Background : With the increasing demand of the mistletoe in larger quantities for cancer therapy, it has been depleted from its natural habitat in the Far East countries including Korea because of overharvesting for high-value products (e.g., lectins and viscotoxins). The rapid multiplication of mistletoe by tissue culture can help this problem and provide the benefits in the phamaceutical industry. Methods and Results : Mistletoe plants growing on oak trees were collected and their leaf and stem segments were inoculated on MS basal medium supplemented with various concentrations of growth regulators. Calli were induced only from stem explants on MS medium containing 2,4-D and transferred onto MS medium supplemented with various concentrations of 2,4-D, NAA and BAP, respectively, for their propagation. The best callus multiplication rate of more than 15 folds (759 mg) was obtained in treatment of 2,4-D (4 mg/L) that produced yellowish-green, white and friable callus on this medium. To compare biochemical characterization, lectins were partially purified from natural mistletoe plants (nML) and in vitro cultured mistletoe calli (cML), respectively. The former was purified by lactose-agarose affinity chromatography and the latter was done by ammonium sulfate precipitation followed Sephadex G-25 chromatography. Both nML and cML showed similar molecular weight on SDS-PAGE and Western blot analyses. In addition, they showed similar carbohydrate-binding specificities and hemagglutination activities. Conclusion : From the above results, we may suggest that nML and cML showed the similarity in biochemical characters.

Background : Lectins were individually isolated from natural Korean mistletoe (nML) and its in vitro cultured calli (cML). Both of the lectins showed the difference in bioactivities such as cytotoxicity and cytokine induction. Methods and Results : Target cells (1 x 104 cells/well) were seeded independently into each well of a 96-well culture plate and incubated with different concentrations of each lectin. Survivability of target cells was determined by CCK-8 kit (Sigma, USA) according to manufacturer’s directions. The nML showed 46, 34 and 5.5 times stronger than cML in cytotoxicity (IC50) to human melanoma cell line (SK-MEL-28), human carcinoma cell line (NCI-H1650) and murine macrophage (RAW 264.7), respectively. In addition, respective lectins directly stimulated macrophage RAW 264.7 but they showed the difference in enhanced productivity of some inflammatory cytokines. Compared with cML, the nML induced both TNF-α and IL-1β at its low concentration. Administration of two kinds of lectins (10-1000 μ g/kg body weight) to ICR mouse did not show any significant changes on the level of alanine transaminase (ALT), glutamate-oxaloacetic transaminase (GOT) and blood urea nitrogen (BUN) in sera. Conclusion : From the above results, we may suggest that nML and cML showed differences in cytotoxic effects and cytokine production due to the difference in amounts from sources.

Background : We studied the anti-oxidant activity and anti-inflammatory effects of Rhododendron lapponicum (L.) Wahlenb. var. parvifolium (Adams) Herder extract (RLE). Methods and Results : The RLE was prepared using methanol. The antioxidant effects of RLE was evaluated for its DPPH (1,1-diphenyl-2-picrylhydrazyl) free-radical scavenging activity, reducing power. Subsequently, using the RAW 264.7 cells, the cell viability of RLE was evaluated with or without LPS (lipopolysaccharide), and the anti-inflammatory effects of RLE was also estimated by nitric oxide (NO) and using real-time polymerase chain reaction (RT-PCR). The extract showed antioxidant activity (DPPH free-radical scavenging activity) with RC50 value of 57.67 ㎍/㎖. The reducing power of the extract was Abs 0.77 at 250 ㎍/ ㎖. The result indicated that RLE would have significantly high anti-oxidative effects. Cell viability was determined using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. To evaluate anti-inflammatory activity, we examined the inhibitory effects on LPS-induced NO production in RAW 264.7 cells. The NO inhibition rate was 85.44% at 200 ㎍/㎖ RLE. At the same concentration, the expression of pro-inflammatory genes such as inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 also decreased. In RLE 50 ㎍/㎖ concentration showed the highest decrease. Conclusion : This result suggest that RLE is a novel resource for the development of foods and drugs that possess anti-oxidant and anti-inflammatory activity. Also, RLE can be developed as an inflammatory agents for cosmetic bases in the future.

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.

Analyzing the 13 sort contents of broccoli leaves by using GC/MS, sulforaphane was found in 11 sort of broccoli leaves for the result. After being grinded by the blinder, amount of sulforaphane in broccoli leaves was rapidly raised after thirty minutes and maintained the amount till sixty minutes have passed. Among the parts of broccoli, the root had the most sulforaphane. In freezing temperature, biosythesized sulforaphane maintained longer than in room temperature. However, even in frozen condition, the amount of sulforaphane was reduced to half or less after 3 weeks.

