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Featured researches published by A. Abdelhadi.


GM crops & food | 2014

Improved regeneration and transformation protocols for three strawberry cultivars.

Hossam Zakaria; Gihan M. Hussein; Abdelhadi A. Abdelhadi; Naglaa A. Abdallah

Strawberry (Fragaria × ananassa) is an economically important soft fruit crop with polyploid genome which makes the breeding of new cultivars difficult. Simple and efficient method for transformation and regeneration is required for cultivars improvement in strawberry. In the present study, adventitious shoot regeneration has been investigated in three cultivated strawberry plants, i.e., Festival, Sweet Charly and Florida via direct organogenesis using the in vitro juvenile leaves as explants. Explants were collected after sub-culturing on a propagation medium composed of MS supplemented with 0.5 mg/l BA; 0.1 mg/l GA3 and 0.1 mg/l IBA. To select the suitable organogenesis, the explants of the three cultivars were cultured on MS medium supplemented with different concentrations of TDZ (1, 2, 3, and 4 mg/l), then incubated at a temperature of 22 °C ± 2. Medium containing 2 mg/l TDZ revealed the best regeneration efficiency with the three cultivars (72% for Festival, and 73% for Sweet Charly and Florida). After 4 weeks, the produced shoots were cultured on MS medium with different concentrations of BA and Kin to enhance shoot elongation. Results showed that the medium containing 1.5 mg/l BA and 0.5 mg/l Kin revealed highest elongation efficiency (88% and 94%) for Festival and Sweet Charly, respectively. On the other hand, medium containing 1.5 mg/l BA and 0.1 mg/l Kin showed highest elongation efficiency (90%) in Florida. Elongated shoots were successfully rooted on MS medium containing 1.5 mg/l NAA. Furthermore, transformation of the two cultivars, Festival and Sweet Charly, has been established via Agrobacterium strain LBA44404 containing the plasmid pISV2678 with gus-intron and bar genes. Three days post co-cultivation, GUS activity was screening using the histochemical assay. The results showed 16% and 18% of the tested plant materials has changed into blue color for Festival and Sweet Charly, respectively. Out of 120 explants only 13 shoots were developed on bialaphos medium for each cultivar, representing 10.8% bialaphos resistant strawberry shoot. The presence of the both genes bar and uid A was detected by PCR and Northern giving a transformation efficiency of 5%.


GM crops & food | 2011

Establishment of regeneration and transformation system of sugarcane cultivar GT54-9 (C9)

Dessoky S. Eldessoky; Roba M. Ismail; Abdelhadi A. Abdelhadi; Naglaa A. Abdallah

Plant regeneration protocols for sugarcane GT54-9(C9) cultivar were developed for direct organogenesis and indirect somatic embryogenesis, using young leaf segments as explants by studying the influence of different concentrations and types of cytokinin and auxin hormones. For the callus formation from young leaves, a medium containing 4mg/l 2,4-D was found very effective. For embryo formation, MS medium supplemented with 1mg/l Kin and 0.5 mg/l 2,4-D was used. While in the case of direct organogenesis protocol, the medium containing 1mg/l BAP and 2mg/l NAA was the best for direct shoot formation. Data showed that the best shoot regeneration and elongation medium for direct organogenesis and indirect somatic embryogenesis was obtained on medium with 2 mg/l Kin and 0.1 mg/l BAP. Root induction was best performed on 2mg/l NAA and complete plantlets were hardened in the greenhouse before transferring to the field for further evaluation. For transformation, young leaf segments of sugarcane from the cultivar GT54-9(C9) were inoculated and co-cultivated with Agrobacterium tumefaciens strain LB4404 harboring the binary vector pISV2678 with the bar and the gus-intron genes. The obtained putative transgenic plantlets were able to grow under bialaphose containing medium. Stable integration of the bar gene into the plant genomes was tested by PCR and Southern blot hybridization. Histochemical assay and leaf painting analysis were carried out to study the expression of the gus and bar genes in transgenic plants, respectively. The results indicated that the direct organogenesis produced a higher yield of regenerated plants (22% more) within shorter time (4 weeks less). Therefore, this method is recommended for sugarcane regeneration and for further use in genetic transformation via A. tumefaciens with desired genes.


