Background : F. fomentarius extracts have been recently reported to process immune-stimulatory and anti-inflammatory effects. In this study, we evaluated the anti-mutagenic effect of F. fomentarius ethanol extracts and the effective chemical components from the extract were analyzed by LC-Q-TOF. Methods and Results : F. fomentarius was extracted with 70% ethanol yielding a crude extract 6.8%. The crude extract was fractionated sequentially to diethyl ether, chloroform, dichloromethane, butanol and water with a yield of 43.9%, 2.958%, 6.8%, 21.6% and 25.5.%, respectively. The anti-mutagenic effect of F. fomentarius extract and its fractions was tested in Ames test by two type of Salmonella typhimurium (TA98 and TA100). Among the five fractions, diethyl ether has shown the highest and significant anti-mutagenic effect (98.3%, at 3,000 ㎍/plate concentration). Moreover, we investigated the chemical constituents of the extract using UPLC-Q-TOF. A total of 10 compounds such as flavonoids (velutin, 3, 4′,5-Trihydroxy-3′,7-dimethoxyflavanone) and fatty acids (γ-linoleic acid, stearic acid) and deterpenoid (kirenol) were tentatively identified with an accurate mass within 10 ppm mass error. Also, flavonoids and fatty acids were detected with higher rate than other compounds based on obtained chromatograms. Conclusion : We demonstrated that F. fomentarius extract and fractions have anti-mutagenic activity through Ames test. Furthermore, we will carry out isolation of each components from its fraction and use them to conduct additional anti-mutagenic test.

Backgoound : This study was conducted to evaluate the quality variation of Ixeris dentata on the antioxidant contents and antioxidant activities according to the different producing area. Methods and Results : The samples were extracted with 70 % EtOH and then analyzed for total flavonoid contents, polypenol contents, DPPH radical scavenging activity, and ABTS radical scavenging activity. Luteinol 7-O-β-D-glucoside, an index component of the Ixeris dentata, was analyzed by HPLC. The leafs of Ixeris dentata in Jinan had the highest concentration of polyphenols (23.91 ㎎/g), followed by Jinbu (22.63 ㎎/g) and Eumseong (21.36 ㎎/g). Flavonoid content was highest in Jinan (15.27 ㎎/g), but there was no significant difference between Jinbu (14.05 ㎎/g) and Eumseong (13.99 ㎎/g). The contents of luteinol 7-O-β-D-glucoside were confirmed in Jinan (0.68%), Jinbu (0.49%) and Eumseong (0.36%), respectively. Conclusion : The comparison of antioxidant contents and antioxidant activities of Ixeris dentata according to the different producing area, Jinan was had the highest concentration, followed by Jinbu and Eumseong. Our results showed that the content of luteoline-7-D-glucoside varied among the different producing area.

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 : 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 : Malonyl ginsenoside content of the Panax ginseng is known to account for 35% to 60% of total ginsenosides content. However, its distribution by ginseng part has not been studied. In this study, four kinds of malonyl ginsenosides were compared in Korean white ginseng part using the purified malonyl ginsenoside standards in our laboratory. Methods and Results : White ginseng was prepared by the air drying (50℃, 48h) or freeze drying (-70℃, 48h) methods form 4-year-old ginseng. Malonyl ginsenoside content in total ginsenosides were similar in air dried and freeze dried white ginseng, 58% and 62%, respectively. Therefore, malonyl ginsenoside contents in main, lateral, and fine root, and in the main root without skin and skin of main root prepared by freeze dried method were compared. Malonyl ginsenosides (m-Rb1, m-Rb2, m-Rc and m-Rd) and total ginsenosides (Rg1, Re, Rf, Rb1, Rc, Rb2, Rb3, Rd, m-Rb1, m-Rb2, m-Rc and m-Rd) were 6.75 and 14.15 mg/g in main root, 14.15 and 26.35 mg/g in lateral root, 46.95 and 84.15 mg/g in fine root. Malonyl ginsenoside contents in skin of main root was 20.08 mg/g, while its contents of the main root without skin was 2.58 mg/g. Conclusion : As a result, the parts each air drying the sample was confirmed that the ratio of the distribution of malonyl ginsenoside (main root : lateral : fine root = 18.7 : 11.1 : 16.2), and distribution ratio of main root, skin of main root, lateral, skin of lateral was found to be (12.2: 14.6 14.3: 3.7). Malonyl ginsenoside content was the highest in fine root, compared to the main or lateral root. Malonyl ginsenoside contents in skin of root was higher than those of the main root without skin. These results is expected to help establish an efficient extraction and standardization. Malonyl ginsenoside analysis of White ginseng using HPLC expects that the standardization process can be established.

