Background : Most of cases in korea, distributed herbal medications are storaged in room temperature and it causes many insect pests during storage. Despite the fact, there is little research about storage conditions. Temperature and humidity in storehouse are highly related with pests’ overwintering, generation length and the number of occurrence in summer. Consequently, in this research, major pests’ overwintering was studied by reviewing facility condition, temperature and humidity in major regional storehouses. The results of this would help to develop how to manage pest occurrence in distributed herbal medications. Methods and Results : For this study, storage facilities was investigated at different regions including Gyeongdong Market in Seoul, the pharmaceutical company in Daegu and farmhouse in Youngcheon by monitering temperature, humidity, ventilator, air conditioning and heating on ground and underground places from June 2014 to July 2016. including Upon investigation, most of building using for storage had simple ventilators. Whereas in case of farmhouse, it is vulnerable to maintaining quality. Because most farmhouse had temporary buildings which are difficult to control temperature and humidity. In addition, among the other surveyed warehouses, Basement in the Gyeongdong Market had most favorable condition for pest overwintering. The lowest temperature of it was seldom below zero maintaining level of about 1.5℃ and average temperature was about 10℃ in underground warehouse during winter. And the ground warehouse of Daegu had similar patterns in temperature and humidity with that of Seoul in that being over 0℃ even in the winter. In Youngcheon, the temperature of farmhouse was below zero degree and the daily range of temperature and humidity was wide. It caused difficulty of overwintering of insect pests but possible to inhabit. Conclusion : According to a study by Han-Mo, Koo(1997), it was impossible to overwinter for Lasioderma serricorne F. which is dominant medicinal plants’ pest in north of the Chungcheong region’s warehouse due to low temperature. But in case of investigated warehouses, it was possible to overwinter because of the high temperature in office building or basement storage area. In order to make prediction of pest occurrence and comprehensive measures, it is urgent to take more investigation by regional groups and types of storage.

Background : Garlic is one of the important vegetables and a source of natural anti-oxidants. This study was carried out to investigate the antioxidant activities including 2,2-diphenyl-1-picr yl-hydrazil (DPPH), 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS), ferric reducing antioxidant power (FRAP), reducing power (RP) and total polyphenol content (TPC) of A. ampel oprasum L. Methods and Results : DPPH, ABTS, FRAP, TPC, and RP were analyzed the extract of aerial plant part in 22 accessions of A. ampeloprasum by spectrophotometry mathod. A. tuberosum Rottler ex Spreng. was used as a control. DPPH was ranged from 0.35 to 3.06 ㎍ ASC ㎎-1 dw. ABTS and FRAP showed wide variation from 4.37 to 29.30 ㎍ Trolox ㎎-1 dw and 3.18 to 10.8 ㎍ ASC ㎎-1 dw, respectively. RP and TPC were ranged from 3.43 to 9.51 ㎍ GAE ㎎-1 dw, and 0.8 to 18.63 ㎍ ASC ㎎-1 dw, respectively. Cluster analysis of A. ampeloprasum germplasm was divided into two major groups. Group Ⅰ (7 accessions) characterized as higher antioxidant activities than the group Ⅱ (15 accessions) and had similar antioxidant activities with A. tuberosum as a control. FRAP value showed a significant strong positive correlation with DPPH (r = 0.868*) and ABTS activity (r = 0.826*). Principal component analysis showed that the first two principal components (PC1 and PC2) cumulatively explained 85.64 % of total variation. Conclusion : From the above results, we may suggest that A. ampeloprasum aerial parts might have useful as a new material for functional food.

