Pluripotent stem cells can be derived from both pre- and post-implantation embryos. Embryonic stem cells (ES cells), derived from inner cell mass (ICM) of blastocyst are naïve pluripotent and epiblast stem cells (EpiSCs) derived from post-implantation epiblast are primed pluripotent. The phenotypes and gene expression patterns of the two pluripotent stem cells are different each other and EpiSCs thought to be in a more advanced pluripotent (primed pluripotent state) than mouse ES cells (naïve pluripotent state). Therefore, we questioned whether EpiSCs are less potential to be differentiated into specialized cell types in vitro. EpiSCs were isolated from 5.5～6.5 day post coitum mouse embryos of the post-implantation epiblast. The EpiSCs could differentiate into all tree germ layers in vivo, and expressed pluripotency markers (Oct4, Nanog). Interestingly, EpiSCs also were able to efficiently differentiate into neural stem cells (NSCs). The NSCs differentiated from EpiSCs (EpiSC-NSCs) expressed NSC markers (Nestin, Sox2, and Musasi), self-renewed over passage 20, and could differentiate into two neural subtypes, neurons, astrocytes and oligodendrocytes. Next, we compared global gene expression patterns of EpiSC-NSCs with that of NSCs differentiated from ES cells and brain tissue. Gene expression pattern of brain tissue derived NSCs were closer to ES cell-derived NSCs than EpiSC-NSCs, indicating that the pluripotent stem cell-derived somatic cells could have different characteristics depending on the origin of pluripotent stem cell types. * This work was supported by the Next Generation Bio-Green 21 Program funded by the Rural Development Administration (Grant PJ 008009).

Neural stem cells (NSCs) are self-renewing tripotent cell populations and have capacity of neuronal (neurons) and glial (astrocytes and oligodendrocytes) differentiation. Many researchers have reported that NSCs have therapeutic effects in neurological disease by transplantation. However, it is not easy to obtain NSCs in vitro. Recently, Yamanaka and colleagues showed that somatic cells could be reprogrammed into pluripotent state by enforcing reprogramming factors. Induced pluripotent stem (iPS) cells undergo unlimited self-renewal and have differentiation potential into various types of cells like embryonic stem cells. Direct differentiation into a specialized cell types from iPS cells hold considerable promise for regenerative medicine as well as basic research. Here, we induced differentiation of iPS cells into NSCs in vitro and in vivo, which were compared with embryonic stem (ES) cell-derived NSCs and brain derived NSCs. NSCs from ES and iPS cells were morphologically indistinguishable from brain derived NSCs and stained positive for NSCs markers Nestin and Sox2. ES cells derived NSCs were transcriptionally distinguishable from brain derived NSCs. However, global gene expression pattern were similar but distinct between iPS derived NSCs and brain derived NSCs. Moreover, iPS derived NSCs were spontaneously aggregated upon passaging, formed ES cell like colonies, and finally reactivated Oct4-GFP. The spontaneously reverted GFP-positive cells (iPS-NSC-iPS) expressed similar levels of pluripotency markers (Oct4,Nanog) to ES and iPS cells, and could form germ line chimera. One possible explanation for this phenomenon is that spontaneously re-reprogramming was associated with transgene re-activation when iPS cells were differentiated into NSCs. However, NSCs from dox-inducible iPScells could not be reprogrammed into pluripotent state without doxycycline. Taken together, iPS derived NSCs were morphologically and similar to brain derived NSCs, but differ in gene expression pattern and maintenance. * This work was supported by the Next Generation Bio-Green21 Program funded by the Rural Development Administration (Grant PJ008009).

Background : Phlomidis radix is known to be effective for pain relief and anti-inflammation. Phlomidis radix (Han-Sok-Dan) is likely to be confused because it is similar in name to Dipsaci Radix (Cheon-Sok-Dan), or is similar in morphology to other Phlomioides species. Therefore, we investigated the origins and morphological characteristics of the origin plant of Phlomidis radix in order to cultivate and consume the correct one. Methods and Results : We reviewed North and South Korean and Chinese Pharmacopoeia and the books on oriental herbal medicine and botanical illustrations. In South Korea, P. umbrosa was defined as the origin plant of Phlomidis radix, whereas in North Korea, P. maximowiczii was defined. In China, it was not used as an oriental medicine. In the past, P. umbrosa, which is native to the Korean peninsula, or P. maximowiczii, which is native to the northern part of the Korea was used instead of Dipsacus asperoides for Dipsaci Radix. However, after the publication of the Korean Pharmacopoeia, Phlomidis radix and Dipsaci Radix have been distinguished, and only P. umbrosa was defined as the origin plant of Phlomidis radix. Therefore, Phlomidis radix and Dipsaci Radix should be distinguished from each other, the origin plant of Phlomidis radix is considered to be P.umbrosa. Furthermore, we investigated the external morphological and growth characteristics of Phlomoides species in Korea. There are three species of Phlomoides in Korea, such as P. umbrosa, P. maximowiczii, P. koraeinsis. P. umbrosa has no basal leaves but only stem leaves, while P. maximowiczii and P. koraeinsis have basal leaves. Also, P. maximowiczii is distinguishable from seeds with hairs while P. koraeinsis from hairless seeds. Conclusion : We considered P. umbrosa as the origins of Phlomidis radix. As a result of investigation of morphological and growth characteristics, P. umbrosa, P. maximowiczii and P.koraeinsis could be distinguished by the presence or absence of basal leaves and the difference in whether the seeds have hair or not. These results could be used as a basis for standard varieties research and cultiviation of P. umbrosa.