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.

The insulin-like growth factor (IGF) pathway is a key signal transduction pathway involved in cell proliferation, migration, and apoptosis. In dairy cows, IGF family proteins and binding receptors, including their intracellular binding partners, regulate mammary gland development. IGFs and IGF receptor interactions in mammary glands influence the early stages of mammogenesis, i.e., mammary ductal genesis until puberty. The IGF pathway includes three major components, IGFs (such as IGF-I, IGF-II, and insulin), their specific receptors, and their high-affinity binding partners (IGF binding proteins [IGFBPs]; i.e., IGFBP1–6), including specific proteases for each IGFBP. Additionally, IGFs and IGFBP interactions are critical for the bioactivities of various intracellular mechanisms, including cell proliferation, migration, and apoptosis. Notably, the interactions between IGFs and IGFBPs in the IGF pathway have been difficult to characterize during specific stages of bovine mammary gland development. In this review, we aim to describe the role of the interaction between IGFs and IGFBPs in overall mammary gland development in dairy cows.

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 : 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: We evaluated the effect of ulinastatin on paw withdrawal threshold (PWT), IL-6, and IL-10 in SD rats after spinal nerve ligation (SNL). Methods: Group C received N/S and Group E received ulinastatin IV for three days following SNL. PWT, IL-6, and IL-10 were measured on the 3rd, 5th, and 7th day. Results: Group E showed higher PWT compared to group C. IL-6 was lower in group E than in group C. No differences in IL-10 were observed between group C and group E. Conclusion: Ulinastatin increased the PWT and its effect appears to be involved with IL-6, not IL-10.

The lumbar discectomy is the surgery with low morbidity itself and need the prone position, which is essential for this surgery. However, the increased intra-abdominal pressure after changing supine to prone position may lead intra-abdominal hypertension or abdominal compartment syndrome, consequently acute mesenteric ischemia. Acute mesenteric ischemia is one of the fatal conditions of acute abdomen with overall mortality of 60% to 100%. Unfortunately, the early detection of mesenteric ischemia in perioperative period of spinal surgery is difficult. We herein report an experience of acute mesenteric ischemia after lumbar discectomy in prone position, and review the relative literatures.

The wild relatives of soybean [Glycine soja Sieb. and Zucc.] have curly/wavy nature whereas cultivated varieties are upright. Such morphological characteristics have agronomic importance too. To investigate the molecular mechanism of development contributing to coiled morphology, screening was carried out to look for Arabidopsis mutants in activation tagging lines obtained by activation T-DNA treatment that have curly/wavy morphology. A mutant named Coiled Branch 1 (cbr1), is found to have a wavy and curly morphology with coiling branches. Plasmid rescue and genomic southern blot analysis revealed the site of T-DNA insertion in the genome. RT-PCR was performed to monitor expression levels of the genes adjacent to the T-DNA integration sites, and showed the activation of an E3 ubiquitin ligase gene. Database search showed that the gene with the RING domain belongs to a family of E3 ubiquitin ligases. Complementation test by overexpression and RNA interference of the gene was also carried out. The complementation test results showed that the novel gene activation tagging affected the cbr1 mutant phenotypes. Ubiquitylation has been linked virtually to every cellular process including plant development. E3 ubiquitin ligase has been reported to recognize target proteins that are to be ubiquinated for further degradation by the proteasome complex. Further, more detailed studies are needed to identify the specific substrate(s) of the novel E3 ubiquitin ligase gene.