GM crops & food | 2015

Expression of synthetic human tropoelastin (hTE) protein in Nicotiana tabacum

Mona Abdelghani; Ghada A. Abu El-Heba; Abdelhadi A. Abdelhadi; Naglaa A. Abdallah

Plant molecular farming (PMF) is an important growing prospective approach in plant biotechnology; it includes production of recombinant pharmaceutical and industrial proteins in large quantities from engineered plants. Elastin is a major protein component of tissues that require elasticity, it helps keep skin smooth as it stretches to allow normal. Elastin is used as a raw material for the cosmetic industry. In this work, we aimed to use plant as a bioreactor for the expression and production of the full human tropoelastin protein. Agrobacterium- mediated transient expression system into Nicotiana tabacum using syringe agroinfiltration was used to provide fast and convenient way to produce recombinant proteins with greater expression overall the plant leaf. This study aimed to establish an efficient and rapid system for transiently expression and production of human recombinant tropoelastin protein in transgenic N. tabacum plants. Modified elastin (ELN) gene was biosynthesized and cloned into pCambia1390 vector to be used into N. tabacum agroinfilteration. Optimization of codon usage for the human tropoelastin gene, without changing the primary structure of the protein was carried out to ensure high expression in tobacco plants. The obtained data proved that the 5(th) day post-infiltration is the optimum interval to obtain the maximum production of our recombinant protein. Southern blot analysis was able to detect 2175 bp fragment length representing the ELN orf (open reding frame). On the other hand, ELN -expression within plants tissue was visualized by RT-PCR during the period 3-10 days post agroinfiltration. At the protein level, western and ELISA confirmed the expression of recombinant tropoelastin protein. Western blot analysis detected the tropoelastin protein as parent band at ∼70 kDa from freshly extracted protein, while two degraded bands of ∼55 and ∼45 kDa, representing a pattern of tropoelastin were appeared with frozen samples. This study showed that biosynthetic ELN gene was successfully expressed into N. tabacum leaves using agroinfiltration technique.


African Journal of Biotechnology | 2011

Molecular markers associated with salt tolerance in Egyptian wheats

Reda E. A. Moghaieb; Abdelhadi A. Abdelhadi; Neveen B. Talaat

Salinity affects plant growth by the osmotic stress of the salt around the roots, as well as by toxicity caused by excessive accumulation of salt in leaves. In the present study, seven common ( Triticum aestivum ) and two durum ( T. turgidum ssp. Durum) wheat genotypes were subjected to salt stress for 2 weeks. Salt stress decreased leaf osmotic potential in all cultivars. The difference in osmotic adjustment between the cultivars was correlated with the concentrations of minerals examined such as Na + and K + . The salt tolerance in the T. aestivum cultivar Gemmiza 10 and in T. durum cultivars Sohag and Beni Sweif was due to higher ability to maintain osmotic potential of the cells than the other cultivars by increase in osmoticum concentration under salt stress. The genetic variation and relationships among different wheat genotypes with different responses to salt stress were also investigated by RAPD and SSR analyses. 82 out of 118 RAPD markers detected were polymorphic (69.5%) and 42 out of 59 SSR alleles were polymorphic (71%), and can be considered as useful markers for the wheat cultivars tested. 18 random amplified polymorphic DNAs (RAPD) markers and 13 simple sequence repeats (SSR) markers generated were found to be genotype-specific. Seven markers distinguished the cultivar Beni Sweif, six markers for the cultivar Sohag and two markers for the cultivar Gemmiza 10. These markers can be verified as being genetic markers associated with salt tolerance in the three wheat genotypes and help in marker-assisted selection breeding program. Key words : Osmotic adjustment, RAPD and SSR marker, salt tolerance, wheat genomes.


3 Biotech | 2017

Molecular identification and genetic diversity among Photorhabdus and Xenorhabdus isolates

Reda E. A. Moghaieb; Abdelhadi A. Abdelhadi; Hanan A. El-Sadawy; Nesreen A. T. Allam; Baiome Abdelmaguid Baiome; Mohamed H. Soliman

Five bacterial strains were isolated from the hemocoel of the greater wax moth larvae (Galleria mellonella) infected with the entomopathogenic nematodes: Heterorhabditis bacteriophora HP88, Heterorhabditis indicus RM1 and Heterorhabditis sp (S1), Steinernema abbasi and Steinernema sp. (S II). Strains were identified as Photorhabdus luminescens HRM1, P. luminescens HS1, P. luminescens HP88, Xenorhabdus indica and X. nematophila ATTC19061 using 16S rDNA sequence analysis. To reveal the genetic diversity among these strains, three molecular markers (RAPD, ISSR and SRAP) were employed. RAPD analysis showed 73.8 and 54.5 polymorphism percentages for the Photorhabdus and Xenorhabdus strains, respectively. ISSR analysis resulted in 70.1 and 75.2 polymorphism percentages among the Photorhabdus and Xenorhabdus strains, respectively. The SRAP analysis indicated that 75.6 and 61.2% genetic polymorphism was detected among Photorhabdus and Xenorhabdus strains, respectively. The cluster analysis grouped the three Photorhabdus strains together in one cluster and the two Xenorhabdus strains together in another cluster indicating the phylogenetic relationships among them. The genotype-specific markers detected from the three molecular markers (RAPD, ISSR and SRAP) were sufficient to distinguish between the different bacterial strains tested and can be used in the future IBM program that could be built on the use of these strains.