Background : The natural stable isotope ratio of common bio-elements like carbon (C), nitrogen (N), oxygen (O), or sulfur (S) varies with diverse isotope fractionation processes in nature. Therefore, measuring the variation of these stable isotope ratios in ginseng roots can be a feasible tool to discriminate the geographical origins of ginseng in Korea. Methods and Results : The 3-year-old six Korean ginseng cultivars were cultivated at the five regions in Korea, and then used for measuring the stable isotope ratios of C, N, O, and S by isotope ratio mass spectrometry (IRMS). The mean C, N, O, and S stable isotope ratio values in the ginseng roots significantly differed according to the cultivation regions (p < 0.05). However, these isotope ratios in ginseng roots had relatively weak discriminative power against to the ginseng cultivars at each cultivation region. The interaction of the cultivation region and ginseng cultivar type also significantly affected to the C, N, O, and S stable isotope ratio in ginseng roots (p < 0.0001). The two-dimensional plots associated with the N stable isotope ratio can effectively separate the ginseng roots in Jinan compared to those in the other regions. The partial least squares-discriminant analysis showed more significant separation between ginseng geographical origins compared to the principal component analysis. Conclusion : Our findings improve our understanding of how the isotope composition of ginseng roots varies with respect to cultivation regions and cultivars, and suggest that the analysis of the stable isotope ratios combined with chemometrics can be used as a feasible tool to discriminate geographical origin of ginseng in Korea.

Background : Adenophora triphylla var. japonica Hara. is one of the important oriental medicinal herbs being used to cure lung/respiratory diseases. It belongs to the five Sam (ginseng) including Panax ginseng, Salvia miltiorrhiza, Sophora flavescens, and Scrophularia buergeriana. In 2015, Korea produced 72 tons of medicinal herbs in a 14-hectare area and imported 24 tons valued at US$95,000. This study was carried out to investigate the effect of several biopesticides to control leaf spot of A. triphylla var. japonica Hara. Methods and Results : One year plants at the Department of Herbal Crop Research, NIHHS, RDA were used. The treatments used biopesticides such as plant extracts (BP), microbial agent (DM), microbial agent (PJ), sulfur powder (HS), and controlled chemicals (ST), while non treatment (NT) was used as control. After the early part of disease outbreak, the plants were treated four times with 7 days interval. The incidence of leaf spot, efficacy, phytotoxicity and control value were investigated. Based on the efficacy test, incidence of leaf spot is lowest at 15.6% in plant extracts (BP). Other biopesticides showed higher incidence - 16.7% for microbial agent (DM), 17.8% for microbial agent (PJ), 18.9% for sulfur powder (HS) and 71.1% for non treatment (NT), compared with 11.1% in controlled chemicals (ST). The microbial agent (PJ), sulfur powder (HS), plant extracts (BP) and microbial agent (DM) were not damaged in the phytotoxicity test. In the test of application dose, time and methods, the control value was highest in plant extracts (BP) at 78.1%, followed by microbial agent (DM) at 76.6%, microbial agent (PJ) at 75.0%, and sulfur powder (HS) at 73.4% , compared with controlled chemicals (ST) at 84.4%. Conclusion : It is possible to use biopesticides at the proper time to prevent leaf spot in Adenophora triphylla var. japonica Hara.

Background : Adenophora triphylla var. japonica Hara. is used to cure respiratory/ lung diseases, which makes it an important oriental medicinal herb. It is one of the five Sam (ginseng), which includes Panax ginseng, Salvia miltiorrhiza, Sophora flavescens, and Scrophularia buergeriana. In 2015, Korea imported 24 tons of medicinal herbs worth US$95,000. This study was carried out to investigate the effect of several mulching materials on the growth and root yield of A. triphylla var. japonica using organic fertilizer. Methods and Result : Seeds of A. triphylla var. japonica were collected at the nursery of the Department of Herbal Crop Research, NIHHS, RDA in 2015. Mulching materials used are black plastic, black woven fabric, and biodegradable plastic. Hand weeding was done to serve as control. The soil was fertilized two months before planting. Seeds of A. triphylla var. japonica were sowed in tray in early March and were allowed to grow for two months. It was planted in early May with planting density of 30 ㎝ x 15 ㎝ and was covered with various mulching materials. Parameters investigated are survival rate, aerial part and root growth characteristics, and yield at harvest time. Results showed that biodegradable plastic is most favorable for plant growth with height of 17.2cm, leaf length of 13.4cm and leaf width of 5.6 ㎝. Dry plant weight was heaviest in biodegradable plastic treatment at 2.8 g, and lightest at 2.5 g when planted without mulch. Biodegradable plastic has the highest dry root weight of 19.3 g, followed by black vinyl (18.4 g), woven fabric (18.3 g) and without mulch (17.9 g). The yield obtained per 10 a was highest in biodegradable plastic treatment at 174 ㎏, while black vinyl and woven fabric treatments produced 169 ㎏ and 160 ㎏, respectively. Plants without mulch had the lowest yield of 157 ㎏. Conclusion : In the above results, biodegradable plastic and woven fabric are considered suitable mulching materials for the cultivation of A. triphylla var. japonica.

