High yield is the most important trait in various agricultural characteristics. Many approaches to improve yield have been tried in conventional agricultural practice and recently biotechnological tools employed for same goal. Genetic transformation of key genes to increase yield is one way to overcome current limitation in the field. We are producing transgenic soybean plants through high efficient transformation method by introducing all gene member with AT-hook binding domain, hoping to obtain manageable delay of senescence. Many transgenic soybean plants are growing in greenhouse and GMO field, and will be evaluated their senescence and any association with yield increase.

Soybean is a crop of importance economically and nutritionally in many parts of the world. Thanks to many new genes brought from genomic research, It is possible to introduce various candidate genes through genetic transformation to see the performance of the genes in field. In our lab, soybean transformations have been tried for last 10 years to probe the possibility of traits improvement by transformation of new gene into soybean. For this purpose, three different genes were transformed into Korean soybean variety, Kwangan. First, the gene that controls early flowering of plant was transformed into Kwangan. Second, a candidate gene for soybean mosaic virus (SMV) resistance was transformed to produce transgenic plants. Third, another candidate gene for drought tolerance was transformed. All the transgenic plants from three genes transformation were produced for their gene insertion and their expression using PCR, qRT-PCR. Further analysis including harvesting seeds is currently undertaken.

As sessile organisms, plants have evolved mechanisms that allow them to adapt and survive periods of various environmental stresses including high salinity and drought. The ubiquitin-proteasome system (UPS) is an integral player in plant response and adaptation to various abiotic stresses. Understanding UPS function has centered mainly on defining the role of E3 ubiquitin ligases, which are the substrate-recruiting component of the ubiquitination pathway. Here, we report on Ring finger E3 ligase, Oryza sativa salt- and drought-induced RING finger protein1 gene (OsSDRFP1) in defense responses to osmotic stresses. Results of qRT-PCR and In vitro ubiquitination assay demonstrated that OsSDRFP1 act as an E3 ligase in response to salt and drought stresses. in this study, Subcellular localizations showed that the OsSDRFP1 was observed in cytosol (66%) and nucleus (34%) under non-treated conditions. However, the florescence signals of rice protoplasts after salt treatments detected in nucleus (60%) higher than in cytosol (30%). The Arabidopsis plants overexpressing OsSDRFP1 clearly exhibited hypersensitive responses to salt stress. whereas, OsSDRFP1-overexpressing plants were more tolerant to both drought- and ABA-stresses than the wild-type plants. These results might suggest that OsSDRFP1 has a dual function as a regulator of high salt- and drought-stresses.

In this study, we report that the development of a multiplex PCR method using species-specific primers for the simultaneous detection of Poaceae family members, including adlay, barley, maize, rice and wheat, based on the sequence polymorphism of the DNA-directed RNA polymerase beta'' chain (rpoC2) genes Species-specific primers were constructed with common forward primer and each reverse primers which have differences on the basis of sequences. Each primer pairs could amplify PCR products of 443 bp for rice, 346 bp for barley, 278 bp for adlay, 221 bp for wheat and 96 for maize, respectively, from the five chloroplast DNAs. The series of template DNA concentrations were identified by the sensitivity of multiplex PCR. The band of products were clearly amplified from the DNA concentration range of 0.01 to 10 ng/μL. In addition, the species-specific primers were examined for the detection of seven commercial flour mixed products. The combination of the sensitivity of a multiplex PCR with the specificity of the primers for the detection of species would allow to be applied in analyses of processed foods.