Background : The small-scale medicinal crops is rapidly decreasing, and there is a risk of collapse of the pharmaceutical production base due to the radical market opening such as the Korea - China FTA. Among them, the Artemisia gmelinii has been used in the oriental medicine, and it is effective in treating the fever, humidity, urination, and scabies. Although it is being grown in small areas such as Gyeongnam and Gangwon Province in Korea, standardization of raw material production is insufficient. Then we were carried out on seed characteristics and growth characteristics by harvest time of A. gmelinii. Methods and Results : We were collected 114 accession from 7 regions from 2017 to 2018. The collected resources were growing to the testing field of Department of Herbal Crop Research. An average length and width of seed in A. gmelinii were 1.05 ㎜, 0.57 ㎜ respectively. A thousand seed weight were 0.42 g. Seed germination rate was highest at 25℃ (76.4%). Plant height was similar, stem diameter was 9 – 15 ㎜ in early growth collected resources. AG1801 had many leaves (60 ea), AG1802 was blooming fast. Conclusion : The above results showed that AG1801, AG1802, and AG1803 were selected good resources. We will analyze a valid ingredient by harvest time.

Background : Licorice has been a medicinal crop which was mainly used as a traditional medicine and food, depends on most imports (99%) in Korea. We have been trying to produce licorice in Korea for a long time. However when it grow in Korea the main ingredients are below the standard value and occur of physiological disorder. Therefore, we evaluated the growth characteristics and major components of genetic resources to develop licorice varieties suitable for the domestic environment. Methods and Results : We collected 19 accession of licorice from 7 regions including China, Mongolia, Uzbekistan, and Russia from 2013 to 2014. After regenerating the resources of licorice in the field we identified and classified the species. Among them, 13 lines of 2 species (G. uralensis Fisch, G. glabra L.) were selected from 2015 to 2016. We evaluated the aerial parts, underground parts, yield and glycyrrhizin content of 13 line of licorice grown one year. Plant length and stem diameter of GLY2014 - 002 was high and thick on the aerial parts. GLY2013 - 005 had many pods. GLY2014-006 had many adventitious roots like taproot (3 ea/plant) and high of root weight (170 g/plant), GLY2013 - 005, GLY2014 - 006 had high glycyrrhizin content (2.1%) in the underground parts. Conclusion : As a result of growth characteristics and glycyrrhizin analysis, GLY2013 - 002, GLY2013 - 005, GLY2013 - 007, GLY2014 - 002, GLY2014 - 006, and GLY2014 - 007 were evaluated as good resources. Therefore of six lines will be used to test productivity and conduct regional testing for developing licorice variety.

Background: The demand of recycling renewable agricultural by-products is increasing. Radiation breeding is a method used to improve saccharification efficiency. Thus, we investigated the effect of gamma ray irradiation on the pretreatment and enzymatic hydrolysis of the stalks of Senna tora, an important medicinal plants. Methods and Results: S. tora seeds were irradiated with gamma ray at doses of 100, 200, 300, and 400 Gy. In the pretreated biomass, glucan and lignin content were higher in the M1 (1st generations of irradiation) S. tora stalks than in the M2 (2nd generations of irradiation) stalks, this can be explained by the higher degradation rate in M1. After oxalic acid pretreatment, the concentration of total phenolic compounds (TPCs) in the hydrolysate increased in the gamma ray treated seeds. The highest relative increase rate in crystallinity in the pretreated biomass was observed in M1-400 Gy and M2-100 Gy. The cellulose conversion rate was higher in M1 than in M2, except for 200 Gy. Conclusions: Gamma ray irradiation at an appropriate dose can be used to improve the efficiency of pretreatment and enzymatic hydrolysis, thereby increasing biomass availability.