Background : Angelica gigas N., one of the herbs has been used most frequently with Glycyrrhiza uralensis. It is usually cultivated in semi-alpine of more than 400m in North Central province of South Korea. However its production is becoming more unstable due to the climate change especially abnormally high temperature. In particular, the environmental change may cause not only a change in production but also a change in active ingredient of Angelica gigas N. This study was performed to investigate these problems. Methods and Results : This study was carried out in temperature gradient tunnel(TGT) in Pyeongchang, Gangwon province. Temperature conditions were maintained the ambient+0~2℃ (T1), ambient+2~4℃(T2), ambient+4~5℃(T3) throughout the year. We planted and grown one year old seedlings of Angelica gigas N. from mid-April to early November. As a result, there was no difference in early growth of Angelica gigas N. among the temperature range but there was a difference between each treatment region from the survey in June. Average length of soot in T1, T2, T3 was the largest(36cm), medium(27cm), smallest(19cm) respectively. This trend was similar to a survey in September and November. Each of death rate was 0%(T1), 19%(T2), 71%(T3). There was no bolting according to the temperature. The yield of a plant of Angelica gigas N. was 476g(T1), 76g(T2), 26g(T3), respectively. Production of Angelica gigas N. is foreseen 84.7% and 94.5% reduction if the temperature rises to 3±1℃ and 4±1℃ more than it is at present based on temperature in Pyeongchang 2015. Total amount of Decursin was reduced at higher temperatures. Conclusion : The results of this study show that quality and production of Angelica gigas N. is expected to decline significantly if the temperature rise than the current. Therefore development of various techniques is required to response to climate change.

Background : The roots of Curcuma longa Radix is used to make herbal medicine, which is registrated in the Korean Pharmacopoeia. This species is a perennial herb, and is distributed throughout tropical and/or sub tropical regions. High temperature and humidity with small difference between daytime and nighttime is the finest environment condition in this species. Recently, demand for C. longa has been expanded to cultivation area as central region from southern region. Therefore, this study aimed to investigate the effect of harvest time on the growth, yield characteristics and main composition contents in C. longa. Methods and Results : Curcuma longa rhizomes were harvested from farmland in Jindo, Jeollanam-do in end October, early November, middle November, end November, early December, middle December and end December 2015. Harvested rhizome were dried in hot air drier, and it was investigated as growth characteristics and yield. Contents of curcumin, demethoxycurcumin, bisdemethoxycurcumin was analyzed from dried powder of rhizome. The analysis column was used to Phenomenex, Kinetex 2.6um F5 100A 100*2.1mm. 10 mg of standard was taken, 80% MeOH was added to dissolve in 10 ㎖ flask. 0.5, 1, 2, 4 ㎖ was taken from dissolved solution and then was added to 10 ㎖ of 80% MeOH. It was filtered through 0.45 ㎛ filter and then 5 μℓ was taken to make standard solution. Analytical calibration curve was measured to diluted solution within sample concentration. 2 g of ground sample was taken, 40㎖ of 80% MeOH was added to extract by ultrasound sonication during 60 minutes and then it was filtered through 0.2 ㎛ filter to analyze component content. Conclusion : The root weight significantly increased from 376.1 g in end October to 695.9 g in end December as 1.8 double. Curcumin contents differed significantly among harvest times in end November and early December. Demethoxycurcumin increased from early November to end November, and Bisdemethoxycurcumin increased to the early November.

