Salinity tolerance of Aegilops cylindrica genotypes collected from hyper-saline shores of Uremia Salt Lake using physiological traits and SSR markers

Uremia Salt Lake, in North West Iran, has a hyper-saline water. A rare highly salinity-tolerant grass species, Aegilops cylindrica grows along its shores. Salinity tolerance of 44 genotypes of Ae. cylindrica, mainly collected from the Lake, was evaluated under control and 400 mM NaCl conditions using the physiological traits of plant height, dry weight, proline content, Na⁺ and K⁺ concentrations as well as K⁺/Na⁺ ratio. To evaluate the association between microsatellite (EST-SSR and SSR) markers and salinity tolerance, 35 primer pairs were used. Results showed a significant variation in the 44 genotypes studied in terms of their traits except for proline content. Ten most salinity-tolerant genotypes were identified based on their ability to survive, to produce the highest dry weight, and to sustain the least leaf Na⁺ concentration under salinity stress. The very high negative correlation found between Na⁺ concentration and salinity tolerance revealed the importance of individual or a combination of Na⁺ exclusion and excretion mechanisms contributing to the hyper-salinity tolerance of these genotypes. Clustering analysis based on marker data divided the 44 studied genotypes into two groups that were consistent with their saline and non-saline geographical areas. Results of molecular markers showed that four microsatellite markers (Xgwm312, Xwmc170, Xgwm291 and Xgwm410) generated a distinguished banding pattern in ten most salinity-tolerant genotypes. These results supported previous reports on their linkage with Na⁺ exclusion genes (HKT1;5 and HKT1;4) in wheat, which provided further evidence of usefulness of both genes and the linked markers to the salinity tolerance of the halophytic grass family species.     

Nrf-2 overexpression in mesenchymal stem cells reduces oxidative stress-induced apoptosis and cytotoxicity

The most prominent capabilities of mesenchymal stem cells (MCSs) which make them promising for therapeutic applications are their capacity to endure and implant in the target tissue. However, the therapeutic applications of these cells are limited due to their early death within the first few days following transplantation. Therefore, to improve cell therapy efficacy, it is necessary to manipulate MSCs to resist severe stresses imposed by microenvironment. In this study, we manipulated MSCs to express a cytoprotective factor, nuclear factor erythroid-2 related factor 2 (Nrf2) to address this issue. Full-length human Nrf2 cDNA was isolated and TOPO cloned into TOPO cloning vector and then transferred to gateway adapted adenovirus expression vector by LR recombination reaction. Afterwards, the Nrf2 bearing recombinant virus was prepared in appropriate mammalian cell line and used to infect MSCs. The viability and apoptosis of the Nrf2 expressing MSCs were evaluated following hypoxic and oxidative stress conditions. Transient expression of Nrf2 by MSCs protected them against cell death and the apoptosis triggered by hypoxic and oxidative stress conditions. Nrf2 also enhanced the activity of SOD and HO-1. These findings could be used as a strategy for prevention of graft cell death in MSC-based cell therapy. It also indicates that management of cellular stress responses can be used for practical applications.     

Ralstonia eutropha as a biocatalyst for desulfurization of dibenzothiophene

The potential of Ralstonia eutropha as a biocatalyst for desulfurization of dibenzothiophene (DBT) was studied in growing and resting cell conditions. The results of both conditions showed that sulfur was removed from DBT which accompanied by the formation of 2-hydroxybiphenyl (2-HBP). In growing cell experiments, glucose was used as an energy supplying substrate in initial concentrations of 55 mM (energy-limited) and 111 mM (energy-sufficient). The growing cell behaviors were quantitatively described using the logistic equation and maintenance concept. The results indicated that 2-HBP production was higher for the energy-sufficient cultures, while the values of the specific growth rate and the maintenance coefficient for these media were lower than those of the energy-limited cultures. Additionally, the kinetic studies showed that the half-saturation constant for the energy-limited cultures was 2 times higher than the energy-sufficient ones where the inhibition constant (0.08 mM) and the maximum specific DBT desulfurization rate (0.002 mmol g_cell ⁻¹ h⁻¹) were almost constant. By defining desulfurizing capacity (D _DBT) including both the biomass concentration and time to reach a particular percentage of DBT conversion, the best condition for desulfurizing cell was determined at 23% g_cell L⁻¹ h⁻¹ which corresponded with the resting cells that were harvested at the mid-exponential growth phase.

     

Selenium ameliorates salinity stress in <Emphasis Type="Italic">Petroselinum crispum</Emphasis> by modulation of photosynthesis and by reducing shoot Na accumulation

This study was performed to understand the mechanisms for Se-enhanced resistance of parsley (Petroselinum crispum L.) plants to salinity stress. Plant growth was negatively affected by salt stress; however, Se treatments at 1 mg/L significantly improved the growth rate and enhanced the salt tolerance of seedlings. This increased tolerance in Se-supplied plants was obtained by reduced damaging effect on maximal quantum yield of photosystem II (PSII) (F _v/F _m) coupled with higher levels of carotenoids and non-photochemical quenching (NPQ). The performance index (PI_ABS), as evidence for modulation of PSII function, was downregulated by salt stress; while Se mitigated this effect. Moreover, analysis of OJIP transients demon-strated that Se reduced salt damaging effect on PSII function through improvement of excitation energy trapping (TR₀/CS) and electron transport (ET₀/CS) per excited cross-section of leaf. The Na concentrations in shoots and roots of parsley seedlings considerably enhanced after NaCl treatment. Interestingly, treatment of salt-stressed plants with Se decreased the Na contents in shoots via the limitation of the root-to-shoot translocation of Na and exclusion of Na from cell sap, as well as the retention of K/Na and Ca/Na ratios. These data provide the first evidence that the Se application alleviates salinity stress by enhancing PSII function and by decreasing Na content in the shoot via binding of Na to the root cell wall.     

