Control of apple blue mould disease with Torulaspora delbrueckii in combination with Silicon

In this study, Torulaspora delbrueckii alone and in combination with silicon were evaluated for the control of apple blue mould disease caused by Penicillium expansum. In vitro, the antagonistic effects of T. delbrueckii in controlling mycelial growth of P. expansum on potato-dextrose-agar (PDA) in dual cultures, and the growth of P. expansum alone with cell-free metabolites and volatile components of T. delbrueckii were assayed. In vitro, to evaluate the direct effect of silicon on mycelial growth of pathogen, silicon at different concentrations (0.2, 0.4, 0.6, 1 and 2% (wt./vol.)) was added to PDA medium. Silicon at 0.6% (wt./vol.) and above concentrations completely inhibited the mycelial growth of P. expansum. However, it had no significant effect on population dynamics of yeast in vitro and in apple wounds. In vivo, silicon at 0.2 and 1% (wt./vol.) in combination with antagonistic yeast (1 × 10⁸ cell/ml) was a more effective approach to reduce the lesion diameter of blue mould decay of apples than the application of silicon or T. delbrueckii alone at 20 and 4°C, respectively.     

The effect of different salts (heavy metals) on the mycelium growth of Nematophagus fungi

Environmental pollution in addition to direct damage on plant growth, with the destruction of biological control agents, causes indirect damage to plants. The aim of this research was to study the effects of different concentrations (0, 500, 1000, 1500 and 2000 ppm) of heavy metals including Ag, Co, Cu, Fe, Hg, Mn, Pb and Zn on the mycelial growth and to assess the fungicidal or fungistatic effects of these salts on five Nematophagus fungi including Trichoderma harzianum (T8), Trichoderma virens (T21), Trichoderma hamatum (T9), Pochonia chlamydosporia var. chlamydosporia and Arthrobotrys oligospora. The results show that Ag, Co, Cu, Fe and Hg could stop the mycelium growth of all fungi, but Mn, Pb and Zn cannot inhibit the growth of these fungi completely. Among the first group, Hg and Cu stopped the growth of fungi even in 500 ppm. Among these metals that inhibit the growth of fungi, Cu has fungistatic effect and others have fungicide effect. The experiment was conducted in vitro condition, using potato dextrose agar (PDA) under complete randomised design with four replications. The data of mycelium growth were recorded at seven days after inoculation at 25 ± 2°C.     

Characterisation of digestive protease in the rose sawfly,Arge rosae Linnaeus (Hymenoptera: Argidae)

Insect midgut proteases are excellent targets for insecticidal agents such as protease inhibitors. These inhibitors are used for producing transgenic plants, resistant to pests. For achieving this goal, it is necessary to find the nature of specific proteases and their properties for adopting possible pest management procedure. Therefore, characterisation of the enzymes in the gut of the rose sawfly, Arge rosae (Hymenoptera: Argidae), responsible for proteolysis, was performed using a range of synthetic substrates and specific inhibitors. The optimum conditions for general proteases and trypsin were achieved at pH 10. The highest activity for general proteases was obtained at a temperature of 45°C. The use of specific inhibitors and SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) provided evidence to suggest that most of the proteases belonged to the serine group because of high inhibitory effect of phenyl methane sulfonyl fluoride on total proteolytic activity. Also, inhibition assays and zymogram analysis showed that metalloproteases are present in A. rosae digestive system. These results indicated that A. rosae larvae mainly used serine proteases for protein digestion, with chymotrypsin as the dominant form. The kinetic parameters of trypsin-like proteases using N-benzoyl-dl-arg-p-nitroanilide as substrate indicated that the K _m and V _max values of trypsin in the gut of the fifth instar larvae were 730 ± 17.3 μM and 456 ± 13.85 nmol min^?1 mg^?1 protein, respectively.     

Biocontrol of blue mold of apple by Candida membranifaciens in combination with silicon

In this study, antagonistic yeast Candida membranifaciens was combined with different concentrations of silicon (Si; 0, 0.1, 0.3 and 0.5% wt/vol) to evaluate the control of blue mold of apple in storage at 20°C and 5°C. Preliminary studies showed that Si at 0.6% or above inhibited mycelial growth of pathogens significantly in vitro. In vitro studies showed that Si at 0.1% had lower effect on yeast growth. In vivo studies showed that combination of different concentrations of Si with C. membranifaciens improved the efficacy of yeast in control of disease better than Si and yeast alone (P < 0.05). Our result showed that the effective concentration of Si is varied based on pathogen isolates and temperature, so that the most effective concentration of Si was 0.5% for isolate P2 at 20°C and 0.5% and 0.1% for isolates P1 and P2 at 5°C.     

