Bunchy top disease is one of the most destructive viral diseases caused by the Banana Bunchy Top Virus (BBTV). Using of virus particles as an antigen for polyclonal and monoclonal antibodies production by common serological techniques has several disadvantages concerning the purity and concentration of the viral particles and by extension the produced antibodies. In this study, the coat protein gene (CP) of BBTV was expressed using baculovirus expression vector system (BEVS) under the control of the Polyhedrin promoter (Polh). Accordingly, the generated cassette consisting of the CP gene and the improved Green Florescent Protein (GFP) gene. The generated recombinant virus (vAc-CPpolh-GFPp10) was proved using PCR analysis. Spreading of the recombinant virus in Sf9 insect cells was successfully detected using GFP protein florescence under inverted fluorescent microscopy. The generated recombinant virus was amplified in Sf9 cells and the expression of CP was tested using the infected cell lysate by Dot blot analysis. The anti-BBTV polyclonal antibodies serologically reacted with the recombinant CP which was expressed in Sf9 cells as well as to the BBTV infected plants. This result suggested the possibility of using the expressed CP of BBTV for further development of antisera that can be implemented for BBTV detection in infected plants.
{"title":"GENERATION OF BACMID-BASED RECOMBINANT BACULOVIRUS FOR BANANA BUNCHY TOP VIRUS COAT PRO-TEIN GENE EXPRESSION IN INSECT CELLS","authors":"W. Elmenofy, I. Ismail, H. N. El-Din","doi":"10.21608/ejgc.2016.9595","DOIUrl":"https://doi.org/10.21608/ejgc.2016.9595","url":null,"abstract":"Bunchy top disease is one of the most destructive viral diseases caused by the Banana Bunchy Top Virus (BBTV). Using of virus particles as an antigen for polyclonal and monoclonal antibodies production by common serological techniques has several disadvantages concerning the purity and concentration of the viral particles and by extension the produced antibodies. In this study, the coat protein gene (CP) of BBTV was expressed using baculovirus expression vector system (BEVS) under the control of the Polyhedrin promoter (Polh). Accordingly, the generated cassette consisting of the CP gene and the improved Green Florescent Protein (GFP) gene. The generated recombinant virus (vAc-CPpolh-GFPp10) was proved using PCR analysis. Spreading of the recombinant virus in Sf9 insect cells was successfully detected using GFP protein florescence under inverted fluorescent microscopy. The generated recombinant virus was amplified in Sf9 cells and the expression of CP was tested using the infected cell lysate by Dot blot analysis. The anti-BBTV polyclonal antibodies serologically reacted with the recombinant CP which was expressed in Sf9 cells as well as to the BBTV infected plants. This result suggested the possibility of using the expressed CP of BBTV for further development of antisera that can be implemented for BBTV detection in infected plants.","PeriodicalId":31811,"journal":{"name":"Egyptian Journal of Genetics and Cytology","volume":"45 1","pages":"63-75"},"PeriodicalIF":0.0,"publicationDate":"2016-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68481322","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Dual culture technique was used to evaluate the effect of three species of Trichoderma that showed a potential control of Fusarium oxysporium. Trichoderma harzianum showed maximum growth inhibition (79.3%) followed by Trichoderma viridi (68.5%) and Trichoderma longibrachiatum (44.3%). β-1,3-glucanases was purified from Trichoderma harzianum to homogeneity by ion exchange chromatography on DEAE-Sephacel and gel filtration on Sephadex G100. A typical procedure provided 20-fold purification with 11.9% yield. The apparent molecular mass was 30 kD and it was active on a broad pH range, however the maximal activity was detected at pH 7.5. The optimum temperature of the β-1,3-glucanase was 55C. Polymerase chain reaction (PCR) was used to amplify a fragment about 600 bp from β-1,3 gluanase gene using specific glu forward and reverse primers. The eluted DNA was ligated into pGEM-T-Easy vector and transformed into competent E. coli JM109. White transformed colony, named T1glu, containing recombinant plasmid was validated by PCR using both glu forward and reverse and M13 forward and reverse primers to confirm the presence of β 1,3 glucanase gene insert in right orientation whereas, the fragment amplified with glu forward and glu reverse primers was 600 bp. Partial sequence of the amplified DNA fragment showed 97% sequence homology with the other published sequences.