When the rice blast fungus attacks rice, fungal proteins are secreted into the plant apoplast to facilitate infection. The rice plant recognizes such secreted proteins, which result in the induction of defense responses. However, the molecular mechanisms of how rice plant recognizes secreted proteins remain elusive. Here, we report that a small, secreted protein, Magnaporthe oryzae snodprot1 homolog (MSP1), is recognized by rice plants and triggers host cell death and defense responses. Furthermore, pre-treatment of rice with Domain II, elicitor-active epitope of MSP1, induces resistance to the pathogen KJ301. We demonstrated that secretion of MSP1 into the apoplast is prerequisite for triggering cell death and activating defense-related gene expression, suggesting that it is recognized by a receptor in the host plasma membrane. Through comprehensively analysis of transcriptional profile in rice leaves and suspension cultured cells (SCCs) in response to exogenous MSP1 and Domain II treatment using 60K Agilent microarray chip, we found that 27 signaling genes, such as F-box(6), MAPK(4), protein kinase(11), transcription factor(6), were up-regulated in leaves and SCCs and six protein kinases were targeted into plasma membrane. Thus, we suggest that some of these genes may act as receptor of MSP1 in response to exogenous MSP1 treatment. Expression pattern of candidate genes was further checked in response to different environment cues using open rice data. These results demonstrate that these genes may be also involved in the signaling in response to cold stress, root-JA treatment and brown plant hopper (BPH) attack.

In this study, genetic diversity of wild Codonopsis lanceolata collected in Korea were analysed using SSR makers. Wild C. lanceolata roots were collected in Jeollanam-do Jangheung-gun Choentae Mountain as in roots. The wild C. lanceolata plants were cultivated in Chungbuk National University greenhouse and the leaves were sampled from 36 plants. The genomic DNA of C. lanceolata was extracted using CTAB. PCR was performed using a program of 35 cycles at 94℃ for 30 sec, 60℃ for 30 sec, and 72℃ for 30 sec with an pre-denaturation of 94℃ for 5 min and a final extension of 72℃ for 30 min. The PCR reaction mixture contains 5 pmole of primers and 20 ng of DNA template in a 20 μL reaction volume. The genotype of the analyzed samples were very different. Therefore, the wild C. lanceolata collected in Korea look genetically diverse.

Angelica 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 using Angelica acutiloba. in Japan to obtain many active constituents such as dercursin, decursinol angelate, nodakenetin, nodakenin, umbelliferone, β-sisterol, or α-pinene. The plants of the Angelica family are used to improve gynecological health. The biggest problem in the cultivation of A. gigas is bolting. If the bolting occurs, A. gigas can not be used as a medicinal component because the roots are lignified. In this study, 11 A. gigas genetic resources in Korea; 1. Hwangje variety, 2. Sungwoo Jongmyo company, 3. Bonghwa No. 1, 4. Bonghwa No. 2, 5. Bonghwa No. 3, 6. Bonghwa No. 4, 7. Jechun local variety, 8. Jirisan local variety, 9. Manchu variety in Eumseong, 10. Manchu variety in Bonghwa, 11. Jinbu local variety, were collected and performed phylogenetic analysis using RAPD molecular markers.

Soybean [Glycine max (L.) Merr.] seeds are abundant in high-quality proteins and fats. In addition, soybean seeds are also rich in secondary metabolites, such as isoflavones, lecithin, and saponins. Triterpene saponins are major components of these physiologically active metabolites in soybean seeds. Soybean saponins are classified as group A and DDMP saponins. Among them group A saponins are undesirable component of food products due to bitterness and astringency and also cause foaming in tofu production. Whereas, DDMP saponins and their derivatives are less bitter and astringent and beneficial to human health when consumed as regular diet. Therefore, reducing the group A saponins or increasing the DDMP saponins are required to improve the food quality. The present study focused to identify and characterize the gene which is encoding a protein responsible for biosynthesis of DDMP saponins. EMS mutant lines (sg-7-1 & sg-7-2) which lack DDMP saponins were developed. The breeding cross has been made with these two mutants with two cultivars, Pungsannamul and Wooram to study the segregation and genetic linkage analysis, respectively. The segregation analysis showed that the mutant phenotype is controlled by single recessive gene. TLC analysis for phenotyping F2 population of Wooram X sg-7-1 showed mutant, wild and heterozygous types. To surprise two more patterns were detected and they were named as strange type1 (ST1) and strange type2 (ST2). Further, SSR marker analysis will be carried out to locate the gene which encoding a protein responsible for biosynthesis of DDMP saponins.

