Viruses of the honeybee, Apis mellifera L. are known to reside at low levels in colonies, typically showing no apparent signs of infection. Chronic paralysis virus(CBPV) is known to induce significant losses in honey bee colonies. The pathology is characterized by clusters of trembling, flightless, crawling bees and by individual bees, sometimes hairless, standing at the hive entrance. A minusstrand-specific RT-PCR was used to assess viral replication. This is the first report on the infection of CBPV in Korea. Using (-)RT-PCR, 27 apiaries in korea were screened for the honeybee viruses, with positive colonies being analysed for viral genetic diversity. We got 550-nt PCR product from CBPV genomic RNA. Nucleotide sequences were aligned to the complete CBPV genomic RNA sequence deposited in the GenBank database and was revealed 96%(AM-CBPV) identity, respectively. Sequence comparison with other CBPV and honeybee virus.

Mortality of honeybees is a serious problem that beekeepers have to face periodically in Korea and worldwide. The presence of RNA viruses, in addition to other pathogens may be one of its possible causes. In this work, we detected Deformed wing virus(DWV), Israle Acute Paralysis Virus (IAPV), Black queen cell virus (BQCV), Cloudy wing virus(CWV), Kashmir bee virus(KBV), Sacbrood virus(SBV), Chronic bee paralysis virus(CBPV) in samples of korea honeybees with or without Varroa destructor and Nosema apis. The detection of viruses in all provinces, simultaneous co-infection of colonies by several viruses and the fact that 96.3% of the samples were infected with one or more virus, indicates they are widely spread in the region. Using uniplex and multiplex RT-PCR we screened honey bee colonies for the presence of several bee viruses, including DWV, IAPV, BQCV, KBV, CWV, and described the detection of mixed virus infections in bees from these colonies. Conclusively, investigated disease of the bee, and confirmed new virus that lead to bee disease, this is thought by valuable thing as data for development of beekeeping industry such as CCD(Colony Collapse Disorder)'s cause searching examination.

본 연구는 4DCT를 이용하여 종양의 움직임을 분석하였다. ITV 여유에 따른 TC, II, CI를 이용하여 적절 한 균일 IM을 확인하였다. 또한 각각의 경우에서 DVH와 NTCP를 비교하였다. 균일 IM 적용 시 종양에 대한 선량 분석 결과 TC, CI, II를 모두 만족하는 최적의 치료계획은 20% 위상에서 2 mm, 40%는 3 mm로 평가되었다. 이를 PTV20, PTV40의 정상조직에 조사되는 선량과 비교하였다. 20% 방사선 조사 영역에서 폐에 대한 V5, V10, V20은 1.49, 1.26, 0.65% 증가하였고 40%는 1.9, 2.41, 1.23% 증가하였다. NTCP 역시 20%와 4 0% 각각 0.57, 0.029% 선량 증가가 있었지만 PTV100 보다 확실한 이점이 있었다. 척수와 심장은 각각 균일 IM에서 선량증가는 있었지만 큰 차이는 없었다. Novalis ExacTrac 시스템을 이용하여 호흡연동방사선치료 시 20% 위상의 ITV설정은 균일 IM 2 mm, 40%는 3 mm를 적용함에 있어서 종양선량의 기하학적 누락 없이 최적의 치료계획을 할 수 있음을 확인하였다.

Roasting has revealed coffee’s potentials as a good source of bioactive compounds. This study was done to investigate the quantitative presence and activity of bioactive compounds including caffeine, chlorogenic acid (CGA), amino acids, and antioxidant capacity on Coffea arabica L. (Guatemala finca San Sebastian) and C. robusta L. (India Azad Hind). Analysis was performed on Green Bean (GB) Medium-Light (ML), Medium (ME) and Medium-Dark (MD) samples of both varieties. From the results, caffeine content was highest in ME samples of both varieties. GB samples of both varieties had high CGA content which decreased after increasing roasting time and temperature. Most amino acids in GB samples was highest, however, glutamic acid, valine, tyrosine, isoleucine, leucine and phenylalanine had highest quantitative increase in ME samples for both varieties. IC50 of DPPH and ABTS radical scavenging activity was highest in ML samples of both varieties. IC50 of reducing power and total phenolic content was highest in GB sample of both varieties but decreased after increasing roasting conditions. Generally Robusta had the highest quantity of bioactive compounds and antioxidant activity. From this study, the optimal roasting condition for coffee is ME above which there is a significant reduction of bioactive compounds and antioxidant activity.

