Somatic cells such as oviduct epithelial cell, uterine epithelial cell, cumulus-granulosa cell and buffalo rat river cell has been used to establish an effective culture system for bovine embryos produced in in vitro. But nitric oxide (NO) metabolites secreted from somatic cells were largely arrested the development of bovine in vitro matured/ in vitro fertilized (IVM/IVF) embryos, suggesting that NO was induced the embryonic toxic substance into culture medium. The objective of this study was to investigate whether BOEC co-culture system can ameliorate the NO-mediated oxidative stress in the culture of bovine IVM/IVF embryos. Therefore, we evaluated the developmental rate of bovine IVM/IVF embryos under BOEC co-culture system in the presence or absence of sodium nitroprusside (SNP), as a NO donor, and also detected the expression of growth factor (TGF-p , EGF and IGFBP) and apoptosis (Caspase-3, Bax and Bcl-2) genes. The supplement of SNP over 5 uM was strongly inhibited blastocyst development of bovine IVM/IVF embryos than in control and 1 uM SNP group (Table 2). The developmental rates beyond morulae stages of bovine IVM/IVF embryos co-cultured with BOEC regardless of SNP supplement (40.4% in 5 uM SNP+ BOEC group and 65.1% in BOEC group) were significantly increased than those of control (35.0%) and SNP single treatment group (23.3%, p<0.05: Table 3). The transcripts of Bax and Caspase-3 genes were detected in all experiment groups (1:Isolated fresh cell (IFC), 2:Primary culture cell (PCC), 3:PCC after using the embryo culture, 4: PCC containing 5 uM SNP and 5: PCC containing 5 uM SNP after using the embryo culture), but Bcl-2 gene was not detected in IFC and PCC (Fig. 1). In the expression of growth factor genes, TGF-p gene was found in all experimental groups, and EGF and IGFBP genes were not found in IFC and PCC (Fig. 2). These results indicate that BOEC co-culture system can increase the development beyond morula stages of bovine IVM/IVF embryos, possibly suggesting the alleviation of embryonic toxic substance like nitric oxide.

The yields of the n-hexane-soluble, ethyl acetate-soluble, methanol-soluble and methanol-insoluble fractions from the powdered water extracts were 8.2%, 10.6%, 32.0% and 49.2%, respectively (Table 1). The antifungal activities of water extracts, n-hexane-soluble fraction, ethyl acetate-soluble fraction, methanol-soluble fraction and methanol-insoluble fraction against L. edodes were 23.5%, 26.2%, 41.5%, 25.4% and 2.5%, respectively, at 1000ppm (Figure3). The ethyl acetate-soluble fraction showed much higher antifungal activity than the other fractions. The antifungal activity of the ethyl acetate-soluble fraction against L. edodes at 1000 ppm showed a statistically significant difference in the fractions of n-hexane-soluble, ethyl acetate-soluble, methanol-soluble and methanol-insolub

The yields of the fractions of n-hexane-soluble, ethyl acetate-soluble and methanol-soluble from the P. densiflora sawdust were obtained 1.36%, 2.21% and 4.03% using organic solvents, respectively (Table 1). The antifungal activity of the n-hexane extracts against L. edodes at concentrations 125, 250, 500, 750 and 1000 ppm were 36.5%, 41.2%, 43.1%, 45.4% and 47.6%, respectively (Table 2). The percent of antifungal activity against L. edodes was the greatest for the n-hexane extracts, ranging from 36.5% to 47.6% at a concentrations of 125 ppm to 1000ppm, with the values for all concentrations significantly different from one another. In this experiment, the antifungal activity by the n-hexane extracts was the highest. This result supposed that the n-hexane extracts have not only many more inhibitory compounds against L. edodes than the ethyl acetate and methanol extracts but also the inhibitory compound may be more easily dissolved by n-hexane than by ethyl acetate or methanol.

Cordyceps has been used in traditional Chinese medicine more than 2000 year ago. In this study, the new Cordyceps militaris was founded and isolated from O-dae mountain in Korea, and was identified its genetic characteristics. The newly isolation strain HB8 was most closet to Cordyceps militaris W141449 (99.82%), Cordyceps militaris JLCY-LI819 (99.82%) and Cordyceps militaris 4642 (99.81%), respectively. the genotypic result was show that train HB8 was belonging to the Cordyceps militaris genus, therefore, Cordycep militaris HB8 proposed with accession number MT835161. This study we find the optimal condition for production of cordycepin from Cordyceps militaris HB8 was 8 ㎎ /g (200 g of pupa, 1 g of KH2PO4, 0.5 g of K2HPO4, 20 g of glucose, 1 g of MgSO4, 0.05 g of vitamin B1, and 1 ㎎ of NAA per liter; light condition 300-700 Lux and day/night was 14 h/10 h) and the optimum condition for the production of adenosine was 2.6 ㎎/g (15 g of skim milk powder, 1 g of KH2PO4, 0.5 g of K2HPO4, 20 g of glucose, 1 g of MgSO4, 0.05 g of vitamin B1, and 1 ㎎ of NAA per liter; light condition 300-700 Lux and day/night was 14 h/10 h).

