A Modified DMT-HFO Technique for Investigating the Kinetics of Phosphorus Desorption from Calcareous Soils and its Relationship with Maize Growth

In this research, phosphorus (P) desorption was investigated using DMT-HFO as an “infinite” sink for 10 calcareous soils over a period of 1–35 days and the kinetic data was related to soil properties and P uptake by maize (Zea mays L.). First- and second-order, and simple Elovich equations adequately described P desorption kinetics, however parabolic, exponential and zero order equations provided the best fits to the data. A systematic relationship exists between the second order rate parameter (k₂) and the clay content of the soils. There are positive relationships between P desorption rate constants (for zero order, Elovich and parabolic equations), and both dry matter and P content of maize shoots and also for roots. These relationships indicate that P desorption rate may be a limiting factor for maize growth in these soils. The DMT-HFO technique may be used to assess the long-term availability of P to plants.     

Isolation and characterization of potassium solubilizing bacteria in some Iranian soils

The ability of a few soil bacteria to transform unavailable forms of potassium (K) to an available form is an important feature in plant growth-promoting bacteria for increasing plant yields of high-K-demand crops. In this research, isolation, screening, and characterization of six isolates of K solubilizing bacteria (KSB) from some Iranian soils were carried out. The ability of all isolates were tested in three treatments including acid-leached soil, biotite, and muscovite by analyzing the soluble K content after 5 days of incubation at 28 ± 2°C. Identification and phylogenetic analyses were also carried out by morphological, biochemical, and 16S rDNA analyses. Among the six efficient isolates, five isolates belonged to Bacillus megaterium (JK3, JK4, JK5, JK6, and JK7), while isolate JK2 belonged to Arthrobacter sp. The soluble K contents in all isolated-treatments were significantly (p < 0.01) higher than the contents in nonbacteria treatment. Herein, isolate JK2 had lower potential for K solubilization (910 mg kg^?1) compared with other isolates in acid-leached soils. The six bacterial strains showed higher solubilized K in biotite treatment than other two treatments. Overall, it can be concluded that the isolates belong to B. megaterium are the most efficient KSB under in vitro condition.     

Wheat ferritins: Improving the iron content of the wheat grain

The characterization of the full complement of wheat ferritins show that the modern hexaploid wheat genome contains two ferritin genes, TaFer1 and TaFer2, each represented by three homeoalleles and placed on chromosome 5 and 4, respectively. The two genes are differentially regulated and expressed. The TaFer1 genes are, except in the endosperm, the most abundantly expressed and regulated by iron and abscisic acid status. The promoter of TaFer1, in contrast to TaFer2, has iron- and ABA-responsive elements, supporting the expression data. The TaFer1 and TaFer2 genes encode two isoforms, probably functional different and acting in heteropolymer structures of ferritin in cereals. Iron biofortification of the wheat grain is possible. Endosperm targeted intragenic overexpressing of the TaFer1-A gene results in a 50–85% higher iron content in the grain.     

Whole cell inactivated autogenous vaccine effectively protects red Nile tilapia (Oreochromis niloticus) against francisellosis via intraperitoneal injection

Francisella noatunensis subsp. orientalis is a pathogen of tilapia and other warm‐water fish for which no vaccines are commercially available. In this study, a whole cell formalin‐inactivated vaccine was developed for the first time using the highly virulent isolate STIR‐GUS‐F2f7 and the oil‐based adjuvant Montanide? ISA 763A VG. The efficacy of the vaccine was assessed in red Nile tilapia via intraperitoneal (i.p.) injection using homologous experimental infection and correlates of protection such as seral antibody production and bacterial loads in the spleen. For immunization, fish were i.p. injected with 0.1 ml of the vaccine, the adjuvant alone or PBS. At 840 degree days post‐vaccination, all fish were i.p. injected with 4.0 × 10³ CFU/fish of pathogenic bacteria. The RPS at the end of the trial was 100% in the vaccinated group with significantly higher survival than in the adjuvant and control groups. The RPS in the adjuvant group was 42%, and no significant difference was seen in survival between this and the PBS group. Moreover, significantly higher antibody titres in the serum and significantly lower bacterial loads in the spleen were detected in the vaccinated fish by ELISA and qPCR, respectively. These findings highlight the potential of autogenous vaccines for controlling francisellosis in tilapia.     

Zygosity identification in transgenic cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis>) by real-time quantitative PCR

Successful identification of homozygous and heterozygous transgenic plant with currently available techniques such as southern blot hybridization, dot blot hybridization, fluorescence in situ hybridization (FISH) and so on, demands tedious and time-consuming procedures with a high proportion of ambiguous results. Real-time PCR is a quantitative and extremely precise method with high throughput that could be applied to the analysis of large number of plants differing only by a factor of two in the amount of target sequences. In the present study, we determined zygosity level of transgenes in cotton [Gossypium hirsutum L.] with two zygosity assays, based on TaqMan technology that uses a fluorogenic probe which hybridizes to a PCR target sequence flanked by primers. TPS, a single copy gene per haploid Gossypium hirsutum genome was used as the endogenous reference to estimate copy number of transgene. Both assays were accurate and reproducible in determination of the number of transgenes present in a cell line. These methods are standard curves and Delta delta C _t method.     

