First Report of Iris yellow spot virus Infection of Garlic and Egyptian Leek in Egypt.

Egyptian leek (Allium ampeloprasum), garlic (A. sativum), and onion (A. cepa) are key vegetables produced by small- and large-scale farmers in Egypt for national and international markets. Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) is an economically important viral pathogen of bulb and seed onion crops in many onion-growing areas of the world (1,3). During February and March of 2011, symptoms of spindle-shaped, straw-colored, irregular lesions with occasional green islands were observed on onion, garlic, and Egyptian leek cultivated on large and small farms in Dakahlia, Gharbia, Kalubia, Menofia, Qena, and Assiut governorates in Egypt. The presence of IYSV was confirmed by specific double antibody sandwich (DAS)-ELISA Flash Kits (Agdia Inc., Elkhart, IN) (2). A survey was carried out by collecting 100 plant samples (10 asymptomatic and 90 symptomatic) of each plant species from fields in the governorates of Dakahlia, Gharbia, Kalubia, Menofia, Qena, and Assiute and testing the plants using DAS-ELISA. For onion and garlic, 45% of the symptomatic samples and 0% of the asymptomatic plants tested positive. For leek, 34% of the symptomatic samples tested positive and 0% of the asymptomatic samples. ELISA-positive samples were tested using a reverse transcription (RT)-PCR assay with primers specific to the S RNA of IYSV (forward primer 5'-TAAAACAAACATTCAAACAA-3' and reverse primer 5'-CTCTTAAACACATTTAACAAGCAC-3') (2). Amplicons of approximately 1,100 bp were obtained from all symptomatic samples that were ELISA positive, but none of the asymptomatic plants nor the sterile water control sample produced PCR amplicons. The amplicons were cloned (at least three clones per plant species) using the TOPO TA Cloning Kit (Invitrogen, Grand Island, NY), and sequenced. The Egyptian onion IYSV isolate (GenBank No. JN541273) had the greatest nucleotide sequence identity (86%) with the corresponding S RNA region of IYSV isolates from India (GenBank Nos. EU310290, EU310284, and EU310276). The Egyptian garlic IYSV isolate (GenBank No. JN541275) showed the strongest identity (93%) with that of a Sri Lankan IYSV isolate (GenBank No. GU901211). The Egyptian leek IYSV isolate (GenBank No. JN541274) exhibited 91% sequence identity with that of the same Sri Lankan isolate (No. GU901211). To our knowledge, this is the first report of IYSV infecting garlic and Egyptian leek in Egypt. IYSV infection of onion was reported previously from the agricultural farm of the Faculty of Agriculture, Cairo University, Giza (4), but to our knowledge, this is the first report of natural infection by the virus in commercial onion production in Egypt. Further surveys and monitoring of IYSV incidence and distribution in the entire Egyptian governorate are under investigation. References: (1) D. H. Gent et al. Plant Dis. 88:446, 2004. (2) H. R. Pappu et al. Arch. Virol. 151:1015, 2006. (3) H. R. Pappu et al. Virus Res. 141:219, 2009. (4) A. Manal et al. Egypt. J. Virol. 3:49, 2006.

Physiological studies on microalgal culture additives to optimize growth rate and oil content.

Insulin, in nature, has a stimulatory effect on microorganisms. These effects include the acceleration of sugar metabolism, triacylglycerol anabolism, growth rate, and formation of oils. We also observed that insulin may cause indirect activation of triacylglycerol lipase by forcing the cell to permanently require an energy source. Thus, cells can consume all of their accumulated internal fuel sources such as lipids, proteins, and carbohydrates. After studying the effects of using two types of insulin (Humulin 70/30, and human insulin expressed in yeast) at different concentrations on microalgae (Chlorella sp.), we found that with certain concentrations of insulin (1:3.3 ml unit Humulin 70/30 per ml; 1:2.6 ml unit yeast insulin per ml), there was an increase in algal growth rate and decrease in cell size. We therefore studied the effect of insulin under conditions of lipase inhibition by Triton WR 1339 (Tyloxapol), which was used at different concentrations with and without insulin. We found strong regression in the growth rate with increasing Triton concentrations. However, we also observed that the cell size under the effect of Triton and Triton-insulin was larger than the cell size under the effect of insulin alone, and also larger than for control cells. Also, the oil content of the Triton-insulin cells was higher than those of the control cells or the cells under the effect of insulin alone.

BMPR-1B, BMP-15 and GDF-9 genes structure and their relationship with litter size in six sheep breeds reared in Egypt.

