Background : This study for the stable production and supply of seeds of Angelica gigas Nakai(Man-chu Korean Angelica), when seeds harvested using nets, seed productivity was investigated. Methods and Results : Planting density is 50 × 25cm, Fertilizer per 10a was sprayed amount of N-P2O5-K2O = 16-17-10kg. And the amount of compost per 10a was sprayed 3000kg. Seed harvesting nets were used for a time formed the endosperm of the seeds (later in mid-August to late). And net for seed production was used for onion nets (at least 13 × 18cm). Shoot growth conditions were as follows. Bolting rate was 89.0% in the untreated, the treated group was 93.1%. The length and thickness of each stem was 129.3 ~ 130.8cm, 1.8cm. The number of nodes per plant was 6.7 ~ 7.5 pieces, and the number of petiole was 14.8 ~ 15.5 per plant. The number of umbel was 10.3 ~ 11.1 piece per plant, and number of deleted umbel was 7.1 ~ 7.2 piece. Seed weight per plant was 24.2g of the net treatment, but ripening seeds 19.6g, 1000 grain weight were all treated and untreated 2.8g. The total seed weight per plant, the net treated was 24.2g, was the weight of the ripening seeds 19.6g. The weight of the ripening seeds were heavier than those of the control. However, the weight of 1000 grain were both treated and untreated 2.8g. When treated nets, the total seed yield per 10a was 88.0kg production, increased by 60.9% compared to untreated. In addition, the ripening seed production per 10a was 71.5kg production, increased by 50.1% compared to untreated. Researching after germination Seed Production, germination rate was 50.8% in the control group and the treatment group was 54.9%. When applying the germination rate, high-quality seed production per 10a was able to produce 39.2kg, compared to control obtain the results increased by 65%. Conclusion : Through the above results, When producing angelica seed, use of net for seed production is thought to be used as a way to prevent early shattering and insect damage.(True bug, etc.).

Background : This study was performed to investigate by antioxidant activity, total phenolic contents, total flavonoid contents, and effective component of Astragalus membranaceus treated with different artificial light Sources (fluorescent lamp, red, blue, green, white, LEP). Methods and Results : We investigated the effects of various artificial light sources on the DPPH radical activity, total phenol and flavonoid contents, tyrosinase activity and main flavonoid compounds contents (formononetin and calycosin) and other biological activities in A. membranaceus. Antioxidant activities were 53.6% as the highest level of activity under LEP light. Growth under LEP light also produced the highest total phenolic and flavonoid contents of 36.05 and 5.94 mg/ml, respectively. Extracts from plants grown under LEP light caused the highest inhibition of tyrosinase activity with inhibition of 35.37, 61.87, and 65.49%, respectively, for extract concentrations of 100 μg/ml, 500 μg/ml, and 1000 μg/ml compared with other artificial light treatments. Conclusion : Little information is available on the influence of LED and LEP light sources on antioxidant production or other biological activities in A. membranaceus. Our goal in this study was to determine the effects of LED and LEP artificial light sources on the production of new functional compounds in A. membranaceus.

Background : Cordyceps militaris has been an wonder drug to anti-aging efficacy and called the three main drugs with ginseng and deer antler from the past. Cordycepin, cordycepic acid (d-mannitol) and adenosine are known as functional ingredients in Cordyceps militaris. Among them, cordycepin, the representative component, has been reported as antimicrobial substance containing immune enhancement, anti-cancer and anti-inflammatory effects. Methods and Results : After Cordyceps militaris produced from different types of medium mixed with 10-fold volume of purified water, the mixture were extracted at 70±5℃ for a hour and that extracts re-extracted using ultrasonics wave for 30 minutes. Qualitative analysis of the index component was determined by using the Q-TOF (A quadrupole time-of-flight mass spectrometer), and quantitative analysis was performed by using HPLC (high-performance liquid chromatography) with Xselect HSS T3 column (2.1 X 100 mm, 2.5㎛, Waters, USA) and ultrapure water and acetonitrile as mobile phase A and B. Detection column temperature, injection volume and the flow rate were 35℃, 2 μL and 0.3 mL / minute respectively. The cordycepin content of Cordyceps militaris produced from medium mixed with vegetable and animal ingredients higher than single ingredient. Moreover, through a variety of analyzes by varying the type and content of the medium additives, the cordycepin in Cordyceps militaris produced from medium mixed with animal ingredients highest. Furthermore, the cordycepin content of a fruit body was higher than those of the a mycelium. Conclusion : These results provide a method for producing an high cordycepin content of Cordyceps militaris as functional food ingredient.