Five caffeoylquinic acids of Aster altaicus var. uchiyamae Kitamura (Compositae) leaves were identified using standard compounds by HPLC and determined as follows: 3,4-di-O-caffeoylquinic acid (4.92 ± 0.06 ㎎/g dried weight), 3,5-di-O-caffeoylquinic acid (3.95 ± 0.13 ㎎/g), 4,5-di-O-caffeoylquinic acid (1.39 ± 0.10 ㎎/g), 5-O-caffeoylquinic acid (chlorogenic acid, 8.05 ± 0.21 ㎎/g), 3-O-caffeoylquinic acid (4.97 ± 0.18 ㎎/g). The total content of five caffeoylquinic acids were calculated as 26.73 ± 0.26 ㎎/g dried weight while the percentage of the five compounds in the MeOH extract was calculated as 25.22 ± 0.25%. The IC50 value of the MeOH extract scavenging peroxynitrite (ONOO - ) was shown as 5.16 ± 0.15 ㎍/㎖.

Background : Irregular meals and insufficient exercise are major modern lifestyle-related risk factors for constipation. This study aimed to examine the effects of the aqueous extract of Dendrobium speciosum var. (DM) on the improvement of intestinal function and prevention of constipation in rats. Methods and Results : Constipation in rat was induced by loperamide (4 ㎎/㎏) injection for 5 days and rats were randomly assigned to the following groups: normal control rats (NOR), constipated rats (LOP induced) and constipated rats supplemented with dulcolax-S (POS), DMSG, 100 ㎎/㎏ (DMSG-100), 300 ㎎/㎏ (DMSG-300) and 500 ㎎/㎏ (DMSG-500). The DMSG groups showed increments in the frequency and amount of stools compared to the LOP group. Loperamide treatment markedly reduced the water content of feces, whereas DMSG administration significantly increased fecal water contents. Moreover, DMSG improved intestinal transit speed in constipated-rats. Alcian blue staining revealed increased mucus production by crypt cells and mucus contents in feces and the mucosal surface. Conclusions : In summary, DM extraction significantly improves intestinal function in rats, which indicates the effectiveness of this extract in the prevention and treatment of constipation.

Humankind has been searching for medicinal materials from various plant sources in an attempt to treat disease. Mistletoe is one indubitable plant source for these materials due to its effectiveness in treating various diseases, but it has almost disappeared from the mountainous areas of Korea due to excessive harvesting. In this study, in order to select host tree species for Korean mistletoe [Viscum album var. coloratum (Kom.) Ohwi] by seed inoculation and to clarify the effect of host specificity among various tree species were conducted for the purpose of gaining basic information for the artificial cultivation of Korean mistletoe. Almost all the seeds of Korean mistletoe germinated in vitro at the temperature of 15℃. Among host trees used in this study, Prunus mume showed the highest parasitic affinity with inoculated Korean mistletoe, compared with any other host plants. However, treatment of hormones could not increase the low survival rate of Korean mistletoe on the host trees.

Background : Aruncus dioicus var. kamtschaticus, functional wild vegetable, is perennial herb and young leaves with soft texture are generally used as edible food. So the demand for the vegetable has increased recently. This study was carried out to determine the effects of temperature and shading on germination characteristics of A. dioicus. Methods and Results : The experiment was performed by temperature and shading treatments. Seed pre-treatment before the germination experiment was carried out by the storage in low temperature (4℃) under wet condition (LTW) for 0, 15, 30, 45 and 60 days and shading treatment were 35%, 50%, and 75% under control, BA (6-benzyladenine) and GA3 (gibberellic acid) condition for 24 hours. Increasing the length of the storage periods led to increases seed germination percent in low temperature (4℃) under wet condition (LTW), germination rate of A. dioicus seed was the highest at 15℃ with 60 days of seed pre-treatment. In the case of seeds pre-treatment with LTW, the more temperature went up, the more days to 50% of Germination of Final Germination Rate (T50) went down. As a result of surveying shading treatment, germination rate was the highest in control of 35% shading and the next higher was in control of 50% shading. Conclusions : It is concluded that the temperature and shading are important factors to produce A. dioicus. Also, We suggest these results as basic data of A. dioicus for sexual propagation.