OBJECTIVE: The aim of this work was to investigate three different mutations; Fec-B, FecXG, Fec-GH at three candidate genes; Bone morphogenetic protein receptor IB, Bone morphogenetic protein 15 and Growth Differentiation Factor 9, respectively, in six sheep breeds reared in Egypt namely; Rahmani, Barki, Rahmani X Barki cross, Awassi, Awassi X Suffolk cross, and Ossimi and their association with litter size. RESULTS: Genomic DNA of 132 sheep was investigated for the Fec-B, FecXG, and Fec-GH mutations by Restriction Fragment Length Polymorphism, Single-Stranded Conformation Polymorphism and DNA sequencing. The results revealed that all breeds did not carry Fec-B mutation. On the other side, the mutations of FecXG, and Fec-GH were detected in Rahmani, and Rahmani X Barki cross which is associated with the high twinning rate/litter size of Rahmani (1.28) and Rahmani X Barki cross (1.22). While, the average litter size for other breeds had almost a constant values rate over six parities, ranging between 1.00 and 1.04.

Sequence heterogeneity of the envelope-like domain in cultivated allotetraploid Gossypium species and their diploid progenitors.

Retroviral envelope (env)-like sequences in 2 cultivated allotetraploid cottons and their diploid progenitors have been identified and characterized in this study. DNA sequence analysis reveals that these sequences are heterogeneous. The observed sequence diversity, however, seems to preserve coding information. This is evidenced by the detection of the transmembrane domain (TM), which is the most conserved feature of the divergent retroviral env genes. The high ratio of synonymous to nonsynonymous changes suggests that these sequences are evolving under purifying selection. Phylogenetic analysis shows that Gossypium sequences closely cluster with a lineage of plant endogenous retroviruses that have an env-like gene. These results provide evidence for the antiquity and the wide diversity of env-like sequences in the Gossypium genome.

Silver nanoparticles enhance the larvicidal toxicity of Photorhabdus and Xenorhabdus bacterial toxins: an approach to control the filarial vector, Culex pipiens.

Mosquito-control is still based mostly on chemical insecticides which are toxic and cause environmental deprivation. This study investigates synthesizing silver bio-nanoparticles (AgNPs) from nematode-symbiotic bacterial toxin complexes as an alternative larvicidal bioinsecticide agent against Culex pipiens larvae. Five species/strains of nematode-symbiotic bacteria, Xenorhabdus indica, Xenorhabdus spp., Photorhabdus luminescens laumondii HP88, Photorhabdus luminescens akhurstii HRM1 and Photorhabdus luminescens akhurstii HS1 were used. AgNPs were characterized by scanning electron microscopy and x-ray diffraction analysis. Larvae were initially exposed to descending concentrations (300, 150, 75, 37.5 and 18.75 µg/ml) of each of the five bacterial toxins (as positive controls) or to the bio-AgNPs synthesized from the same bacterial toxins (200, 100, 50, 25, 12.5, 6.25, 3.12 and 1.5 µg/ml) for 48 hours. Results of toxicity bioassays showed that mortality of treated larvae was concentration-dependent, toxins from X. indica, P. luminescens laumondii HP88 and P. luminescens akhurstii HS1 showed LC50 of 29, 28 and 2002 µg/ml, respectively. While, toxins from P. luminescens akhurstii HRM1 and Xenorhabdus sp. showed LC50 of 199, 318 µg/ml, respectively. Bio-AgNPs synthesized from, X. indica or Xenorhabdus sp. toxins have significantly increased their larvicidal activities (LC50 of 1.6, 3.7 µg/ml ) at 48h post-treatment. Moreover, bio-AgNPs synthesized from P. luminescens laumondii HP88, P. luminescens akhurstii HRM1 or P. luminescens akhurstii HS1 toxins significantly increased their larvicidal activities (LC50 of 2.1, 1.5, 13.9 µg/ml, respectively) at 48h post treatment. In conclusion, the highest larval toxicity was observed when larvae were treated with bio-AgNPs synthesized from P. luminescens akhurstii HRM1 and X. indica, followed by P. luminescens laumondii HP88 and Xenorhabdus sp. Subsequently, data of the present study suggest these bio-AgNPs toxin complexes as potentially effective bio-control candidates in the battle against mosquito. However, testing other types of bio-synthesized nanomaterials, and their synergistic combinations against different mosquito species still under investigation.

Design, synthesis, molecular modeling and anti-proliferative evaluation of novel quinoxaline derivatives as potential DNA intercalators and topoisomerase II inhibitors.

New series of [1,2,4]triazolo [4,3-a]quinoxaline and bis([1,2,4]triazolo)[4,3-a:3',4'-c]quinoxaline derivatives have been designed, synthesized and biologically evaluated for their cytotoxic activities against three tumor cell lines (HePG-2, Hep-2 and Caco-2). Compounds 16e, 21, 25a and 25b exhibited the highest activities against the examined cell lines with IC50 values ranging from 0.29 to 0.90 µM comparable to that of doxorubicin (IC50 ranging from 0.51 to 0.73 µM). The most active members were further evaluated for their topoisomerase II (Topo II) inhibitory activities and DNA intercalating affinities as potential mechanisms for their anti-proliferative activities. Interestingly, the results of Topo II inhibition and DNA binding assays were consistent with that of the cytotoxicity data, where the most potent anti-proliferative derivatives exhibited good Topo II inhibitory activities and DNA binding affinities, comparable to that of doxorubicin. Moreover, the most active compound 25a caused cell cycle arrest at G2/M phase and induced apoptosis in Caco-2 cells. In addition, Furthermore, molecular docking studies were performed for the novel compounds against DNA-Topo II complex to investigate their binding patterns. Based on these studies, it was concluded that DNA binding and/or Topo II inhibition may contribute to the observed cytotoxicity of the synthesized compounds.