Background : Platycodon grandiflorum root(PGR) was one of the primary herbs used in a phlegm-relieving herb from the past. Purified platycoside compounds from the roots of PGR may exhibit neuroprotective, antimicrobial, anti-inflammatory, anti-cancer, anti-allergy, improved insulin resistance, and cholesterol-lowering properties. To evaluate preference and functionality of PGR extracts, PGR was fermented by several lactic acid bacteria. Lactic acid bacteria used were Leuc. mesenteroides N12-4 and N58-5, L. plantarum N76-10 and 56-12, L. brevis N70-9 and E3-8. Methods and Results : This study was performed in order to investigate the changes of platycosides, as well as the antimicrobial activities on bronchus diseases inducing bacteria(C. diphtheriae, K. pnneumoniae, S. aureus, S. pyogenes) of Platycodon grandiflorum root(PGR) fermented by using lactic acid bacteria(Leuc. mesenteroides N12-4, Leuc. mesenteroides N58-5, L. plantarum N76-10, L. plantarum N56-12, L. brevis N70-9, L. brevis E3-8). Growth of L. plantarum on PGR was the most active during lactic acid fermentation by some different strains. Total platycoside, platycoside E, platycodin A, polygalacin D2, polygalacin D and diapioplatycoside E contents of PGR fermented for 96 hours at 37℃ by Leuc. mesenteroides and L. plantarum were increased, while platycodin D and platycodin D3 were decreased. The antimicribial activity on PGR fermented by L. plantarum N56-12 exhibited a strong microbial proliferation in all four kinds of bronchus diseases inducing bacteria and was higher than non-fermented PGR extract. Conclusion : Thus, this results showed antimicrobial activities on bronchus diseases inducing bacteria and platycosides content of PGR by L. plantarum N56-12 were higher than non-fermented PGR extract.

Background : Recently, there is a urgent demand for development of new varieties with enhanced resistance to various biotic/abiotic stresses. In order to develop ginseng varieties with such traits, systematic breeding program and comprehensive field studies are prerequisite. Methods and Results : ‘Cheonmyeong' seeds were collected in 2008 from the farmer field of Buyeo. Physiological investigation and propagation were conducted from 2009 to 2011. It was given the name 'Eumseong No. 8 through the observed yield trial from 2012 to 2013 and local adaptability was carried out from 2014 to 2015. All phenotypes including agronomic characteristics, seed yield, and physiological response to biotic/abiotic stresses were investigated according to the ginseng GAP and UPOV guidelines. Yellow-red leaf and pink berry at maturing stage were observed. The time of emergence, flowering and berry maturity of the ‘Cheonmyeong’ were faster than those of ‘Chunpoong’. Stem length of ‘Cheonmyeong’ was shorter than that of ‘Chunpoong’, whereas stem diameter was thicker than that of ‘Chunpoong’. Main root length was shorter but main root diameter is thicker than that of ‘Chunpoong’. Number of seeds of ‘Cheonmyeong’ was fewer than that of ‘Chunpoong’ but 1,000-seeds weight and stratification rate were higher than those of ‘Chunpoong’. The yield performance of this variety was 661 kg/10 a in local adaptability test for two years, which is 22% higher than that of ‘Chunpoong’. ‘Cheonmyeong’ showed strong resistance to phytophthora blight, mulberry mealybug and nematode and moderate resistance to alternaria blight. ‘Cheonmyeong’ did almost not occur yellow spot of aerial part and rusty skin of root, show moderate resistance at high temperatures. Conclusion : Our study demonstrated that ‘Cheonmyeong’ is an ideal variety with heavier root weight and enhanced stress resistance and contribute will enhance biotic/abiotic stress resistance and increase the farmers' income.