Antioxidant and Neuroprotective Effects of Scrophularia striata Extract Against Oxidative Stress-Induced Neurotoxicity

In this study, the neuroprotective effect of Scrophularia striata Boiss (Scrophulariaceae) extract, a plant growing in northeastern of Iran, against oxidative stress-induced neurocytotoxicity in PC12 was evaluated. The PC12 cell line pretreated with different concentrations (10, 50, 100, and 200 μg/ml) of the extract and then treated with H₂O₂ to induce oxidative stress and neurotoxicity. Survival of the cells, reactive oxygen species (ROS) generation, and apoptosis were measured using MTT assay, fluorescent probe 2′,7′-dichlorofluorescein diacetate, and annexin V/propidium iodide, respectively. Moreover, the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) was used to evaluate the antioxidant capacity of the plant extract. Phytochemical assay by thin layer chromatography showed that the main components, including phenolic compounds, phenyl propanoids and flavonoids, were presented in the S. striata extract. The extract in concentrations of 50–200 μg/ml protected PC12 cells from H₂O₂-induced toxicity. The survival of the cells at concentration of 200 μg/ml was 64 % compared to that of H₂O₂ alone-treated cells (48 %) (p < 0.001). The extract also dose-dependently reduced intracellular ROS production (p < 0.001). Moreover, the extract showed antioxidative effects and decreased apoptotic cells. Collectively, these findings indicated the ability of S. striata to decrease ROS generation and cell apoptosis and also suggest the presence of the neuroprotective agents in this plant.     

Change in membrane fatty acid compositions and cold-induced responses in chickpea

Plant cells often increase cold tolerance by reprogramming their genes expression which results in adjusted metabolic alternations, a process enhanced under cold acclimation (CA) phase. In present study, we assessed the changes of membrane fatty acid compositions and defense machine (like antioxidative enzymes) along with damage indexes like electrolyte leakage index (ELI) and malondialdehyde (MDA) during CA, cold stress (CS) and recovery (R) phases in chickpea (Cicer arietinum L.). Results showed an increase in unsaturated fatty acids ratio compare to saturated ones which is a sign of cold tolerance especially after CA phase. Antioxidant enzymes had an important role during CA and R phases while CS affected their activity which can be a sign for associating other metabolites or enzymes activities to create cold tolerance in plants. To investigation of enzymes assay under experimental treatments, the expression pattern of some enzymes including superoxide dismutase (sod), catalase (cat) and lipoxygenase (lox) was studied using quantitative real time PCR. LOX activity has shown a bilateral behavior: a positive relation with membrane damage index in CA and an interesting link with double bond index (DBI) in CS indicating probably its role in secondary metabolites like jasmonic acid signaling pathway. It was suggested that increased DBI and low LOX activity under CS could be a reason for plant cold tolerance.     

Cytokine gene polymorphism and graft-versus-host disease: a survey in Iranian bone marrow transplanted patients

Graft versus host disease (GVHD) is a major complication of bone marrow transplantation (BMT). Numerous studies have shown the potential role of cytokine genotypes in the occurrence of GVHD. In this retrospective, case–control study we aimed to investigate the association between 13 cytokine genes and acute GVHD (aGVHD) after HLA-identical sibling BMT in 91 Iranian subjects. Negative association was found between aGVHD and donor IL-10/GCC haplotype or donor IL-4Ra-A allele in the population study. When compared within the leukemia subgroup, we observed positive association between recipient IL-1α ?889/C allele and aGVHD. Also there were negative association between recipient IL-10/CAA haplotype and donor IL-4Ra/A allele and development of aGVHD. Among the different genotypes only donor IL-4Ra and donor IL-12 showed significant association. We conclude that several cytokine polymorphisms are positively and negatively associated with aGVHD in Iranian HLA matched siblings, of which IL-4Ra and IL-12 may play important roles.     

ATR kinase activation in G1 phase facilitates the repair of ionizing radiation-induced DNA damage

The kinase ATR is activated by RPA-coated single-stranded DNA generated at aberrant replicative structures and resected double strand breaks. While many hundred candidate ATR substrates have been identified, the essential role of ATR in the replicative stress response has impeded the study of ATR kinase-dependent signalling. Using recently developed selective drugs, we show that ATR inhibition has a significantly more potent effect than ATM inhibition on ionizing radiation (IR)-mediated cell killing. Transient ATR inhibition for a short interval after IR has long-term consequences that include an accumulation of RPA foci and a total abrogation of Chk1 S345 phosphorylation. We show that ATR kinase activity in G1 phase cells is important for survival after IR and that ATR colocalizes with RPA in the absence of detectable RPA S4/8 phosphorylation. Our data reveal that, unexpectedly, ATR kinase inhibitors may be more potent cellular radiosensitizers than ATM kinase inhibitors, and that this is associated with a novel role for ATR in G1 phase cells.