Background : Adenophora triphylla var. japonica Hara. is used to cure internal organ, which is called to important one of the five “Sam (ginseng)” oriental medicinal herbs included Panax ginseng, Salvia miltiorrhiza, Sophora flavescens, Scrophularia buergeriana. Korea imported 24 ton 129 thousand dollar for medicinal herbs and 23 ton 29 thousand dollar for food at 2013. This study was carried out to investigate the effect of several organic compost on the growth and root yield of A. triphylla var japonica with organic fertilizer. Methods and Results : Seeds were collected from Adenophora triphylla var. japonica Hara. at last year in nursery of Department of Herbal Crop Research, NIHHS, RDA. Four types organic fertilizer (germiculture (GG), granular mix oil cake (CJ), mixed organic (DJ), plant oil cake (TG)), chemical fertilizer (CF) and non-fertilizer (NF) were treated. It was fertilized on soil before 2 month of planting. Seeds A. triphylla var. japonica were sowed in tray at early March and then were grown during 2 month. It were planted in furrowed nursery at early with 30 × 10 cm planting distance and black vinyl covering condition. Soil chemical property, survival rate, aerial part and root growth characteristics and yield were investigated at harvest time. For main composition analysis, 10 mg of standard (b-sitosterol, lupenone) was taken, 80% MeOH was added to dissolve in 10 ㎖ flask. 0.5, 1, 2, 4 ㎖ was taken from dissolved solution and then was added to 10 ㎖ of 80% MeOH. It was filtered through 0.45 ㎛ filter and then 5 μℓ was taken to make standard solution. Analytical calibration curve was measured to diluted solution within sample concentration. 2 g of ground sample was taken, 40㎖ of 80% MeOH was added to extract by ultrasound sonication during 60 minutes and then it was filtered through 0.2 ㎛ filter to analyze component content. Conclusion : GG and DJ were the most suitable for organic cultivation of denophora triphylla var. japonica. Content of beta-sitosterol showed the highest value in DJ treatment and the lowest value in CF. Content of lupenone showed the highest value in DJ and the lowest value in CJ.

Although Sophorae Flos (SF) has been reported to exert an anti-cancer activity, molecular targets and mechanisms associated with anti-cancer activity of SF have been unclear. Because cyclin D1 has been regarded as an important regulator in the cell proliferation, we focused cyclin D1 and investigated the effect of SF on the cyclin D1 regulation in light of elucidating the molecular mechanism for SF’s anti-cancer activity. The treatment of SF decreased cellular accumulation of cyclin D1 protein. However, SF did not change the level of cyclin D1 mRNA. Inhibition of proteasomal degradation by MG132 attenuated SF-mediated cyclin D1 downregulation and the half-life of cyclin D1 was decreased in the cells treated with SF. In addition, a point mutation of threonine-286 to alanine attenuated SF-mediated cyclin D1 downregulation. Inhibition of ERK1/2 by a selective inhibitor, PD98059 suppressed cyclin D1 downregulation by SF. From these results, we suggest that SF-mediated cyclin D1 downregulation may result from proteasomal degradation through its threonine-286 phosphorylation via ERK1/2. SF-induced proteasomal degradation of cyclin D1 might inhibit proliferation in human colorectal cancer cells. The current study provides information on molecular events for an anti-cancer activity of SF

The flower buds of Sophora japonica L (SF), as a well-known traditional Chinese medicinal herb, have been used to treat bleeding-related disorders such as hematochezia, hemorrhoidal bleeding, dysfunctional uterine bleeding, and diarrhea. However, no specific anti-cancer effect and its molecular mechanism of SF have been described. Thus, we performed in vitro study to investigate if treatment of SF affects activating transcription factor 3 (ATF3) expression and ATF3-mediated apoptosis in human colorectal cancer cells. The effects of SF on cell viability and apoptosis were measured by MTT assay and Western blot analysis against cleaved poly (ADP-ribose) polymerase (PARP). ATF3 activation induced by SF was evaluated using Western blot analysis, RT-PCR and ATF3 promoter assay. SF treatment caused decrease of cell viability and increase of apoptosis in a dose-dependent manner in HCT116 and SW480 cells. Exposure of SF activated the levels of ATF3 protein and mRNA via transcriptional regulation in HCT116 and SW480 cells. Inhibition of extracellular signal-regulated kinases (ERK) 1/2 by PD98059 and p38 by SB203580 attenuated SF-induced ATF3 expression and transcriptional activation. Ectopic ATF3 overexpression accelerated SF-induced cleavage of PARP. These findings suggest that SF-mediated apoptosis may be the result of ATF3 expression through ERK1/2 and p38-mediated transcriptional activation.