The β-carotene biofortified transgenic soybean was developed recently through Agrobacterium -mediated transformation using the recombinant PAC (Phytoene synthase-2A-Carotene desaturase) gene in Korean soybean (Glycine max L. cv. Kwangan). GM crops prior to use as food or release into the environment required risk assessments to environment and human health in Korea. Generally, transgenic plants containing a copy of T-DNA were used for stable expression of desirable trait gene in risk assessments. Also, information about integration site of T-DNA can be used to test the hypothesis that the inserted DNA does not trigger production of unintended transgenic proteins, or disrupt plant genes, which may cause the transgenic crop to be harmful. As these reasons, we selected four transgenic soybean lines expressing carotenoid biosynthesis genes with a copy of T-DNA by using Southern blot analysis, and analyzed the integration sites of their T-DNA by using flanking sequence analysis. The results showed that, T-DNA of three transgenic soybean lines (7-1-1-1, 9-1-2, 10-10-1) was inserted within intergenic region of the soybean chromosome, while T-DNA of a transgenic soybean line (10-19-1) located exon region of chromosome 13. This data of integration site and flanking sequences is useful for the biosafety assessment and for the identification of the β-carotene biofortified transgenic soybean.

A variety of genetically modified (GM) crops have been developed in Korea. In these crops, the resveratrol-enriched transgenic rice plant has moved ahead to generate the dossier for regulatory review process required for commercialization of GM crop. The resveratrol-enriched transgenic rice plant could be released to farmers for cultivation after national regulators have determined that it is safe for the environment and human health. Here we developed a PCR-based DNA marker based on flanking sequences of transgene for the discrimination of zygosity in resveratrol-enriched transgenic rice plant. This DNA marker will be useful for identifying of resveratrol-enriched transgenic rice plant, and can also be use to estimate transgene movement occurred by pollen transfer or seed distribution.

Clubroot is a devastating disease caused by Plasmodiophora brassicae and results in severe losses of yield and quality in Brassica crops including Brassica oleracea. Therefore, it is important to identify resistance gene for CR disease and apply it to breeding of Brassica crops. In this study, we applied genotyping-by-sequencing (GBS) technique to construct high resolution genetic map and mapping of clubroot resistance (CR) genes. A total of 18,187 GBS markers were identified between two parent lines resistant and susceptible to the disease, of which 4,103 markers were genotyped in all 78 F2 plants generated from crossing of both parent lines. The markers were clustered into nine linkage groups spanning 879.9 cM, generating high resolution genetic map enough to refine reported reference genome of cabbage. In addition, through QTL analysis using 78 F2:3 progenies and mapping based on the genetic map, two and single major QTLs were identified for resistance of race 2 and race 9 of P. brassicae, respectively. These QTLs did not show collinearity with CR loci found in Chinese cabbage (Brassica rapa) but roughly overlapped with CR loci identified in cabbage for resistance to race 4. Taken together, genetic map and QTLs obtained in this study will provide valuable information to improve reference genome and clubroot resistance in cabbage.

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.

Resveratrol rice Iksan526 was developed by overexpession of T-DNA (RB::P-Ubi::RS::T-NOS::P-35S::PAT::T-35S::LB) in rice variety Dongjin. To confirm one locus insertion of T-DNAs, Mendelian genetic analysis was carried out on selection marker bar gene and objective RS gene separately by using a F2 population derived from a cross of Dongjin/Iksan526 (T6). A total of 450 four-leaf-old plants from F2 population were treated by 0.3% basta, and a phenotypic separation ratio of 3:1 (321 survival: 129 dead, p>0.90) complied with Mendelian inheritance indicating one locus insertion of bar gene. Genotypic separation was analyzed by using PCR with specific primers for 300 plants, which were selected from 321 survival plants after phenotypic separation. Results revealed a ratio 1:2 of homologous to heterozygous (92:208, p>0.90), which further confirmed one locus insertion of RS gene. In addition, comparison on agronomic traits and resveratrol contents between transgenic rice and the donor variety were launched to evaluate the phenotypic performance over multi-generations (years).

The C-repeat/dehydration-responsive element binding transcription factors (CBF/DREBs) are involved in an important pathway for abiotic stress-response in plants. We have identified CBF/DREB1 gene family from Brassica rapa whole genome sequence and designated them as BrDREB1s. They contain conserved nucleus localization signal, AP2/EREBP domain, and CBF/DREB1 signature, as other known plant CBF/DREB1s. By comparative genomics, we found that nine of ten BrDREB1 genes were present in seven macro-synteny blocks co-linear to four Arabidopsis counterpart blocks and also genomic organizations of their flanking regions were very similar to those for co-linear Arabidopsis CBF/DREB1 genes. In particular, three genes, BrDREB1A, BrDREB1B1, and BrDREB1C1, were closely located within a 59 kb genomic sequence, which was similar to that of their Arabidopsis counterpart genes. However, the genomic regions of those BrDREB1 genes contained additional sequences, compared to their co-linear regions in A. thaliana. The expression of BrDREB1 genes under abiotic stresses were examined by searching microarray database and by RT-PCR analysis. All of eight genes tested were highly up-regulated during cold treatment and some of them were also responsive to salt, drought, and ABA treatment. Taken together, these results indicate that CBF/DREB1-mediated stress signaling pathway is also functioning in B. rapa. On the other hand, differences in genomic organization and gene number for CBF/DREB1 are thought to cause different response to stress between B. rapa and A. thaliana. In this presentation, we will introduce more detailed results for CBF/DREB1 gene family in B. rapa.