Background : Ixeris genus has been used in traditional medicines as stomachics, sedatives, and diuretics. Ixeris dentata var albiflora is a kind of perennial herbaceous plant and one of the plants of the genus Ixeris (Asteraceae). It is well-known for edible wild vegetable in Korea, China, Japan, and Mongolia. Specially, Korean has its root and young leaf with appetizing vegetable due to bitter taste. Methods and Results : We isolated 8 genes that are involved in carotenoid biosynthesis using the Illumina/Solexa HiSeq2000 platform. In this study, a full-length cDNA clone encoding phytoene synthase (IdPSY), phytoene desaturase (IdPDS), ξ-carotene desaturase (IdZDS), lycopene β-cyclase (IdLCYB), and zeaxanthin epoxidase (IdZEP) and partial-length cDNA clones encoding lycopene ε-cyclase (IdLCYE), ε-ring carotene hydroxylase (IdCHXE), and β-ring carotene hydroxylase (IdCHXB2) were identified in I. dentata. The theoretical molecular weight (MW) and isoelectric point values of 8 genes were investigated. Sequence analyses revealed that these proteins shared high identity and conserved domains with their orthologous genes. IdPSY, IdPDS, IdZDS, IdLCYB, IdCHXB2, and IdZEP were constitutively expressed in the roots, stems, leaves, and flowers of I. dentata. Conclusion : Our study on the biosynthesis of carotenoids in I. dentata will provide basic data for elucidating the contribution of carotenoids to the considerable medicinal properties of I. dentata.

Background : In previous studies, adlay seeds showed a prevalence of diversified fungal flora with the predominant fungal genera being Fusarium (45.6%) and In vitro test showed that fungal toxins like Fumonisin and Zearalenone were produced by Fusarium fujikuroi, F. asiaticum and F. graminearum. Because of this we performed experiments to selecting disinfective chemicals for controlling the Fusarium contamination in the adlay seed. Methods and Results : We carried out the chemical efficacy test such as seed disinfectants selection test appling before planting and pesticides selection test using in the earing season. In the present study, eleven different commercially available seed disinfectant were applied to the adlay seeds. Among 11 seed disinfectants, Hexaconazole+Prochloraz emulsifiable concentrate (EC) and Prochloraz emulsifiable concentrate (EC) had control value of 80% or above against Fusarium species tested. In the pesticides selection test, seven different commercially available pesticides for Fusarium blight (Scab) control were applied and we observed that Metconazole suspension concentrate(SC) strongly inhibited the mycelial growth of 10 Fusarium species all. Conclusion : From the above results, we selected Hexaconazole+Prochloraz EC and Prochloraz EC as a seed disinfectants and Metconazole SC as a pesticide using in the earing season for Scab control.

Background : This experiment was conducted to select GAP applying seed disinfectants in Astragalus membranaceus and Platycodon grandiflorum. Methods and Results : We carried out the chemical efficacy and injury test. For the efficacy test, we investigated fungal detection rate by seed disinfectants and for the chemical injury, we investigated germination rate and emergence rate by seed disinfectants in reference amount and fold amount. These experiments carried out two times. The results obtained are as follows. In the experiment for seed disinfectants selection of Astragalus membranaceus, all tested chemicals such as Tebuconazole emulsifiable concentrate(EC), Thiophanate-methyl + Triflumizole wettable powder(WP), Prochloraz copper chloride complex+Tebuconazole suspension concentrate(SC), Prochloraz emulsifiable concentrate(EC), Fludioxonil wetting liquid(WL) and Hexaconazol+Prochloraz emulsifiable concentrate(EC) had control value of 80% or above against seed contaminated fungi. However two chemicals such as Tebuconazole EC and Prochloraz copper chloride complex+Tebuconazole SC and two chemicals such as Prochloraz EC and Hexaconazol+Prochloraz EC exhibited chemical injury significantly in reference amount and in fold amount respectively, compared to non treated control. In the case of seed disinfectants selection of Platycodon grandiflorum, Prochloraz copper chloride complex+Tebuconazole SC, Prochloraz EC and Hexaconazol+Prochloraz EC had control value of above 80% against seed contaminated fungi except Thiophanate-methyl+Triflumizole WP and Fludioxonil WL. However Hexaconazol+Prochloraz EC and Prochloraz copper chloride complex+Tebuconazole SC exhibited chemical injury significantly in reference amount and in fold amount respectively, compared to non treated control. Conclusion : From the above results, we finally selected three items of seed disinfectants including Thiophanate-methyl+Triflumizole WP and Fludioxonil WL in Astragalus membranaceus and Prochloraz emulsion in Platycodon grandiflorum.

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.

Through earlier breeding efforts, portions of the genome of the wild species Oryza grandiglumis (2n=48) have been introgressed into the rice cultivar, Hwaseongbyeo (Oryza sativa spp. japonica). Hwaseongbyeo was used as recurrent parent and O. grandiglumis