{"title":"PURIFICATION, CHARACTERISATION AND CLONING OF -1,3 GLUCANASE GENE FROM Trichoderma harzinum","authors":"Noha F. El-Badawy, R. Yehia","doi":"10.21608/EJGC.2016.9703","DOIUrl":"https://doi.org/10.21608/EJGC.2016.9703","url":null,"abstract":"Dual culture technique was used to evaluate the effect of three species of Trichoderma that showed a potential control of Fusarium oxysporium. Trichoderma harzianum showed maximum growth inhibition (79.3%) followed by Trichoderma viridi (68.5%) and Trichoderma longibrachiatum (44.3%). β-1,3-glucanases was purified from Trichoderma harzianum to homogeneity by ion exchange chromatography on DEAE-Sephacel and gel filtration on Sephadex G100. A typical procedure provided 20-fold purification with 11.9% yield. The apparent molecular mass was 30 kD and it was active on a broad pH range, however the maximal activity was detected at pH 7.5. The optimum temperature of the β-1,3-glucanase was 55C. Polymerase chain reaction (PCR) was used to amplify a fragment about 600 bp from β-1,3 gluanase gene using specific glu forward and reverse primers. The eluted DNA was ligated into pGEM-T-Easy vector and transformed into competent E. coli JM109. White transformed colony, named T1glu, containing recombinant plasmid was validated by PCR using both glu forward and reverse and M13 forward and reverse primers to confirm the presence of β 1,3 glucanase gene insert in right orientation whereas, the fragment amplified with glu forward and glu reverse primers was 600 bp. Partial sequence of the amplified DNA fragment showed 97% sequence homology with the other published sequences.","PeriodicalId":31811,"journal":{"name":"Egyptian Journal of Genetics and Cytology","volume":"45 1","pages":"147-161"},"PeriodicalIF":0.0,"publicationDate":"2016-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68481582","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Salt stress is one of the major environmental factor that affecting plant growth and development. High salt content in the soil causes accumulated stress to the cultivated plants in some areas. Plants may be modified genetically to tolerate these effects that lead to some physiological and morphological modification too (Shanon, 1986; Fisher and Turner, 1978). These interacted changes and modifications are not that simple and a lot of response pathways are required to overcome the unfavorable conditions (Neumann, 1997; Yao, 1998; Hasegewa et al., 2000; Munns, 2002). In salty habitat, some plants tolerate these effects and grow in full capacity. These plants have a unique genetic profile tolerating the crucial environment. Bad drainage and high temperature are affecting the agricultural lands that lead to accumulation of salt in the soil (Zhu et al., 2005). These problems are common in most of eastern and southern Mediterranean Sea countries including Syria, Lebanon, Jordan, Egypt, Libya, Tunisia, Algeria and Morocco. In Egypt, bad drainage has bad environmental impacts in the north of Nile delta and the west north coast (FAO, 2005).
