In a survey conducted in 2002 and 2003, Citrus tristeza virus (CTV) was detected in the Jordan Va... more In a survey conducted in 2002 and 2003, Citrus tristeza virus (CTV) was detected in the Jordan Valley. The direct tissue blot immunoassay (DTBIA) indicated that 12.7 and 15.2% of samples tested in the central and northern Jordan Valley respectively were infected with CTV. Similar results showed that all citrus species grown in the Jordan Valley were susceptible to CTV. DAS-ELISA analysis of samples from a citrus orchard in the Dir Alla area with severe CTV symptoms indicated that 49% of samples were infected with CTV. Using a CTV specific primer pair (CTV1/CTV10), the coat protein gene of the virus was successfully amplified from leaf extracts obtained from CTVinfected trees by IC-RT-PCR. After cloning and sequencing the coat protein gene, the sequence of the amplified product was deposited in the GenBank.
Four crop-infecting begomoviruses were found naturally affecting Charlock mustard (<i>Sinap... more Four crop-infecting begomoviruses were found naturally affecting Charlock mustard (<i>Sinapis arvensis</i> L.) plants in Jordan and characterized at the molecular level. Symptoms included leaf curling and stunting. PCR analysis showed the presence of <i>Tomato yellow leaf curl virus</i> (TYLCV), <i>Tomato yellow leaf curl Sardinia virus</i> (TYLCSV), <i>Squash leaf curl virus</i> (SLCV) and <i>Watermelon chlorotic stunt virus</i> (WmCSV). These viruses were associated with Cotton leaf curl Gezira betasatellite (CLCuGβ). Sequence analysis revealed that TYLCV and TYLCSV had 98.5% and 99.3% nucleotide (nt) sequence identities with a previously described TYLCV strain from Florida and TYLCSV from Sardinia, respectively. The other two viruses, SLCV and WmCSV, shared 99.7% and 99.5% nt identities with previously described strains from Jordan. Sequence analysis also showed that CLCuGβ had 98.8% nt identity with a strain from Israel...
Powdered, dried olive (Olea europaea) cake was extracted with hexane, methanol and butanol. Six p... more Powdered, dried olive (Olea europaea) cake was extracted with hexane, methanol and butanol. Six phenolic compounds, coumaric acid, ferulic acid, oleuropein, caffeic acid, protocatechuic acid and cinnamic acid, were isolated from these extracts after fractionation. The fractions were tested for their antifungal activity against Verticillium sp., Fusarium oxysporum, Rhizopus sp., Penicillium italicum, Rhizoctonia solani, Stemphylium solani, Cladosporium sp., Mucor sp., Colletotrichum sp. and Pythium sp. Strongest activity was reported against Fusarium oxysporum and Verticillium sp. No effect was observed against Alternaria sp.
Inoculation of the two bottom leaves (1 and 2) of tomato plants (Lycopersicon esculentum) cv. Vol... more Inoculation of the two bottom leaves (1 and 2) of tomato plants (Lycopersicon esculentum) cv. Vollendung with tobacco necrosis virus (TNV) induced systemic resistance against late blight disease caused by Phytophthora infestans SDS PAGE of acid soluble proteins extracted from TNV-inoculated plants showed accumulation of six pathogenesis-related proteins in inoculated leaves (1 and 2) and in upper leaf tissue (leaf 3). Western blot analysis with antisera raised against different pathogenesis-related proteins of tomato and tobacco revealed that four of these proteins were P14, AP24, chitinase and β-(1,3)-glucanase. An in itro study of the antifungal activity of these pathogenesis-related proteins showed that only the basic fraction has direct antifungal activity toward Phytophthora infestans. The induction of systemic resistance was accompanied by an increase of peroxidase activity in inoculated leaves as well as in upper leaf tissue (leaf 3). On the other hand, significant increase in the activity of β-(1,3)-glucanase was observed only in leaves inoculated with tobacco necrosis virus. Leaf tissue analysis of induced plants showed that inoculation with TNV resulted in the induction of three new peroxidase and one β-(1,3)-glucanase isozymes.