Background : Schizandra chinensis Baillon have five tastes and lately it is using a beverage broadly. Schizandra chinensis is one of the top producing medicinal plant in Korea. Mungyeong of Gyongbuk province produce almost of Schizandra chinensis. Maturity of Schizandra chinensis get 3 years and proliferation of Schizandra chinensis was not a manual. It is needed that a new cultivar has a big fruit and high quality chracteristics using processed food and beverage. Methods and Results : 105 lines of Schizandra chinensis were collected on Mungyeong, Yeongwol, Jinan. It were studied it’ characteristics especially it’s fruit trait. Fruit traits of Schizandra chinensis were researched on fruit length, fruit weight, maturity, number of fruit, male and female ratio, powdery mildew. Fruit length of Schizandra chinensis is relation of fruit weight. It were founded 15 lines of long fruit length. 5 lines were studied high fruit weight and it’s weight were 32 to 41g. Number of fruit has relation with fruit weight and high fruit weight gets many fruits. it’s numer of fruits were 3 to 41. Male and female ratio were very impotant characteristic. High level of female ratio has quantity of fruit. High level of female ratio were founded 2 lines. Finally It was selected 3 good breed lines of Schizandra chinensis. Conclusion : 105 lines of Schizandra chinensis Baillons were collected on Mungyeong, Yeongwol, Jinan. It were founded 15 lines of long fruit length and 5 lines were studied high fruit weight. High level of female ratio were founded 2 lines. 3 good breed lines of Schizandra chinensis were selected.

To overcome the risk of the ovarian hyperstimulation syndrome (OHSS) in patients have polycystic ovarian syndrome (PCOS) and to prepare emergency fertility preservation in patients undergoing anticancer treatment, several researchers have reported IVM of oocytes retrieved from ovaries exposed by only hCG priming. However, the maturation rate and the developmental potential of embryos from IVM oocytes are significantly lower than those of oocytes matured in vivo. Here, we investigated the optimal time point for immature oocyte collection at post hCG only injection for in vitro maturation, in vitro fertilization and blastocyst formation. Immature GV oocytes were collected from 25 days old B6D2F1 female mouse at 12 hr, 14 hr, 16 hr or 24 hr post hCG injection. Oocytes were collected from antral or late secondary follicle by puncturing with 26 G needle. Collected oocytes were cultured in G2 medium with 10% FBS, FSH, estradiol, and hCG for 16 hr in vitro and subjected in vitro fertilization and further embryonic development. To examine follicular maturation, we estimated the numbers of primordial, primary, secondary follicle and antral follicle on ovaries of each time point post hCG. To confirm the optimal time point post hCG injection for collecting immature oocytes, we recovered the oocytes from each time point. There is no difference in the number of oocytes per mice. Oocytes collected at 14 hr post hCG injection were shown higher maturation rate to MII stage and blastocyst formation compare to other three groups (p<0.01). However, there is no difference in the maturation rate on the other three groups. Also, apoptotic signal with TUNEL assay or anti-PARP staining was not change in ovaries from all experimental groups. Granulosa cell proliferation test with anti Ki-67 or anti AMH was not show any difference. According to these results, there are no significant differences in four different time points at 12 hr, 14 hr, 16 hr or 24 hr of collection of immature oocytes in hCG primed mouse. However, oocytes from 14 hr post hCG injection showed higher percentages of maturation rate, in vitro fertilization rate, blastocyst formation.