Functional Production and Characterization of a Fibrin-Specific Single-Chain Antibody Fragment from Bacillus subtilis: Effects of Molecular Chaperones and a Wall-Bound Protease on Antibody Fragment Production

To develop an ideal blood clot imaging and targeting agent, a single-chain antibody (SCA) fragment based on a fibrin-specific monoclonal antibody, MH-1, was constructed and produced via secretion from Bacillus subtilis. Through a systematic study involving a series of B. subtilis strains, insufficient intracellular and extracytoplasmic molecular chaperones and high sensitivity to wall-bound protease (WprA) were believed to be the major factors that lead to poor production of MH-1 SCA. Intracellular and extracytoplasmic molecular chaperones apparently act in a sequential manner. The combination of enhanced coproduction of both molecular chaperones and wprA inactivation leads to the development of an engineered B. subtilis strain, WB800HM[pEPP]. This strain allows secretory production of MH-1 SCA at a level of 10 to 15 mg/liter. In contrast, with WB700N (a seven-extracellular-protease-deficient strain) as the host, no MH-1 SCA could be detected in both secreted and cellular fractions. Secreted MH-1 SCA from WB800HM[pMH1, pEPP] could be affinity purified using a protein L matrix. It retains comparable affinity and specificity as the parental MH-1 monoclonal antibody. This expression system can potentially be applied to produce other single-chain antibody fragments, especially those with folding and protease sensitivity problems.     

Isolation and structure of d-xylans from pericarp seeds of Opuntia ficus-indica prickly pear fruits

Xylans were isolated from the pericarp of prickly pear seeds of Opuntia ficus-indica (OFI) by alkaline extraction, fractionated by precipitation and purified. Six fractions were obtained and characterized by sugar analysis and NMR spectroscopy. They were assumed to be (4-O-methyl-d-glucurono)-d-xylans, with 4-O-α-d-glucopyranosyluronic acid groups linked at C-2 of a (1→4)-β-d-xylan. The sugar composition and the ¹H and ¹³C NMR spectra showed that their chemical structures were very similar, but with different proportions of d-Xyl and 4-O-Me-d-GlcA. Our results showed that, on average, the water soluble xylans have one nonreducing terminal residue of 4-O-methyl-d-glucuronic acid for every 11 to 14 xylose units, whereas in the water non-soluble xylans, xylose units can varied from 18 to 65 residues for one nonreducing terminal residue of 4-O-methyl-d-glucuronic acid.     

Study of Pyrex and quartz insulators contamination effect on the X-ray intensity in a 4-kJ plasma focus device

The variation of the X-ray intensity has been investigated with the Pyrex and quartz insulators surface contamination in a 4-kJ plasma focus device with argon gas at 11.5-kV charging voltage. Elemental analysis (EDAX) showed that the Cu evaporated from the electrode material and was deposited on the sleeve surface improves the breakdown conditions. A small level of sleeve contamination by copper is found to be essential for good focusing action and high HXR intensity. The SEM imaging showed the grain-type structure of Cu formed on the surface and it changed the surface property. Resistance measurements of original and coated Pyrex surface proved that the copper deposition on the sleeve surface will reduce its resistance as compared to the almost infinitely large resistance of the uncontaminated sleeve. As the contamination is surpassed to some critical level, the HXR intensity from the device is deteriorated.     

First report of occurrence of two viruses on pea field in Iran

An intensive survey was conducted to identify virus diseases affecting pea crops in Tehran province of Iran. A total of 270 pea samples were collected randomly from pea fields. samples were tested by Double Antibody Sandwich Enzyme Linked Immunosorbent Assay (DAS-ELISA) using polyclonal antisera prepared against PSBMV (AS-0129, DSMZ, Braunschweig, Germany) and TSWV (AS-0580, DSMZ, Braunschweig, Germany). Virus disease incidence in pea samples was followed by PSBMV (33%) TSWV (24.4%) and PSBMV+TSWV (17.77). The positive samples with PSBMV were extracted in 0.05M phosphate buffer pH 6.5-7 containing 2% pvp and inoculated on Pisum sativum, Vicia faba, Chenopodium quinoa, Chenopodium amaranticolor. That produced in Pisum sativum; leaflets roll downwards, shoots curl, internodes shorten and plants are rosetted. Early infections reduce flower and fruit formation or eliminate their development. Broad bean has symptoms accompanied by a certain margin rolling and leaflet distortion. In Chenopodium amaranticolor necrotic local lesions and Chenopodium quinoa chlorotic local lesions had produced. The positive samples with TSWV were extracted in 0.01 M phosphate buffer containing 1% Na2 SO3 and inoculated on Petunia hybrida, Pisum sativum. TSWV causes several symptoms in infected peas, including brown leaf petiole and stem coloration, leaflet spotting, vein necrosis. In petunia hybrida after approximately 5 days showed local necrotic lesion. Biological purification in TSWV with chlorotic local lesions in Petunia hybrida and in PSBMV; chlorotic local lesions in Chenopodium quinoa were done. In PSBMV, back inoculated on Pisum sativum and Vicia faba also tested with DAS-ELISA. RT-PCR confirmed the results. This is the first report of PSBMV and TSWV naturally infecting pea in Iran.