盐胁迫是影响植物生长发育的主要环境因子之一。在一些地区，土壤含盐量高对栽培植物造成了积累性的胁迫。植物可以通过基因改造来耐受这些影响，从而导致一些生理和形态上的改变(Shanon, 1986;Fisher and Turner, 1978)。这些相互作用的变化和修改并不是那么简单，需要很多的反应途径来克服不利的条件(Neumann, 1997;姚,1998;Hasegewa et al.， 2000;穆恩一家,2002)。在含盐的生境中，有些植物能忍受这些影响，并能充分生长。这些植物有一种独特的基因谱，可以耐受这种关键的环境。排水不良和高温影响着农田，导致土壤盐分积累(Zhu et al.， 2005)。这些问题在大多数地中海东部和南部国家都很常见，包括叙利亚、黎巴嫩、约旦、埃及、利比亚、突尼斯、阿尔及利亚和摩洛哥。在埃及，恶劣的排水系统对尼罗河三角洲北部和西北海岸的环境造成了恶劣的影响(FAO, 2005)。
{"title":"DETERMINATION AND QUANTIFICATION OF SALT STRESS RELATED GENES IN FABA BEAN (Vicia faba)","authors":"Z. Husseini, H. F. William, O. Hassan","doi":"10.21608/EJGC.2016.9704","DOIUrl":"https://doi.org/10.21608/EJGC.2016.9704","url":null,"abstract":"Salt stress is one of the major environmental factor that affecting plant growth and development. High salt content in the soil causes accumulated stress to the cultivated plants in some areas. Plants may be modified genetically to tolerate these effects that lead to some physiological and morphological modification too (Shanon, 1986; Fisher and Turner, 1978). These interacted changes and modifications are not that simple and a lot of response pathways are required to overcome the unfavorable conditions (Neumann, 1997; Yao, 1998; Hasegewa et al., 2000; Munns, 2002). In salty habitat, some plants tolerate these effects and grow in full capacity. These plants have a unique genetic profile tolerating the crucial environment. Bad drainage and high temperature are affecting the agricultural lands that lead to accumulation of salt in the soil (Zhu et al., 2005). These problems are common in most of eastern and southern Mediterranean Sea countries including Syria, Lebanon, Jordan, Egypt, Libya, Tunisia, Algeria and Morocco. In Egypt, bad drainage has bad environmental impacts in the north of Nile delta and the west north coast (FAO, 2005).","PeriodicalId":31811,"journal":{"name":"Egyptian Journal of Genetics and Cytology","volume":"45 1","pages":"163-170"},"PeriodicalIF":0.0,"publicationDate":"2016-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68481724","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Neveen Abdel Fatah, Mroog A. Abouarab, A. Amin, A. Diab
Fig (Ficus carica L.) is a deciduous tree that belongs to the Moraceae family, and one of the most suitable species for cultivation in semiarid environments found in regions of the Mediterranean and Middle-East; it is considered to be one of the important crop plants grown in Egypt. Many of the species are currently threatened; continuously vulnerable to loss and genetic transmutation due to the absence of safe longterm preservation. In vitro preservation of vegetative propagated genetic resources aided in providing an effective conservation system for the guarantee of food supplies. The present study used shoot tip cultures that were obtained from the black fig (Ficus carica) from Siwa. Shoot tip explants were cultured on conservation media composed of full strength MS medium with 0.8% (w/v) agar and different concentrations of different osmotic agents. The study investigated the use of osmotic stabilizers (mannitol and sorbitol, with concentration of [40 g/L, 50 g/L and 60 g/L]) in the media at two different temperatures (5C and 10C) through a 3 months period in order to determine which osmotic stabilizer, concentration, and temperature would display an eminent effect on the in vitro short-term storage of fig shoot cultures. A (30 g/L) concentration of sucrose was used as the control media. Results were in favor of sorbitol (50 g/L) at a temperature of 5C. In addition Inter Simple Sequence Repeats (ISSR) marker was performed to assess molecular characterization of genetic identity and stability; results illustrated that no genomic instability and mutations were found in the propagated fig (Ficus carica L.) cultures.