Cryopreservation has become a powerful method of the assisted reproduction technology and supports fertility preservation of cancer and other indication patients. After controlled ovarian hyperstimulation, surplus oocytes and embryos were recommended to store using cryopreservation. Recently, vitrification is replaced with traditional slow freezing protocol, because of improved survival rates and clinical outcomes. Vitrification requires a high concentration of CPAs that may induce significant osmotic and metabolic damage to cells including oocytes even in a short exposure of a few minutes. Generally, MPF plays a crucial role in the cell cycle regulation and maintaining the meiotic arrest of oocytes. In fact, it has been observed to decline in MII ovine oocytes after vitrification and would be suggested that one of the main causes of low fertilization rate and developmental competence derived from cryoinjury during vitrification. Therefore, the aim of this study was to evaluate the effect of caffeine treatment on the activity of MPF, MAPK level in vitrified/warmed mouse mature eggs. Caffeine, Phosphataseinhibitor, may maintain active form of MPF. We evaluated their survival after warming procedure, fertilization, cleavage, and developmental rates. Ovulated MII eggs were retrieved from 6 weeks old B6D2F1 female mouse at 14hr post hCG injection. Collected MII eggs were maintained in HTF medium containing 10% KSR with or without caffeine for 1hr. Eggs were vitrified in 7.5%EG +7.5%DMSO equilibrium solution, 15%EG + 15%DMSO + 0.5M sucrose vitrification solution with or without caffeine. Also warming solution contained sucrose (0.5M, 0.25M, 0.125M, and 0M) with or without caffeine. After warming, eggs were cultured in HTF medium with or without caffeine for 2 hr then fertilized with epididymal sperm in vitro and cultured in KSOM for 5 days to analyze embryonic development. Survival rates were similar in all experimental groups. However, fertilization rate was higher in with caffeine group compare to without caffeine significantly (80% vs. 85%, p<0.05). 2-cell and blastocyst formation were increased in caffeine group (p<0.05). MPF activity and MAP kinase activity were recovered in with caffeine group after vitrification/warming process. In conclusion, Caffeine may maintain MPF and MAPK level in vitrified/warmed MII eggs, and enhance fertilization and further embryonic development.

Reactive oxygen species (ROS) are produced in organisms as the natural products of oxidative metabolism by environmental stress and pathogen invasion. ROS, such as superoxide anion and hydrogen peroxide, can be toxic to cells and tissues to cause oxidative stress. Recent study revealed that olive flounder (Paralichthys olivaceus) superoxide dismutase (SOD) has been identified as a partial gene and strongly induced to benzoin[a]pyrene and it was deduced indicator of aquatic oxidative stress responses, but its transcriptional response against viral infection has not been investigated. In the present study, spatial and temporal expression profile was analyzed to investigate the function of Of-SOD in the anti-viral response. Of-SOD transcripts were ubiquitously detected in diverse tissues with variable levels using a real-time PCR. The expression of Of-SOD was significantly higher in the muscle, liver and brain, but extremely low in the stomach and spleen. Following VHSV challenge, the expression of Of-SOD increased within 3 hours and subsequently decreased to the original level at 2 days post-challenge in kidney. Although expression pattern and induction time are slight differences depending on the tissue, the transcript of Of-SOD was consistently increased in acute infection response, but expression is low in the chronic response. Collectively, Of-SOD expressions were inducible after VHSV infection and they were probably involved in the immune response against viral challenge. These results suggest that SODs may play important roles in the immune defense system of P. olivaceus and perhaps contribute to the protective effects against oxidative stress in this flounder.

Sequence diversity was accumulated through evolution and breeding process. A set of 595 PCR-based novel insertion/deletion (InDel) markers was designed in order to widen the genetic basis for national rice breeding programs. The markers were generated by analyzing of 40 Korean cultivars and published genome sequences of rice(Oryza sativa L. spp japonica). We selected 112 markers spread across all rice chromosomes among the 595 InDel markers, and they showed polymorphic between rice cultivars, which are 284 Korean japonica and Tongil varieties. Due to their simplicity in design and robustness in genotyping, these InDel markers have been routinely used in quantitative trait loci (QTL) mapping studies and marker assisted selection programs for rice. Moreover, the PCR amplification type of InDel markers was converged to digital code, 0 or 1 and then finally represented as one- and two dimensional bar-code system, which could easily differentiate genetically highly homologous japonica rice cultivars. The developed InDel markers uniquely discriminated among each of the Korean cultivars. Therefore, the systems we developed may be valuable tools in discrimination from cultivars

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.