Immunocytochemical study on microtubule reorganization in HL-60 leukemia cells undergoing apoptosis

BACKGROUND: Microtubules (MT) are important components of cell cytoskeleton and play key roles in cell motility mitosis and meiosis. They are also the targets of several anticancer agents which indicating their importance in maintaining cell viability. Microtubular reorganization contributing to apoptotic morphology occurs in normal and neoplastic cells undergoing apoptosis induced by cytotoxic drugs. The aim of this study was to correlate the changes in the MT with behavior of the gamma-tubulin in apoptotic cell, and to see if apoptitic MT showed biochemical characteristics of stable MT. METHODS: Apoptosis was induced in the human leukemia cells (HL-60) by treatment with 1 microM of all-trans retinoic acid over a 5-day period. The time course of changes was assessed using flow cytometry, DNA fragmentation and immunocytochemistry in cells labeled for alpha-tubulins, acetylated alpha-tubulin and gamma-tubulin. RESULTS: The results indicated that gamma-tubulin content is increased after cells have gone through the apoptosis with a diffuse cytoplasmic pattern. Alpha-tubulin did not reveal any specific pattern of polymerization in apoptotic cells and acetylated alpha-tubulin content was also decreased in comparison with non-apoptotic cells. CONCLUSION: Our results support the idea that microtubule reorganization is an important factor of the mammalian cells response to apoptosis, and the altered properties of the MT did not reflect changes in function as apoptosis progresses.     

Sunflower Response to Application of L-Ascorbate Under Thermal Stress Associated with Different Sowing Dates

Unlike edaphic factors, it is difficult to control climatic conditions and their changes that affect plant growth and development. To deal with such posture, farmers have to cultivate their crops at the right time. From this point, over two–year of 2014 and 2015, a field experiment was performed at El-Nubaria region, El-Behaira Governorate, Egypt, to assess the response of sunflower to different combinations between sowing dates (early, mid and delayed) and ascorbic acid treatments (with ascorbic acid and without ascorbic acid). Ascorbic acid was sprayed at 30, 40 and 50 days after sowing. The results showed that sowing sunflower in mid sowing (May 21) achieved the highest values of cumulative heat units utilization efficiency as well as all growth and yield traits, while the lowest values were recorded under the delayed sowing date (June 21). Application of ascorbic acid was effective for promoting growth and yield traits under all studied sowing dates. Ascorbic acid achieved 11.6, 10.1, 10.7 and 12.9% increases under early sowing, and 9.5, 6.6, 8.6 and 10.3% increases under delayed sowing in head diameter, seed weight plant^?1, seed yield and oil yield, respectively. Comparing to mid sowing?×?without ascorbic acid, application of ascorbic acid alleviated seed yield losses associated early sowing from 15.7–6.6% and from 23.0–16.4% with delayed one. In conclusion, for remediating the thermal stressful impacts of early or late sowing of sunflower, farmers are advised to treat crop plants with ascorbic acid to avoid yield losses.

     

Cloning,Expression and Characterization of Recombinant Exotoxin A-Flagellin Fusion Protein as a New Vaccine Candidate against Pseudomonas aeruginosa Infections

Background: Infections due to Pseudomonas aeruginosa are among the leading causes of morbidity and mortality in patients who suffer from impaired immune responses and chronic diseases such as cystic fibrosis. At present, aggressive antibiotic therapy is the only choice for management of P. aeruginosa infections, but emergence of highly resistant strains necessitated the development of novel alternative therapeutics including an effective vaccine. Several P. aeruginosa antigens have been tested for vaccine development, including lipopolysaccharide alone, polysaccharides alginate, extracellular proteins, exotoxin A (exo A) and killed whole cell. However, none of them are currently available clinically. Methods: In this research, recombinant exoA-flagellin (fliC) fusion protein as a cocktail antigen was expressed and purified and its antigenic characteristics were evaluated. Results: Expression of recombinant fusion protein by E. coli using pET22b vector resulted in production of exoA-fliC fusion protein in high concentration. Based on Western-blotting results, recombinant fusion protein showed a good antigenic interaction with sera from patients with various P. aeruginosa infections. Conclusion: These results suggested that recombinant exoA-fliC fusion protein can be produced in the laboratory, and tested as a candidate vaccine in P. aeruginosa infections. Key Words: Pseudomonas aeruginosa, Exotoxin A (exoA), Flagellin (fliC), Vaccines     

Healing potential of mesenchymal stem cells cultured on a collagen-based scaffold for skin regeneration

Background: Wound healing of burned skin remains a major goal in public health. Previous reports showed that the bone marrow stem cells were potent in keratinization and vascularization of full thickness skin wounds. Methods: In this study, mesenchymal stem cells were derived from rat adipose tissues and characterized by flowcytometry. Staining methods were used to evaluate their differentiation ability. A collagen-chitosan scaffold was prepared by freeze-drying method and crosslinked by carbodiimide-based crosslinker. Results: The results of immunecytochemistry and PCR experiments confirmed the adipose-derived stem cells (ASC) in differentiation to the keratinocytes under the treatment of keratinocyte growth factor. The isolated ASC were seeded on the scaffolds and implanted at the prepared wounds. The scaffolds without cells were considered as a control and implanted on the other side of the rat. Histopathological analyses confirmed the formation of new tissue on the scaffold-cell side after 14 days with the formation of dermis and epidermis. Conclusion: These results indicated the capacity of ASC in differentiation to keratinocytes and also wound healing in vivo. Key Words: Tissue engineering, Keratinocytes, Mesenchymal stem cells