African Journal of Biotechnology | 2011

Induction, characterization and genetic analysis of Aspergillus flavus resistant mutants in Arachis hypogaea

Ahmed Nagib Sharaf; Ahmed Gaber; Abdelhadi A. Abdelhadi; Walid Ahmed Korani

The Aspergillus flavus infection of peanut ( Arachis hypogaea ) results in the accumulation of aflatoxins in seeds, which are very harmful to humans and animals. Mutation breeding programs are an effective way of inducing resistant mutants. In this study, we induced a genetic variation by using ethyl methanesulfonate (EMS) and gamma rays treatment for four peanut cultivars (32 mutants from Giza 6, 22 mutants from Gregory, 15 mutants from Giza 4 and 15 mutants from Giza 5). The resistant mutants for A. flavus were identified by analyzing β-1-3-glucanases activities of the controls and infected mutants using polyacrylamide gel electrophoresis (PAGE). Two, four and four mutants derived from Giza 6, Gregory and Giza 4, respectively, showed high activities of β-1-3-glucanases and therefore more resistant to the infection of A. flavus . The genetic similarity of these mutants and their controls was also tested using random amplified polymorphic DNA (RAPD) approach. Although natural polymorphism among peanut cultivars was very low, RAPD patterns showed high polymorphism percentage of DNA fragments (37.13%). Key words : Aspergillus flavus, aflatoxins, Arachis hypogaea, peanut, gamma rays, ethyl methanesulfonate (EMS), pathogenesis related (PR) proteins


Applied Biochemistry and Biotechnology | 2017

Designing of a Recombinant Agip Bacmid Construct with Infectious Properties Against Black Cutworm Agrotis ipsilon Larvae

Naglaa A. Abdallah; Wael El-Menofy; Abdelhadi A. Abdelhadi

In this study, Agrotis ipsilon nucleopolyhedrovirus bacmid has been constructed as an infectious bacmid in an attempt to allow genome recombination and generation of virus mutants. Since the FseI, a unique restriction site, is located in a viral coding region (ORF_119), PCR was performed to partially amplify the ORF_119 fragment containing the FseI site to facilitate the bacmid construction in a proper way without interrupting the ORF expression. Construction with repeated fragments at the end of the cloned viral was carried out in an attempt to facilitate circulation during infection in insect cells. The amplified gp_119 fragment was cloned into the BAC_Bsu361 plasmid derived from the AcMNPV Bac-to-Bac® system. Recombinant plasmid was used to subclone the Agrotis ipsilon nucleopolyhedrovirus (AgipNPV)-linearized genome using the FseI unique site. The Agip bacmid DNA extracted from Escherichia coli was used to transfect A. ipsilon third instar larvae by injection into the hemolymph. The produced occlusion bodies were purified from infected larvae and used to feed healthy larvae for amplifying the virus, and infectivity was recorded. Using bacmid technology will facilitate manipulation of the AgipNPV genome and help in determining the genetic factors involved in virus virulence and biology.


AMB Express | 2017

Boosting isoprene production via heterologous expression of the Kudzu isoprene synthase gene ( kIspS ) into Bacillus spp. cell factory

Lamis Gomaa; Michael E. Loscar; Haggag S. Zein; Nahed Abdel-Ghaffar; Abdelhadi A. Abdelhadi; Ali S. Abdelaal; Naglaa A. Abdallah


Biotechnology(faisalabad) | 2016

Identification, Characterization and Genetic Improvement of Bacteriocin Producing Lactic Acid Bacteria

Abdelhadi A. Abdelhadi; Nagwa I. Elarabi; Rasha Salim; Ahmed Naguib Sharaf; Nivien A. Abosereh


Microbiology and Biotechnology Letters | 2018

Molecular Characterization of Some Antilisterial Bacteriocin Genes from Enterococcus faecium and Pediococcus pentosaceus

Nagwa I. Elarabi; Rasha Salim; Nivien A. Abosereh; Abdelhadi A. Abdelhadi

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