L-Arginase, hydrolyzing L-arginine to L-ornithine
and urea, is a powerful anticancer, L-arginine-... more L-Arginase, hydrolyzing L-arginine to L-ornithine and urea, is a powerful anticancer, L-arginine-depleting agent, against argininosuccinate synthase expressing tumors. Otherwise, the higher antigenicity and lower thermal stability of this enzyme was the main biochemical hurdles. Since, the intrinsic thermal stability of enzymes follow the physiological temperature of their producer, thus, characterization of Larginase from thermotolerant Penicillium chrysogenum was the objective of this study. L-Arginase (Arg) was purified to its homogeneity from P. chrysogenum by 10.1-fold, with 37.0 kDa under denaturing PAGE, optimum reaction at 50 C, pH stability (6.8–7.9), with highest molar ratio of constitutional arginine, glutamic acid, lysine and aspartic acid. The purified enzyme was PEGylated and immobilized on chitosan, with 41.9 and 22.1 % yield of immobilization. At 40 C, the T1/2 value of free-Arg, PEG-Arg and Chit-Arg was 10.4, 15.6, 20.5 h, respectively. The free-Arg and Chit- Arg have a higher affinity to L-arginine (Km 4.8 mM), while, PEG-Arg affinity was decreased by about 3 fold (Km 15.2 mM). The inhibitory constants to the free and PEG-Arg were relatively similar towards HA and PPG. The IC50 for the free enzyme against HEPG-2 and A549 tumor cells was 0.136 and 0.165 U/ml, comparing to 0.232 and 0.496 U/ml for PEG-Arg, respectively. The in vivo T1/2 to the free Arg and PEG-Arg was 16.4 and 20.4 h, respectively as holoenzyme. The residual L-arginine level upon using free Arg was 156.9 and 144.5 lM, after 6 and 8 h, respectively, regarding to initials at 253.6 lM, while for Peg-Arg the level of L-arginine was nil till 7 h of initial dosing. The titer of IgG was induced by 10–15 % in response to free-Arg after 28 days comparing to IgG titer for PEG-Arg.
Durum wheat (Triticum turgidum ssp. durum) is an economically and nutritionally important cereal ... more Durum wheat (Triticum turgidum ssp. durum) is an economically and nutritionally important cereal crop in the Mediterranean region and its production is largely influenced by environmental stresses, such as drought, salinity, heat and nutrient deficiency. The objective of this study was to dissect quantitative trait loci (QTL) controlling grain yield, yield components and drought tolerance in durum. A molecular genetic linkage map for F2 durum mapping population derived from an intraspecific cross between Baniswif-1 x Sohag-2 was constructed using 114 DNA markers (9 SSRs, 14 SCoTs, 90 AFLPs and 1 RAPDs) distributed over the 14 linkage groups and spanning 2040.9 cM of the durum wheat genome. The size of linkage groups varied greatly from 6.8cM for LG11 to 317.5cM for LG4 with an average length of 145.8cM. Based on the used anchor SSR markers, only eight linkage groups were assigned to chromosomes, where LG1, LG3, LG5, LG6, LG7, LG9, LG13 and LG14 were assigned to chromosomes 1B, 3B, 5B, 6A, 6B, 7A, 3A and 2B, respectively. Single point analysis was used to identify genomic regions controlling eleven morpho-physiological traits related to grain yield, yield components and drought tolerance. A total of 74 QTL were identified for the eleven traits on all linkage groups except (LG10 and LG11). These included 3 QTL for root length (RL), 11 QTL for plant height (PH), 7 QTL for spike length (SL), 3 QTL for number of branches/plant (NBP), 3 QTL for number of spike/plant (NSP), 8 QTL for number of spikelets/spike (NSS), 15 QTL for number of kernel/spike (NKS), 10 QTL for thousand-kernel weight (TKW), 4 QTL for fresh weight (FW), 5 QTL for dry weight (DW) and 5 QTL for total amino acids (TAA). This work represents the first genetic linkage map for durum wheat population derived from an intraspecific cross between ‗Baniswif-1‘ and ‗Sohag-2‘ showing chromosomal regions associated with 11 morpho-physiological traits related to grain yield, yield components and drought tolerance in durum wheat.
Molecular markers provide a rapid approach to breeding for desired traits. To use them, it is nec... more Molecular markers provide a rapid approach to breeding for desired traits. To use them, it is necessary to determine the linkage between quantitative trait loci (QTL) and such markers. This study was conducted to investigate the genetic basis of salinity responses in Egyptian bread wheat (Triticum aestivum L.). In this context, a doubled haploid (DH) population (SGDH) of 139 individuals was produced from the cross between two Egyptian breeding cultivars (Sakha 93, salt tolerant) and (Gemmeza 7, salt susceptible). The DH population was tested under saline hydroponics culture and various plant responses were measured. A molecular genetic map of the SGDH population covering 3645.3 cM, was constructed using Restriction Fragment Length Polymorphism (RFLP), Microsatellite or Simple Sequence Repeats (SSR), and Amplified Fragment Length Polymorphism (AFLP) markers. In total, 325 loci along the 21 wheat chromosomes were mapped. The B genome showed the highest number of mapped markers followed by the A and the D genomes respectively. Interval and composite interval mapping (using QTL cartographer) were used to identify the genomic regions controlling traits related to salt tolerance with a threshold of LOD 3. Analysis of QTLs has revealed the approximate location of the significant markers associated with 12 traits related to salt tolerance traits across the A, B and D genomes. Fifty five significant QTL were detected on 15 of the 21 chromosomes mapped in this study, for some of these more than one QTL was identified. In many cases QTL were mapped very close to each other, indicating possible gene clustering or pleiotropy. For some traits, the total percentage of phenotypic variation explained by all QTL exceeded 50-60% but on average it was in the region of 15%. Chromosomes of homologues groups 2 and 5 exerted the biggest effect on most phenotypic traits, especially chromosomes 2B and 5B. The results indicated that there is considerable potential for improving salt tolerance of hexaploid wheat by using marker-assisted selection.
Net blotch, caused by Pyrenophora teres f. teres, is one of the most devastating diseases causing... more Net blotch, caused by Pyrenophora teres f. teres, is one of the most devastating diseases causing significant losses in barley yield and quality. In the present investigation, two barley verities “AT4” (net blotch resistant) and “Femina” (net blotch susceptible) were used to develop a segregating F2 population. Linkage analysis and map construction were performed using Map Manager. The constructed genetic linkage map consisted of 85 markers including 45 AFLP , 11 SSR, 6 CAPS, 5 SCoT, 3 STS and 6 NBL. Linkage groups were assigned to individual barley chromosomes using the published map locations of the SSR markers as reference point. The produced map showed 7 linkage groups with 85 markers covered a total length of 1644.8 cM. The average length of linkage groups ranged from 77.7 to 739.4. Single point analysis was used to identify the genomic regions associated with net blotch resistance in barley. A total of 14 QTL with a significance ranging from 0.01% to 5% were identified on 4 linkage groups (2, 4, 5 and 6). The most significant QTL was found on chromosome 6H. This QTL presents a promising opportunity for the strategic improvement of barley resistance to net blotch using marker assisted selection.
Trehalose is a non-reducing disaccharide which consists of two glucose units that functions as a ... more Trehalose is a non-reducing disaccharide which consists of two glucose units that functions as a compatible solute to stabilize the membrane structures under heat and desiccation stress. Trehalose-6-phosphate synthase (TPS) and trehalose-6- phosphate phosphatase (TPP) are the key enzymes for trehalose biosynthesize in the plant kingdom. On the basis of bioinformatics prediction, fragment containing an open reading frame of 945 bp was cloned from durum wheat. Sequence comparison and analysis of conserved domains revealed the presence of a TPP domain. Full length of the gene was isolated using gene race technology. Semi-quantitative RT-PCR and real time quantitative PCR indicated that the expression of this gene is up-regulated in response to drought stress. The biochemical assay of the trehalase activity showed that the enzyme's activity decreased under the dehydration stress. The obtained phylogenic tree showed that the isolated TPP protein forms a distinct clad close to the Oryza sativa trehalose-6- phosphate phosphatase. In silico and comparative mapping indicated that the isolated TPP gene is localized on rice chromosome 8, durum wheat chromosome 20, bread wheat chromosome 3B, oat linkage group E, sorghum chromosome 4 and barley 5H.
Cotton is the world’s leading natural fiber and second most important oilseed crop and has been a... more Cotton is the world’s leading natural fiber and second most important oilseed crop and has been a focus of genetic, systematic and breeding research. The genetic and physiological bases of some important agronomic traits in cotton were investigated by QTL mapping through constructing of genetic map with chromosomal assignment. A segregating F2 population derived from an interspecific cross (G. barbadense x G. hirsutum) between two genotypes, cvs. “Giza 83” and “Deltapine” was used in this study. Different molecular markers including SS R, ES T, ES T-SS R, AFLP and RAPD were employed to identify markers that reveal differences between the parents. In total 42 new markers were merged with 140 previously mapped markers to produce a new map with 182 loci covering a total length of 2370.5 cM. Among these new markers, some of them were used to assign chromosomes to the produced 26 linkage groups. The LG2, LG3, LG11 and LG26 were assigned to chromosomes 1, 6, 5 and 20 respectively. Single point analysis was used to identify genomic regions controlling traits for plant height, number of nodes at flowering time, bolling date, days to flowering and number of bolls. In total 40 significant QTL were identified for the five traits on 11 linkage groups (1, 2, 3, 4, 5, 10, 11, 12, 18, 19 and 23). This work represents an improvement of the previously constructed genetic map in addition to chromosomal assignment and detection of new significant QTL for the five traits in Egyptian cotton. The Significant QTLs detected in this study can be employed in marker assisted selection for molecular breeding programs aiming at developing cotton cultivars with improved agronomic traits
Efficient regeneration methods are a priority for the successful application of genetic engineeri... more Efficient regeneration methods are a priority for the successful application of genetic engineering to vegetatively propagated plants such as grape (Vitis vinifera L.). An efficient in vitro shoot micropropagation protocol leading to the regeneration from shoot tip explants was successfully developed for mass production. Callus was induced from shoot tips and axillary buds on C2D medium supplemented with 4 mg/l 6-benzyladenine (BA). Bud differentiation from callus was possible on C2D medium supplemented with 1 mg/l gibberellic acid and 0.1 mg/l kinetin. Bud initials were elongated by sub-culturing on elongation medium containing C2D salts supplemented with 0.1 mg/l BA and resulting shoots could be mass micropropagated on C2D medium supplemented with 1 mg/l BA. Shoots that were regenerated via organogenesis or micropropagation were rooted on C2D medium supplemented with 1 mg/l indole-3-butyric acid. In this study, regeneration and micropropagation of grape cv. ‘Spero’ was successfully achieved through shoot tip and axillary bud culture. Such a protocol would be useful for future use in the strategic improvement of grape using genetic engineering
Banana is one of the most important fruit crop worldwide, as its production reaches approximately... more Banana is one of the most important fruit crop worldwide, as its production reaches approximately 70 million tones per year. By tissue culture, triploid and seedless bananas can now be produced in a high quantity, in a less time and at any time of the year without being limited to a season. The aim of this study is to substitute the artificial hormones used in the tissue culture such as, cytokinins and auxins by natural materials such as, Pineapple, Coconut milk, Cacao, Coffee, and Charcoal. Five different media composed of these natural materials were prepared in different concentrations, Pineapple (5,10 and 20cm), Coconut milk (5,10 and20cm,), Cacao (1,2.5 and 5 g), Coffee (1,2.5 and 5 g) and Charcoal (1,2.5 and 5 g) to study their effect on the development and browning of the bananas' shoots and roots in vitro, with the aid of various statistical analyses. The results indicated that the pineapple and coconut milk produced the highest number and length of both shoots and roots, as well as the soluble sugars contents, while the cocoa and coconut milk showed the highest protein content. The charcoal reduced significantly the phenolic content. On the other hand, there was no significant difference between the coffee, cacao. This study has highlighted the benefits and advantages of using natural materials in producing high quality banana.
