Effect of NaCl stress on dihaploid tobacco lines tolerant to Potato virus Y

Salinity is an important abiotic factor that limits plant growth and development. The influence of salt stress induced by sodium chloride on plant growth, proline content, level of lipid peroxidation and activities of antioxidative enzymes was studied in F1 hybrid DH10 and four dihaploid lines (207B, 238C, 239K, 244B) of tobacco (Nicotiana tabacum L.). Dihaploids were obtained from anther-derived haploids of hybrid DH10 and were previously proved to be tolerant to Potato virus Y (PVY). In our study, plants were grown in vitro and exposed to NaCl (100 and 200 mM) for 33 days. All dihaploids and hybrid DH10 showed reduced growth after NaCl treatment. They accumulated significant amounts of sodium and proline in response to salt stress as have already been observed in tobacco and other plant species. In tobacco exposed to NaCl the lipid peroxidation level did not increase and activities of superoxide dismutase (SOD), guaiacol peroxidase (POD), ascorbate peroxidase and catalase (CAT) mostly did not change significantly. The exception was line 239K where salt induced higher activities of SOD, CAT and POD. Two (238C and 244B) out of four dihaploids appeared more susceptible to salt stress as they showed weak growth in correlation with high proline and sodium content. Therefore, it seems that salt tolerance is not associated with tolerance to PVY. Variations in malondialdehyde and proline content as well as in enzymes activities observed among tobacco lines imply that dihaploids have different genetic properties which might result in different sensitivity to NaCl.     

Induction by α,ά-trehalose derivatives of α,ά trehalase fromSaccharomyces cerevisiae andAureobasidium pullulans

Monodeoxy and dideoxy derivatives of α,?-trehalose increased the level of intracellular α, ?-trehalaee in the yeastSaccharomyces cerevisiae and the yeast-likeAureobasidium pullulans.     

Cellular localization of dual positional specific maize lipoxygenase-1 in transgenic rice and calcium-mediated membrane association

The dual positional maize lipoxygenase-1 was introduced into rice and T2 transgenic plants were produced. Cellular location of maize lipoxygenase-1 in transgenic rice and effects of calcium ion on membrane association in vitro were analyzed. Localization study by confocal microscopic analysis indicated that the maize lipoxygenase-1 was localized in cytoplasm. Sucrose-density fractionation experiment and in vitro protein transport to chloroplast showed that the maize lipoxygenase-1 can be associated with chloroplast. Secondary structure alignment revealed putative calcium binding sites in the PLAT domain of maize lipoxygenase-1 and the association of the maize lipoxygenase-1 with membranes was mediated by calcium ion in vitro. Our results provide evidences for calcium-mediated translocation of dual positional LOX without chloroplast targeting sequence from cytoplasm to chloroplast in plants for the first time.     

Stabilization of Bacillus subtilis Lipase A by increasing the residual packing

Introduction of well-packed residues to the interior of a protein structure could be considered as a stabilization strategy since the reduction of buried cavities might stabilize protein structure. In this study, the less-packed residues with no water-contact were selected as target sites for increasing residual packing. When Lipase A from Bacillus subtilis (179 amino acids) was used as a model system, 43 less-packed residues were initially considered by analyzing their residual packing value and residual exposure ratio. Among the 43 residues, small amino acids such as GLY and ALA were chosen as target sites. Packing increases of ALA to VAL and GLY to ALA were estimated, by molecular modeling, to give 0.5368∼0.7433 kcal mol^-1 stabilization. Mutants of Lipase A such as A38V, A75V, G80A, A105V A146V, and G172A were obtained via protein engineering. Thermostability assays revealed that A38V, G80A and G172V were the most stable mutants. This procedure for selecting the target residues for improved thermostability of Lipase A could be applied for improving the thermostability of other proteins and enzymes.     

Effect of Elastic Network of Ceramic Fillers on Thermal Cycle Stability of a Solid Oxide Fuel Cell Stack

Glass‐based seals for planar solid‐oxide fuel‐cell (SOFC) stacks are open to uncontrolled deformation and mechanical damages, limiting both sealing integrity and stack reliability, particularly in thermal cycle operations. If the glass‐based seals work like an elastomer‐based compressive seal, SOFC stacks may survive unprecedented numbers of thermal cycles. A novel composite sealing gasket is successfully developed to mimic the unique features of the elastomer‐based compressive seal by controlling the composition and packing behavior of binary ceramic fillers. A single‐cell SOFC stack undergoes more than 100 thermal cycles with little performance loss, during which the sealing integrity is lost/recovered repeatedly upon cooling and reheating, corresponding to unloading/loading of the elastomer‐based compressive seal. The thermal‐cycle responses of the SOFC stack are explained in sequence by the concurrent events of elastic deformation/recovery of ceramic filler network and corresponding redistribution of sealing glass.