{"title":"SHORT TERM PRESERVATION FOR FIG (Ficus carica cv. Black fig) BY DIFFERENT OSMOTIC STABILIZERS","authors":"Neveen Abdel Fatah, Mroog A. Abouarab, A. Amin, A. Diab","doi":"10.21608/EJGC.2016.9594","DOIUrl":"https://doi.org/10.21608/EJGC.2016.9594","url":null,"abstract":"Fig (Ficus carica L.) is a deciduous tree that belongs to the Moraceae family, and one of the most suitable species for cultivation in semiarid environments found in regions of the Mediterranean and Middle-East; it is considered to be one of the important crop plants grown in Egypt. Many of the species are currently threatened; continuously vulnerable to loss and genetic transmutation due to the absence of safe longterm preservation. In vitro preservation of vegetative propagated genetic resources aided in providing an effective conservation system for the guarantee of food supplies. The present study used shoot tip cultures that were obtained from the black fig (Ficus carica) from Siwa. Shoot tip explants were cultured on conservation media composed of full strength MS medium with 0.8% (w/v) agar and different concentrations of different osmotic agents. The study investigated the use of osmotic stabilizers (mannitol and sorbitol, with concentration of [40 g/L, 50 g/L and 60 g/L]) in the media at two different temperatures (5C and 10C) through a 3 months period in order to determine which osmotic stabilizer, concentration, and temperature would display an eminent effect on the in vitro short-term storage of fig shoot cultures. A (30 g/L) concentration of sucrose was used as the control media. Results were in favor of sorbitol (50 g/L) at a temperature of 5C. In addition Inter Simple Sequence Repeats (ISSR) marker was performed to assess molecular characterization of genetic identity and stability; results illustrated that no genomic instability and mutations were found in the propagated fig (Ficus carica L.) cultures.","PeriodicalId":31811,"journal":{"name":"Egyptian Journal of Genetics and Cytology","volume":"45 1","pages":"47-61"},"PeriodicalIF":0.0,"publicationDate":"2016-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68481259","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hypertension, a well-known epidemic health disease, is risk factor for various cardiovascular, peripheral vascular and renal diseases. Renin angiotensin system (RAS) being the most important pathogenic mechanism of hypertension is mediated by a key component; the angiotensin converting enzyme (ACE). Moreover, Mutations in the methylenetetrahydrofolate reductase gene (MTHFR) have been established to be associated with the risk of cardiovascular disease as well as hypertension. This case-control study was conducted out to investigate the potential relationship of ACE (I/D) and MTHFR (C677T) gene polymorphisms with hypertension susceptibility and the responsive ability of antihypertensive drugs in Egyptian hypertensive patients. Thirty six patients suffering of high blood pressure were compared with age and sex matching 14 control cases. MTHFR (C677T) and ACE (I/D) polymorphisms were genotyped by polymerase chain reaction (PCR). The demographic and clinical features of patients and control showed no particular significance (p > 0.05) except for the consanguinity and the obesity. For MTHFR polymorphism frequency, total hypertensive cases showed significantly higher frequency rate for the mutant allele 677T compared to controls with a lower frequency of the wild type 677CC genotype. Whereas the mutant 677TC+TT genotypes were not significantly associated with the hypertension risk when compared to the wild genotype among the case group. For ACE gene polymorphism, also showed only higher frequency rates of DD allele. Interestingly, ACE DD genotype showed significant association with blood pressure, obesity and diabetes. Finally in response of the antihypertensive drugs, we found that, the best responsive group is DD genotype group when treated with the ACE inhibitors. These result suggested that the MTHFR polymorphism was not associated with hypertension while the ACE DD genotype may be associated with essential hypertension and considered as a potent risk factor for hypertension and moreover it is the best responsive group when treated with ACE inhibitors in Egyptian patients.
