This study was aimed to evaluate the dose-response the effects of nano-encapsulated conjugated linoleic acids(CLAs) on in vitro ruminal fermentation profiles. A fistulated Holstein cow was used as a donor of rumen fluid. Nano-encapsulated CLAs(LF, 5% of nano-encapsulated CLA-FFA; HF, 10% of nano-encapsulated CLA-FFA; LT, 5% of nano-encapsulated CLA-TG; HT, 10% of nano-encapsulated CLA-TG) were added to the in vitro ruminal fermentation experiment. In the in vitro ruminal incubation test, the total gas production on incubation with nano-encapsulated CLAs was increased significantly according to the incubation time, compared with the control(p<0.05). The tVFA concentrations on addition of LF and HT were significantly higher than that of the control(p<0.05). Thus, nano-encapsulated CLAs might improve the ruminal fermentation characteristics without adverse effects on the incubation process. In addition, the population of Butyrivibrio fibrisolvens which is closely related to ruminal biohydrogenation was increased by adding HT, while decreased by adding LF at 12 h incubation. These results showed that nano-encapsulated CLA-FFA could be applied to enhance CLA levels in ruminants by maintaining the stability of CLA without causing adverse effects on ruminal fermentation profiles considering the optimal dosage.

The wheat-rye translocations are world widely used in wheat breeding. Among the various forms of wheat-rye translocation, the 1RS (short arm of rye chromosome 1) translocations are the most widely used because of the valuable genes (e.g. biotic / abiotic resistance genes) introduced from rye. We have developed a new wheat-rye cultivar ‘TRANS’ which have freezing resistance and high yield in this study. ‘TRANS’ is a new wheat-rye translocation cultivar developed by crossing between common wheat ‘Keumkangmil’ and ‘951188-G3-G1’, a 1AL.1RS translocation derived from ‘Fleming’ with the aim of high yield and resistant genes to various unfavorable environments carried by 1RS. ‘TRNAS’ have clearly different genetic and agronomic traits to the control cultivar ‘Keumkangmil’. The heading date and maturity of ‘TRANS’ are later than that of ‘Keumkangmil’. ‘TRANS’ has 1023 spike number per square meters and grain yield of 541 kg/10a, which are higher than ‘Keumkangmil’ (904 spike number/m2 and 504 kg/10a yield). ‘TRANS’ showed winter hardiness and powdery mildew resistance in artificial infection test and field evaluation. ‘TRANS’ gives lower flour yield than ‘Keumkangmil’ but ash and protein content were similar to that of ‘Keumkangmil’. The color of flour and noodle dough of ‘TRANS’ were darker than ‘Keumkangmil’. Quality parameters related to milling, flour quality, noodle dough and end-use quality of ‘TRANS’ indicates that ‘TRANS’ is soft wheat suitable for noodle making. ‘TRANS’ can be cultivated in entire part of Korea. Registration Grant No.: 4695

Lepisorus thunbergianus (Kaulf.) Ching has been used in folk medicine in Korea. In this study, a L. thunbergianus methanol extract and its fractions were investigated for their antioxidant properties. The results showed that the ethyl acetate and butanol fractions of L. thunbergianus possess potent DPPH radical scavenging activities. Both fractions also possessed reducing power and inhibited reactive oxygen species formation. In addition, the cytotoxic activity of the L. thunbergianus n-hexane fraction (HF) was investigated. The results suggested that the HF remarkably suppressed proliferation of human breast, liver and colon cancer cells. These results demonstrate, for the first time, that L. thunbergianus extract induces apoptosis in SW620 cells, suggesting that L. thunbergianus may have potential as a therapeutic agent for colon cancer.

Nutritious and functional foods from crop have received great attention in recent years. Colored-grain wheat contains high phenolic compound and a large number of flavonoid. One of plant pigments, wheat anthocyanin is increasingly emerging as natural compounds for consumer´s health and condition. Red grains and white grains with different antioxidant activity was used to conduct germination assay. Antioxidant enzyme assay of POD, APX, CAT, GST, GR and GPx was conducted during the imbibitional phase of mature seeds. Malondialdehyde (MDA) content was analyzed to assess the activity of ROS during imbibition phase of mature seeds and alpha-amylase contents were quantified for 3 days during dark imbibition. Additionally, sprouting rates of developing seeds in spikelet after anthesis with damp condition were measured in each red grain groups for two weeks to evaluate sprout ability affected by phytochemical of red grain wheat. In summary, we identified that red grain wheat showed higher antioxidant enzyme activity involved in ROS scavenging during imbibition. Sprouting rate during dark imbibition in developmental spikelet of four groups classified by color suggest that phytochemicals in dark red grain wheat caused negative effects to sprouting.