Recently, the increased consumption of mixed-grain flour products have led to improved human health in busy modern life. For this reason, the verification of commercial food authenticity is one of important subjects. The development of DNA techniques such as real-time PCR has led to the increasing efficiency of illegal food product detection. Here, we have developed a comprehensive method for detecting the grain flour of various rice cultivars in commercial food products derived from different plant species. In the genetic variation analysis of different protein coding genes on various chloroplast genomes, we found the high numbers of segregating sites in rpoB and rpoC2 more than in other genes. Thus, we have attempted to develop chloroplast DNA (cpDNA) markers, which were Os_m_rpoB in rpoB, and Os_m1_rpoC2 and Os_m2_rpoC2 in rpoC2. To assess the applicability of three cpDNA markers, we have identified the appropriate statistical measurements of each marker in various mixed-grain flour samples derived from rice cultivars and different plant species by real-time PCR, In addition, the three cpDNA markers successfully applied for detecting of nonexistent rice flour in different commercial food products.

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.

DNA barcoding is the use of short DNA sequences of the genome for large scale species identification. The Consortium for the Barcode of Life (CBOL) plant-working group recommended the 2-locus combination as the standard plant barcode. The evolutions of the chloroplast regions combine with nuclear gens are sufficiently rapid to allow discrimination between closely related species. We evaluated the efficacy of the proposed plant barcoding loci matK along with ITS2 for barcoding Vigna species. To assess the discrimination ability of barcoding loci to resolve Vigna species, we sampled 52 of the taxonomically best known groups in the genus. Topologies of the phylogenetic trees based on ITS2 and matK analyses were similar but a few accessions were placed into distant phylogenetic groups. Neither ITS2 nor matK analyses were able to discriminate some closely related Vigna species alone. Thus, we used concatenated data to increase the resolving power of ITS2 and used matK as an additional tool for phylogenetic analysis in Vigna because characterization of the nucleotide sequences of matK region was easier to recover and more cost-effective than those of the ITS region.

Exposure to ionizing radiation is regarded as a kind of abiotic stresses that can change the expression of genes in living organisms. This study aimed on investigating the variations in gene expressions induced by two different types of irradiations with different doses, which were low linear energy transfer (LET) gamma rays (100, 200, and 400 Gy) and high LET ion-beams (20, 40, and 80 Gy) on rice. RNA sequencing was carried out using the Illumina HiSeq-2500 platform. The average amount of reads were 4.8 Gb per individual, and 5 to 8% of the reads were removed after quality control. More than 90% of the RNA-seq reads were mapped to the rice reference genome sequence (IRGSP-1.0). A total of 247 differentially expressed genes (DEGs) were identified by comparison of the gene expression levels between the wildtype and the irradiated individuals. The 247 DEGs were divided into five modules and 27 intra-modular hub genes were found using the weighted correlation network analysis (WGCNA) method. The MEturquiose module had the most number of genes with 75 related to carbohydrate and small molecule metabolic processes. The co-expression network reconstructed using ARACNE (algorithm for reconstruction of accurate cellular networks) showed specific up- or down-regulation of the genes in each module according to the types and doses of radiation. This study will contribute to understanding the gene expression responses to ionizing irradiation.

본 연구는 3,7-diarylpyrazolo [1,5-α]pyrimidines의 효과적인 one-pot regioselective 합성을 보여준다. 더욱이, 그 유도체는 뛰어난 CB1R 저해 활성을 나타냈다. 3,7-position에 diaryl group이 치환된 pyrazolo [1,5-α]pyrimidine은 CB1R 후보로서 가능성 있는 pharmacophore이다.

Slaughter of cattle, pigs, and chickens has increased continuously. In particular, slaughter of chickens has been grown up about 150% in 2010 than that in 2003, that is approximately 120,000 tons. All of them are underwent consigned treatment even though those can be used as a resource and an energy source. With this regards, THR (Thermal Hydrolysis Reaction) leads to reduce water content drastically (<30% in sludge cakes). In addition, Dehydrated solid would be re-used as solid fuels (SRF) as well. In this study, We have applied THR to a plant (10 ton/day) on the basis of our lab and pilot results. Water content of sludge cakes showed with a ranges of 30 to 40% after solid-liquid separation. Dairy SRF produced 1.5 ton/day and its heat capacity for SRF has 6,500 kcal/kg. This gave the steam produced about 12 ton/day throughout the plant operation, suggesting that THR system would expect energy savings.