Drought is a major planetary stress problem that places greater limits on cereal productivity. Cr... more Drought is a major planetary stress problem that places greater limits on cereal productivity. Crop species belonging to the tribe Triticale represent the main foodstuff sources for humans. Because durum wheat (Triticum turgidum L.) shows better adaptation to semi-arid climates than bread wheat, its genome should contain a pool of genes that are expressed for wide environmental adaptability. To investigate and understand the genetic response of drought tolerance in durum wheat, differentially expressed sequence tags (dESTs) were detected, characterized and cloned from durum wheat under different regimes of drought stress and the functions of some of these dESTs were determined. In this study, genes involved in drought stress at early stages of growth were profiled by a differential display technique from Egyptian durum wheat (Sohag 3). One, three, six, and 10 hours of drought stress were applied to produce 15 expressed cDNA fragments. This will highlight the genes that are expressed under drought stress, which might help in the understanding of the plant’s defense mechanism under drought conditions. This will also give a chance to identify gene(s) for the future use in transgenic research to enhance drought tolerance in wheat and other commercially important crops. The cloned dESTs might also be used as molecular markers for genetic and QTL mapping for wheat and other cereals under drought stress conditions.
Drought is a major planetary stress problem that places greater limits on cereal productivity. Cr... more Drought is a major planetary stress problem that places greater limits on cereal productivity. Crop species belonging to the tribe Triticale represent the main foodstuff sources for humans. Because durum wheat (Triticum turgidum L.) shows better adaptation to semi-arid climates than bread wheat, its genome should contain a pool of genes that are expressed for wide environmental adaptability. To investigate and understand the genetic response of drought tolerance in durum wheat, differentially expressed sequence tags (dESTs) were detected, characterized and cloned from durum wheat under different regimes of drought stress and the functions of some of these dESTs were determined. In this study, genes involved in drought stress at early stages of growth were profiled by a differential display technique from Egyptian durum wheat (Sohag 3). One, three, six, and 10 hours of drought stress were applied to produce 15 expressed cDNA fragments. This will highlight the genes that are expressed under drought stress, which might help in the understanding of the plant’s defense mechanism under drought conditions. This will also give a chance to identify gene(s) for the future use in transgenic research to enhance drought tolerance in wheat and other commercially important crops. The cloned dESTs might also be used as molecular markers for genetic and QTL mapping for wheat and other cereals under drought stress conditions.
Morphogenetic potential of seed and explants culturing of three capsicum annuum L. ( pepper) geno... more Morphogenetic potential of seed and explants culturing of three capsicum annuum L. ( pepper) genotypes (Gedion, Moaz, and Mohand) were studied to evaluate different plant regeneration protocols and develop a reliable system for plant propagation. An efficient procedure of in vitro plant regeneration through seed and direct shoot bud induction was tested from different explants of capsicum annuum L. Several methods of media preparation with combinations of growth regulators were used, and 2 combinations were found ideal for seed propagation with 7 mg/l 2,4-D or 5 mg/l IBA respectively. Additionally, it was indicated that seed dormancy can effect its propagation also the positioning of the seed or explants onto the media. Propagated seeds demonstrated shoot and root elongation. Regeneration of the explants did not show satisfactory results because most of the explants did not develop into normal shoots but instead developed into calli after 15 days of culture. HPLC Analysis of cultivars demonstrated that fructose suger percentage was higher in the three different types of pepper, followed by sucrose then glucose. Also the HPLC analysis drew out that media supplemented with 7mg/l 2,4-D generally had the highest effect of capsaicinoid sugar content in the following order i.e. cultivar Mohand was showed high significant in sugar content of capsaicinoid followed by Gedion and Moaz. Application of DPPH method illustrated that extracts obtained from Moaz leaves characterized the most proactive antioxidant (vitamin C) properties than Gedion and Mohand. A were noticed. The results of these studies indicated that antioxidant activities of the extract prepared from pepper leaves depended mainly on phenolic compounds.
The objective of this investigation was to assess the genetic diversity among three Egyptian duru... more The objective of this investigation was to assess the genetic diversity among three Egyptian durum wheat i.e. Beni Suif 4, Beni Suif 5 and Beni Suif 6 and one bread wheat i.e. Sids 12 cultivars using sodium dodecyle sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and randomly amplified polymorphic (RAPD) markers. Protein electrophoreses showed that Beni Suif 4 was characterized by the absence of band-3 with 41.56 kDa. RAPD analysis showed that the number of polymorphic amplicons was 56 out of a total of 93 amplicons, thus revealing a level of 60.0 % polymorphism. The highest genetic similarity revealed by RAPD analysis (95.0%) was between Beni Suif 5 and Beni suif 6. While, lowest similarity (65 %) was found between Beni Suif 4 and Beni Suif 5. The dendrogram separated Sids 12 from all the other genotypes, thus demonstrating the distinctiveness of the genetic background of this genotype from all the other genotypes. The three genotypes constituted a subcluster divided into two groups, one group composed of Beni Suif 5 and Beni Suif 6 , while the second group comprised Beni Suif 4.
The main objective of this investigation was to assess the genetic diversity among five Egyptian
... more The main objective of this investigation was to assess the genetic diversity among five Egyptian bread wheat genotypes (Misr1 and Sids 13 cutlivars and the promising lines No.1, No.2 and No.11 ) using SDS-PAGE and RAPD markers. The total number of SDS- bands was seven. Six SDS- bands were monomorphic while the other was polymorphic. Line-1 was characterized by the presence of band -3 with a molecular weight of 41.56 kDa. RAPD analysis showed that the number of polymorphic amplicons was 66 out of a total of 93 amplicons, thus revealing a level of 70.97 % polymorphism. The highest genetic similarity revealed by RAPD analysis (93.1%) was between Misr1 and Line 2 genotype. While, the lowest similarity (85.2 %) was between Line 1 and Line 2. The dendrogram separated Line1 from all the other genotypes. The four genotypes constituted a subcluster divided into two groups, one group composed of Misr 1 and Line 2, while the second group comprised Sids 13 and Line 11.
Carboplatin is a synthetic antineoplastic agent used for cancer treatment and is considered to
be... more Carboplatin is a synthetic antineoplastic agent used for cancer treatment and is considered to be the analogous of cisplatin. Nigella sativa oil is a herbaceous plant, it has been used for thousands of years for culinary and medical purposes. This study aimed to evaluate the effect of carboplatin and Nigella sativa oil alone and in combination together on human breast cancer cells (MCF-7) in vitro and Ehrlich ascites tumor bearing female mice (in vivo). The in vitro experiment on MCF-7 cells illustrated that IC50 of carboplatin was 11.8 μg/ml, also IC50 of Nigella sativa oil was 39 μg/ml on MCF-7 cells. In addition, IC50 of the combination between carboplatin and Nigella sativa oil was found to be 3.78 and 40 μg/ml, respectively. The in vivo experiment illustrated that carboplatin (10mg/kg) increased the enzyme activity of aspartate amino transferase (GOT) and aniline amino transferase (GPT)by 56.52% and 51.14%, respectively as compared to both healthy control (nontumor transplanted mice) and negative control. Also, the activity of GOT and GPT was increased by 14.75% and 19.84%, respectively as compared to healthy control under the effect of Nigella sativa oil (12ml/kg). While the activity of GOT and GPT was decreased as compared to negative control. The combination of carboplatin and Nigella sativa oil appeared to increase the enzyme activity of GOT and GPT by 62.41 and 49.39%, respectively as compared to both healthy control and negative control. Also, carboplatin induced DNA damage of liver tissue was performed by agarose gel electrophoresis and comet assay, while Nigella sativa oil showed intact DNA without any damage. The combination of carboplatin and Nigella sativa oil appeared to decrease the DNA damage as compared to carboplatin alone.
Squash is considered as one of the most important crops in Egypt and worldwide. Zucchini
yellow m... more Squash is considered as one of the most important crops in Egypt and worldwide. Zucchini yellow mosaic potyvirus (ZYMV) is considered as one of the most important viruses infects squash. We used the biological, serological and cytopathological studies to identify an Egyptian isolate of ZYMV. Results showed that the Egyptian isolate of ZYMV showed systemic symptoms in the form of severe mosaic and vein banding on Cucurbita pepo cv. Eskandarani under open field conditions. Direct antigen coated (DAC)-enzyme-linked immunosorbent assay (ELISA) using polyclonal antibodies was used as a diagnostic tool for detecting ZYMV in the virus-infected C. pepo samples. It showed positive reactions with ZYMV antiserum. Filamentous virus-like particles measuring 750X13 nm was successfully purified from ZYMV-infected squash plants, based on the use of polyethylene glycol and ultracentrifugation. The electron microscope of ultrathin sections of virusinfected leaf tissues revealed the presence of cylindrical inclusions as pinwheels, laminated aggregates and scrolls in the cytoplasm of cells infected with ZYMV. In addition, disorganization of plastids, nucleus and mitochondria in the virus-cells infected was also observed. In addition, the nucleotide of CI gene of the viral isolate under investigation was determined and its similarities to some overseas isolates were addressed.
The present investigation studied the effect of amino acids (Glutamine, Spermidine and Asparagine... more The present investigation studied the effect of amino acids (Glutamine, Spermidine and Asparagine) with different concentration (50, 250.500 mg/l) used as precursors to produce secondary metabolites (steroids) and growth development during different stages (callus, embryoids and shooting) of date palm (Malakaby cv.). In Embryogenic callus stage, callus volume was the highest (4.00) when treated with any of the three amino acids, 50 mg/l of Glutamine or Asparagine showed no effect compared to the control giving the lowest callus volume (3.00). Total steroids in callus tissues clearly showed that using Glutamine 250 mg/l in medium gave the highest steroid content 0.662 mg/g and percentage (336% of control), while the lowest (0.111mg/g) was found with Asparagine (500mg/l) and 56.35% of control. Glutamine at 250 mg/l resulted in the highest weight of embryos (2.100 gm). As well as, 500 mg/l Spermidine seemed to be the best amino acid used in order to stimulate steroid biosynthesis resulting in 202.1% of control (0.782 mg/g). In shooting stage, according to the number of shoots, the highest number of shoots (2.33) was achieved with Glutamine and Spermidine at 500 mg/l. In shoot weight, the highest weight (7.267 gm) was achieved by using 500mg/l Glutamine , as to steroid biosynthesis in shooting stage, the best result obtained, were by using Glutamine at 500 mg/l which gave highest steroid biosynthesis (0.534mg/g), 206.0% of control.