Soybean germplasm have diverse accessions with great variation in their ability to survive and reproduce under salt stress conditions. In general, cultivated soybeans are more sensitive to salt stress than their wild relatives, however exceptions are found in both the groups. These variations in response to salt stress makes soybean germplasm an interesting collection of genetic resources to be explored for the identification of salt-tolerance genes, and their mechanism of action. Here, in this report we presented a data showing differential response of selected accessions of both cultivated and wild soybeans to salt stress. Two modes of salt treatment; gradual salt stress (GS) as well as salt shock (SS) were used in this study. The GS was found more effective in finding the difference in response of soybean accessions to salt stress. Various genetic marker based methods are in use to identify and isolate the potential genes contributing to the salt tolerance in soybean. Even then there is a paucity of knowledge on the key genes contributing to the salt tolerance in soybean. We expect that a recently developed functional screen based method, like yeast based functional screen, using cDNA library generated from different salt tolerant accessions of soybean could lead to identification of novel genes responsible for salt tolerance in soybean. Also, we propose for the use of RNA isolated from different stages of GS and SS for making cDNA library to be used for functional screening.

Soybean [Glycine max (L.) Merr.] have a variety of flower colors which are controlled by six different genes (W1,W2,W3,W4,Wm, and Wp). Among these genes, mutation in W3 gene causes near white flowers in the background of w4 genotype whereas the genotype W3w4 does purple throat flowers. Earlier studies showed that dihydroflavonol 4-reductase1 (DFR1) gene was closely linked to the flower color variants for W3 locus. In order to find out the W3 gene responsible for w3 phenotype, we first, studied the candidate gene Glyma14g07940 (DFR1) which is having 100% similarity with DFR probe sequence. Sequence analysis of DFR1 between W3 and w3 soybeans showed one base substitution in exon 6 of w3 mutant soybean resulting in one amino acid change in the amino acid sequence. However, comparison of amino acid sequences of DFR proteins from various crop plants showed that there is no functional change in the protein. Besides, the promoter analysis showed that, 311 bp of indel was traced in 5’-upstream promoter region of DFR1 gene in the w3 mutant. Here, we show that the near white or purple throat phenotypes in G. max is associated with existence or nonexistence of indel at 5’- upstream promoter region and low or high expression of DFR1, respectively. These results suggest that w3 phenotype may be caused by certain regulator of DFR1 gene located near or distant from DFR1 in G. max. In further study, we need to check the correlation between promoter indel with W3 expression level through GUS analysis.

Codonopsis lanceolata is a perennial climber. The roots are used as medicinal materials or vegetables. Recently, demand for C. lanceolata is increasing as a healthy food. C. lanceolata is distributed in India and East Asia such as China, Japan as well as Korea. In South Korea, this plant is widely cultivated in Gangwon-do province. No C. lanceolata varieties were developed in Korea. The objective of this study is to analyze genetic diversity of C. lanceolata cultivated in Korea using SSR makers. C. lanceolata roots were collected in each region were cultivated in Chungbuk National University greenhouse. Samples were obtained from fresh leaves of 5 plants from each collection region. The genomic DNA was extracted using CTAB. Genetic diversity was analysed using 4 sets of C. lanceolata SSR makers. PCR was performed in total 20 μL reaction volume containing 20 ng of DNA template, 5 pmole of primers. The genotypes of the analyzed samples were very similar. That means that the genetic diversity of C. lanceolata cultivated in Korea is very low.