UGT72E3/2 gene encodes UDP-glycosyltransferase shown to glucosylate several phenylpropanoids such as syringin and coniferin. Syringin has effect of anti-stress and anti-fatigue. Korean soybean variety Kwangan was transformed with UGT72E3/2 gene. This gene was transformed into Kwangan using highly efficient soybean transformation system. This study used two promoters, beta-conglycinin promoter for seed-specific expression and 35s promoter for total expression. Transgenic plants were confirmed for gene introduction and their expression using PCR and RT-PCR. The analysis of syringin in transgenic plants was performed using HPLC. Currently, the confirmation of stable gene introduction with UGT72E3/2 gene is also performing by Southern blot analysis.

Soybean mosaic virus (SMV), a member of Potyviridae family, is one of the most typical viral diseases and results in yield and quality loss of cultivated soybean. Due to the depletion of genetic resources for resistance breeding, a trial of genetic transformation to improve disease resistance has been performed by introducing SMV-CP and HC-Pro gene by RNA interference (RNAi) method via Agrobacterium-mediated transformation. Transgenic plants were infected with SMV strain G5 and investigated the viral response. As a result, two lines (3 and 4) of SMV-CP(RNAi) transgenic plants and three lines (2, 5 and 6) of HC-Pro(RNAi) transgenic plants showed viral resistance. In genomic Southern blot analysis, most of lines contained at least one T-DNA insertion in both SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. Subsequent investigation confirmed that no viral CP and HC-Pro gene expression was detected in two SMV-resistant lines of SMV-CP(RNAi) and three lines of HC-Pro(RNAi) transgenic plants, respectively. On the other hand, non-transgenic plants and other lines showed viral RNA expression. Viral symptoms affected seed morphology, and clean seeds were harvested from SMV-resistant line of SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants. In addition, strong viral gene expression was detected from seeds of SMV-susceptible non-transgenic plants and SMV-susceptible transgenic lines. When compared the viral resistance between SMV-CP(RNAi) and HC-Pro(RNAi) transgenic plants, soybean transgenic plants with the HC-Pro gene using RNAi strategy showed much stronger and higher frequency of viral resistance.

Bacillus thuringiensis(Bt) crystal protein (Cry1Ac) genes encode insecticidal δ-endotoxins that are widely used for the development of insect-resistant crops. Common soybean is a crop of economic and nutritious importance in many parts of the world. Korea soybean variety Kwangan was transformed with Bacillus thuringiensis(Bt) crystal protein genes. We transformed three difference Cry1Ac (Cry1Ac and two modified Cry1Ac) genes into Kwangan using highly efficient soybean transformation system. Transgenic plants with Bt crystal protein genes were confirmed for gene introduction and their expression using PCR, real-time PCR, and RT-PCR. We generated 30 independent lines of transgenic soybean plants. Analysis of the flanking sequences isolated by Inverse PCR revealed complex T-DNA insertion patterns and preferential integration of T-DNA into the intergenic spacer region of the soybean genome. We found 5 different intergenic transgenic soybean lines of soybean genome. Currently, the confirmation of stable gene introduction with Bt genes is also performing by southern blot analysis, physiology test, and agronomic characters are investigating.

Twenty two common millet (Panicum miliaceum L.) varieties collected from Korea, China and Russia were investigated for their phylogenetic relationship using 5S ribosomal DNA sequences with a hope to provide the basic information on their exact origin. Sequences of 5S rDNA were isolated by PCR. The primers, 5s-rRNA1 and 5s-rRNA2, were designed to isolate the complete NTS. Genomic DNA amplification produced two fragments with different length, 900 bp and 400 bp fragments, confirming the presence of two types of 5S rDNA repeats that differed from each other in the length of the NTS region. Amplified DNAs of 400 bp fragment were subcloned and used for further investigation. The obtained NTS sequences ranged from 200 to 300 bp and homology of sequences among plant materials was much higher than long repeat. CLUSTALW multiple aligment of 5S rDNA sequences from 22 different common millets revealed the clear difference by their origin. And critically different areas with insert or deletion were also confirmed. Those sequence difference seems to be used for discrimination of cultivars from different origin and use as molecular markers for origin identification. In phylogenic tree construction, the clear classification was shown where the genotypes from China and Russia is positioned together and stay away from domestic genotypes.