This study was aimed to investigate the genotoxicity and hepatotoxicity of the anticancer
drugs “... more This study was aimed to investigate the genotoxicity and hepatotoxicity of the anticancer drugs “cis-diamminedichloro platinum (CDDP) (8 mg/kg b.w.) and Doxorubicin (DOX) (10 mg/kg b.w)” alone and the protective action of Curcuma longa (Turmeric) (250 mg/kg b.w.) in combination with CDDP and DOX in Ehrlich ascites tumor bearing male mice. The results illustrated that the effect of CDDP and DOX alone decreased total protein, albumin and globulin concentrations in serum. Also, there are no significant changes in serum protein banding patterns between treatments on SDS-PAGE. On the other hand, the lysosomal enzymaic activities of β- Nacetyl glucosaminidase (β-NAG) and acid phosphatase (ACP) as well as glutathione-S-transferase (GST) in liver homogentae were increased. Total protein content in liver homogenate was reduced as compared to the control. The percentage of structural aberrations of chromosomes of bone marrow cells after 24 hours of drugs administration were increased by 261.0% and 149.1% for CDDP and DOX, respectively . On the other hand, the turmeric extract were increased in mixture with each drug resulted in increasing the total protein, albumin and globulin concentrations in serum. Also, the enzymes activity of ACP, β-NAG and GST were decreased. Total protein contents was increased in liver homogenate as compared to the control. In addition to, the percentage of structural and numerical aberrations of chromosomes of bone marrow cells after 24 h of curcumin administration were decreased, CDDP and DOX showed damage and smear bands of genomic DNA as compared with negative control group, while the mixture of turmeric showed less damage of genomic DNA bands.
Drought stress is one of the most important environmental factors reducing cereal yields. The gen... more Drought stress is one of the most important environmental factors reducing cereal yields. The genetic and physiological bases of drought tolerance in durum wheat was investigated by QTL mapping and by mapping candidate genes derived from differentially expressed genes and previous investigations. A recombinant inbred population derived from a cross between two durum (Triticum turgidum L. var durum) parents Jennah Khetifa and Cham1 that exhibit contrasting traits for drought tolerance was employed. Twelve known genes and 103 differentially expressed sequence tags (dESTs) were surveyed and 55 detected polymorphism between the two parental lines. In total, 162 loci including 6 known genes, 37 dESTs, and an additional 119 markers that were unlinked in the previous map have been merged with 306 previously mapped markers to produce a new map with 468 loci. Single point analysis and composite interval mapping were used to identify the genomic regions controlling traits related to drought stress. Significant QTL were identified for canopy temperature, photosynthesis-related parameters and water status index. One hundred and seventy eight markers, including 6 candidate genes and 19 differentially expressed sequences were associated with QTL for drought tolerance traits. The results indicate that there is considerable potential for improving drought tolerance of durum wheat by using marker-assisted selection.
L-Arginase, hydrolyzing L-arginine to L-ornithine
and urea, is a powerful anticancer, L-arginine-... more L-Arginase, hydrolyzing L-arginine to L-ornithine and urea, is a powerful anticancer, L-arginine-depleting agent, against argininosuccinate synthase expressing tumors. Otherwise, the higher antigenicity and lower thermal stability of this enzyme was the main biochemical hurdles. Since, the intrinsic thermal stability of enzymes follow the physiological temperature of their producer, thus, characterization of Larginase from thermotolerant Penicillium chrysogenum was the objective of this study. L-Arginase (Arg) was purified to its homogeneity from P. chrysogenum by 10.1-fold, with 37.0 kDa under denaturing PAGE, optimum reaction at 50 C, pH stability (6.8–7.9), with highest molar ratio of constitutional arginine, glutamic acid, lysine and aspartic acid. The purified enzyme was PEGylated and immobilized on chitosan, with 41.9 and 22.1 % yield of immobilization. At 40 C, the T1/2 value of free-Arg, PEG-Arg and Chit-Arg was 10.4, 15.6, 20.5 h, respectively. The free-Arg and Chit- Arg have a higher affinity to L-arginine (Km 4.8 mM), while, PEG-Arg affinity was decreased by about 3 fold (Km 15.2 mM). The inhibitory constants to the free and PEG-Arg were relatively similar towards HA and PPG. The IC50 for the free enzyme against HEPG-2 and A549 tumor cells was 0.136 and 0.165 U/ml, comparing to 0.232 and 0.496 U/ml for PEG-Arg, respectively. The in vivo T1/2 to the free Arg and PEG-Arg was 16.4 and 20.4 h, respectively as holoenzyme. The residual L-arginine level upon using free Arg was 156.9 and 144.5 lM, after 6 and 8 h, respectively, regarding to initials at 253.6 lM, while for Peg-Arg the level of L-arginine was nil till 7 h of initial dosing. The titer of IgG was induced by 10–15 % in response to free-Arg after 28 days comparing to IgG titer for PEG-Arg.
Durum wheat (Triticum turgidum ssp. durum) is an economically and nutritionally important cereal ... more Durum wheat (Triticum turgidum ssp. durum) is an economically and nutritionally important cereal crop in the Mediterranean region and its production is largely influenced by environmental stresses, such as drought, salinity, heat and nutrient deficiency. The objective of this study was to dissect quantitative trait loci (QTL) controlling grain yield, yield components and drought tolerance in durum. A molecular genetic linkage map for F2 durum mapping population derived from an intraspecific cross between Baniswif-1 x Sohag-2 was constructed using 114 DNA markers (9 SSRs, 14 SCoTs, 90 AFLPs and 1 RAPDs) distributed over the 14 linkage groups and spanning 2040.9 cM of the durum wheat genome. The size of linkage groups varied greatly from 6.8cM for LG11 to 317.5cM for LG4 with an average length of 145.8cM. Based on the used anchor SSR markers, only eight linkage groups were assigned to chromosomes, where LG1, LG3, LG5, LG6, LG7, LG9, LG13 and LG14 were assigned to chromosomes 1B, 3B, 5B, 6A, 6B, 7A, 3A and 2B, respectively. Single point analysis was used to identify genomic regions controlling eleven morpho-physiological traits related to grain yield, yield components and drought tolerance. A total of 74 QTL were identified for the eleven traits on all linkage groups except (LG10 and LG11). These included 3 QTL for root length (RL), 11 QTL for plant height (PH), 7 QTL for spike length (SL), 3 QTL for number of branches/plant (NBP), 3 QTL for number of spike/plant (NSP), 8 QTL for number of spikelets/spike (NSS), 15 QTL for number of kernel/spike (NKS), 10 QTL for thousand-kernel weight (TKW), 4 QTL for fresh weight (FW), 5 QTL for dry weight (DW) and 5 QTL for total amino acids (TAA). This work represents the first genetic linkage map for durum wheat population derived from an intraspecific cross between ‗Baniswif-1‘ and ‗Sohag-2‘ showing chromosomal regions associated with 11 morpho-physiological traits related to grain yield, yield components and drought tolerance in durum wheat.
Molecular markers provide a rapid approach to breeding for desired traits. To use them, it is nec... more Molecular markers provide a rapid approach to breeding for desired traits. To use them, it is necessary to determine the linkage between quantitative trait loci (QTL) and such markers. This study was conducted to investigate the genetic basis of salinity responses in Egyptian bread wheat (Triticum aestivum L.). In this context, a doubled haploid (DH) population (SGDH) of 139 individuals was produced from the cross between two Egyptian breeding cultivars (Sakha 93, salt tolerant) and (Gemmeza 7, salt susceptible). The DH population was tested under saline hydroponics culture and various plant responses were measured. A molecular genetic map of the SGDH population covering 3645.3 cM, was constructed using Restriction Fragment Length Polymorphism (RFLP), Microsatellite or Simple Sequence Repeats (SSR), and Amplified Fragment Length Polymorphism (AFLP) markers. In total, 325 loci along the 21 wheat chromosomes were mapped. The B genome showed the highest number of mapped markers followed by the A and the D genomes respectively. Interval and composite interval mapping (using QTL cartographer) were used to identify the genomic regions controlling traits related to salt tolerance with a threshold of LOD 3. Analysis of QTLs has revealed the approximate location of the significant markers associated with 12 traits related to salt tolerance traits across the A, B and D genomes. Fifty five significant QTL were detected on 15 of the 21 chromosomes mapped in this study, for some of these more than one QTL was identified. In many cases QTL were mapped very close to each other, indicating possible gene clustering or pleiotropy. For some traits, the total percentage of phenotypic variation explained by all QTL exceeded 50-60% but on average it was in the region of 15%. Chromosomes of homologues groups 2 and 5 exerted the biggest effect on most phenotypic traits, especially chromosomes 2B and 5B. The results indicated that there is considerable potential for improving salt tolerance of hexaploid wheat by using marker-assisted selection.
Net blotch, caused by Pyrenophora teres f. teres, is one of the most devastating diseases causing... more Net blotch, caused by Pyrenophora teres f. teres, is one of the most devastating diseases causing significant losses in barley yield and quality. In the present investigation, two barley verities “AT4” (net blotch resistant) and “Femina” (net blotch susceptible) were used to develop a segregating F2 population. Linkage analysis and map construction were performed using Map Manager. The constructed genetic linkage map consisted of 85 markers including 45 AFLP , 11 SSR, 6 CAPS, 5 SCoT, 3 STS and 6 NBL. Linkage groups were assigned to individual barley chromosomes using the published map locations of the SSR markers as reference point. The produced map showed 7 linkage groups with 85 markers covered a total length of 1644.8 cM. The average length of linkage groups ranged from 77.7 to 739.4. Single point analysis was used to identify the genomic regions associated with net blotch resistance in barley. A total of 14 QTL with a significance ranging from 0.01% to 5% were identified on 4 linkage groups (2, 4, 5 and 6). The most significant QTL was found on chromosome 6H. This QTL presents a promising opportunity for the strategic improvement of barley resistance to net blotch using marker assisted selection.
Trehalose is a non-reducing disaccharide which consists of two glucose units that functions as a ... more Trehalose is a non-reducing disaccharide which consists of two glucose units that functions as a compatible solute to stabilize the membrane structures under heat and desiccation stress. Trehalose-6-phosphate synthase (TPS) and trehalose-6- phosphate phosphatase (TPP) are the key enzymes for trehalose biosynthesize in the plant kingdom. On the basis of bioinformatics prediction, fragment containing an open reading frame of 945 bp was cloned from durum wheat. Sequence comparison and analysis of conserved domains revealed the presence of a TPP domain. Full length of the gene was isolated using gene race technology. Semi-quantitative RT-PCR and real time quantitative PCR indicated that the expression of this gene is up-regulated in response to drought stress. The biochemical assay of the trehalase activity showed that the enzyme's activity decreased under the dehydration stress. The obtained phylogenic tree showed that the isolated TPP protein forms a distinct clad close to the Oryza sativa trehalose-6- phosphate phosphatase. In silico and comparative mapping indicated that the isolated TPP gene is localized on rice chromosome 8, durum wheat chromosome 20, bread wheat chromosome 3B, oat linkage group E, sorghum chromosome 4 and barley 5H.