{"title":"ASSOCIATION OF MTHFR (C677T) AND ACE (I/D) POLYMOR-PHISMS WITH HYPERTENSION AND RESPONSE TO TREAT-MENT AMONG EGYPTIAN PATIENTS","authors":"A. Shafik, K. Hemida, M. Seif-ElNasr, H. Ismail","doi":"10.21608/EJGC.2016.9597","DOIUrl":"https://doi.org/10.21608/EJGC.2016.9597","url":null,"abstract":"Hypertension, a well-known epidemic health disease, is risk factor for various cardiovascular, peripheral vascular and renal diseases. Renin angiotensin system (RAS) being the most important pathogenic mechanism of hypertension is mediated by a key component; the angiotensin converting enzyme (ACE). Moreover, Mutations in the methylenetetrahydrofolate reductase gene (MTHFR) have been established to be associated with the risk of cardiovascular disease as well as hypertension. This case-control study was conducted out to investigate the potential relationship of ACE (I/D) and MTHFR (C677T) gene polymorphisms with hypertension susceptibility and the responsive ability of antihypertensive drugs in Egyptian hypertensive patients. Thirty six patients suffering of high blood pressure were compared with age and sex matching 14 control cases. MTHFR (C677T) and ACE (I/D) polymorphisms were genotyped by polymerase chain reaction (PCR). The demographic and clinical features of patients and control showed no particular significance (p > 0.05) except for the consanguinity and the obesity. For MTHFR polymorphism frequency, total hypertensive cases showed significantly higher frequency rate for the mutant allele 677T compared to controls with a lower frequency of the wild type 677CC genotype. Whereas the mutant 677TC+TT genotypes were not significantly associated with the hypertension risk when compared to the wild genotype among the case group. For ACE gene polymorphism, also showed only higher frequency rates of DD allele. Interestingly, ACE DD genotype showed significant association with blood pressure, obesity and diabetes. Finally in response of the antihypertensive drugs, we found that, the best responsive group is DD genotype group when treated with the ACE inhibitors. These result suggested that the MTHFR polymorphism was not associated with hypertension while the ACE DD genotype may be associated with essential hypertension and considered as a potent risk factor for hypertension and moreover it is the best responsive group when treated with ACE inhibitors in Egyptian patients.","PeriodicalId":31811,"journal":{"name":"Egyptian Journal of Genetics and Cytology","volume":"63 1","pages":"77-88"},"PeriodicalIF":0.0,"publicationDate":"2016-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68481382","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sixteen Simple Sequence Repeats (SSR) and three Inter Simple Sequence Repeats (ISSR) primers were used to estimate the genetic diversity and its distribution in twenty garlic clones. A high level of polymorphism amongst studied clones was found with both SSR and ISSR markers. The total number of bands that were detected by all used primers was 75 including 6 monomorphic, 5 unique and 64 polymorphic. The percentage of polymorphism identified by SSR primers were varied between 33.3 and 100. However, all of the studied ISSR primers were polymorphic conferring a 100% of polymorphism. Results showed that each of the Asa14, Asa17, Asa18 and Asa59 primers generated one monomorphic band of 77, 120, 102 and 113 bp, respectively, in all of the studded garlic clones. Two monomorphic bands of 104 and 177 bp were generated by using Asa24 primer. Asa17 and Asa59 SSR primers produced only one unique band of 154 (Egaseed 2) and 646 bp (EGA 1), respectively. Two unique bands of 225 and 250 bp were detected for Egaseed 2 (ft) by using HB 13 ISSR primer. The highest similarity value (0.969) was found between AZO 2 and AZO 3, while the lowest value (0.482) was found between AZO 4 and EGA 5 clones. Dendrogram of genetic distances amongst all tested clones showed two distinct major clusters with overlapping. In general, the present results reveal the importance of using molecular markers to assess genetic diversity among such closely related genotypes which were difficult to distinguish with other markers.
