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.

The composition and distribution of seed storage proteins are important factors for eating quality such as grain flavor and quality in rice (OryzasativaL.) Rice protein disulfide isomerase (OsPDI) and binding protein (OsBIP) regulate synthesis, stability and sorting of storage proteins. We thus have tried to develop a marker protein for selection of rice cultivars which have different eating quality. Immunoblot analysis revealed that protein levels of OsPDI and OsBIP have no direct correlation with eating quality, suggesting that they may indirectly participate in control of eating quality through their-interacting partners or other regulatory mechanism.

The flowers of Buddleja officinalis are used to treat sore and damaged eyes, a condition which is similar to skin wounds. However, whether it has any protective effect on oxidative DNA damage and cell death induced by hydroxyl radical remains unclear. In this study, we evaluated the protective effects of the extracts against oxidative DNA and cell damage caused by hydroxyl radical. DPPH radical, hydroxyl radical, hydrogen peroxide and intracellular ROS scavenging assay, and Fe2+ chelating assay were used to evaluate the antioxidant properties. phi X 174 RF I plasmid DNA and intracellular DNA migration assay were used to evaluate the protective effect against oxidative DNA damage. Lastly, MTT assay and lipid peroxidation assay were used to evaluate the protective effect against oxidative cell damage. It was found to prevent intracellular DNA and the normal cells from oxidative damage caused by hydroxyl radical via antioxidant activities. These results suggest that Buddleja officinalis may exert the inhibitory effect on ROS-induced carcinogenesis by blocking oxidative DNA damage and cell death.