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.

Fusarium head blight (FHB), also known as scab, caused mainly by Fusarium graminearum is a devastating disease of wheat in regions that are warm and humid during flowering. In addition to significant yield and quality losses, the mycotoxin deoxynivalenol produced by the pathogen in infected wheat kernels is a serious problem for food and feed safety. Twenty- three Korean cultivars and "Sumai 3", which is a FHB-resistant Chinese cultivar were tested for Type I, Type II resistances of FHB. Three cultivars were identified as resistant in Type I assessment, and two cultivars were resistant in Type II assessment. Genetic variation and relationship among the cultivars were evaluated on the basis of 11 Simple Sequence Repeat (SSR) and 29 Sequence Tagged Site (STS) markers that were linked to FHB resistance Quantitative Trait Loci (QTL) on chromosome 3BS. One SSR and 7 STS markers detected polymorphisms. Especially, using a STS marker (XSTS3B-57), 32.4% of the variation for Type II FHB resistance could be explained. Genetic relationship among Korean wheat cultivars was generally consistent with their released year. These markers on chromosome 3BS have the potential for accelerating the development of Korean wheat cultivars with improved Fusarium head blight resistance through the use of marker-assisted selection.

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.

Glutamine synthetase (GS) plays important roles in plants like assimilation of ammonium and detoxification of the ammonium released from many metabolic processes such as amino acid degradation or photorespiration. Using ATP, ammonia is combined with glutamate to yield glutamine by the action of GS. Phosphinothricin (PPT) is widely used as a herbicide because it competes with glutamate to bind the active site of GS. PPT has been used to produce transgenic Brachypodium distachyon callus and plants as a selectable agent. PPT treatment raises ammonium content and induces toxicity in non-transformed cells. To find out efficient condition for selecting transformed callus, ammonium content were measured in this study. Non-transformed callus were derived from mature seeds of Brachypodium distachyon (Bd21). The callus were cultured on the callus inducing media (CIM) or regeneration media (RM) containing serial dilutions of the PPT (2, 5, 10 and 15 mg/l) with or without light. Ammonium content was measured 2 weeks after PPT application. Ammonium toxicity associated with PPT treatment was dose-dependent on RM whereas PPT treatment was not significantly influenced on CIM. There is no influence on dark or light condition. Additionally, callus were cultured on the media containing phytohormones combined with PPT (5 mg/l) and the most affecting element causing increased ammonium content has been identified. Acknowledgements: This work was supported by a grant (No. 20070301-034-016-007) from BioGreen 21 Program, RDA, Republic of Korea.

Rye (Secale cereale L.) chromatins have been used to introduce agronomically important traits into wheat (Triticum aestivum L.). Wheat-rye translocations in the form of 1RS.1AL, 1RS.1BL, 2BS.2RL have been developed for an important genetic source of disease and pest resistance. The long arm of rye chromosome 2 (2RL) has valuable genes that confer resistance to pests such as biotype L of Hessian fly, powdery mildew, leaf and stem rust. Here, we report the generation and analysis of expressed sequence tags from Hessian fly infested wheat-rye translocation. RNAs were isolated from young seedlings infested by Hessian fly. cDNA library was constructed using Clontech cDNA library construction kit. Random sequencing of candidate clones were performed. The EST clones might be useful to clone target gene sequences and would provide clues on molecular interaction between wheat and Hessian fly.

barley grain and malt is highly related to beer quality, especially hordein is known to be a more significant factor in malting process than albumin. In this study, we proposed selection criteria for high quality malting barley with aid of grain and malt quality parameter scores and storage protein subunit profile informations. Albumin and hordein were extracted and denatured protein subunits were evaluated with malt and grain quality parameters. Total 13 local adaptability test (LAT) lines were planted in four locations (Naju, Iksan, Jeju, and Jinju) and evaluated for malt and beer making qualities. Seventeen germplasms (world collections for high or low seed storage protein content) were also evaluated for biochemical genetic marker. Denatured seed storage protein subunits of albumin and hordein of all tested lines and germplasms were evaluated using 12% 1D SDS-PAGE. Scored data of protein subunit's presence or absence was applied to Agglomerative Hierarchical Clustering (AHC) for statistical analysis. Subunits fractionated within specific molecular weight ranges (97.4-31.0, 66.2-31.0, and 45.0-31.0 kDa) were highly correlated with agricultural characteristics. Several LAT lines showing good performance in agricultural characteristics were clustered in dendrogram constructed by biochemical-genetic assay using XLSTAT. Specific band pattern showed in good performance LAT lines were also observed in some germplasms of world collection having low protein contents which are known to have superior quality in malting. The results would provide selection criteria for high quality malting barley in the malting barley breeding program.