利用16个SSR引物和3个ISSR引物对20个大蒜无性系的遗传多样性及其分布进行了分析。在所研究的无性系中，SSR和ISSR标记均存在较高的多态性。所有引物共检测到75个条带，其中单态条带6个，唯一条带5个，多态条带64个。SSR引物鉴定的多态性百分比在33.3% ~ 100%之间。然而，所有研究的ISSR引物都是多态性的，多态性为100%。结果表明，Asa14、Asa17、Asa18和Asa59引物分别在所有大蒜克隆中产生一条单态条带，长度分别为77、120、102和113 bp。用Asa24引物分别获得了104和177 bp的单态条带。Asa17和Asa59引物仅产生一条独特的条带，分别为154 bp (Egaseed 2)和646 bp (Egaseed 1)。利用hb13 ISSR引物对Egaseed 2 (ft)进行了225和250 bp的特异条带检测。azo2与azo3的相似性值最高，为0.969,azo4与ega5的相似性值最低，为0.482。所有被测无性系的遗传距离树状图显示出两个明显重叠的主簇。总的来说，目前的结果表明，使用分子标记来评估这些密切相关的基因型之间的遗传多样性的重要性，这些基因型很难与其他标记区分。
{"title":"ASSESMENT OF GENETIC DIVERSITY IN GARLIC CLONES USING SSR AND ISSR MARKERS","authors":"G. Anwar, R. K. Helmey, Y. Mostafa","doi":"10.21608/EJGC.2016.9585","DOIUrl":"https://doi.org/10.21608/EJGC.2016.9585","url":null,"abstract":"Sixteen Simple Sequence Repeats (SSR) and three Inter Simple Sequence Repeats (ISSR) primers were used to estimate the genetic diversity and its distribution in twenty garlic clones. A high level of polymorphism amongst studied clones was found with both SSR and ISSR markers. The total number of bands that were detected by all used primers was 75 including 6 monomorphic, 5 unique and 64 polymorphic. The percentage of polymorphism identified by SSR primers were varied between 33.3 and 100. However, all of the studied ISSR primers were polymorphic conferring a 100% of polymorphism. Results showed that each of the Asa14, Asa17, Asa18 and Asa59 primers generated one monomorphic band of 77, 120, 102 and 113 bp, respectively, in all of the studded garlic clones. Two monomorphic bands of 104 and 177 bp were generated by using Asa24 primer. Asa17 and Asa59 SSR primers produced only one unique band of 154 (Egaseed 2) and 646 bp (EGA 1), respectively. Two unique bands of 225 and 250 bp were detected for Egaseed 2 (ft) by using HB 13 ISSR primer. The highest similarity value (0.969) was found between AZO 2 and AZO 3, while the lowest value (0.482) was found between AZO 4 and EGA 5 clones. Dendrogram of genetic distances amongst all tested clones showed two distinct major clusters with overlapping. In general, the present results reveal the importance of using molecular markers to assess genetic diversity among such closely related genotypes which were difficult to distinguish with other markers.","PeriodicalId":31811,"journal":{"name":"Egyptian Journal of Genetics and Cytology","volume":"45 1","pages":"333-345"},"PeriodicalIF":0.0,"publicationDate":"2016-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68480792","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Somatic embryogenesis and plant regeneration at high frequency have been restricted to few sweet potato varieties. For enhancing and accelerating somatic embryogenesis from stem segments of the Egyptian sweet potato cultivar Abees were investigated using three different phytohormones; 2,4-dichlorophenoxy-acetic acid (2,4-D), benzyleaminopurine (BAP) and indole acetic acid (IAA). The phytohormone BAP was found to be the best for the induction of embryogenic calli and most studied traits. Data analysis showed a significant variation in three different tissue culture media for all parameters, expect root induction percentage. Two different isozymes; peroxidase (PRX) and α naphthyl acetate esterase (EST) were used and analyzed to determine the genetic variability among the regenerated plants. The two analyzed isozymes successively showed polymorphic variations among the parent and 98 sweet potato plants regenerated from the three different callus induction media. Peroxidase isozyme produced seven polymorphic bands showing genetic variation as compared to the control (Abees cultivar), while esterase isozyme produced only three polymorphic bands. The regenerated plants exhibited somaclonal variations that can be utilized for selection of desired traits in sweet potato. On the other hand, five RAPD primers were used for assessment of genetic diversity in the somaclonal variants compared with control. A total of 68 RAPD loci were amplified with molecular size range of 300–3000 bp with 13.6 loci per each primer. Out of the 68 loci scored, 26 loci (38.24%) were found to be polymorphic and the polymorphism% ranged between 18.18% for (OPB-05) and 75% for (OPB-07). Moreover, all primers produced positive and negative unique DNA bands, except OPB-05 for negative unique bands and OPB-07 for positive unique bands. The same result was confirmed by the cluster and principal coordinate analyses for the positions of somaclonal variant no. 4 which showed high diversity from the parental cultivar.