Cotton is the world’s leading natural fiber and second most important oilseed crop and has been a... more Cotton is the world’s leading natural fiber and second most important oilseed crop and has been a focus of genetic, systematic and breeding research. The genetic and physiological bases of some important agronomic traits in cotton were investigated by QTL mapping through constructing of genetic map with chromosomal assignment. A segregating F2 population derived from an interspecific cross (G. barbadense x G. hirsutum) between two genotypes, cvs. “Giza 83” and “Deltapine” was used in this study. Different molecular markers including SS R, ES T, ES T-SS R, AFLP and RAPD were employed to identify markers that reveal differences between the parents. In total 42 new markers were merged with 140 previously mapped markers to produce a new map with 182 loci covering a total length of 2370.5 cM. Among these new markers, some of them were used to assign chromosomes to the produced 26 linkage groups. The LG2, LG3, LG11 and LG26 were assigned to chromosomes 1, 6, 5 and 20 respectively. Single point analysis was used to identify genomic regions controlling traits for plant height, number of nodes at flowering time, bolling date, days to flowering and number of bolls. In total 40 significant QTL were identified for the five traits on 11 linkage groups (1, 2, 3, 4, 5, 10, 11, 12, 18, 19 and 23). This work represents an improvement of the previously constructed genetic map in addition to chromosomal assignment and detection of new significant QTL for the five traits in Egyptian cotton. The Significant QTLs detected in this study can be employed in marker assisted selection for molecular breeding programs aiming at developing cotton cultivars with improved agronomic traits
Efficient regeneration methods are a priority for the successful application of genetic engineeri... more Efficient regeneration methods are a priority for the successful application of genetic engineering to vegetatively propagated plants such as grape (Vitis vinifera L.). An efficient in vitro shoot micropropagation protocol leading to the regeneration from shoot tip explants was successfully developed for mass production. Callus was induced from shoot tips and axillary buds on C2D medium supplemented with 4 mg/l 6-benzyladenine (BA). Bud differentiation from callus was possible on C2D medium supplemented with 1 mg/l gibberellic acid and 0.1 mg/l kinetin. Bud initials were elongated by sub-culturing on elongation medium containing C2D salts supplemented with 0.1 mg/l BA and resulting shoots could be mass micropropagated on C2D medium supplemented with 1 mg/l BA. Shoots that were regenerated via organogenesis or micropropagation were rooted on C2D medium supplemented with 1 mg/l indole-3-butyric acid. In this study, regeneration and micropropagation of grape cv. ‘Spero’ was successfully achieved through shoot tip and axillary bud culture. Such a protocol would be useful for future use in the strategic improvement of grape using genetic engineering
Banana is one of the most important fruit crop worldwide, as its production reaches approximately... more Banana is one of the most important fruit crop worldwide, as its production reaches approximately 70 million tones per year. By tissue culture, triploid and seedless bananas can now be produced in a high quantity, in a less time and at any time of the year without being limited to a season. The aim of this study is to substitute the artificial hormones used in the tissue culture such as, cytokinins and auxins by natural materials such as, Pineapple, Coconut milk, Cacao, Coffee, and Charcoal. Five different media composed of these natural materials were prepared in different concentrations, Pineapple (5,10 and 20cm), Coconut milk (5,10 and20cm,), Cacao (1,2.5 and 5 g), Coffee (1,2.5 and 5 g) and Charcoal (1,2.5 and 5 g) to study their effect on the development and browning of the bananas' shoots and roots in vitro, with the aid of various statistical analyses. The results indicated that the pineapple and coconut milk produced the highest number and length of both shoots and roots, as well as the soluble sugars contents, while the cocoa and coconut milk showed the highest protein content. The charcoal reduced significantly the phenolic content. On the other hand, there was no significant difference between the coffee, cacao. This study has highlighted the benefits and advantages of using natural materials in producing high quality banana.
Drought is a major planetary stress problem that places greater limits on cereal productivity. Cr... more Drought is a major planetary stress problem that places greater limits on cereal productivity. Crop species belonging to the tribe Triticale represent the main foodstuff sources for humans. Because durum wheat (Triticum turgidum L.) shows better adaptation to semi-arid climates than bread wheat, its genome should contain a pool of genes that are expressed for wide environmental adaptability. To investigate and understand the genetic response of drought tolerance in durum wheat, differentially expressed sequence tags (dESTs) were detected, characterized and cloned from durum wheat under different regimes of drought stress and the functions of some of these dESTs were determined. In this study, genes involved in drought stress at early stages of growth were profiled by a differential display technique from Egyptian durum wheat (Sohag 3). One, three, six, and 10 hours of drought stress were applied to produce 15 expressed cDNA fragments. This will highlight the genes that are expressed under drought stress, which might help in the understanding of the plant’s defense mechanism under drought conditions. This will also give a chance to identify gene(s) for the future use in transgenic research to enhance drought tolerance in wheat and other commercially important crops. The cloned dESTs might also be used as molecular markers for genetic and QTL mapping for wheat and other cereals under drought stress conditions.
Drought is a major planetary stress problem that places greater limits on cereal productivity. Cr... more Drought is a major planetary stress problem that places greater limits on cereal productivity. Crop species belonging to the tribe Triticale represent the main foodstuff sources for humans. Because durum wheat (Triticum turgidum L.) shows better adaptation to semi-arid climates than bread wheat, its genome should contain a pool of genes that are expressed for wide environmental adaptability. To investigate and understand the genetic response of drought tolerance in durum wheat, differentially expressed sequence tags (dESTs) were detected, characterized and cloned from durum wheat under different regimes of drought stress and the functions of some of these dESTs were determined. In this study, genes involved in drought stress at early stages of growth were profiled by a differential display technique from Egyptian durum wheat (Sohag 3). One, three, six, and 10 hours of drought stress were applied to produce 15 expressed cDNA fragments. This will highlight the genes that are expressed under drought stress, which might help in the understanding of the plant’s defense mechanism under drought conditions. This will also give a chance to identify gene(s) for the future use in transgenic research to enhance drought tolerance in wheat and other commercially important crops. The cloned dESTs might also be used as molecular markers for genetic and QTL mapping for wheat and other cereals under drought stress conditions.
Morphogenetic potential of seed and explants culturing of three capsicum annuum L. ( pepper) geno... more Morphogenetic potential of seed and explants culturing of three capsicum annuum L. ( pepper) genotypes (Gedion, Moaz, and Mohand) were studied to evaluate different plant regeneration protocols and develop a reliable system for plant propagation. An efficient procedure of in vitro plant regeneration through seed and direct shoot bud induction was tested from different explants of capsicum annuum L. Several methods of media preparation with combinations of growth regulators were used, and 2 combinations were found ideal for seed propagation with 7 mg/l 2,4-D or 5 mg/l IBA respectively. Additionally, it was indicated that seed dormancy can effect its propagation also the positioning of the seed or explants onto the media. Propagated seeds demonstrated shoot and root elongation. Regeneration of the explants did not show satisfactory results because most of the explants did not develop into normal shoots but instead developed into calli after 15 days of culture. HPLC Analysis of cultivars demonstrated that fructose suger percentage was higher in the three different types of pepper, followed by sucrose then glucose. Also the HPLC analysis drew out that media supplemented with 7mg/l 2,4-D generally had the highest effect of capsaicinoid sugar content in the following order i.e. cultivar Mohand was showed high significant in sugar content of capsaicinoid followed by Gedion and Moaz. Application of DPPH method illustrated that extracts obtained from Moaz leaves characterized the most proactive antioxidant (vitamin C) properties than Gedion and Mohand. A were noticed. The results of these studies indicated that antioxidant activities of the extract prepared from pepper leaves depended mainly on phenolic compounds.
The objective of this investigation was to assess the genetic diversity among three Egyptian duru... more The objective of this investigation was to assess the genetic diversity among three Egyptian durum wheat i.e. Beni Suif 4, Beni Suif 5 and Beni Suif 6 and one bread wheat i.e. Sids 12 cultivars using sodium dodecyle sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and randomly amplified polymorphic (RAPD) markers. Protein electrophoreses showed that Beni Suif 4 was characterized by the absence of band-3 with 41.56 kDa. RAPD analysis showed that the number of polymorphic amplicons was 56 out of a total of 93 amplicons, thus revealing a level of 60.0 % polymorphism. The highest genetic similarity revealed by RAPD analysis (95.0%) was between Beni Suif 5 and Beni suif 6. While, lowest similarity (65 %) was found between Beni Suif 4 and Beni Suif 5. The dendrogram separated Sids 12 from all the other genotypes, thus demonstrating the distinctiveness of the genetic background of this genotype from all the other genotypes. The three genotypes constituted a subcluster divided into two groups, one group composed of Beni Suif 5 and Beni Suif 6 , while the second group comprised Beni Suif 4.
The main objective of this investigation was to assess the genetic diversity among five Egyptian
... more The main objective of this investigation was to assess the genetic diversity among five Egyptian bread wheat genotypes (Misr1 and Sids 13 cutlivars and the promising lines No.1, No.2 and No.11 ) using SDS-PAGE and RAPD markers. The total number of SDS- bands was seven. Six SDS- bands were monomorphic while the other was polymorphic. Line-1 was characterized by the presence of band -3 with a molecular weight of 41.56 kDa. RAPD analysis showed that the number of polymorphic amplicons was 66 out of a total of 93 amplicons, thus revealing a level of 70.97 % polymorphism. The highest genetic similarity revealed by RAPD analysis (93.1%) was between Misr1 and Line 2 genotype. While, the lowest similarity (85.2 %) was between Line 1 and Line 2. The dendrogram separated Line1 from all the other genotypes. The four genotypes constituted a subcluster divided into two groups, one group composed of Misr 1 and Line 2, while the second group comprised Sids 13 and Line 11.
Carboplatin is a synthetic antineoplastic agent used for cancer treatment and is considered to
be... more Carboplatin is a synthetic antineoplastic agent used for cancer treatment and is considered to be the analogous of cisplatin. Nigella sativa oil is a herbaceous plant, it has been used for thousands of years for culinary and medical purposes. This study aimed to evaluate the effect of carboplatin and Nigella sativa oil alone and in combination together on human breast cancer cells (MCF-7) in vitro and Ehrlich ascites tumor bearing female mice (in vivo). The in vitro experiment on MCF-7 cells illustrated that IC50 of carboplatin was 11.8 μg/ml, also IC50 of Nigella sativa oil was 39 μg/ml on MCF-7 cells. In addition, IC50 of the combination between carboplatin and Nigella sativa oil was found to be 3.78 and 40 μg/ml, respectively. The in vivo experiment illustrated that carboplatin (10mg/kg) increased the enzyme activity of aspartate amino transferase (GOT) and aniline amino transferase (GPT)by 56.52% and 51.14%, respectively as compared to both healthy control (nontumor transplanted mice) and negative control. Also, the activity of GOT and GPT was increased by 14.75% and 19.84%, respectively as compared to healthy control under the effect of Nigella sativa oil (12ml/kg). While the activity of GOT and GPT was decreased as compared to negative control. The combination of carboplatin and Nigella sativa oil appeared to increase the enzyme activity of GOT and GPT by 62.41 and 49.39%, respectively as compared to both healthy control and negative control. Also, carboplatin induced DNA damage of liver tissue was performed by agarose gel electrophoresis and comet assay, while Nigella sativa oil showed intact DNA without any damage. The combination of carboplatin and Nigella sativa oil appeared to decrease the DNA damage as compared to carboplatin alone.