Occurrence of iris yellow spot virus from onion crops in Egypt.

Iris yellow spot virus (IYSV) is a infects onion bulb and seed crops in many countries including Egypt. Results of the mechanical inoculation reveled that, small chlorotic lesions and systemic necrosis were observed on both Nicotiana benthamiana and Datura stramonium after 10 days, while there were no symptoms were appeared on the onion plant. The viral biological transmission with Thrips tabaci was highly reported to be efficiently for virus transmitted. Our results confirmed the presentence of virus-like particles of a Tospovirus infected onion leaf using transmission electron microscopy. Both of sequence and phylogenetic analysis of N gene revealed that our viral isolate is IYSV with 95 % identity with reported Israel isolate. The sequence of N gene had three motifs: casein kinase II Phosphorylation site, N-myristoylation site and protein kinase C phosphorylation site. These motifs are involved in regulation, activity and stability of IYSV. To our knowledge, this is the first molecular characterization of IYSV in Egypt.

Biochemical and molecular aspects of aluminium chloride-induced neurotoxicity in mice and the protective role of Crocus sativus L. extraction and honey syrup.

Aluminium has been proposed as an environmental factor that may affect several enzymes and other biomolecules related to neurotoxicity and Alzheimer's disease (AD). The promising protective effect of aqueous saffron extract and honey syrup on neurotoxicity induced by aluminuim chloride (AlCl(3)) may be derived from their own antioxidant properties. Balb/c and C57BL/6 mice (35-40 g) were injected with AlCl(3), 40 mg/kg/day for 45 days. Each mice strain was divided into four groups: AlCl(3) treated group, AlCl(3) plus water saffron extract group (administered with saffron extract at 200 mg/kg b.w. once a day for the experimental period), AlCl(3) plus honey syrup group (administered with honey syrup at 500 mg/kg b.w. for 45 days). The control group received no treatment. Oxidative stress and antioxidant status were estimated in the brain and differential display was performed for both mice strains to scan the mRNA in the treated and non treated groups. In addition, the up and down regulated genes were isolated, cloned and sequenced. The sequence analysis was performed and compared with the other genes cited on GenBank. The results show that there was a decrease in the activity of the antioxidant enzymes (P≤0.001) such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in the AlCl3 groups of both mice strains. The level of brain thiobarbituric acid reactive substances (TBARS) showed a significant increase (P≤0.001) of lipid peroxidation (LPO) in the AlCl(3) groups. There was an indication of carcinogenicity in the AlCl(3) treated group representing an increase in serum tumor markers such as arginase and a-l-fucosidase. More than 350 band patterns were obtained and about 22 different up-down regulated genes were observed. The sequence analysis of the three selected up-regulated genes revealed that they are similar to B-cell lymphoma 2 (Bcl-2), R-spondin and the inositol polyphosphate 4-phosphatase genes (INPP4B), respectively. The R-spondin gene was up-regulated in all examined animals except the control ones but the other two genes were only induced in the animals treated with AlCl(3) and honey syrup. We conclude that the biochemical and molecular studies showed the neurotoxicity of AlCl(3) in the brains of mice. In addition, there was an ameliorative change with saffron extract and honey syrup against AlCl(3) neurotoxicity. The obtained molecular results suggest that AlCl(3) made induction for BCL-W gene, which is an anticancer gene or belongs to the DNA repair system in the brain cells, as well as for R-spondin and inositol polyphosphate 4-phosphatase genes, which help in cell proliferation.

Induction of defense responses in common bean plants by arbuscular mycorrhizal fungi.

Interaction between arbuscular mycorrhizal fungi as a bio-agent and Rhizoctonia root rot disease of common bean plant was investigated in this study under natural conditions in pot experiment. A mixture of Egyptian formulated AM (Multi-VAM) in suspension form (1 × 10(6) unit L(-1) in concentration) was used at dilution of 5 ml L(-1) water. The results demonstrated that colonization of bean plants with AM fungi significantly increased growth parameters, yield parameters and mineral nutrient concentrations and reduced the negative effects on these parameters as well as both disease severity and disease incidence. Different physical and biochemical mechanisms have been shown to play a role in enhancement of plant resistance against Rhizoctonia solani, namely, improved plant nutrition, improved plant growth, increase in cell wall thickening, cytoplasmic granulation, and accumulation of some antimicrobial substances (phenolic compounds and defense related enzymes).