Muhammad Shahzad Iqbal

Transformation and Evaluation of Cry1Ac+Cry2A and GTGene in Gossypium hirsutum L.

More than 50 countries around the globe cultivate cotton on a large scale. It is a major cash crop of Pakistan and is considered “white gold” because it is highly important to the economy of Pakistan. In addition to its importance, cotton cultivation faces several problems, such as insect pests, weeds, and viruses. In the past, insects have been controlled by insecticides, but this method caused a severe loss to the economy. However, conventional breeding methods have provided considerable breakthroughs in the improvement of cotton, but it also has several limitations. In comparison with conventional methods, biotechnology has the potential to create genetically modified plants that are environmentally safe and economically viable. In this study, a local cotton variety VH 289 was transformed with two Bt genes (Cry1Ac and Cry2A) and a herbicide resistant gene (cp4 EPSPS) using the Agrobacterium mediated transformation method. The constitutive CaMV 35S promoter was attached to the genes taken from Bacillus thuringiensis (Bt) and to an herbicide resistant gene during cloning, and this promoter was used for the expression of the genes in cotton plants. This construct was used to develop the Glyphosate Tolerance Gene (GTGene) for herbicide tolerance and insecticidal gene (Cry1Ac and Cry2A) for insect tolerance in the cotton variety VH 289. The transgenic cotton variety performed 85% better compared with the non-transgenic variety. The study results suggest that farmers should use the transgenic cotton variety for general cultivation to improve the production of cotton.

Prediction of Host-Derived miRNAs with the Potential to Target PVY in Potato Plants

Potato virus Y has emerged as a threatening problem in all potato growing areas around the globe. PVY reduces the yield and quality of potato cultivars. During the last 30 years, significant genetic changes in PVY strains have been observed with an increased incidence associated with crop damage. In the current study, computational approaches were applied to predict Potato derived miRNA targets in the PVY genome. The PVY genome is approximately 9 thousand nucleotides, which transcribes the following 6 genes:CI, NIa, NIb-Pro, HC-Pro, CP, and VPg. A total of 343 mature miRNAs were retrieved from the miRBase database and were examined for their target sequences in PVY genes using the minimum free energy (mfe), minimum folding energy, sequence complementarity and mRNA-miRNA hybridization approaches. The identified potato miRNAs against viral mRNA targets have antiviral activities, leading to translational inhibition by mRNA cleavage and/or mRNA blockage. We found 86 miRNAs targeting the PVY genome at 151 different sites. Moreover, only 36 miRNAs potentially targeted the PVY genome at 101 loci. The CI gene of the PVY genome was targeted by 32 miRNAs followed by the complementarity of 26, 19, 18, 16, and 13 miRNAs. Most importantly, we found 5 miRNAs (miR160a-5p, miR7997b, miR166c-3p, miR399h, and miR5303d) that could target the CI, NIa, NIb-Pro, HC-Pro, CP, and VPg genes of PVY. The predicted miRNAs can be used for the development of PVY-resistant potato crops in the future.

In silico MCMV Silencing Concludes Potential Host-Derived miRNAs in Maize

Maize Chlorotic Mottle Virus (MCMV) is a deleterious pathogen which causes Maize Lethal Necrosis Disease (MLND) that results in substantial yield loss of Maize crop worldwide. The positive-sense RNA genome of MCMV (4.4 kb) encodes six proteins: P32 (32 kDa protein), RNA dependent RNA polymerases (P50 and P111), P31 (31 kDa protein), P7 (7 kDa protein), coat protein (25 kDa). P31, P7 and coat protein are encoded from sgRNA1, located at the 3′end of the genome and sgRNA2 is located at the extremity of the 3′genome end. The objective of this study is to locate the possible attachment sites of Zea mays derived miRNAs in the genome of MCMV using four diverse miRNA target prediction algorithms. In total, 321 mature miRNAs were retrieved from miRBase (miRNA database) and were tested for hybridization of MCMV genome. These algorithms considered the parameters of seed pairing, minimum free energy, target site accessibility, multiple target sites, pattern recognition and folding energy for attachment. Out of 321 miRNAs only 10 maize miRNAs are predicted for silencing of MCMV genome. The results of this study can hence act as the first step towards the development of MCMV resistant transgenic Maize plants through expression of the selected miRNAs.

Salt Tolerance of Potato: Genetically Engineered with Atriplex canescens BADH Gene Driven by 3 Copies of CAMV35s Promoter

Biochemical effect of tannic acid and curcumin on female mice experimentally induced Ehrlich Ascitis Carcinoma (EAC) was investigated. This study was carried out on 220, 12-14 weeks old female mice and weighted 25-30 g. Mice were classified into two main large experiments. Experiment 1: Non-tumor bearing mice (NTB) Included 100 of animals and divided into four groups each one comprised 25 mice. Group 1: NTBcontrol saline treated .Group 2:NTB-treated with curcumin orally (350 mg/kg/day) for 6 weeks. Group 3:NTB-treated with tannic acid orally (160 mg/kg/day) for 6 weeks. Group 4:NTB-treated with curcumin and tannic acid orally at ratio (50% : 50%) for 6 weeks. Experiment 2: Tumor bearing (TB) mice. Included 120 of animals and divided into four groups each one comprised 30 mice. Group 1:TBM-control saline treated. Group 2: TBM-treated with curcumin orally (350 mg/kg/day) for 6 weeks. Group 3:TBM-treated with tannic acid orally (160 mg/kg/day) for 6 weeks. Group 4:TBM-treated with curcumin and tannic acid orally at ratio (50%: 50%) for 6 weeks. Blood samples were collected from all animals groups after 2, 4 and 6 weeks from treatment. Serum were separated and processed directly for glucose, insulin, total cholesterol, triacylglycerol, total protein determination. The obtained results revealed that, a highly significant decrease in serum glucose, total cholesterol, total protein concentration. Mean while, a highly significant increase in serum triacylglycerol concentration. But a non significant decrease in serum insulin levels were observed in tumor bearing mice when compared with control. The results of this study indicated that curcumin, tannic acid and their combination treatment have potential benefits in cancer treatment. Kew words: Curcumin, tannic acid, triacylglycerols, tumor and anticancer. Corresponding author: Mohammed F. EL-Shiekha, Department of Biochemistry, Faculty of Veterinary Medicine, Benha University, Moshtohor, Qalioubeya, Egypt. E-mail : drmohchem@gmail.com