Squash is considered as one of the most important crops in Egypt and worldwide. Zucchini
yellow m... more Squash is considered as one of the most important crops in Egypt and worldwide. Zucchini yellow mosaic potyvirus (ZYMV) is considered as one of the most important viruses infects squash. We used the biological, serological and cytopathological studies to identify an Egyptian isolate of ZYMV. Results showed that the Egyptian isolate of ZYMV showed systemic symptoms in the form of severe mosaic and vein banding on Cucurbita pepo cv. Eskandarani under open field conditions. Direct antigen coated (DAC)-enzyme-linked immunosorbent assay (ELISA) using polyclonal antibodies was used as a diagnostic tool for detecting ZYMV in the virus-infected C. pepo samples. It showed positive reactions with ZYMV antiserum. Filamentous virus-like particles measuring 750X13 nm was successfully purified from ZYMV-infected squash plants, based on the use of polyethylene glycol and ultracentrifugation. The electron microscope of ultrathin sections of virusinfected leaf tissues revealed the presence of cylindrical inclusions as pinwheels, laminated aggregates and scrolls in the cytoplasm of cells infected with ZYMV. In addition, disorganization of plastids, nucleus and mitochondria in the virus-cells infected was also observed. In addition, the nucleotide of CI gene of the viral isolate under investigation was determined and its similarities to some overseas isolates were addressed.
The present investigation studied the effect of amino acids (Glutamine, Spermidine and Asparagine... more The present investigation studied the effect of amino acids (Glutamine, Spermidine and Asparagine) with different concentration (50, 250.500 mg/l) used as precursors to produce secondary metabolites (steroids) and growth development during different stages (callus, embryoids and shooting) of date palm (Malakaby cv.). In Embryogenic callus stage, callus volume was the highest (4.00) when treated with any of the three amino acids, 50 mg/l of Glutamine or Asparagine showed no effect compared to the control giving the lowest callus volume (3.00). Total steroids in callus tissues clearly showed that using Glutamine 250 mg/l in medium gave the highest steroid content 0.662 mg/g and percentage (336% of control), while the lowest (0.111mg/g) was found with Asparagine (500mg/l) and 56.35% of control. Glutamine at 250 mg/l resulted in the highest weight of embryos (2.100 gm). As well as, 500 mg/l Spermidine seemed to be the best amino acid used in order to stimulate steroid biosynthesis resulting in 202.1% of control (0.782 mg/g). In shooting stage, according to the number of shoots, the highest number of shoots (2.33) was achieved with Glutamine and Spermidine at 500 mg/l. In shoot weight, the highest weight (7.267 gm) was achieved by using 500mg/l Glutamine , as to steroid biosynthesis in shooting stage, the best result obtained, were by using Glutamine at 500 mg/l which gave highest steroid biosynthesis (0.534mg/g), 206.0% of control.
This study was aimed to investigate the genotoxicity and hepatotoxicity of the anticancer
drugs “... more This study was aimed to investigate the genotoxicity and hepatotoxicity of the anticancer drugs “cis-diamminedichloro platinum (CDDP) (8 mg/kg b.w.) and Doxorubicin (DOX) (10 mg/kg b.w)” alone and the protective action of Curcuma longa (Turmeric) (250 mg/kg b.w.) in combination with CDDP and DOX in Ehrlich ascites tumor bearing male mice. The results illustrated that the effect of CDDP and DOX alone decreased total protein, albumin and globulin concentrations in serum. Also, there are no significant changes in serum protein banding patterns between treatments on SDS-PAGE. On the other hand, the lysosomal enzymaic activities of β- Nacetyl glucosaminidase (β-NAG) and acid phosphatase (ACP) as well as glutathione-S-transferase (GST) in liver homogentae were increased. Total protein content in liver homogenate was reduced as compared to the control. The percentage of structural aberrations of chromosomes of bone marrow cells after 24 hours of drugs administration were increased by 261.0% and 149.1% for CDDP and DOX, respectively . On the other hand, the turmeric extract were increased in mixture with each drug resulted in increasing the total protein, albumin and globulin concentrations in serum. Also, the enzymes activity of ACP, β-NAG and GST were decreased. Total protein contents was increased in liver homogenate as compared to the control. In addition to, the percentage of structural and numerical aberrations of chromosomes of bone marrow cells after 24 h of curcumin administration were decreased, CDDP and DOX showed damage and smear bands of genomic DNA as compared with negative control group, while the mixture of turmeric showed less damage of genomic DNA bands.
Drought stress is one of the most important environmental factors reducing cereal yields. The gen... more Drought stress is one of the most important environmental factors reducing cereal yields. The genetic and physiological bases of drought tolerance in durum wheat was investigated by QTL mapping and by mapping candidate genes derived from differentially expressed genes and previous investigations. A recombinant inbred population derived from a cross between two durum (Triticum turgidum L. var durum) parents Jennah Khetifa and Cham1 that exhibit contrasting traits for drought tolerance was employed. Twelve known genes and 103 differentially expressed sequence tags (dESTs) were surveyed and 55 detected polymorphism between the two parental lines. In total, 162 loci including 6 known genes, 37 dESTs, and an additional 119 markers that were unlinked in the previous map have been merged with 306 previously mapped markers to produce a new map with 468 loci. Single point analysis and composite interval mapping were used to identify the genomic regions controlling traits related to drought stress. Significant QTL were identified for canopy temperature, photosynthesis-related parameters and water status index. One hundred and seventy eight markers, including 6 candidate genes and 19 differentially expressed sequences were associated with QTL for drought tolerance traits. The results indicate that there is considerable potential for improving drought tolerance of durum wheat by using marker-assisted selection.
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Papers by Ayman Diab
and urea, is a powerful anticancer, L-arginine-depleting
agent, against argininosuccinate synthase expressing tumors.
Otherwise, the higher antigenicity and lower thermal stability
of this enzyme was the main biochemical hurdles. Since, the
intrinsic thermal stability of enzymes follow the physiological
temperature of their producer, thus, characterization of Larginase
from thermotolerant Penicillium chrysogenum was
the objective of this study. L-Arginase (Arg) was purified to
its homogeneity from P. chrysogenum by 10.1-fold, with
37.0 kDa under denaturing PAGE, optimum reaction at
50 C, pH stability (6.8–7.9), with highest molar ratio of
constitutional arginine, glutamic acid, lysine and aspartic
acid. The purified enzyme was PEGylated and immobilized
on chitosan, with 41.9 and 22.1 % yield of immobilization.
At 40 C, the T1/2 value of free-Arg, PEG-Arg and Chit-Arg
was 10.4, 15.6, 20.5 h, respectively. The free-Arg and Chit-
Arg have a higher affinity to L-arginine (Km 4.8 mM), while,
PEG-Arg affinity was decreased by about 3 fold (Km
15.2 mM). The inhibitory constants to the free and PEG-Arg
were relatively similar towards HA and PPG. The IC50 for
the free enzyme against HEPG-2 and A549 tumor cells was
0.136 and 0.165 U/ml, comparing to 0.232 and 0.496 U/ml
for PEG-Arg, respectively. The in vivo T1/2 to the free Arg and PEG-Arg was 16.4 and 20.4 h, respectively as holoenzyme.
The residual L-arginine level upon using free Arg
was 156.9 and 144.5 lM, after 6 and 8 h, respectively,
regarding to initials at 253.6 lM, while for Peg-Arg the level
of L-arginine was nil till 7 h of initial dosing. The titer of IgG
was induced by 10–15 % in response to free-Arg after
28 days comparing to IgG titer for PEG-Arg.
Mediterranean region and its production is largely influenced by environmental stresses, such as drought, salinity, heat and
nutrient deficiency. The objective of this study was to dissect quantitative trait loci (QTL) controlling grain yield, yield
components and drought tolerance in durum. A molecular genetic linkage map for F2 durum mapping population derived
from an intraspecific cross between Baniswif-1 x Sohag-2 was constructed using 114 DNA markers (9 SSRs, 14 SCoTs, 90
AFLPs and 1 RAPDs) distributed over the 14 linkage groups and spanning 2040.9 cM of the durum wheat genome. The
size of linkage groups varied greatly from 6.8cM for LG11 to 317.5cM for LG4 with an average length of 145.8cM. Based
on the used anchor SSR markers, only eight linkage groups were assigned to chromosomes, where LG1, LG3, LG5, LG6,
LG7, LG9, LG13 and LG14 were assigned to chromosomes 1B, 3B, 5B, 6A, 6B, 7A, 3A and 2B, respectively. Single point
analysis was used to identify genomic regions controlling eleven morpho-physiological traits related to grain yield, yield
components and drought tolerance. A total of 74 QTL were identified for the eleven traits on all linkage groups except
(LG10 and LG11). These included 3 QTL for root length (RL), 11 QTL for plant height (PH), 7 QTL for spike length (SL),
3 QTL for number of branches/plant (NBP), 3 QTL for number of spike/plant (NSP), 8 QTL for number of spikelets/spike
(NSS), 15 QTL for number of kernel/spike (NKS), 10 QTL for thousand-kernel weight (TKW), 4 QTL for fresh weight
(FW), 5 QTL for dry weight (DW) and 5 QTL for total amino acids (TAA). This work represents the first genetic linkage
map for durum wheat population derived from an intraspecific cross between ‗Baniswif-1‘ and ‗Sohag-2‘ showing
chromosomal regions associated with 11 morpho-physiological traits related to grain yield, yield components and drought
tolerance in durum wheat.
determine the linkage between quantitative trait loci (QTL) and such markers. This study was conducted to investigate the
genetic basis of salinity responses in Egyptian bread wheat (Triticum aestivum L.). In this context, a doubled haploid (DH)
population (SGDH) of 139 individuals was produced from the cross between two Egyptian breeding cultivars
(Sakha 93, salt tolerant) and (Gemmeza 7, salt susceptible). The DH population was tested under saline hydroponics
culture and various plant responses were measured. A molecular genetic map of the SGDH population covering 3645.3
cM, was constructed using Restriction Fragment Length Polymorphism (RFLP), Microsatellite or Simple Sequence
Repeats (SSR), and Amplified Fragment Length Polymorphism (AFLP) markers. In total, 325 loci along the 21 wheat
chromosomes were mapped. The B genome showed the highest number of mapped markers followed by the A and the D
genomes respectively. Interval and composite interval mapping (using QTL cartographer) were used to identify the
genomic regions controlling traits related to salt tolerance with a threshold of LOD 3. Analysis of QTLs has revealed the
approximate location of the significant markers associated with 12 traits related to salt tolerance traits across the A, B and
D genomes. Fifty five significant QTL were detected on 15 of the 21 chromosomes mapped in this study, for some of these
more than one QTL was identified. In many cases QTL were mapped very close to each other, indicating possible gene
clustering or pleiotropy. For some traits, the total percentage of phenotypic variation explained by all QTL exceeded
50-60% but on average it was in the region of 15%. Chromosomes of homologues groups 2 and 5 exerted the biggest effect
on most phenotypic traits, especially chromosomes 2B and 5B. The results indicated that there is considerable potential for
improving salt tolerance of hexaploid wheat by using marker-assisted selection.