Miniature inverted- repeat transposable elements are expected to play vital role in evolution of genes and genome of major eukaryotic organisms. However, there have been little reports on MITEs in B. rapa, a polyploidy model genome. We identified 13 novel MITE families in B. rapa genome by computational approach. Out of 13 MITEs families three, eight and two were classified under stowaway-like, tourist-like and hAT super families based on their unique structural characteristics. We characterized the members of 13 MITE families from the available 256 Mbp from whole genome draft sequences of B. rapa. We found ech MITE has high copiy number ranges from 14 to 977 which are distributed randomly along all the chromosomes. We also found more than 40% of the MITE members were associated with genes and gene rich regions. Furthermore, the polymorphism due to insertion and non-insertion of MITEs analysis suggest that MITEs are active in the genome. As, such the newly identified MITEs will provide a foundation for the further analysis of roles of MITEs in gene and genome evolution.

Ginseng (Panax ginseng) is frequently used in Asian countries as a traditional medicine. The major components of ginseng are ginsenosides. Among these, ginsenoside compound K has been reported to prevent the formation of malignancy and metastasis of cancer by blocking the formation of tumor and suppressing the invasion of cancer cells. In this study, ginsenoside Rb1 was converted into compound K, via secreted β-glucosidase enzyme from the Leuconostoc lactis DC201 isolated, which was extracted from Kimchi. The strain DC201 was suspended and cultured in MRS broth at 37℃. Subsequently, the residue from the cultured broth supernatant was precipitated with EtOH and then dissolved in 20 mM sodium phosphate buffer (pH 6.0) to obtain an enzyme liquid. Meanwhile, the crude enzyme solution was mixed with ginsenoside Rb1 at a ratio of 1:4 (v/v).The reaction was carried out at 30℃ and 190 rpm for 72 hours, and then analyzed by TLC and HPLC. The result showed that ginsenoside Rb1 was transformed into compound K after 72 hours post reaction.

family in the Brassica genome sequences by computational approach. The MITE family showed a total of 264bp length including 36bp terminal inverted repeats and remained 2bp (TA) targets it eduplication by its insertion. By searching the genome database of Brassica species, 516, 227, and 15 members were identified from 470Mbp of Brassica oleraceae, 154Mbp of B.rapa and 15Mbp of B.napus, respectively, indicating that there are approximately 692, 760, 1235 copies in B.oleracea, B.rapa and B.napus genomes,respectively. A total of 225 relatively intact MITE members, 146,68, and 11 members, which showed >80% sequence similarity and sequence coverage were identified and retrieved for MITE analysis from B.oleracea, B.rapa and B.napus genomes, respectively. Out of 225 MITE family members 159 having full structure of MITE and 66 having the truncated end either in right TIR or left TIR. Insertion polymorphism due to insertion or non-insertion of MITEs showed high level of polymorphism among accessions intra and inter species of Brassica. The new MITE would provide abetter tool for study molecular breeding in Brassica species and also helpful to understand their contribution in evolution and diversification of the highly duplicated Brassica genome.

Perilla is a genus as a member of the mint family Lamiaceae which is known to contain lots of volatile metabolite. Perilla has been called as ‘deulkae’ indicating ‘wild sesame’ that means it has been maintained in Korea with long history. It has been very friendly used as edible oil and as fresh leaf vegetable. Perilla oil is valued for its medicinal benefit because it contains best amounts of unsaturated fatty acids, especially for the alpha-linolenic acid, known to omega-3 fatty acid, among all of the plant oils. It also include many beneficial phytochemicals. However, little study is conducted on their genetics. Here, we announce construction of well normalized and full length enriched-perilla cDNA library from a whole plant of one cultivar ‘Youngho-deulkae’ and their sequence characterization to provide useful resources for genetics, breeding and metabolite engineering. By sequencing of 5,760 cDNA clones, we 5,438 high quality EST sequences. Sequence trimming and assembly resulted 3,995 unigenes which consists 1,004 contigs and 2,991 singletones. Unigenes that showed little homology at the DNA sequence level with known genes in other plants even though they showed similarity at the protein domain level based on BLASTN, BLASTX, and TBLASTX. This study may provide good resources for initiation of further genomics, comparative genomics, functional genomics such as metabolic engineering and molecular breeding.