World Journal of Microbiology & Biotechnology, 2003
The plasmid profile of two thermophilic bacterial strains isolated from recreation thermal spring... more The plasmid profile of two thermophilic bacterial strains isolated from recreation thermal springs in Jordan has been investigated. These strains are Streptococcus thermophilus and Bacillus sp1, which have been isolated from Zerka – Maeen and Himma hot springs respectively. Supercoiled and circular plasmid forms were detected, explaining the effect of DNA conformation on the mobility of the plasmid in the
Yellowing symptoms similar to coconut yellow decline phytoplasma disease were observed on lipstic... more Yellowing symptoms similar to coconut yellow decline phytoplasma disease were observed on lipstick palms (Cyrtostachys renda) in Selangor state, Malaysia. Typical symptoms were yellowing, light green fronds, gradual collapse of older fronds and decline in growth. Polymerase chain reaction assay was employed to detect phytoplasma in symptomatic lipstick palms. Extracted DNA was amplified from symptomatic lipstick palms by PCR using phytoplasma-universal primer pair P1/P7 followed by R16F2n/R16R2. Phytoplasma presence was confirmed, and the 1250 bp products were cloned and sequenced. Sequence analysis indicated that the phytoplasmas associated with lipstick yellow frond disease were isolates of 'Candidatus Phytoplasma asteris' belonging to the 16SrI group. Virtual RFLP analysis of the resulting profiles revealed that these palm-infecting phytoplasmas belong to subgroup 16SrI-B and a possibly new 16SrI-subgroup. This is the first report of lipstick palm as a new host of aster yellows phytoplasma (16SrI) in Malaysia and worldwide.
In a survey conducted in 2002 and 2003, Citrus tristeza virus (CTV) was detected in the Jordan Va... more In a survey conducted in 2002 and 2003, Citrus tristeza virus (CTV) was detected in the Jordan Valley. The direct tissue blot immunoassay (DTBIA) indicated that 12.7 and 15.2% of samples tested in the central and northern Jordan Valley respectively were infected with CTV. Similar results showed that all citrus species grown in the Jordan Valley were susceptible to CTV. DAS-ELISA analysis of samples from a citrus orchard in the Dir Alla area with severe CTV symptoms indicated that 49% of samples were infected with CTV. Using a CTV specific primer pair (CTV1/CTV10), the coat protein gene of the virus was successfully amplified from leaf extracts obtained from CTVinfected trees by IC-RT-PCR. After cloning and sequencing the coat protein gene, the sequence of the amplified product was deposited in the GenBank.
A genomic strategy for PPV identification has been recently developed (Pasquini et al., 2008). Th... more A genomic strategy for PPV identification has been recently developed (Pasquini et al., 2008). The method is based on using a 70-mer oligonucleotide DNA microarray chip capable of simultaneously detecting and genotyping PPV strains. Universal and specific probes have been identified and used with a sensitive protocol of hybridization using an indirect fluorescent labelling of cDNA product with cyanine able to enhance the sensitivity of the virus detection avoiding the use of the PCR amplification step. In order to evaluate the protocol fitness for diagnostic use, about 30 samples belonging to a PPV isolates collection, including M, D, EA and C strains, have been used for its validation, that was determined, estimating the performance criteria that include the following parameters: diagnostic sensitivity (D-SN), diagnostic specificity (D-SP) and diagnostic accuracy (D-AC). Keywords : oligonucleotides chip, PPV, sensitivity, specificity, accuracy, performance criteria
In 2013, 23 tomato samples were collected from plants grown at Homret Al-Sahen region exhibiting ... more In 2013, 23 tomato samples were collected from plants grown at Homret Al-Sahen region exhibiting typical Tomato yellow leaf curl disease (TYLCD) symptoms. After total nucleic acids were extracted, all samples were subjected to PCR analysis using primers previously reported to detect Tomato yellow leaf curl Sardinia virus (TYLCSV), Tomato yellow leaf curl virus (TYLCV), recombinant type A viruses (named RecA, a recombinant between TYLCV/TYLCSV) and betasatellites. PCR analysis showed that 22 samples were infected with a mixture of TYLCV, TYLCSV and betasatellites, whereas one sample was infected with TYLCV, TYLCSV and RecA virus. The full-length genome (2763 bp) of RecA type virus, was amplified, cloned and sequenced. Sequence analysis showed that the RecA virus had the highest nucleotide identity (96%) with the Italian strain of Tomato yellow leaf curl Axarquia virus (TYLCAxV-Sic2-[IT-Sic2/5-04], EU734832). Recombination analysis revealed that RecA virus detected in this study (here...