barley yield and quality. In the present investigation, two barley verities “AT4” (net blotch resistant) and “Femina”
(net blotch susceptible) were used to develop a segregating F2 population. Linkage analysis and map construction
were performed using Map Manager. The constructed genetic linkage map consisted of 85 markers including 45
AFLP , 11 SSR, 6 CAPS, 5 SCoT, 3 STS and 6 NBL. Linkage groups were assigned to individual barley
chromosomes using the published map locations of the SSR markers as reference point. The produced map showed
7 linkage groups with 85 markers covered a total length of 1644.8 cM. The average length of linkage groups ranged
from 77.7 to 739.4. Single point analysis was used to identify the genomic regions associated with net blotch
resistance in barley. A total of 14 QTL with a significance ranging from 0.01% to 5% were identified on 4 linkage
groups (2, 4, 5 and 6). The most significant QTL was found on chromosome 6H. This QTL presents a promising
opportunity for the strategic improvement of barley resistance to net blotch using marker assisted selection.
systematic and breeding research. The genetic and physiological bases of some important agronomic traits in cotton
were investigated by QTL mapping through constructing of genetic map with chromosomal assignment. A segregating
F2 population derived from an interspecific cross (G. barbadense x G. hirsutum) between two genotypes, cvs. “Giza 83”
and “Deltapine” was used in this study. Different molecular markers including SS R, ES T, ES T-SS R, AFLP and RAPD were
employed to identify markers that reveal differences between the parents. In total 42 new markers were merged with
140 previously mapped markers to produce a new map with 182 loci covering a total length of 2370.5 cM. Among these
new markers, some of them were used to assign chromosomes to the produced 26 linkage groups. The LG2, LG3, LG11
and LG26 were assigned to chromosomes 1, 6, 5 and 20 respectively. Single point analysis was used to identify genomic
regions controlling traits for plant height, number of nodes at flowering time, bolling date, days to flowering and number
of bolls. In total 40 significant QTL were identified for the five traits on 11 linkage groups (1, 2, 3, 4, 5, 10, 11, 12, 18, 19
and 23). This work represents an improvement of the previously constructed genetic map in addition to chromosomal
assignment and detection of new significant QTL for the five traits in Egyptian cotton. The Significant QTLs detected in
this study can be employed in marker assisted selection for molecular breeding programs aiming at developing cotton
cultivars with improved agronomic traits
vegetatively propagated plants such as grape (Vitis vinifera L.). An efficient in vitro shoot
micropropagation protocol leading to the regeneration from shoot tip explants was successfully
developed for mass production. Callus was induced from shoot tips and axillary buds on C2D medium
supplemented with 4 mg/l 6-benzyladenine (BA). Bud differentiation from callus was possible on C2D
medium supplemented with 1 mg/l gibberellic acid and 0.1 mg/l kinetin. Bud initials were elongated by
sub-culturing on elongation medium containing C2D salts supplemented with 0.1 mg/l BA and
resulting shoots could be mass micropropagated on C2D medium supplemented with 1 mg/l BA. Shoots
that were regenerated via organogenesis or micropropagation were rooted on C2D medium
supplemented with 1 mg/l indole-3-butyric acid. In this study, regeneration and micropropagation of
grape cv. ‘Spero’ was successfully achieved through shoot tip and axillary bud culture. Such a protocol
would be useful for future use in the strategic improvement of grape using genetic engineering
million tones per year. By tissue culture, triploid and seedless bananas can now be produced in a high quantity, in a
less time and at any time of the year without being limited to a season. The aim of this study is to substitute the
artificial hormones used in the tissue culture such as, cytokinins and auxins by natural materials such as, Pineapple,
Coconut milk, Cacao, Coffee, and Charcoal. Five different media composed of these natural materials were prepared
in different concentrations, Pineapple (5,10 and 20cm), Coconut milk (5,10 and20cm,), Cacao (1,2.5 and 5 g),
Coffee (1,2.5 and 5 g) and Charcoal (1,2.5 and 5 g) to study their effect on the development and browning of the
bananas' shoots and roots in vitro, with the aid of various statistical analyses. The results indicated that the
pineapple and coconut milk produced the highest number and length of both shoots and roots, as well as the soluble
sugars contents, while the cocoa and coconut milk showed the highest protein content. The charcoal reduced
significantly the phenolic content. On the other hand, there was no significant difference between the coffee, cacao.
This study has highlighted the benefits and advantages of using natural materials in producing high quality banana.
(Gedion, Moaz, and Mohand) were studied to evaluate different plant regeneration protocols and develop a reliable
system for plant propagation. An efficient procedure of in vitro plant regeneration through seed and direct shoot bud
induction was tested from different explants of capsicum annuum L. Several methods of media preparation with
combinations of growth regulators were used, and 2 combinations were found ideal for seed propagation with 7 mg/l
2,4-D or 5 mg/l IBA respectively. Additionally, it was indicated that seed dormancy can effect its propagation also
the positioning of the seed or explants onto the media. Propagated seeds demonstrated shoot and root elongation.
Regeneration of the explants did not show satisfactory results because most of the explants did not develop into
normal shoots but instead developed into calli after 15 days of culture. HPLC Analysis of cultivars demonstrated
that fructose suger percentage was higher in the three different types of pepper, followed by sucrose then glucose.
Also the HPLC analysis drew out that media supplemented with 7mg/l 2,4-D generally had the highest effect of
capsaicinoid sugar content in the following order i.e. cultivar Mohand was showed high significant in sugar content
of capsaicinoid followed by Gedion and Moaz. Application of DPPH method illustrated that extracts obtained from
Moaz leaves characterized the most proactive antioxidant (vitamin C) properties than Gedion and Mohand. A were
noticed. The results of these studies indicated that antioxidant activities of the extract prepared from pepper leaves
depended mainly on phenolic compounds.
Beni Suif 4, Beni Suif 5 and Beni Suif 6 and one bread wheat i.e. Sids 12 cultivars using sodium dodecyle sulfate
polyacrylamide gel electrophoresis (SDS-PAGE) and randomly amplified polymorphic (RAPD) markers. Protein
electrophoreses showed that Beni Suif 4 was characterized by the absence of band-3 with 41.56 kDa. RAPD analysis
showed that the number of polymorphic amplicons was 56 out of a total of 93 amplicons, thus revealing a level of 60.0
% polymorphism. The highest genetic similarity revealed by RAPD analysis (95.0%) was between Beni Suif 5 and
Beni suif 6. While, lowest similarity (65 %) was found between Beni Suif 4 and Beni Suif 5. The dendrogram
separated Sids 12 from all the other genotypes, thus demonstrating the distinctiveness of the genetic background of
this genotype from all the other genotypes. The three genotypes constituted a subcluster divided into two groups,
one group composed of Beni Suif 5 and Beni Suif 6 , while the second group comprised Beni Suif 4.
bread wheat genotypes (Misr1 and Sids 13 cutlivars and the promising lines No.1, No.2 and No.11 )
using SDS-PAGE and RAPD markers. The total number of SDS- bands was seven. Six SDS- bands
were monomorphic while the other was polymorphic. Line-1 was characterized by the presence of
band -3 with a molecular weight of 41.56 kDa. RAPD analysis showed that the number of
polymorphic amplicons was 66 out of a total of 93 amplicons, thus revealing a level of 70.97 %
polymorphism. The highest genetic similarity revealed by RAPD analysis (93.1%) was between
Misr1 and Line 2 genotype. While, the lowest similarity (85.2 %) was between Line 1 and Line 2.
The dendrogram separated Line1 from all the other genotypes. The four genotypes constituted a
subcluster divided into two groups, one group composed of Misr 1 and Line 2, while the second
group comprised Sids 13 and Line 11.
be the analogous of cisplatin. Nigella sativa oil is a herbaceous plant, it has been used for thousands
of years for culinary and medical purposes. This study aimed to evaluate the effect of carboplatin
and Nigella sativa oil alone and in combination together on human breast cancer cells (MCF-7) in
vitro and Ehrlich ascites tumor bearing female mice (in vivo). The in vitro experiment on MCF-7
cells illustrated that IC50 of carboplatin was 11.8 μg/ml, also IC50 of Nigella sativa oil was 39 μg/ml
on MCF-7 cells. In addition, IC50 of the combination between carboplatin and Nigella sativa oil was
found to be 3.78 and 40 μg/ml, respectively. The in vivo experiment illustrated that carboplatin
(10mg/kg) increased the enzyme activity of aspartate amino transferase (GOT) and aniline amino
transferase (GPT)by 56.52% and 51.14%, respectively as compared to both healthy control (nontumor
transplanted mice) and negative control. Also, the activity of GOT and GPT was increased by
14.75% and 19.84%, respectively as compared to healthy control under the effect of Nigella sativa
oil (12ml/kg). While the activity of GOT and GPT was decreased as compared to negative control.
The combination of carboplatin and Nigella sativa oil appeared to increase the enzyme activity of
GOT and GPT by 62.41 and 49.39%, respectively as compared to both healthy control and negative
control. Also, carboplatin induced DNA damage of liver tissue was performed by agarose gel
electrophoresis and comet assay, while Nigella sativa oil showed intact DNA without any damage.
The combination of carboplatin and Nigella sativa oil appeared to decrease the DNA damage as
compared to carboplatin alone.
yellow mosaic potyvirus (ZYMV) is considered as one of the most important viruses infects squash.
We used the biological, serological and cytopathological studies to identify an Egyptian isolate of
ZYMV. Results showed that the Egyptian isolate of ZYMV showed systemic symptoms in the form
of severe mosaic and vein banding on Cucurbita pepo cv. Eskandarani under open field conditions.
Direct antigen coated (DAC)-enzyme-linked immunosorbent assay (ELISA) using polyclonal
antibodies was used as a diagnostic tool for detecting ZYMV in the virus-infected C. pepo samples.
It showed positive reactions with ZYMV antiserum. Filamentous virus-like particles measuring
750X13 nm was successfully purified from ZYMV-infected squash plants, based on the use of
polyethylene glycol and ultracentrifugation. The electron microscope of ultrathin sections of virusinfected
leaf tissues revealed the presence of cylindrical inclusions as pinwheels, laminated
aggregates and scrolls in the cytoplasm of cells infected with ZYMV. In addition, disorganization of
plastids, nucleus and mitochondria in the virus-cells infected was also observed. In addition, the
nucleotide of CI gene of the viral isolate under investigation was determined and its similarities to
some overseas isolates were addressed.
different concentration (50, 250.500 mg/l) used as precursors to produce secondary metabolites (steroids) and
growth development during different stages (callus, embryoids and shooting) of date palm (Malakaby cv.). In
Embryogenic callus stage, callus volume was the highest (4.00) when treated with any of the three amino acids, 50
mg/l of Glutamine or Asparagine showed no effect compared to the control giving the lowest callus volume (3.00).