{"title":"MOLECULAR GENETIC DIVERSITY AND EFFICIENT PLANT REGENERATION SYSTEM VIA SOMATIC EMBRYOGENESIS IN SWEET POTATO (Ipomoea batatas (L.) Lam.)","authors":"A. A. Aboulila","doi":"10.21608/ejgc.2016.9586","DOIUrl":"https://doi.org/10.21608/ejgc.2016.9586","url":null,"abstract":"Somatic embryogenesis and plant regeneration at high frequency have been restricted to few sweet potato varieties. For enhancing and accelerating somatic embryogenesis from stem segments of the Egyptian sweet potato cultivar Abees were investigated using three different phytohormones; 2,4-dichlorophenoxy-acetic acid (2,4-D), benzyleaminopurine (BAP) and indole acetic acid (IAA). The phytohormone BAP was found to be the best for the induction of embryogenic calli and most studied traits. Data analysis showed a significant variation in three different tissue culture media for all parameters, expect root induction percentage. Two different isozymes; peroxidase (PRX) and α naphthyl acetate esterase (EST) were used and analyzed to determine the genetic variability among the regenerated plants. The two analyzed isozymes successively showed polymorphic variations among the parent and 98 sweet potato plants regenerated from the three different callus induction media. Peroxidase isozyme produced seven polymorphic bands showing genetic variation as compared to the control (Abees cultivar), while esterase isozyme produced only three polymorphic bands. The regenerated plants exhibited somaclonal variations that can be utilized for selection of desired traits in sweet potato. On the other hand, five RAPD primers were used for assessment of genetic diversity in the somaclonal variants compared with control. A total of 68 RAPD loci were amplified with molecular size range of 300–3000 bp with 13.6 loci per each primer. Out of the 68 loci scored, 26 loci (38.24%) were found to be polymorphic and the polymorphism% ranged between 18.18% for (OPB-05) and 75% for (OPB-07). Moreover, all primers produced positive and negative unique DNA bands, except OPB-05 for negative unique bands and OPB-07 for positive unique bands. The same result was confirmed by the cluster and principal coordinate analyses for the positions of somaclonal variant no. 4 which showed high diversity from the parental cultivar.","PeriodicalId":31811,"journal":{"name":"Egyptian Journal of Genetics and Cytology","volume":"45 1","pages":"347-356"},"PeriodicalIF":0.0,"publicationDate":"2016-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68481028","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Genetic diversity across Tilapia species is important key fordevelopment aquaculture strains, protection of endangered populations and biogeographical inferences. Total soluble protein and esterases isozymes were extracted from flesh (muscles) and liver of all individuals from the three populations (Ryad, Bahr El-Baqar and Motobs) from each species under study to estimate the genetic diversity. With comparing the obtained bands from the three Oreochromis niloticus, it was found that 13 bands out of the 20 were common bands among the three populations. The three T. zilli populations exhibited 8 bands out of the 21 were common bands among those three populations. The three O. aurea populations exhibited 10 bands were common between three populations, while the rest of bands appeared in some of population and disappeared in the other. Ryad and Motobs individuals of O. niloticus showed four isozymes. Meanwhile, Bahr El-Baqar exhibited only three. Ryad and Bahr El-Baqar individuals of T. zilli showed three isozymes. Meanwhile, Motobs exhibited four ones. Maximum two isozymes were detected in population of O. aurea. Band 1 was dark in Ryad individuals and ranged from very faint, faint and dark in Bahr El-Baqar and Motobs. The phylogenetic relationship within the studied populations of the three locations concerning O. niloticus, T. zilli and O. aurea was conducted and different variations were detected. Population from Ryad was highly differentiated than other populations.
{"title":"MUSCLE PROTEIN AND LIVER ESTERASES BANDING PAT-TERNS AS BIOCHEMICAL MARKERS TO DETERMINE GENETIC DIVERSITY IN EGYPTIAN POPULATIONS OF Tilapia SPECIES","authors":"G. El-Fadly, M. Rehan, I. Khatab, A. Kalboush","doi":"10.21608/ejgc.2016.9575","DOIUrl":"https://doi.org/10.21608/ejgc.2016.9575","url":null,"abstract":"Genetic diversity across Tilapia species is important key fordevelopment aquaculture strains, protection of endangered populations and biogeographical inferences. Total soluble protein and esterases isozymes were extracted from flesh (muscles) and liver of all individuals from the three populations (Ryad, Bahr El-Baqar and Motobs) from each species under study to estimate the genetic diversity. With comparing the obtained bands from the three Oreochromis niloticus, it was found that 13 bands out of the 20 were common bands among the three populations. The three T. zilli populations exhibited 8 bands out of the 21 