Whiteflies are economically important plant pests that cause damage to crops worldwide. This stud... more Whiteflies are economically important plant pests that cause damage to crops worldwide. This study aimed to update the status of whiteflies in Jordan by combining the classical morphological identification and the DNA markers using the mitochondrial cytochrome oxidase I (mtCOI) gene. Over the course of three consecutive years, 111 whiteflies were collected from different geographical regions and different plant hosts in Jordan. The results showed that, in addition to Bemisia tabaci, another nine different whitefly species were identified, including two species that were recorded for the first time in Jordan: Africaleurodes coffeacola, and Tetraleurodes neemani. A special focus has been given to economically important plant pests like the B. tabaci species complex. Three different diagnostic techniques were used to identify B. tabaci putative species based on mtCOI gene. All the collected samples of B. tabaci species complex were identified as Middle East–Asia Minor 1 (MEAM1) putativ...
Tomato yellow leaf curl disease (TYLCD) is a major threat affecting tomato production in both tro... more Tomato yellow leaf curl disease (TYLCD) is a major threat affecting tomato production in both tropical and subtropical areas worldwide. The disease is caused by the Tomato yellow leaf curl virus and other related viruses that belong to the genus Begomovirus of the family Geminiviridae, which are transmitted by the whitefly (Bemisia tabaci). Since mechanical transmission of the disease is difficult and impractical, an efficient and easy method is needed. In this study, an agrodrench inoculation method was utilized where the suspension of Agrobacterium, harbouring a plasmid with the DNA of the virus, is drenched into the soil near the roots. This method proved to be easy and can be used to overcome several problems associated with other inoculation methods. In this study, the effectiveness of the agrodrench as an inoculation method for TYLCV was found to be influenced by the virus strain, agrobacterium strain and concentration, plant age, stem-wounding and host plant species.The metho...
The non-linear transition to adulthood of today’s youth signaled by numerous theoreticians has co... more The non-linear transition to adulthood of today’s youth signaled by numerous theoreticians has consequences on the forms of political participation favoured by young people. Nevertheless, citizenship education continues to play a significant role in shaping the political behaviour of future adults. After highlighting the characteristics of citizenship education in Europe as outlined in the most recent Eurydice study (2012) and briefly presenting the alternative forms of political and social participation embraced by the young generation, the present paper aims to determine the level of interest and involvement of Romanian teenagers in political and social activities at both national and European level. The research method employed is a quantitative one, pupils from the 11 and 12grade being the subjects of a survey conducted in six Romanian high schools. Findings reveal that although teenagers are willing to make their voice heard by various means, including alternative forms of part...
A protocol for production of virus-free Vitis vinifera using meristem culture was developed. Meri... more A protocol for production of virus-free Vitis vinifera using meristem culture was developed. Meristems (0.1-0.2 mm) of V. vinifera infected with grapevine fanleaf virus and grape leafroll associated viruses were excised from 1-year-old growing vines. Shoot tips were cultured on half-strength Murashige and Skoog (MS) medium, supplemented with 0.02 mg L-1 of benzylaminopurine (BAP) and 0.01 mg L-1 of naphthalene acetic acid (NAA). BAP and kinetin resulted in differences in the number of new shoots per explant, shoot height, and number of new leaves per explant. BAP at 0.8 mg L-1 gave the highest in vitro multiplication rate, with 5.25 shoots per explant, whereas elongation was greatest in the presence of 0.2 mg L-1 of kinetin. Root initiation was tested on an MS medium supplemented with 0.0, 0.2, 0.4, 0.6, 0.8, or 1.0 mg L-1 of IBA (indole-3-butyric-acid), IAA (indole-3-acetic-acid), or NAA. Maximum root number was achieved using 0.6 mg L-1 of IBA. A survival rate of 95% was achieved ...