Citrus is one of the major fruits produced in Korea. There are about 20 species mainly grown in Jeju Island, Korea. Four representative species, which are quite different in the shape of leaf and the taste of fruit, were selected and were used to profile the transcriptomes. These species are ‘Miyagawa Wase’ (C. unshiu Marcov.) satsuma mandarin, ‘Kiyomi’ (C. unshiu Marcov. × C. sinensis) mandarin hybrid, ‘Dangyuja’ (C. grandis) and ‘Natsudaidai’ (C. natsudaidai). Classification of the up-regulated and down-regulated genes using the Cluster of Orthologous Groups of proteins (COG) database reveals that the number of genes included in each group differed significantly among the four species. Several genes that showed significant differences in expression on the microarray were selected and their expression patterns were examined by reverse transcription- ploymerase chain reaction. Metabolic genes such as tyrosine decarboxylase and β-glucosidase ligase were found to be highly expressed in Miyagawa Wase, relative to other species. On the other hand, the expression level of mannose phosphate isomerase was lower in Miyagawa Wase. An efflux pump gene was found to be up-regulated in Kiyomi, whereas cinnamyl-alcohol dehydrogenase was down-regulated. β-carotene 15,15’-dioxygenase, which is involved in the vitamin metabolism, was up-regulated in Natsudaidai. Interspecific differentiations of gene expression are analyzed in terms of the metabolic pathways and their possible roles in citrus species.

Rye has important genes for biotic and abiotic stress resistance. Introduction of these genes to wheat by breeding wheat-rye translocation have been intensively used in wheat breeding program. Rye chromatin 1RS and/or 2RL show superior performance in unfavorable environments. In order to develop high yielding wheat, we applied various molecular breeding strategies. To develop EST-derived 1RS specific markers, we used comparative genomics with public sequence databases of Poaceae family. Putative rye chromatin specific sequences were used to design 1RS specific markers. To identify genes related to water deficiency, cDNA AFLP analysis was used in PEG treated seedlings of 1RS RILs. For functional analysis of identified genes and markers, we used Brachypodium distachyon, as a new model plant of temperate grasses. B. distachyon were recently applied for transformation and we constructed Agrobacterium-mediated transformation system. Integration of those strategies and conventional breeding method would enhance the usefulness of rye chromatins for wheat improvement.

Wild rice might have previously unidentified genes important for disease resistance and stress tolerance in response to biotic and abiotic stresses. A set of subtractive library was constructed both from leaves of wild rice plants, Oryza grandiglumis (CCDD, 2n=48), treated with fungal elicitor and from wounded leaves. A partial fragment that was homologous to PR10 genes from other plant species was identified via suppression subtractive hybridization and cDNA macroarray. The obtained full-length cDNA sequence (OgPR10) contains an open reading frame of 480 bp nucleotide, encoding 160 amino acids with a predicted molecular mass of 16.944 kDa and an isoelectric point (pI) of 4.91. The multiple alignment analyses showed the higher sequence homology of OgPR10 with PR10 genes identified in rice plants at amino acid level. The OgPR10 mRNA was not expressed by treatment with wounding, jasmonic acid, and salicylic acid, but markedly expressed in leaves treated with protein phosphatase inhibitors cantharidin and endothall, and yeast extract. In addition, the expression of OgPR10 mRNA was induced within 72 h after treatment with probenazole, one of well-known chemical elicitors, and reached the highest level at 144 h. Heterologous expression of OgPR10 caused growth inhibition and seedling lethality in E. coli and Arabidopsis, respectively. Chemically induced OgPR10 expression with glucocorticoid-mediated transcriptional induction system further reconfirmed its lethality on Arabidopsis seedling. In addition, OgPR10-expressing rice plants, Oryzae sativar were resistant against the infection of rice blast fungus, Magnaporthe grisea. These results indicate that OgPR10 is involved in probenazole- and microbe associated molecular patterns-mediated disease resistance responses in plants and is a potential gene for developing disease resistance crop plants.