Truncated hemoglobins (trHbs) constitute a familkrof small oxygen-binding heme proteins distributed in eubacteria, cyanobacteria, protozoa, and plants. Three distinct types of trHbs (HbN, HbO and HbP) have been identified within the mycobacterial genome. The soluble trHb of the cyanobacterium Nostoc commune is localized on the cytoplasmic face of the cell membrane and is expressed only under anaerobic conditions. In addition, the gene encoding trHb is coexpressed with genes of the nitrogen fixation complex. The trHb of the unicellular green alga C. eugametos is induced in response to active photosynthesis and is localized, in part, along the chloroplast thylakoid membranes. Yeast II hybridization screening was performed using TatrHb as a bait protein to elucidate putative role of TatrHb. cDNA Library for Yeast II hybridization screening was constructed with mRNA from young seedling treated 0% oxygen for 48 hours. 12 positive clones were selected by colony PCR and sequenced. Among them, 4 clones are related in photosynthesis and show highly homologe to Hordeum vulgare chloroplast photosystem I PSK-I subunit mRNA (393 bp), Zea mays photosystem II subunit PsbS1 mRNA (900 bp), Triticum aestivum ribulose-1,5-bisphosphate carboxylase/oxygenase(RubisCo) large subunit mRNA (1434 bp), Triticum aestivum ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit mRNA (525 bp).

UNUSUAL FLORAL ORGAN (UFO), a novel gene, is involved in controlling flowering initiation and development. In Arabidopsis, UFO is required for floral organ identity in the second and third whorls. However, the mode of expression and function of TaUFO have not been studied yet. The cDNA sequence of TaUFO is comprised of 1344 bp open reading frame which encodes 50.82 KDa polypeptide consisting amino acid residues. F-box protein, the components of TaUFO, plays an important regulatory role in a wide diversity of developmental and physiological responses. In almost all F box proteins, the N terminus of the protein contains the F-box motif, and the rest of the protein contains the protein-protein interaction domains required for target protein binding. In order to elucidate the function of the TaUFO, various phytohormones and abiotic stresses were applied on young seedlings (14 day after germination) and its transcripts were evaluated. TaUFO:GFP fusion construct was transformed into onion epidermal cells by particle bombardment to elucidate the subcellular localization of the TaUFO protein. The function of the F-box protein is to interact with target proteins. With the use of a yeast two-hybrid screen to isolate proteins interacting with the TaUFO (F box protein), we identified potential TaUFO interactive protein in wheat spikelet library.

To investigate the mutagenecity of sodium azide (SA) and to select mutants having various agronomic characteristics in rice (Oryza sativa L. ssp. japonica), dry seeds of rice variety 'Suweon 472' were treated with 0, 0.5, 1.0, 2.0, and 4.0 mM SA solutions prepared in 0.1 M phosphate buffer (pH 3.0). Germination rate, seedling height and sterility were investigated in M1 generation and chlorophyll mutations were observed in M3 generation. Germination rate and seedling height decreased as the increase of SA concentration in M1 generation, the maximum seed sterility (40.8%) was found at 4.0 mM SA concentration. Chlorophyll mutants were occurred in M3 generation and the frequency calculated on a line basis was 13.5% at the same treatment. Many kinds of mutations for morphological and agronomic characters were observed and mutations with short culm and glabrous leaf were frequently found in M3 generation. Interestingly, five mutant lines resistant to blast or bacterial blight (BB) were selected and evaluated with several isolates in M3 generation although Suweon 472 has been known to be susceptible to blast and BB. These mutants showed all resistance to seven isolates of blast and a total of 76 lines among 2,567 lines evaluated showed resistant to race K1 of bacterial blight. Two mutant lines (440172 and 41272) showed different reaction to BB isolates from the other resistant mutants. A few kinds of endosperm mutants were also identified and most of them were waxy mutants.