In a survey conducted in 2002 and 2003, Citrus tristeza virus (CTV) was detected in the Jordan Va... more In a survey conducted in 2002 and 2003, Citrus tristeza virus (CTV) was detected in the Jordan Valley. The direct tissue blot immunoassay (DTBIA) indicated that 12.7 and 15.2% of samples tested in the central and northern Jordan Valley respectively were infected with CTV. Similar results showed that all citrus species grown in the Jordan Valley were susceptible to CTV. DAS-ELISA analysis of samples from a citrus orchard in the Dir Alla area with severe CTV symptoms indicated that 49% of samples were infected with CTV. Using a CTV specific primer pair (CTV1/CTV10), the coat protein gene of the virus was successfully amplified from leaf extracts obtained from CTVinfected trees by IC-RT-PCR. After cloning and sequencing the coat protein gene, the sequence of the amplified product was deposited in the GenBank.
Four crop-infecting begomoviruses were found naturally affecting Charlock mustard (<i>Sinap... more Four crop-infecting begomoviruses were found naturally affecting Charlock mustard (<i>Sinapis arvensis</i> L.) plants in Jordan and characterized at the molecular level. Symptoms included leaf curling and stunting. PCR analysis showed the presence of <i>Tomato yellow leaf curl virus</i> (TYLCV), <i>Tomato yellow leaf curl Sardinia virus</i> (TYLCSV), <i>Squash leaf curl virus</i> (SLCV) and <i>Watermelon chlorotic stunt virus</i> (WmCSV). These viruses were associated with Cotton leaf curl Gezira betasatellite (CLCuGβ). Sequence analysis revealed that TYLCV and TYLCSV had 98.5% and 99.3% nucleotide (nt) sequence identities with a previously described TYLCV strain from Florida and TYLCSV from Sardinia, respectively. The other two viruses, SLCV and WmCSV, shared 99.7% and 99.5% nt identities with previously described strains from Jordan. Sequence analysis also showed that CLCuGβ had 98.8% nt identity with a strain from Israel...
Powdered, dried olive (Olea europaea) cake was extracted with hexane, methanol and butanol. Six p... more Powdered, dried olive (Olea europaea) cake was extracted with hexane, methanol and butanol. Six phenolic compounds, coumaric acid, ferulic acid, oleuropein, caffeic acid, protocatechuic acid and cinnamic acid, were isolated from these extracts after fractionation. The fractions were tested for their antifungal activity against Verticillium sp., Fusarium oxysporum, Rhizopus sp., Penicillium italicum, Rhizoctonia solani, Stemphylium solani, Cladosporium sp., Mucor sp., Colletotrichum sp. and Pythium sp. Strongest activity was reported against Fusarium oxysporum and Verticillium sp. No effect was observed against Alternaria sp.
Inoculation of the two bottom leaves (1 and 2) of tomato plants (Lycopersicon esculentum) cv. Vol... more Inoculation of the two bottom leaves (1 and 2) of tomato plants (Lycopersicon esculentum) cv. Vollendung with tobacco necrosis virus (TNV) induced systemic resistance against late blight disease caused by Phytophthora infestans SDS PAGE of acid soluble proteins extracted from TNV-inoculated plants showed accumulation of six pathogenesis-related proteins in inoculated leaves (1 and 2) and in upper leaf tissue (leaf 3). Western blot analysis with antisera raised against different pathogenesis-related proteins of tomato and tobacco revealed that four of these proteins were P14, AP24, chitinase and β-(1,3)-glucanase. An in itro study of the antifungal activity of these pathogenesis-related proteins showed that only the basic fraction has direct antifungal activity toward Phytophthora infestans. The induction of systemic resistance was accompanied by an increase of peroxidase activity in inoculated leaves as well as in upper leaf tissue (leaf 3). On the other hand, significant increase in the activity of β-(1,3)-glucanase was observed only in leaves inoculated with tobacco necrosis virus. Leaf tissue analysis of induced plants showed that inoculation with TNV resulted in the induction of three new peroxidase and one β-(1,3)-glucanase isozymes.