Lonicera caerulea var. edulis is a rare species found in some alpine region of Korea. Genetic variation in L. caerulea var. edulis has been investigated by examining 161 individuals from six natural populations: Mt. Seorak 1, Mt. Seorak 2, Mt. Jeombong, Mt. Bangtae, Mt. Gyebang, Mt. Halla. The mean genetic diversity for all the six populations was 0.25 (S.I.). The highest genetic diversity was found in Mt. Seorak (S.I.=0.3158) and the lowest was in Mt. Gyebang (S.I.=0.1047). Comparatively low level of genetic diversity was observed (Ae=1.25, P= 64.6%, S.I.=0.25), which is a typical pattern for rare tree species. AMOVA showed exceptionally large proportion of genetic variations both for among populations (34.69%) and within populations (65.31%). Excluding Mt. Gyebang, the genetic variation among and within population was 18.71% and 81.29% respectively. The UPGMA dendrogram based on genetic distance is not suitable for geographic relationship. Genetic distance of Mt. Gyebang was most distant from the other populations. Excluding Mt. Gyebang, the genetic identities among the five populations were 0.95 to 0.97, which is very high similarity level of genetic identity. This low level of genetic variations and the lack of site in nature indicates that L. caerulea var. edulis demanded a serious conservation.

Adenophora triphylla var. japonica HARA is a herbaceous plant belongs to Campanulaceae. Adenophora root is mainly used for medicinal purpose. It is effective for lung cleaning, sputum remove, viscera strengthening, cough stopping and cancer treatments. Adenophora has about 70 species in the world and 17 of the species are distributed in Korea. Genetic resources of A. triphylla var. japonica HARA are valuable as the habitat is concentrated in East Asia. The intraspecies variation is very high according to the environmental conditions. A new A. triphylla var. japonica HARA variety, ‘Harang’, was developed through polyploid breeding in 2011. But, low domestic production and passive studies caused our country to rely on imports for almost all amount of the A. triphylla var. japonica HARA demands. In this experiment, genetic diversity between the collections were analyzed using 32 RAPD primers. Through this study, limit of morphologic classification could be solved and genetic diversity of this plant could be assured.

Cabbage head splitting can greatly affect both the quality and commercial value of cabbage (Brassica oleracea). To detect the genetic basis of head-splitting resistance, a genetic map was constructed using an F2 population derived by crossing “748” (head-splitting-resistant inbred line) and “747” (head-splitting-susceptible inbred line). The map spans 830.9cM and comprises 270 markers distributed in nine linkage groups, which correspond to the nine chromosomes of B. oleracea. The average distance between adjacent markers was 3.6cM. A total of six quantitative trait loci (QTLs) conferring resistance to head splittingwere detected in chromosome 2, 4, and 6. Two QTLs, SPL-2-1 and SPL-4-1, on chromosomes 2 and 4, respectively, were detected in the experiments over 2 years, suggesting that these two potential loci were important for governing the head-splitting resistance trait. Markers BRPGM0676 and BRMS137, which were tightly linked with head-splitting resistance, were detected in the conserved QTL SPL-2-1 region using bulked segregant analysis. Synteny analysis showed that SPL-2-1 was anchored to a 3.18Mb genomic region of the B. oleracea genome, homologous to crucifer ancestral karyotype E block in chromosome 1 of Arabidopsis thaliana. Moreover, using a field emission scanning electron microscope, significant differences were observed between the two parental lines in terms of cell structures. Line “747” had thinner cell wall, lower cell density, larger cell size, and anomalous cell wall structure compared with the resistant line “748”. The different cell structures can provide a cytological base for assessing cabbage head splitting.

The oriental melon (C. melo var. makuwa), called ‘Chamoe’ in Korean, is a popular fruit crop cultivated mainly in Asia and a high–market value crop in Korea. To provide a genomic resource as a reference genome for the Cucurbitaceae crop improvement, we performed whole genome sequencing of Korean landrace, Gotgam chamoe. We used Illumina HiSeq2000 sequencing platform to generate 89 Gb (205X) of paired and mate pair sequence reads. The pre-processed reads were de novo assembled resulting in 4,764 scaffolds with a N50 scaffold length of 249kb. This assembly represented 379.8Mb which was 84.7% of the 448Mb of the whole genome. The assembled draft was predicted 26,634 genes of which 80% were predicted by known protein or C. melo unigene homology. Approximately 20% of predicted genes were hypothetical. A total of 1,885 non-coding RNA was detected including rRNA. The transposable elements were accounted for 21% (71.6Mb) of the total assembly. All the marker candidates including SSR, INDEL, SNP were mined and presented. The draft genome will provide a useful platform for genomic research and improvement for Cucurbitacea crops.