Total steroids in callus tissues clearly showed that using Glutamine 250 mg/l in medium gave the highest steroid
content 0.662 mg/g and percentage (336% of control), while the lowest (0.111mg/g) was found with Asparagine
(500mg/l) and 56.35% of control. Glutamine at 250 mg/l resulted in the highest weight of embryos (2.100 gm). As
well as, 500 mg/l Spermidine seemed to be the best amino acid used in order to stimulate steroid biosynthesis
resulting in 202.1% of control (0.782 mg/g). In shooting stage, according to the number of shoots, the highest
number of shoots (2.33) was achieved with Glutamine and Spermidine at 500 mg/l. In shoot weight, the highest
weight (7.267 gm) was achieved by using 500mg/l Glutamine , as to steroid biosynthesis in shooting stage, the best
result obtained, were by using Glutamine at 500 mg/l which gave highest steroid biosynthesis (0.534mg/g), 206.0%
of control.
drugs “cis-diamminedichloro platinum (CDDP) (8 mg/kg b.w.) and Doxorubicin (DOX) (10 mg/kg
b.w)” alone and the protective action of Curcuma longa (Turmeric) (250 mg/kg b.w.) in
combination with CDDP and DOX in Ehrlich ascites tumor bearing male mice. The results
illustrated that the effect of CDDP and DOX alone decreased total protein, albumin and globulin
concentrations in serum. Also, there are no significant changes in serum protein banding patterns
between treatments on SDS-PAGE. On the other hand, the lysosomal enzymaic activities of β- Nacetyl
glucosaminidase (β-NAG) and acid phosphatase (ACP) as well as glutathione-S-transferase
(GST) in liver homogentae were increased. Total protein content in liver homogenate was reduced
as compared to the control. The percentage of structural aberrations of chromosomes of bone
marrow cells after 24 hours of drugs administration were increased by 261.0% and 149.1% for
CDDP and DOX, respectively . On the other hand, the turmeric extract were increased in mixture
with each drug resulted in increasing the total protein, albumin and globulin concentrations in
serum. Also, the enzymes activity of ACP, β-NAG and GST were decreased. Total protein contents
was increased in liver homogenate as compared to the control. In addition to, the percentage of
structural and numerical aberrations of chromosomes of bone marrow cells after 24 h of curcumin
administration were decreased, CDDP and DOX showed damage and smear bands of genomic
DNA as compared with negative control group, while the mixture of turmeric showed less damage
of genomic DNA bands.
and physiological bases of drought tolerance in durum wheat was investigated by QTL mapping and by
mapping candidate genes derived from differentially expressed genes and previous investigations. A
recombinant inbred population derived from a cross between two durum (Triticum turgidum L. var
durum) parents Jennah Khetifa and Cham1 that exhibit contrasting traits for drought tolerance was
employed. Twelve known genes and 103 differentially expressed sequence tags (dESTs) were surveyed
and 55 detected polymorphism between the two parental lines. In total, 162 loci including 6 known
genes, 37 dESTs, and an additional 119 markers that were unlinked in the previous map have been
merged with 306 previously mapped markers to produce a new map with 468 loci. Single point analysis
and composite interval mapping were used to identify the genomic regions controlling traits related to
drought stress. Significant QTL were identified for canopy temperature, photosynthesis-related parameters
and water status index. One hundred and seventy eight markers, including 6 candidate genes
and 19 differentially expressed sequences were associated with QTL for drought tolerance traits. The
results indicate that there is considerable potential for improving drought tolerance of durum wheat by
using marker-assisted selection.
and urea, is a powerful anticancer, L-arginine-depleting
agent, against argininosuccinate synthase expressing tumors.
Otherwise, the higher antigenicity and lower thermal stability
of this enzyme was the main biochemical hurdles. Since, the
intrinsic thermal stability of enzymes follow the physiological
temperature of their producer, thus, characterization of Larginase
from thermotolerant Penicillium chrysogenum was
the objective of this study. L-Arginase (Arg) was purified to
its homogeneity from P. chrysogenum by 10.1-fold, with
37.0 kDa under denaturing PAGE, optimum reaction at
50 C, pH stability (6.8–7.9), with highest molar ratio of
constitutional arginine, glutamic acid, lysine and aspartic
acid. The purified enzyme was PEGylated and immobilized
on chitosan, with 41.9 and 22.1 % yield of immobilization.
At 40 C, the T1/2 value of free-Arg, PEG-Arg and Chit-Arg
was 10.4, 15.6, 20.5 h, respectively. The free-Arg and Chit-
Arg have a higher affinity to L-arginine (Km 4.8 mM), while,
PEG-Arg affinity was decreased by about 3 fold (Km
15.2 mM). The inhibitory constants to the free and PEG-Arg
were relatively similar towards HA and PPG. The IC50 for
the free enzyme against HEPG-2 and A549 tumor cells was
0.136 and 0.165 U/ml, comparing to 0.232 and 0.496 U/ml
for PEG-Arg, respectively. The in vivo T1/2 to the free Arg and PEG-Arg was 16.4 and 20.4 h, respectively as holoenzyme.
The residual L-arginine level upon using free Arg
was 156.9 and 144.5 lM, after 6 and 8 h, respectively,
regarding to initials at 253.6 lM, while for Peg-Arg the level
of L-arginine was nil till 7 h of initial dosing. The titer of IgG
was induced by 10–15 % in response to free-Arg after
28 days comparing to IgG titer for PEG-Arg.
Mediterranean region and its production is largely influenced by environmental stresses, such as drought, salinity, heat and
nutrient deficiency. The objective of this study was to dissect quantitative trait loci (QTL) controlling grain yield, yield
components and drought tolerance in durum. A molecular genetic linkage map for F2 durum mapping population derived
from an intraspecific cross between Baniswif-1 x Sohag-2 was constructed using 114 DNA markers (9 SSRs, 14 SCoTs, 90
AFLPs and 1 RAPDs) distributed over the 14 linkage groups and spanning 2040.9 cM of the durum wheat genome. The
size of linkage groups varied greatly from 6.8cM for LG11 to 317.5cM for LG4 with an average length of 145.8cM. Based
on the used anchor SSR markers, only eight linkage groups were assigned to chromosomes, where LG1, LG3, LG5, LG6,
LG7, LG9, LG13 and LG14 were assigned to chromosomes 1B, 3B, 5B, 6A, 6B, 7A, 3A and 2B, respectively. Single point
analysis was used to identify genomic regions controlling eleven morpho-physiological traits related to grain yield, yield
components and drought tolerance. A total of 74 QTL were identified for the eleven traits on all linkage groups except
(LG10 and LG11). These included 3 QTL for root length (RL), 11 QTL for plant height (PH), 7 QTL for spike length (SL),
3 QTL for number of branches/plant (NBP), 3 QTL for number of spike/plant (NSP), 8 QTL for number of spikelets/spike
(NSS), 15 QTL for number of kernel/spike (NKS), 10 QTL for thousand-kernel weight (TKW), 4 QTL for fresh weight
(FW), 5 QTL for dry weight (DW) and 5 QTL for total amino acids (TAA). This work represents the first genetic linkage
map for durum wheat population derived from an intraspecific cross between ‗Baniswif-1‘ and ‗Sohag-2‘ showing
chromosomal regions associated with 11 morpho-physiological traits related to grain yield, yield components and drought
tolerance in durum wheat.
determine the linkage between quantitative trait loci (QTL) and such markers. This study was conducted to investigate the
genetic basis of salinity responses in Egyptian bread wheat (Triticum aestivum L.). In this context, a doubled haploid (DH)
population (SGDH) of 139 individuals was produced from the cross between two Egyptian breeding cultivars
(Sakha 93, salt tolerant) and (Gemmeza 7, salt susceptible). The DH population was tested under saline hydroponics
culture and various plant responses were measured. A molecular genetic map of the SGDH population covering 3645.3
cM, was constructed using Restriction Fragment Length Polymorphism (RFLP), Microsatellite or Simple Sequence
Repeats (SSR), and Amplified Fragment Length Polymorphism (AFLP) markers. In total, 325 loci along the 21 wheat
chromosomes were mapped. The B genome showed the highest number of mapped markers followed by the A and the D
genomes respectively. Interval and composite interval mapping (using QTL cartographer) were used to identify the
genomic regions controlling traits related to salt tolerance with a threshold of LOD 3. Analysis of QTLs has revealed the
approximate location of the significant markers associated with 12 traits related to salt tolerance traits across the A, B and
D genomes. Fifty five significant QTL were detected on 15 of the 21 chromosomes mapped in this study, for some of these
more than one QTL was identified. In many cases QTL were mapped very close to each other, indicating possible gene
clustering or pleiotropy. For some traits, the total percentage of phenotypic variation explained by all QTL exceeded
50-60% but on average it was in the region of 15%. Chromosomes of homologues groups 2 and 5 exerted the biggest effect
on most phenotypic traits, especially chromosomes 2B and 5B. The results indicated that there is considerable potential for
improving salt tolerance of hexaploid wheat by using marker-assisted selection.
barley yield and quality. In the present investigation, two barley verities “AT4” (net blotch resistant) and “Femina”
(net blotch susceptible) were used to develop a segregating F2 population. Linkage analysis and map construction
were performed using Map Manager. The constructed genetic linkage map consisted of 85 markers including 45
AFLP , 11 SSR, 6 CAPS, 5 SCoT, 3 STS and 6 NBL. Linkage groups were assigned to individual barley
chromosomes using the published map locations of the SSR markers as reference point. The produced map showed
7 linkage groups with 85 markers covered a total length of 1644.8 cM. The average length of linkage groups ranged
from 77.7 to 739.4. Single point analysis was used to identify the genomic regions associated with net blotch
resistance in barley. A total of 14 QTL with a significance ranging from 0.01% to 5% were identified on 4 linkage
groups (2, 4, 5 and 6). The most significant QTL was found on chromosome 6H. This QTL presents a promising
opportunity for the strategic improvement of barley resistance to net blotch using marker assisted selection.
systematic and breeding research. The genetic and physiological bases of some important agronomic traits in cotton
were investigated by QTL mapping through constructing of genetic map with chromosomal assignment. A segregating
F2 population derived from an interspecific cross (G. barbadense x G. hirsutum) between two genotypes, cvs. “Giza 83”
and “Deltapine” was used in this study. Different molecular markers including SS R, ES T, ES T-SS R, AFLP and RAPD were
employed to identify markers that reveal differences between the parents. In total 42 new markers were merged with
140 previously mapped markers to produce a new map with 182 loci covering a total length of 2370.5 cM. Among these
new markers, some of them were used to assign chromosomes to the produced 26 linkage groups. The LG2, LG3, LG11
and LG26 were assigned to chromosomes 1, 6, 5 and 20 respectively. Single point analysis was used to identify genomic
regions controlling traits for plant height, number of nodes at flowering time, bolling date, days to flowering and number
of bolls. In total 40 significant QTL were identified for the five traits on 11 linkage groups (1, 2, 3, 4, 5, 10, 11, 12, 18, 19
and 23). This work represents an improvement of the previously constructed genetic map in addition to chromosomal
assignment and detection of new significant QTL for the five traits in Egyptian cotton. The Significant QTLs detected in
this study can be employed in marker assisted selection for molecular breeding programs aiming at developing cotton
cultivars with improved agronomic traits
vegetatively propagated plants such as grape (Vitis vinifera L.). An efficient in vitro shoot
micropropagation protocol leading to the regeneration from shoot tip explants was successfully
developed for mass production. Callus was induced from shoot tips and axillary buds on C2D medium
supplemented with 4 mg/l 6-benzyladenine (BA). Bud differentiation from callus was possible on C2D
medium supplemented with 1 mg/l gibberellic acid and 0.1 mg/l kinetin. Bud initials were elongated by
sub-culturing on elongation medium containing C2D salts supplemented with 0.1 mg/l BA and
resulting shoots could be mass micropropagated on C2D medium supplemented with 1 mg/l BA. Shoots
that were regenerated via organogenesis or micropropagation were rooted on C2D medium
supplemented with 1 mg/l indole-3-butyric acid. In this study, regeneration and micropropagation of
grape cv. ‘Spero’ was successfully achieved through shoot tip and axillary bud culture. Such a protocol
would be useful for future use in the strategic improvement of grape using genetic engineering
million tones per year. By tissue culture, triploid and seedless bananas can now be produced in a high quantity, in a
less time and at any time of the year without being limited to a season. The aim of this study is to substitute the
artificial hormones used in the tissue culture such as, cytokinins and auxins by natural materials such as, Pineapple,
Coconut milk, Cacao, Coffee, and Charcoal. Five different media composed of these natural materials were prepared
in different concentrations, Pineapple (5,10 and 20cm), Coconut milk (5,10 and20cm,), Cacao (1,2.5 and 5 g),
Coffee (1,2.5 and 5 g) and Charcoal (1,2.5 and 5 g) to study their effect on the development and browning of the
bananas' shoots and roots in vitro, with the aid of various statistical analyses. The results indicated that the
pineapple and coconut milk produced the highest number and length of both shoots and roots, as well as the soluble
sugars contents, while the cocoa and coconut milk showed the highest protein content. The charcoal reduced
significantly the phenolic content. On the other hand, there was no significant difference between the coffee, cacao.
