study using the MicroSAtellite identification tool (MISA). We selected 949 A. membranaceus genomic SSR markers that were showed variation for the five collections in silico screening with CLC genomics workbench program. The genetic diversity of all A. membranaceus resources was analyzed using 17 SSR markers employing the DNA fragment analysis method. Based on the genetic diversity analysis, these lines were classified into four distinct groups. Conclusion : These findings could be used for further research on cultivar development using molecular breeding techniques and for conservation of the genetic diversity of A. membranaceus. Furthermore, the markers could be used for marker-assisted selection for crop breeding.

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.

Background : Angelica gigas is a monocarpic biennial or short lived perennial plant. A. gigas, also called Dang Gui or Korean Angelica, is a major medicinal herb used in Asian countries such as Korea, Japan and China. In Korea, we are using the roots of A. gigas, but, they are using Angelica sinensis in China and Angelica acutiloba in Japan to obtain many active constituents. The biggest problem in the using of A. gigas would be the confusion with A. acutiloba or A. sinensis. These three plants can't be distinguished by appearance. And the constituent ratios of the three plants are different. This confusion can cause an accident or the pharmaceutical effects do not meet the expectations. In this study, we developed chloroplast SSR markers that can distinguish A. gigas, A. acutiloba and A. sinensis. Methods and Results : We collected A. gigas, A. acutiloba and A. sinensis. and extrated DNA using CTAB method. The DNA was diluted to 10 ng/㎕ and kept -20℃. We designed the primer sets using CLC Main Workbench based on chloroplast DNA SSR region of A. gigas. PCR were performed on the three angelica plant samples (in 5 repeat). Conclusion : We made five primer sets from five SSR regions of A. gigas cpDNA. All of the primer sets amplified the amplicon effectively. Two of the 5 primer sets had polymorphism. We can distinguish A. gigas, A. acutiloba, and A. sinensis using the 2 primer sets

Plant breeding requires the collection of genetically diverse genetic resources. Studies on the characteristics of Platycodon grandiflorum resources have not been carried out so far. The present study was carried out to discriminate P. grandiflorum based on morphological characteristics and genetic diversity using simple sequence repeat (SSR) markers. Methods and Results :We collected 11 P. grandiflorum cultivars: Maries II, Hakone double white, Hakone double blue, Fuji white, Fuji pink, Fuji blue, Astra white, Astra pink, Astra blue, Astra semi-double blue and Jangbaek. Analyses of the morphological characteristics of the collection were conducted for aerial parts (flower, stem and leaf) and underground parts (root). Next, the genetic diversity of all P. grandiflorum resources was analyzed using SSR markers employing the DNA fragment analysis method. We determined that the 11 P. grandiflorum cultivars analyzed could be classified by plant length, leaf number and root characteristic. Based on the genetic diversity analysis, these cultivars were classified into four distinct groups. Conclusions : These findings could be used for further research on cultivar development using molecular breeding techniques and for conservation of the genetic diversity of P. grandiflorum. Moreover, the markers could be used for genetic mapping of the plant and marker-assisted selection for crop breeding.

Background : A series of studies were conducted to optimize adventitious root induction in vitro from explants of Astragalus membranaceus using various nutrient media supplemented with plant hormones. Methods and Results : Levels of active components were analyzed from adventitious roots induced under different media conditions. Among the different media conditions, Murashige and Skoog medium supplemented with 1.0㎎• ℓ −1 indole-3-butyric acid resulted in the greatest adventitious root induction rate. The amount of the major active component of the adventitious roots of Ama1, calycosin-7-O-β-D-glucopyranoside was higher than that of other adventitious root samples. Conclusions : These results suggest that the adventitious roots of A. membranaceus could be used for the commercial production of medicines.