The control of flowering, transition from vegetative to reproductive stage, is crucial for significant success during plant development. Multiple environmental and developmental signals are transmitted to the shoot apical meristem and converted to local cue to process developmental phage. These crucial process are delicately controlled and regulated by expression of tissue specifically expressed genes involved in inflorescence development. Therefore, it is necessary that molecular mechanism associated with inflorescence development is revealed to understand control of flowering by genome-wide expression pattern of inflorescence specific genes. In this study we used Affymetrix GeneChip Wheat Genome Array for genome-wide analysis of the expressed genes of inflorescence development including apical meristem and developing spikelet to understand the mechanism of floral development in early stage of wheat inflorescence. Moreover, meta-analysis of 1479 microarray dataset of GPL 3802 provided by Gene Expression Omnibus (GEO) was conducted to determine expression pattern of each probe throughout whole life cycle. Based on meta-analysis, we demonstrate inflorescence specific expressed genes in wheat inflorescence including apical meristem, spikelet meristem to understand the mechanism of floral development of wheat inflorescence.

Pre-harvest sprouting (PHS) is the precocious germination condition of grains while the spike is still in the mother plant. Because PHS in wheat drastically reduced the quality and economic value of wheat grain, the improving PHS wheat is one of the most important breeding goal in Korean wheat breeding program In this study, we evaluated PHS and germination index (GI) in 33 Korean wheat cultivars, and performed transcriptome analysis between Keumkang (susceptible) and Woori (tolerance). A total of 33 Korean wheat cultivars were used for PHS (28 cultivars) and GI assessment in greenhouse. The DAF (Day After Fertilization) 35 of keumkang and Woori spikes were harvested to perform transcriptome analysis using RNA-sequencing. Each transcriptome was compared with PHS or ABA treated DAF 35 Keumkang and Woori spikes. The PHS in 28 Korean cultivars and GI in 33 cultivars were ranged from 1.33% to 87.44% and from 0.01% to 2.41%, respectively. Woori was demonstrated the second lowest PHS and the lowest GI, however, Keumkang was 23th of 28 cultivars in PHS and 13th of 33 cultivars in GI analysis. Six cDNA library from the DAF 35 of Keumkang and Woori wheat grain, PHS treated DAF 35 of Keumkang and Woori, and ABA treated DAF 35 of Keumkang and Woori were constructed and sequenced. A total of 53.37 Gb of high-quality reads were obtained using HiSeq 2500. The average mapping rate of assembled transcripts were 88.98%. The differentially expressed genes (DEG) revealed total 332 DEG (105 annotated) were upregulated in DAF 35 Woori library, total 5694 DEG (4623 annotated) were upregulated in PHS treated DAF 35 Keumkang library in comparison with DAF 35 Keumkang library. A total of 86 DEG (51 annotated) were upregulated in PHS treated DAF 35 Woori library in comparison with PHS treated DAF 35 Keumkang library. The Gene ontology and further analysis will be discussed.

Grain color distinguishes between the pigmentation of the outer layer of the kernel. It is known that environmental factors affects the production of anthocyanins and abiotic stresses like high light intensity, low temperature, high salinity and/or drought stress, and others increase their amounts. After 7 days the germination rate between yellow and dark-purple seeds were almost the same with and without stress (100% yellow seeds under stress and without stress germinated, 93.3% under stress and 96.6% without stress of purple seeds germinated), even though at the final stage the germination was almost the same, we can conclude base on our observations that the germination takes place at a different rate. We think that this might be related to the seed color, since the germination of purple seeds under salt stress started earlier than the yellow ones, until both reached the same point. The antioxidant activity was higher in seedlings from dark-purple seeds than the yellow ones, and they were higher under salt stress than without it, supporting our hypothesis that the purple color in wheat seeds works as a protection under salt stress. Furthermore, the qRT-pCR showed that some genes related to the flavonoid pathway were expressed or had more expression in the seedlings from dark-purple seeds than yellow ones.