This study has highlighted the benefits and advantages of using natural materials in producing high quality banana.
(Gedion, Moaz, and Mohand) were studied to evaluate different plant regeneration protocols and develop a reliable
system for plant propagation. An efficient procedure of in vitro plant regeneration through seed and direct shoot bud
induction was tested from different explants of capsicum annuum L. Several methods of media preparation with
combinations of growth regulators were used, and 2 combinations were found ideal for seed propagation with 7 mg/l
2,4-D or 5 mg/l IBA respectively. Additionally, it was indicated that seed dormancy can effect its propagation also
the positioning of the seed or explants onto the media. Propagated seeds demonstrated shoot and root elongation.
Regeneration of the explants did not show satisfactory results because most of the explants did not develop into
normal shoots but instead developed into calli after 15 days of culture. HPLC Analysis of cultivars demonstrated
that fructose suger percentage was higher in the three different types of pepper, followed by sucrose then glucose.
Also the HPLC analysis drew out that media supplemented with 7mg/l 2,4-D generally had the highest effect of
capsaicinoid sugar content in the following order i.e. cultivar Mohand was showed high significant in sugar content
of capsaicinoid followed by Gedion and Moaz. Application of DPPH method illustrated that extracts obtained from
Moaz leaves characterized the most proactive antioxidant (vitamin C) properties than Gedion and Mohand. A were
noticed. The results of these studies indicated that antioxidant activities of the extract prepared from pepper leaves
depended mainly on phenolic compounds.
Beni Suif 4, Beni Suif 5 and Beni Suif 6 and one bread wheat i.e. Sids 12 cultivars using sodium dodecyle sulfate
polyacrylamide gel electrophoresis (SDS-PAGE) and randomly amplified polymorphic (RAPD) markers. Protein
electrophoreses showed that Beni Suif 4 was characterized by the absence of band-3 with 41.56 kDa. RAPD analysis
showed that the number of polymorphic amplicons was 56 out of a total of 93 amplicons, thus revealing a level of 60.0
% polymorphism. The highest genetic similarity revealed by RAPD analysis (95.0%) was between Beni Suif 5 and
Beni suif 6. While, lowest similarity (65 %) was found between Beni Suif 4 and Beni Suif 5. The dendrogram
separated Sids 12 from all the other genotypes, thus demonstrating the distinctiveness of the genetic background of
this genotype from all the other genotypes. The three genotypes constituted a subcluster divided into two groups,
one group composed of Beni Suif 5 and Beni Suif 6 , while the second group comprised Beni Suif 4.
bread wheat genotypes (Misr1 and Sids 13 cutlivars and the promising lines No.1, No.2 and No.11 )
using SDS-PAGE and RAPD markers. The total number of SDS- bands was seven. Six SDS- bands
were monomorphic while the other was polymorphic. Line-1 was characterized by the presence of
band -3 with a molecular weight of 41.56 kDa. RAPD analysis showed that the number of
polymorphic amplicons was 66 out of a total of 93 amplicons, thus revealing a level of 70.97 %
polymorphism. The highest genetic similarity revealed by RAPD analysis (93.1%) was between
Misr1 and Line 2 genotype. While, the lowest similarity (85.2 %) was between Line 1 and Line 2.
The dendrogram separated Line1 from all the other genotypes. The four genotypes constituted a
subcluster divided into two groups, one group composed of Misr 1 and Line 2, while the second
group comprised Sids 13 and Line 11.
be the analogous of cisplatin. Nigella sativa oil is a herbaceous plant, it has been used for thousands
of years for culinary and medical purposes. This study aimed to evaluate the effect of carboplatin
and Nigella sativa oil alone and in combination together on human breast cancer cells (MCF-7) in
vitro and Ehrlich ascites tumor bearing female mice (in vivo). The in vitro experiment on MCF-7
cells illustrated that IC50 of carboplatin was 11.8 μg/ml, also IC50 of Nigella sativa oil was 39 μg/ml
on MCF-7 cells. In addition, IC50 of the combination between carboplatin and Nigella sativa oil was
found to be 3.78 and 40 μg/ml, respectively. The in vivo experiment illustrated that carboplatin
(10mg/kg) increased the enzyme activity of aspartate amino transferase (GOT) and aniline amino
transferase (GPT)by 56.52% and 51.14%, respectively as compared to both healthy control (nontumor
transplanted mice) and negative control. Also, the activity of GOT and GPT was increased by
14.75% and 19.84%, respectively as compared to healthy control under the effect of Nigella sativa
oil (12ml/kg). While the activity of GOT and GPT was decreased as compared to negative control.
The combination of carboplatin and Nigella sativa oil appeared to increase the enzyme activity of
GOT and GPT by 62.41 and 49.39%, respectively as compared to both healthy control and negative
control. Also, carboplatin induced DNA damage of liver tissue was performed by agarose gel
electrophoresis and comet assay, while Nigella sativa oil showed intact DNA without any damage.
The combination of carboplatin and Nigella sativa oil appeared to decrease the DNA damage as
compared to carboplatin alone.
yellow mosaic potyvirus (ZYMV) is considered as one of the most important viruses infects squash.
We used the biological, serological and cytopathological studies to identify an Egyptian isolate of
ZYMV. Results showed that the Egyptian isolate of ZYMV showed systemic symptoms in the form
of severe mosaic and vein banding on Cucurbita pepo cv. Eskandarani under open field conditions.
Direct antigen coated (DAC)-enzyme-linked immunosorbent assay (ELISA) using polyclonal
antibodies was used as a diagnostic tool for detecting ZYMV in the virus-infected C. pepo samples.
It showed positive reactions with ZYMV antiserum. Filamentous virus-like particles measuring
750X13 nm was successfully purified from ZYMV-infected squash plants, based on the use of
polyethylene glycol and ultracentrifugation. The electron microscope of ultrathin sections of virusinfected
leaf tissues revealed the presence of cylindrical inclusions as pinwheels, laminated
aggregates and scrolls in the cytoplasm of cells infected with ZYMV. In addition, disorganization of
plastids, nucleus and mitochondria in the virus-cells infected was also observed. In addition, the
nucleotide of CI gene of the viral isolate under investigation was determined and its similarities to
some overseas isolates were addressed.
different concentration (50, 250.500 mg/l) used as precursors to produce secondary metabolites (steroids) and
growth development during different stages (callus, embryoids and shooting) of date palm (Malakaby cv.). In
Embryogenic callus stage, callus volume was the highest (4.00) when treated with any of the three amino acids, 50
mg/l of Glutamine or Asparagine showed no effect compared to the control giving the lowest callus volume (3.00).
Total steroids in callus tissues clearly showed that using Glutamine 250 mg/l in medium gave the highest steroid
content 0.662 mg/g and percentage (336% of control), while the lowest (0.111mg/g) was found with Asparagine
(500mg/l) and 56.35% of control. Glutamine at 250 mg/l resulted in the highest weight of embryos (2.100 gm). As
well as, 500 mg/l Spermidine seemed to be the best amino acid used in order to stimulate steroid biosynthesis
resulting in 202.1% of control (0.782 mg/g). In shooting stage, according to the number of shoots, the highest
number of shoots (2.33) was achieved with Glutamine and Spermidine at 500 mg/l. In shoot weight, the highest
weight (7.267 gm) was achieved by using 500mg/l Glutamine , as to steroid biosynthesis in shooting stage, the best
result obtained, were by using Glutamine at 500 mg/l which gave highest steroid biosynthesis (0.534mg/g), 206.0%
of control.
drugs “cis-diamminedichloro platinum (CDDP) (8 mg/kg b.w.) and Doxorubicin (DOX) (10 mg/kg
b.w)” alone and the protective action of Curcuma longa (Turmeric) (250 mg/kg b.w.) in
combination with CDDP and DOX in Ehrlich ascites tumor bearing male mice. The results
illustrated that the effect of CDDP and DOX alone decreased total protein, albumin and globulin
concentrations in serum. Also, there are no significant changes in serum protein banding patterns
between treatments on SDS-PAGE. On the other hand, the lysosomal enzymaic activities of β- Nacetyl
glucosaminidase (β-NAG) and acid phosphatase (ACP) as well as glutathione-S-transferase
(GST) in liver homogentae were increased. Total protein content in liver homogenate was reduced
as compared to the control. The percentage of structural aberrations of chromosomes of bone
marrow cells after 24 hours of drugs administration were increased by 261.0% and 149.1% for
CDDP and DOX, respectively . On the other hand, the turmeric extract were increased in mixture
with each drug resulted in increasing the total protein, albumin and globulin concentrations in
serum. Also, the enzymes activity of ACP, β-NAG and GST were decreased. Total protein contents
was increased in liver homogenate as compared to the control. In addition to, the percentage of
structural and numerical aberrations of chromosomes of bone marrow cells after 24 h of curcumin
administration were decreased, CDDP and DOX showed damage and smear bands of genomic
DNA as compared with negative control group, while the mixture of turmeric showed less damage
of genomic DNA bands.
and physiological bases of drought tolerance in durum wheat was investigated by QTL mapping and by
mapping candidate genes derived from differentially expressed genes and previous investigations. A
recombinant inbred population derived from a cross between two durum (Triticum turgidum L. var
durum) parents Jennah Khetifa and Cham1 that exhibit contrasting traits for drought tolerance was
employed. Twelve known genes and 103 differentially expressed sequence tags (dESTs) were surveyed
and 55 detected polymorphism between the two parental lines. In total, 162 loci including 6 known
genes, 37 dESTs, and an additional 119 markers that were unlinked in the previous map have been
merged with 306 previously mapped markers to produce a new map with 468 loci. Single point analysis
and composite interval mapping were used to identify the genomic regions controlling traits related to
drought stress. Significant QTL were identified for canopy temperature, photosynthesis-related parameters
and water status index. One hundred and seventy eight markers, including 6 candidate genes
and 19 differentially expressed sequences were associated with QTL for drought tolerance traits. The
results indicate that there is considerable potential for improving drought tolerance of durum wheat by
using marker-assisted selection.