were common bands among those three populations. The three O. aurea populations exhibited 10 bands were common between three populations, while the rest of bands appeared in some of population and disappeared in the other. Ryad and Motobs individuals of O. niloticus showed four isozymes. Meanwhile, Bahr El-Baqar exhibited only three. Ryad and Bahr El-Baqar individuals of T. zilli showed three isozymes. Meanwhile, Motobs exhibited four ones. Maximum two isozymes were detected in population of O. aurea. Band 1 was dark in Ryad individuals and ranged from very faint, faint and dark in Bahr El-Baqar and Motobs. The phylogenetic relationship within the studied populations of the three locations concerning O. niloticus, T. zilli and O. aurea was conducted and different variations were detected. Population from Ryad was highly differentiated than other populations.","PeriodicalId":31811,"journal":{"name":"Egyptian Journal of Genetics and Cytology","volume":"45 1","pages":"187-203"},"PeriodicalIF":0.0,"publicationDate":"2016-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68480321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Salinity is a major abiotic stress which affecting all crops in Egypt especially in the northern part of Nile Delta. More than 30% of the total cultivated areas are irrigated by mixed or saline water. This study was amid to evaluate and clarify the adaptive response in agro-physiological and molecular aspects of 15 Egyptian available barley cultivars. The experiment was conducted during two seasons 2012/2013 and 2013/2014 in randomized complete block design with three replications under both saline and normal conditions. The results showed that Giza 123, Giza 131 and Giza 136 had the highest number of grains spike-1, grain yield, flag leaf area and chlorophyll content under both normal and saline, which were considered as tolerant cultivars. Moreover, proline content, catalase and peroxidase activities were higher in these cultivars than activities in the sensitive cultivars under salt stress. Based on molecular analysis using informative SSR markers, the data represents in total 13 fragments with high polymorphism (100%) ranged from one to four fragments per locus with fragment sizes ranged from (120 to 290 bp). Bmag0770 primer amplified specific fragment in most tolerant cultivars, which was absent in susceptible cultivars with higher PIC value (0.79%). Dendrogram based on SSR marker successfully discriminated the barley cultivars for salt stress.
{"title":"PHYSIOLOGICAL AND MOLECULAR CHARACTERIZATION OF SOME EGYPTIAN BARLEY (Hordeum vulgare L.) CULTI-VARS FOR SALT TOLERANCE","authors":"S. Mariey, M. Farid, I. Khatab","doi":"10.21608/EJGC.2016.9588","DOIUrl":"https://doi.org/10.21608/EJGC.2016.9588","url":null,"abstract":"Salinity is a major abiotic stress which affecting all crops in Egypt especially in the northern part of Nile Delta. More than 30% of the total cultivated areas are irrigated by mixed or saline water. This study was amid to evaluate and clarify the adaptive response in agro-physiological and molecular aspects of 15 Egyptian available barley cultivars. The experiment was conducted during two seasons 2012/2013 and 2013/2014 in randomized complete block design with three replications under both saline and normal conditions. The results showed that Giza 123, Giza 131 and Giza 136 had the highest number of grains spike-1, grain yield, flag leaf area and chlorophyll content under both normal and saline, which were considered as tolerant cultivars. Moreover, proline content, catalase and peroxidase activities were higher in these cultivars than activities in the sensitive cultivars under salt stress. Based on molecular analysis using informative SSR markers, the data represents in total 13 fragments with high polymorphism (100%) ranged from one to four fragments per locus with fragment sizes ranged from (120 to 290 bp). Bmag0770 primer amplified specific fragment in most tolerant cultivars, which was absent in susceptible cultivars with higher PIC value (0.79%). Dendrogram based on SSR marker successfully discriminated the barley cultivars for salt stress.","PeriodicalId":31811,"journal":{"name":"Egyptian Journal of Genetics and Cytology","volume":"45 1","pages":"367-382"},"PeriodicalIF":0.0,"publicationDate":"2016-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68481440","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"INTERNATIONAL JOURNAL DEVOTED TO GENETICAL AND CYTOLOGICAL SCIENCES","authors":"A. Fahmy, K. E. Mangoury, E. A. Qaid","doi":"10.21608/ejgc.2016.9574","DOIUrl":"https://doi.org/10.21608/ejgc.2016.9574","url":null,"abstract":"","PeriodicalId":31811,"journal":{"name":"Egyptian Journal of Genetics and Cytology","volume":"45 1","pages":"171-185"},"PeriodicalIF":0.0,"publicationDate":"2016-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"68480207","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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