Recently, the importance of food safety is increasing due to numerous junk food. Junk food means to violate the law in stage such as production, manufacture, distribution, and sale of food. Many crop plants are processing as foods including bread, noodle, and other foods for supporting nutrition to human. For example, rice is one of the most well-known food crops in the world, and processed rice is being mixed with other processed crops to health food. The object of this study is to detect amount of specific grains, i.e. rice from processed foods mixed with other cereals. This experiment was performed to the following two steps: 1) designed the specific primer sets based on chloroplast DNAs, 2) amplified products using real-time PCR. We designed eleven primer sets within chloroplast DNA of rice, and then the confirmation of primer efficiency was to amplified with rice genomic DNA using real-time PCR. In addition, these primer sets were applied in other crops such as wheat, maize, and adlay to confirm specificity to rice. The rice specific primer sets were determined by the number of amplification and the melting peak through real-time PCR. As a result, five primer sets were confirmed to uniqueness in the rice genome. In conclusion, the specific primer sets would be useful for identifying rice grain from the processed foods to eliminate junk foods and for contribution of food safety.

Brassinosteroids (BRs) play important roles in many aspects of plant growth and development. BR-induced AtBEE3 (brassinosteroid enhanced expression 3) is required for a proper BR response in Arabidopsis. Here, we identified a poplar (Populus alba x P. glandulosa) BEE3 homolog encoding a putative basic helix-loop-helix (bHLH)-type transcription factor through microarray analysis. Transcripts of PagBEE3 were mainly detected in stems, with the internode having a low level of the transcripts and the node having a relatively higher level. The function of the PagBEE3 gene was investigated through the phenotypic analyses with PagBEE3-overexpressing (ox) transgenic lines. This work mainly focused on a potential role of PagBEE3 in stem growth and development of polar. The PagBEE3-ox poplar showed thicker and longer stems than wild-type plants. The xylem cells from the stems of PagBEE3-ox plants revealed remarkably enhanced proliferation, resulting in an earlier thickening growth than wild-type plants. Microarray analysis revealed that the expression of many genes involved in xylem cell proliferation and development was altered in the PagBEE3-ox plants. Therefore, this work suggests that xylem development of poplar is accelerated in PagBEE3-ox plants and PagBEE3 plays a role in the stem growth by increasing the proliferation of xylem cells to promote the initial thickening growth of poplar stems.

Rice is a staple food for over one-half of the world population, especially in Asian countries. Recently, the growth and yield of crop plants was affected by various abiotic stresses, such as salt, drought, and high temperature due to change of climate environment. To study molecular functions of Oryza sativa nuclear-targeted RING Finger Proteins (OsNRFPs) in response to abiotic stresses, we selected 44 OsNRFPs genes, whose subcelluar localizations are predicted to the nucluear, on the basis of expression patterns of a microarray dataset. A total of 44 OsNRFPs were grouped into two types such as RING-HC and RING-H2 via phylogeneitc analysis of their RING domains structures. Subsequently, we surveyed the expression patterns of 44 genes in response to salt stress via qRT-PCR in roots. We found 10 salt stress-induced OsNRFPs and then examined their subcellular localizations. These genes were clearly localized to the nucleus (OsNRFPHC-10), cytoplasm (OsNRFPHC-17 and OsNRFPH2-16) and microtuble (OsNRFPHC-23, OsNRFPH2-17 and OsNRFPH2-05), respectively. These results might provide a key clue for understanding moleuclar functions of the OsNRFP genes associated with salt stress-related signaling pathway