Primary oocytes that are arrested in first meiotic prophase for years enter maturation process to meet a critical precondition for successful fertilization. During maturation, oocyte finishes meiosis I and progresses to the metaphase II stage, achieving meiotic maturity. Although importance of oocyte maturation for oocyte quality has been recognized, it is not fully understood for molecular mechanisms underlying oocyte maturation. Here, we found that dexamethasone-induced Ras-related protein 1 (RASD1), a member of RAS superfamily of small GTPases, was expressed in the mouse ovary. Immunohistochemical analysis revealed that Rasd1 expression was dominant in oocyte cytoplasm. Real-time PCR and RT-PCR analyses showed that Rasd1 mRNA was steadily expressed in germinal vesicle (GV), germinal vesicle break down (GVBD), metaphase I (MI) oocytes, but decreased in metaphase II(MII) oocytes during oocyte maturation. Konckdown of Rasd1 using RNAi system in the GV oocytes suppressed oocyte maturation through disruption of meiotic spindle and formation of misarranged chromosomes. Taken together, Rasd1 is a critical factor for MI-MII transition of oocyte and is involved in the regulation of spindle formation during oocyte maturation. Further study is needed to examine relationship between Rasd1 and spindle formation in MI-MII transition.

Rearing experiments were conducted using several regimes combined with different photoperiods and water temperatures to investigate the role of temperature and photoperiod as environmental cues regulating reproduction in red marbled rockfish, Sebastiscus tertius. The initial gonadosomatic index (GSI) was 1.62±0.32% and that in the 15˚C control was 7.01±0.46% at the end of the experiment. The highest GSI was 9.10±0.35% in the 12L:12D photoperiod-treated fish. The highest GSI at 19˚C was 15.03±0.86% in the 12L:12D photoperiod treatment. The initial hepatosomatic index (HSI) was 3.09±0.72% and that in the 15˚C control was 3.88±0.45%. The highest HSI was 4.00±0.56% in the 15L:9D treated fish. The highest HSI at 19˚C was 4.05±0.63% in the 12L:12D treated fish.

We investigated the effect of light spectra on circadian rhythm by exogenous prolactin (PRL) by using light emitting diodes (LEDs): red, green, and purple. We injected PRL into live fish or treated cultured brain cells with PRL. We measured changes in the expressions of period 2 (Per2), cryptochrome 1 (Cry1), melatonin receptor 1 (MT1) mRNAs, and MT1 proteins, and in the plasma PRL, serotonin, and melatonin levels. After PRL injection and exposure to green LED light, MT1 expression and plasma melatonin levels were significantly lower, but the expressions of Per2 and Cry1 were significantly higher than others. Plasma serotonin after PRL injection and exposure to red LED light was significantly lower than others. These results indicate that injection of high concentration PRL inhibits melatonin, and inhibited melatonin regulates circadian rhythm via clock genes and serotonin. Thus, exogenous PRL regulates the circadian rhythm and light spectra influence the effect of PRL in goldfish.

Fluorescence in situ hybridization (FISH) is a powerful tool for the detection of DNA sequences in the specific region of the chromosomes. As well as for the integrated physical mapping, FISH karyotype analysis has to be preceded. The detailed karyotypes of two onion cultivars, which are resources for onion genome sequencing project (‘Eumginara’ and ‘Sinsunhwang’), were constructed based on triple color fluorescence in situ hybridization (FISH) using 5S rDNA, 45S rDNA, and tandem repeat sequence. All used our materials showed 2n=2x=16 with x=8 as basic chromosome number. 5S rDNAs were located on 4 loci in one pair of interstitial region of short arm chromosome in both onion cultivars. Two pairs of 45S rDNAs were positioned in distal region of short arm chromosome in ‘Eumginara’. Otherwise 5 loci of 45S rDNAs were located in distal region of two pairs of short arm chromosome in ‘Sinsunhwang’. Among them, two signals of 45S rDNAs were co-localized in distal part of short arm and long arm chromosome, respectively. In case of tandem repeat sequence was detected on telomeric region of 8 pairs of chromosomes except on 45S ribosomal DNA sites. These results will provide a valuable background for physical mapping and help to further more understand the genome sequencing project in Allium cepa.