Soil salinity limits crop productivity in many regions. This problem would be more serious as the global climate changes and worldwide water shortages would accelerate soil salinization. This study is fulfilled with aim on resolve crop cultivation in dry/saline land as an international joint research project with Tunisia. Total 48 lines of wheat cultivars including 32 common wheat (16 Korean wheat, 16 Tunisian common wheat) and 16 Tunisian durum wheat were incorporated in this study. Salt stress was applied for 2 weeks by submerging the pots into 500 mM NaCl at 3-leaf stage followed by re-watering for restoration in greenhouse. Numerous agronomic/growth parameters were scored for tolerence. SSR primers that have been known to be related to salt tolerance were applied to explain selected population. The correlation between PCR-based length polymorphism of selected lines and their resistance were evaluated. The obtained information will aid selection for salt tolerance hexa/tetraploid wheats. Acknowledgement: This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (No. 2012K1A3A1A09028123) and carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project title: Development of high yielding wheat with stress tolerance via molecular breeding strategies, Project No. PJ008031)”, Rural Development Administration, Republic of Korea.

Brachypodium distachyon is a temperate annual grass that has a short life cycle, a small genome size, self fertility, and a small physical stature. The relationship with major cereal crop including wheat, Brachypodium is considered as a monocot model plant. Recently, the cell wall composition of Brachypodium is reported closely related with maize and Miscanthus giganteus. Therefore, Brachypodium is emrging as a powerful model plant for bioethanol production. Here, Brachypodium was chronically irradiated with the doses of 50 Gy, 100 Gy, 150 Gy, 200Gy, 250Gy, and 300 Gy. Plant height and fresh weight were observed dosage-dependent negative effect. However, tiller number and internode diameter were found to be increased their value as compared to control. The cell wall yield showed a decreased tendency with dosage-dependent negative, but cell wall yield of 50 Gy and 200 Gy were detected higher than control. The lignin content of irradiated Brachypodium stem was reduced with dosage incease The ratios of lignin content to control were 97.6% (50 Gy), 91.9 (100 Gy), 87.3% (150 Gy), 89.4% (200 Gy), 81.6% (250 Gy), 85.2% (300 Gy). SEM image analysis demonstrated that cell size of 300 Gy plant was decreased by 45% of control. RT-PCR was performed to analyze transcript accumulation of lignin pathway related genes with irradiated Brachypodium stem. CCR, PAL, C4H, and 4CL were detected at least 2 times higher expression than control at 150 Gy, 200 Gy, 250 Gy. The preteatment and enzyme hydrolysis will be discussed for bioethanol production.

Over the last decades, increasing natural disasters and climate change are considered as the major environmental problems facing the globe. Numerous studies have been indicated it would cause huge losses on agriculture, especially in the grain productivity. Therefore, several alternatives are suggested for boosting up productivity of wheat as one of the main human food crop. One of important strategy is proper management of inflorescence development and DELLA proteins have been elucidated to play pivotal roles in growth of many plant organs. In this study, putative negative regulator of DELLA protein, GAST (Gibberellic acids stimulated transcript) have been isolated to identify their role in the developing spike of wheat. Four genes were isolated from its gene family and designated as TaGAST1, 2, 3, 4. Genomic structure was analyzed to demonstrate chromosomal localization of TaGAST genes and evolutionary relationships were also verified with GAST genes in other plant species. RT-PCR was conducted to detect transcriptional changes of TaGAST genes on external phytohormone. Each of TaGAST genes showed considerable changes in transcription level after GA, ABA, PAC treatment, respectively. Through Yeast two-hybrid assay, one protein for TaGAST1, and four proteins for TaGAST2 was isolated as putative interactive proteins in wheat spikes just before and after emergence.

Drought tolerance is the ability of a plant to live, grow, and reproduce properly with limited water supply or under periodic conditions of water deficit. However, the climate changes and worldwide water shortages would result in the loss of applied water to irrigated land, increasing soil water deficit. To control the situation, we have carried out the international joint research project for the aim of developing that drought tolerance common wheat and durum wheat in Korea and Tunisia. Total 79 (41 common wheat, 39 durum wheat) Tunisian lines and 33 Korean wheat cultivars were incorporated in this study. Drought stress was applied for 25 days of stopping irrigation from the 3-leaf stage followed by re-watering for restoration in greenhouse. We selected top 13 (5 Korean line, 8 Tunisian line) tolerant lines and 11 (5 Korean, 6 Tunisian) susceptible lines based on growth parameter analysis. Primers (Operon primers and wheat Dreb1 gene) that have been known to be related drought resistance were applied to explain selected population. The correlation between PCR-based length polymorphism of selected lines and their resistance were evaluated. The obtained primer information will aid selection for drought tolerance durum as well as hexaploid common wheat.