In order to select a rice population with useful trait such as arsenic tolerance for crop improvement, we have developed 3000 M7 Targeting Induced Local Lesions IN Genomes (TILLING) lines by gamma ray (GR) irradiation treatment to a rice variety (cv. Donganbyeo). A total of 2 M7 lines exhibited the arsenic (AsV) tolerant phenotype (hereafter, named Arsenic Tolerant TILLING line 1 and 2, and designed as ATT1 and 2), in which the shoots and roots length of ATT lines were significantly longer than those of wild type (WT) during As(V) treatment. To survey the DNA polymorphism of these plants, we conducted the Whole genome resequencing with 10x coverage in ATT lines. By comparative analysis among ATT lines, we have identified the common DNA polymorphism such as 11,817 SNPs (49.83% in ATT1 and 48.35% in ATT2) and 30,618 InDels (86.72% in ATT1 and 86.23% in ATT2). Also, these mutants were showed the close relationships more than WT. To further study the changed amino acids of genes, we commonly identified the 758 genes for non-synonymous SNPs and 249 genes for changed codon InDels. These genes were mainly exhibited the enriched GO functions such as catalytic activity, nucleic acid binding and transferring phosphorus-containing groups. To determine the genes associated with arsenic-related mechanism in DNA polymorphism of ATT lines, we have retrieved the two structurally altered genes (Os11g47870 and Os03g19900) for metalloid As(V) detoxification toward induced genes in response to arsenic treatments by public microarray datasets. We suggest that As(V) tolerant phenotypes of ATT lines are certainly affected by structurally altered genes associated with phosphorus transferring and As(V) detoxification during GR treatment

Chrysanthemum white rust, caused by Puccinia horiana, is one of the most destructive fungal diseases in chrysanthemum cultivation worldwide. For increasing efficiency of resistant breeding, molecular markers linked to chrysanthemum white rust resistance gene were developed in pseudo F1 cross population between ‘Puma White’ as susceptible and ‘Dancer’ as resistant using bulked segregant analysis (BSA). Of 280 RAPD primers (Operon 10 mer), 18 primers found to be polymorphic. After screening of these primers in 20 individual lines, only OPI-13520 was selected as closely linked marker to white rust disease resistance. Based on correspondence between phenotypic resistant level and marker in 187 segregation population, the genetic distance between white rust resistance gene and OPI-13520 marker assumed to be 3.8 cM. For OPI-13520 marker conversion into sequence characterized amplified region (SCAR) marker, the amplified fragment of OPI-13520 was purified, cloned and sequenced. Based on the DNA sequence of OPI-13520, SCAR maker was generated and verified in 20 individual lines used in BSA-RAPD.The results showed SCAR marker could be used to identify white rust resistance in chrysanthemum.

The metalloid arsenic (As) and the hevy metal cadmium (Cd) are ubiquitously found at low concentrations in the earth, while high concentrations of the both elements in soil and crop are severe dangerous to human health. We have tried to retrieve RING E3 ligase gene, which is believed to regulate substrate proteins in As or Cd uptake via ubiquitin 26S proteasome pathway, related to inhibit metal ion transport system. A total of 48 rice RING E3 ligases were randomly selected and then conducted semi-quantitative RT-PCR for their expression patterns as exposed to As and Cd treatments. We discovered one gene, Oryza sativa heavy metal induced RING E3 ligase 1 (OsHIR1) that was significantly up-regulated against both treatments. A total of 31 positive interaction clones with OsHIR1 were screened depending on their strong α-galactosidase activity via yeast-two hybrid screen. Bimolecular fluorescence complementation analysis evidenced that the OsHIR1 protein was clearly interacted with each of six partner protein including aquaporin tonoplast intrinsic protein 4;1 (OsTIP4;1) in the plasma membrane. Protein degradation assay showed that OsHIR1 strongly degraded the protein level of OsTIP4;1 via ubiquitin 26S proteasome system. Heterogeneous overexpression of OsHIR1 in Arabidopsis showed As- and Cd-insensitive phenotype. In addition, the transgenic plant showed low levels of As and Cd accumulation than the control plant in leaf and root. Here, we report the novel finding that OsHIR1 E3 ligase positively regulates OsTIP4;1 related to As and Cd uptake.