Genome sequencing researches for considerable numbers of crops and wild plants are being developed. Cytogenetic researches according to chromosome number and size are essential to confirm and comprehend ploidy level and genome size before genome sequencing project is actually conducted. Cytogenetic researches on six food crop plants were carried out by DAPI staining and fluorescence in situ hybridization (FISH) method. Fagopyrum esculentum Moench showed 2n=2x=16, each chromosome length of 1.42㎛ to 1.77㎛, total chromosome length of 13.31㎛, and karyotypic formula of 2n=8m; Phaseolus angularis W.F. Wight, 2n=2x=22, 2.01㎛ to 3.84㎛, total 28.03㎛, 2n=9m+2sm, Perilla frutescens var. japonica Hara, 2n=2x=40, 1.73㎛ to 2.76㎛, total 44.36㎛, 2n=5m+13sm+2st. Chromosome sizes of the other three species such as, Panicum miliaceum L., 2n=2x=36, total chromosome length of 30.83㎛, Sesamum indicum L., 2n=2x=26, 27.39㎛, lpomoea batatas L., 2n=2x=30, total 33.51㎛ were too small for each chromosome type to be identified and analyzed. The result of FISH analysis using 5S and 45S rDNA probe showed species-specific chromosome locations in the genome. These preliminary analyses were carried out to decide which food crop to prioritize for genome sequencing. This work was supported by the “Cooperative Research Program for Agriculture Science & Technology Development (No.PJ009837), Rural Development Administration, Republic of Korea.

Mungbean (Vigna radiata) is a fast-growing, warm-season legume crop that is primarily cultivated in developing countries of Asia. We constructed a draft genome sequence of mungbean to facilitate genome research into the subgenus Ceratotropis and to enable a better understanding of the evolution of leguminous species. The draft genome sequence covers 80% of the estimated genome, of which 50.1% consists of repetitive sequences. In total, 22,427 high confidence protein-coding genes were predicted. Based on the de novo assembly of additional wild mungbean species, the divergence of what was eventually domesticated and the sampled wild mungbean species appears to have predated domestication. Moreover, the de novo assembly of a tetraploid Vigna species (Vigna reflexo-pilosa var. glabra) provided genomic evidence of a recent allopolyploid event. To further study speciation, we compared de novo RNA-seq assemblies of 22 accessions of 18 Vigna species and protein sets of Glycine max and Cajanus cajan. The species tree was constructed by a Bayesian Markov chain Monte Carlo method using highly confident orthologs shared by all 24 accessions. The present assembly of V. radiata var. radiata will facilitate genome research and accelerate molecular breeding of the subgenus Ceratotropis.

Rice blast (Magnaporthe oryza B.) is one of the most widespread and devastating diseases of rice. Screening of valuable genetic resources harboring resistance genes is one of the most efficient approaches against blast disease. Because the bioassay to rice blast in the field shows high variations, this study has performed to provide DNA profiles in the accessions of diverse countries using major blast resistant genes linked markers, identified and mapped in different genotypes of rice. Because durable resistance to blast is controlled by a combination of major resistance genes, we surveyed the distribution of blast resistant genes in the 1,500 accessions using major 12 blast resistance genes linked markers. These resistant genes found that the frequency distribution of Pi-39 (66.9%), Pik-m (41.9%), Pit (40.5%), Pii (21%), Pib (19.3%), Pi-d(t)2 (12.7%), but Pita, Pita/Pita-2, Pik, Piz-t, Pi5 genes were identified in less than 10% frequency. Most of accessions contain from 1 to 4 different resistant genes. Pi39 and Pik-m genes amplified in the 69.1% and 51.7% among 356 Korean accessions, Pi39 (79.6%) and Pib (55.8%) in 113 China, Pit (80.6%) and Pib (32%) in 103 Philippines, respectively. In this study, we evaluated the blast resistance degree and the information about the distribution of rice blast resistant genes in rice germplasm. This study will help to develop effective strategies for managing rice blast disease in rice germplasm.