Brachypodium has been focused as new model plant for grass species. Like small size, small room requirement, and fast growth, Brachypodium shows numerous advantages as a model plant. Brachypodium is a typical grass at the genome level, which also exhibits an overall similarity of gene content and gene families when compared with rice (Oryza sativa) and sorghum (Sorghum bicolor) genomes. Brachypodium is an excellent material for structural and functional genomic studies in grass species. Targeting-Induced Local Lesions IN Genomes (TILLING) is a high-throughput technique and an approach for reverse genetics study. Moreover, it has been wildly utilized to find induced mutation. Bradi3g45515 is orthologue of the cellulose synthase-like HvCslF8 in barley. For TILLING library construction, 384 M2 Brachypodium mutants induced by chronic-gamma irradiation were used. Single nucleotide polymorphysm (SNP) and small deletion in Bradi3g45515 were searched through TILLING analysis. Template DNA for PCR reaction were prepared according to two dimensional pooling (eightfold) strategy. Heteroduplex DNA was digested by SURVEYOR nuclease (TRANSGENOMIC) and the DNA fragment was detected using polyacrylamide gel electrophoresis. Positive signal appeared at polyacrylamide gel from more than 4 lines and their Bradi3g45515 region were sequenced. SNP(s) were identified in 509-2 and 677-3 mutant line. Cellulose content and/or cell wall materials content will be measured using these mutants.

Mutant analysis is one of most optimized genome-wide approach towards acquiring utile phenotypes and defining related genes. Gamma-irradiation, an acknowledged way of mutant-generating method, was applied to gain sets of mutant line in Brachypodium distachyon. B. distachyon is a model plant, commonly used in genus of Gramineae for the research of structure genomics and functional genomics. B. distachyon contribute to rapid and easy analysis because of its small size and quick growth. Mutant population was generated by different doses of gamma-irradiation (0, 50, 100, 150, 200, 250 Gy) in the gamma field phytotron. Distance from the source gives same irradiation duration for each plant. Plant growth parameters such as plant height, tiller number, leaf length & width, internode number & diameter, maturity and yield components (ear number biomass) were scored on M0 plants. Plant responses to different doses of radiation are evaluated and the effective radiation dosages to generate mutant using gamma-phytotron are suggested. Chronic irradiation using gamma-phytotron is useful tool to generate mutants for genomic variations such as SNP or INDEL as well as suitable for functional study of genes in Gramineae.

Rye has important genes for biotic and abiotic stress resistance. Introduction of these genes to wheat by breeding wheat-rye translocation have been intensively used in wheat breeding program. Rye chromatin 1RS and/or 2RL show superior performance in unfavorable environments. In order to develop high yielding wheat, we applied various molecular breeding strategies. To develop EST-derived 1RS specific markers, we used comparative genomics with public sequence databases of Poaceae family. Putative rye chromatin specific sequences were used to design 1RS specific markers. To identify genes related to water deficiency, cDNA AFLP analysis was used in PEG treated seedlings of 1RS RILs. For functional analysis of identified genes and markers, we used Brachypodium distachyon, as a new model plant of temperate grasses. B. distachyon were recently applied for transformation and we constructed Agrobacterium-mediated transformation system. Integration of those strategies and conventional breeding method would enhance the usefulness of rye chromatins for wheat improvement.

Brachypodium distachyon is rapidly emerged in biological study and has been currently used as a model system for genetics and functional studies for crop improvement and biofuel production. Phosphinothricin (PPT) has been widely used as a selectable agent, which raises ammonium content and induces toxicity in non-transformed plant cells. However PPT selection is not much effective on Brachypodium callus consequently reducing transformation efficiency. In order to identify the efficient conditions of PPT selection, calli obtained from mature seeds of Brachypodium (PI 254867) were cultured on the callus inducing medium (CIM) or regeneration medium (ReM) containing serial dilutions of the PPT (0, 2, 5, 10, and 15 mg/l) in dark or light condition. Callus growth and ammonium content of each treatment were measured 2 weeks after the treatment. Although callus growth and ammonium content did not show much difference in CIM, slow callus growth and increased ammonium accumulation were found in ReM. No significant difference of ammonium accumulation in response to PPT was found between dark and light conditions. In order to identify major factors affecting increased ammonium accumulation, callus was cultured on the media in combined with phytohormones (2,4-D or kinetin) and carbon sources (sucrose or maltose) containing with PPT (5 mg/l). The highest ammonium content in callus was found in the kinetin and maltose media.