Chloroplast (cp) genome sequences provide a valuable source for DNA barcoding. Molecular phylogenetic studies have concentrated on DNA sequencing of conserved gene loci. However, this approach is time consuming and more difficult to implement when gene organization differs among species. Here we report the complete re-sequencing of the cp genome of Capsicum pepper (Capsicum annuum var. glabriusculum) using the Illumina platform. The total length of the cp genome is 156,817 bp with a 37.7% overall GC content. A pair of inverted repeats (IRs) of 50,284 bp were separated by a small single copy (SSC; 18,948 bp) and a large single copy (LSC; 87,446 bp). The number of cp genes in C. annuum var. glabriusculum is the same as that in other Capsicum species. Variations in the lengths of LSC, SSC and IR regions were the main contributors to the size variation in the cp genome of this species. A total of 125 simple sequence repeat (SSR) and 48 insertions or deletions variants were found by sequence alignment of Capsicum cp genome. These findings provide a foundation for further investigation of cp genome evolution in Capsicum and other higher plants.

Chloroplast DNA sequences are a versatile tool for species identification and phylogenetic reconstruction of land plants. Different chloroplast loci have been utilized for phylogenetic classification of plant species. However, there is no evidence for a short sequence that can distinguish all plant species from each other. Molecular markers derived from the complete chloroplast genome can provide effective tools for species identification and phylogenetic resolution. Thus, the complete chloroplast genome sequence of Korean landrace “Subicho” pepper (Capsicum annuum var. annuum) has been determined here. The total length of the chloroplast genome is 156,878 bp, with 37.7% overall GC content. A pair of IRs (inverted repeats) of 25,801 bp was separated by a small single copy (SSC) region of 17,929 bp and a large single copy (LSC) region of 87,347 bp. The chloroplast genome harbors 132 known genes, including 87 protein-coding genes, 8 ribosomal RNA genes, and 37 tRNA genes. A total of seven of these genes are duplicated in the inverted repeat regions, nine genes and six tRNA genes contain one intron, while two genes and a ycf have two introns. Analysis revealed 144 simple sequence repeat (SSR) loci and 96 variants, mostly located in the non-coding regions. The types and abundances of repeat units in Capsicum species were relatively conserved and these loci will be useful for developing molecular markers.

The chloroplast (cp) is an organelle with its own genome that encodes a number of cp-specific components. Resequencing technology via next-generation sequencing has recently been successfully applied to cp genome characterization. The field of cp characterization is rapidly growing due to its wide versatility and two complete chloroplast (cp) genome sequences of Capsicum species have been reported. We herein report the complete chloroplast genome sequence of Capsicum baccatum var. baccatum, a wild Capsicum species. The total length of the chloroplast genome is 157,145 bp with 37.7% overall GC content. One pair of inverted repeats, 25,910 bp in length, was separated by a small single-copy region (17,974 bp) and large single-copy region (87,351 bp). This region contains 86 protein-coding genes, 30 tRNA genes, and 4 rRNA genes. Eleven genes contain one or two introns. Pair-wise alignments of cp genome were performed for genome-wide comparison. Analysis revealed a total of 134 simple sequence repeat (SSR) motif and 282 insertions or deletions variants in the C. baccatum var. baccatum cp genome.

The authors calculate the long-term predictability of GDP, domestic demand, investment, and net exports for Guangdong province, P.R. China from 2000 to 2013. A vector autoregressive (VAR) model with quarterly data for this period is first co-integrated then the Granger causality test is applied to empirically assess the relationships among gross domestic product (GDP), consumption, investment, and net exports. There is a strong causality effect between investment and net exports in Guangdong province. However, the variance decomposition results indicate that exports respond to foreign shocks rather than domestic ones, making their impact on the Guangdong economy to predict. Results show the stimulating effect of domestic demand on GDP is larger than the stimulating effect of net exports and much larger than even the stimulating effect of investment. The analysis suggests that there are dynamic influences with various levels of persistence between GDP, consumption, investment, and net exports. Macroeconomic policy adjustments are urgently required to expand domestic demand and thereby stimulate economic growth in Guangdong province.