The excessive and indiscriminate use of chemical fertilizers in the past has brought serious soil and other environmental problems so alternatives over this agrochemical are being searched. Our study focuses on the effects of expanded rice hull inoculated with selected beneficial microorganisms on growth (through agronomic characters), yield and yield components, and grain quality indices of rice. Results showed that favorable effects of different expanded rice hull preparations were not readily apparent at vegetative stage and only treatments with supplemental chemical fertilizer application were comparable with the conventional practice. Expanded rice hull combined with 50% rate of chemical fertilizer exhibited a significantly higher yield (6,471 kg ha-1) over conventional practice (5,719 kg ha-1). Good milling quality indices were observed in treatments having 50% chemical fertilizers plus alternatives from expanded rice hull. Finally, we demonstrated that chemical fertilizer rate can potentially be reduced into 50% if combined with expanded rice hull, and show even better output than chemical fertilizer alone.

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.

Mutagenesis approach in combination with whole genome sequencing has become an import role in genetic and molecular biological study and breeding of crop plants. In this study, we screened the fast neutron M4 10,000 soybean mutant plants based on morphological phenotypes of agronomically important traits and characterized the mutant of interest using resequencing. Fast neutron radiation has been known to be a very effective mutagen to cause large deletion in genome. The screened mutant showed abnormal phenotypes in plant heights, seed sizes, color of leaves, number of leaves, maturity and number of branches etc. Among them, the mutant displaying short plant height and bush type of growth habit was selected for identification of the altered genomic regions. Analysis of deletion sites of genome in interesting soybean mutant was performed using next generation sequencer Illumina Hi-seq. Mutant sequence reads generated by paired-end shotgun library were mapped on a draft soybean reference soybean (G. max cv. Williams 82). The paired-end DNA sequences of 21.6 Gb produced by Illumina Hi-seq produced 21 fold sequence depth. Among the predicted deletion sites, total 3 deletion regions confirmed by PCR. Glyma03g02390 gene and Glyma03g03560 gene were involved in the deletion regions. Glyma03g02390 gene was related to AMP binding, catalytic activity, cofactor binding and metabolic process of cell growth and Glyma03g03560 gene was concerned to oxygen binding, defense response to bacterium, and especially process of indole acetic acid (IAA) biosynthesis. These genes detected in this mutant will be studied about their molecular function in stunted phenotype.

The amount of genetic variability of a species is essential for its survival and adaptation in different environments, and studies of genetic diversity using molecular markers are necessary to understand the genetic structure of a population and to orientate effective strategies of germplasm conservation. The aim of current study was to determine the SSR markers that can be used rapidly and reliably to evaluated the pepper of Bulgaria landraces, and applied the markers to assement of introduce genetic diversity of the pepper germplasm. We used 22 polymorphic microsatellite markers to analysis of genetic diversity within 61 pepper collection of Bulgaria landraces germplasm, all SSR primers pairs produced 80 polymorphic and reproducible amplification fragments. An average value of polymorphic information contents (PIC) were 0.334 with a range of 0.061 to 0.63. The mean values of observed (HO) and expected heterozygosity (HE) were 0.383 and 0.154, respectively, indicating a considerable amount of polymorphism within this collection. A genetic distance-based phylogeny grouped into three distinct groups, which was the landrace, moderate and wilde type, genetic distance (GD) value was 0.540. An average day of flowering time was 53 days with a range of 45 to 60 days. The everage od fruit length and width were 9.38cm with a range 2.1 to 23.6cm, and 3.51cm with a range 0.6 to 8.9cm, respectively. Molecular data were complemented with morphological measurements according to the descriptor list for the pepper collection of Bulgaria landraces germplasm.