Olive flounder Paralichthys olivaceus is one of the most widely cultured fish species in Korea. Although olive flounder receive attention from aquaculture and fisheries and extensive research has been conducted eye morphological change in metamorphosis, but little information was known to molecular mechanism and gene expression of eye development- related genes during the early part of eye formation period. For the reason of eyesight is the most important sense in flounder larvae to search prey, the screening and identification of expressed genes in the eye will provide useful insight into the molecular regulation mechanism of eye development in olive flounder. Through the search of an olive flounder DNA database of expressed sequence tags (EST), we found a partial sequence that was similar to crystallin beta A1 and gamma S. Microscopic observation of retinal formation correspond with the time of expression of the crystallin beta A1 and gamma S gene in the developmental stage, these result suggesting that beta A1 and gamma S play a vital role in the remodeling of the retina during eye development. The expression of crystallin beta A1 and gamma S were obviously strong in eye at all tested developing stage, it is also hypothesized that crystallin acts as a molecular chaperone to prevent protein aggregation during maturation and aging in the eye.

Understanding the host defense mechanisms in response to brown leaf spot disease caused by Cochliobolus miyabeanus is very important for production of resistant plant. In this study, two-dimensional gel electrophoresis (2-DGE) in conjunction with mass spectrometry was utilized to unravel changes of stress inducible proteins in rice leaves infected with C. miyabeanus. For this purpose, we firstly observed disease developmental process of C. miyabeanus in rice using trypan blue, anilin blue, acid fuchsin staining, and DAB staining for ROS detection and expressional abundance of ROS related proteins in rice leaves inoculated was confirmed by Western blotting. Proteins were extracted by PEG fractionation and their expression patterns were analyzed by 2-DGE and subjected to image analysis using the ImageMaster 6.0 2D Platinum software, resulting in the identification of 86 differentially expressed protein spots with significantly changes (p<0.05) compared with control. MALDI-TOFTOF-MS analysis revealed that 69 proteins including 42 and 27 significantly up- and down-regulated proteins, respectively, were identified. Based on gene ontology analysis, identified proteins were classified according to their functional groups: metabolism (20%), oxygen-detoxifying (13%), protein stress/defense (24%). Thus, these results for the first time suggest that differentially induced proteins may play a key role for understanding host defense mechanisms during rice -C. miyabeanus interaction.

The rice blast disease caused by Magnaporthe oryzae (M. oryzae) is one of the most serious diseases of cultivated rice (Oryza sativa L.) in most rice-growing regions of the world. In order to investigate early responsible genes in rice in response to M. oryzae, we analyzed transcriptomics analysis using 300 K tilling microarray chip. The quality of RNA samples was initially validated by 4 defense related genes and phytoalexins measurement using RT-PCR and HPLC, respectively, which are well known defense markers. We determined that accumulation of 608 genes showed statistically significant changes in the level of transcription (>2 fold change, P<0.05). Among them, 261 genes were more up-regulated in incompatible interaction than that of compatible one. We further analyzed GO enrichment analysis of the 41 and 231 which were 2 fold up-regulated genes at 12h and 48h in incompatible interaction, respectively, using Rice Oligo nucleotide Array Database (http://ricearray.org). Furthermore, MapMan analysis (http://mapman.gabipd.org/) revealed that 21 and 85 genes including 18 receptor-like genes which were more induced in incompatible interaction compared to compatible interaction were found to be involved in biotic stress. Thus, this study suggests that early inducible genes including receptor-like protein kinases in incompatible interaction may play a key role in disease resistance against M. oryzae attacks.

R genes are a key component of genetic interactions between plants and biotrophic bacteria and are known to regulate resistance against bacterial invasion. The most common R proteins contain a nucleotide-binding site and a leucine-rich repeat (NBS-LRR) domain. Some soybean NBS-LRR genes have also been reported to function in disease resistance. A total of 319 genes were determined to be putative NBS-LRR genes in the soybean genome. The number of NBS-LRR genes on each chromosome was highly correlated with the number of disease resistance QTL in the 2-Mb flanking regions of NBS-LRR genes. In addition, the recently duplicated regions contained duplicated NBS-LRR genes and duplicated disease resistance QTL, and possessed either an uneven or even number of NBS-LRR genes on each side. The significant difference in NBS-LRR gene expression between a resistant near-isogenic line (NIL) and a susceptible NIL after inoculation of Xanthomonas axonopodis pv. glycines supports the conjecture that NBS-LRR genes have disease resistance functions in the soybean genome. The number of NBS-LRR genes and disease resistance QTL in the 2-Mb flanking regions of each chromosome was significantly correlated, and several recently duplicated regions that contain NBS-LRR genes harbored disease resistance QTL for both sides. In addition, NBS-LRR gene expression was significantly different between the BLP-resistant NIL and the BLP-susceptible NIL in response to bacterial infection. From these observations, NBS-LRR genes are suggested to contribute to disease resistance in soybean. Moreover, we propose models for how NBS-LRR genes were duplicated, and apply Ks values for each NBS-LRR gene cluster.

The genus Rubus belongs to the Rosaceae family and is comprised of 600-800 species distributed worldwide. Understanding the genetic relationships and genetic structure in Rubus species is important for enabling efficient management, conservation, characterization and utilization of the species. However, as a minor crop, genetic research foundation was limited to explore genetic diversity and relationships in Rubus species. The present study shows the results of application SSR markers that were developed from SSR-enriched libraries of the one Rubus species (Rubus coreanus Mique.) in our previous study. We used 34 polymorphic microsatellite markers to analysis of genetic diversity within the Rubus species, including redraspberry, blackraspberry, blackberry and mountainberry. All the 34 SSR primers pairs produced 483 polymorphic and reproducible amplification fragments. The largest number of alleles per primer pair was confirmed at GB-RC-167, GB-RC-100, GB-RC-076 and GB-RC-245, which contained 26, 25, 23 and 21, respectively. An average value of polymorphic information contents (PIC) were 0.74 with a range of 0.36 to 0.92. Population structure and phylogenetic analyses showed that all Rubus species formed three largely distinct clusters, which were confirmed by principal coordinate analysis (PCoA). We obtained the results that the developed SSR markers showed a substantial degree of genetic diversity in the various Rubus species distributed in Korea.

Recent release of whole genome draft sequences in legume species have led comparative genome studies among legume plants including Glycine max, G. soja, Cajanus cajan and Medicago truncatula. The majority of comparative genomic researches have been conducted based on synteny of coding sequences and coding sequence variations may be one of major forces for speciation and evolution. However, non-coding sequences have been also reported to be important phenotypic regulators. Especially, since short sequence motifs in the promoter regions are highly conserved, they are suggested to be another resources of interests in comparative studies. In this study, we predicted the conserved short sequence motifs by BLASTN algorithm using dicot promoter database from Softberry (http://www.softberry.com). A total of 37,396 conserved short sequence motifs were identified onto 2 kb upstreams of 46,367 high confident gene model of G. max (cv. Williams 82). Meanwhile, whole genome of 7 soybean landraces (G. max) and 7 wild soybean genotypes (G. soja) were sequenced at low depth of less than ten using Illumina Hiseq 2000. Among these genotypes, nucleotide variations were identified in predicted conserved regulatory motifs by mapping of short reads to the reference genome sequence using the Samtools program (http://samtools.sourceforge.net/). Fifteen and two genes, which have SNPs in regulatory motifs and no SNP in coding sequence, were identified by comparisons of inter-species and intra-species, respectively. qRT-PCR experiments are in progress for investigating differences of these 17 genes expressions at transcriptional level.

Here, we first demonstrate that identification of rice brown spot disease fungus (Cochliobolus miyabeanus, C. miyabeanus) proteins is possible in infected tissues using in planta apoplastic proteome with non-destructive tissues. In planta apoplastic proteins from rice leaves inoculated with C. miyabeanus, CM2 (compatible race), were isolated by vacuum infiltration with CaCl2/Na-acetateextractionbuffer, separated by SDS-PAGE, and identified by MudPIT. Of the 529 proteins that were identified by MudPIT, a large proportion (490) was from the rice. Numerous carbohydrate metabolic process (48), oxidation and reduction (44), response to oxidative stress (20%) were identified and confirmed their expression at RNA levels using microarray. Bioinformatic analysis showed that 176 and 39 of these proteins have a signal peptide in rice and rice brown spot fungus, respectively, using Signal P. The large proportion of proteins interestingly identified from the in planta apoplast were involved inprotease, hydrophobin, and host cell wall hydrolysis (Xylanase, beta-glucosidase) derived from pathogen. Thus, we suggest that in planta rice apoplastic secretome will be an important clue to understand the rice-rice brown spot fungus interactions.