World Journal of Microbiology & Biotechnology, 2003
The plasmid profile of two thermophilic bacterial strains isolated from recreation thermal spring... more The plasmid profile of two thermophilic bacterial strains isolated from recreation thermal springs in Jordan has been investigated. These strains are Streptococcus thermophilus and Bacillus sp1, which have been isolated from Zerka – Maeen and Himma hot springs respectively. Supercoiled and circular plasmid forms were detected, explaining the effect of DNA conformation on the mobility of the plasmid in the
Yellowing symptoms similar to coconut yellow decline phytoplasma disease were observed on lipstic... more Yellowing symptoms similar to coconut yellow decline phytoplasma disease were observed on lipstick palms (Cyrtostachys renda) in Selangor state, Malaysia. Typical symptoms were yellowing, light green fronds, gradual collapse of older fronds and decline in growth. Polymerase chain reaction assay was employed to detect phytoplasma in symptomatic lipstick palms. Extracted DNA was amplified from symptomatic lipstick palms by PCR using phytoplasma-universal primer pair P1/P7 followed by R16F2n/R16R2. Phytoplasma presence was confirmed, and the 1250 bp products were cloned and sequenced. Sequence analysis indicated that the phytoplasmas associated with lipstick yellow frond disease were isolates of 'Candidatus Phytoplasma asteris' belonging to the 16SrI group. Virtual RFLP analysis of the resulting profiles revealed that these palm-infecting phytoplasmas belong to subgroup 16SrI-B and a possibly new 16SrI-subgroup. This is the first report of lipstick palm as a new host of aster yellows phytoplasma (16SrI) in Malaysia and worldwide.
In a survey conducted in 2002 and 2003, Citrus tristeza virus (CTV) was detected in the Jordan Va... more In a survey conducted in 2002 and 2003, Citrus tristeza virus (CTV) was detected in the Jordan Valley. The direct tissue blot immunoassay (DTBIA) indicated that 12.7 and 15.2% of samples tested in the central and northern Jordan Valley respectively were infected with CTV. Similar results showed that all citrus species grown in the Jordan Valley were susceptible to CTV. DAS-ELISA analysis of samples from a citrus orchard in the Dir Alla area with severe CTV symptoms indicated that 49% of samples were infected with CTV. Using a CTV specific primer pair (CTV1/CTV10), the coat protein gene of the virus was successfully amplified from leaf extracts obtained from CTVinfected trees by IC-RT-PCR. After cloning and sequencing the coat protein gene, the sequence of the amplified product was deposited in the GenBank.
A genomic strategy for PPV identification has been recently developed (Pasquini et al., 2008). Th... more A genomic strategy for PPV identification has been recently developed (Pasquini et al., 2008). The method is based on using a 70-mer oligonucleotide DNA microarray chip capable of simultaneously detecting and genotyping PPV strains. Universal and specific probes have been identified and used with a sensitive protocol of hybridization using an indirect fluorescent labelling of cDNA product with cyanine able to enhance the sensitivity of the virus detection avoiding the use of the PCR amplification step. In order to evaluate the protocol fitness for diagnostic use, about 30 samples belonging to a PPV isolates collection, including M, D, EA and C strains, have been used for its validation, that was determined, estimating the performance criteria that include the following parameters: diagnostic sensitivity (D-SN), diagnostic specificity (D-SP) and diagnostic accuracy (D-AC). Keywords : oligonucleotides chip, PPV, sensitivity, specificity, accuracy, performance criteria
In 2013, 23 tomato samples were collected from plants grown at Homret Al-Sahen region exhibiting ... more In 2013, 23 tomato samples were collected from plants grown at Homret Al-Sahen region exhibiting typical Tomato yellow leaf curl disease (TYLCD) symptoms. After total nucleic acids were extracted, all samples were subjected to PCR analysis using primers previously reported to detect Tomato yellow leaf curl Sardinia virus (TYLCSV), Tomato yellow leaf curl virus (TYLCV), recombinant type A viruses (named RecA, a recombinant between TYLCV/TYLCSV) and betasatellites. PCR analysis showed that 22 samples were infected with a mixture of TYLCV, TYLCSV and betasatellites, whereas one sample was infected with TYLCV, TYLCSV and RecA virus. The full-length genome (2763 bp) of RecA type virus, was amplified, cloned and sequenced. Sequence analysis showed that the RecA virus had the highest nucleotide identity (96%) with the Italian strain of Tomato yellow leaf curl Axarquia virus (TYLCAxV-Sic2-[IT-Sic2/5-04], EU734832). Recombination analysis revealed that RecA virus detected in this study (here...
Whiteflies are economically important plant pests that cause damage to crops worldwide. This stud... more Whiteflies are economically important plant pests that cause damage to crops worldwide. This study aimed to update the status of whiteflies in Jordan by combining the classical morphological identification and the DNA markers using the mitochondrial cytochrome oxidase I (mtCOI) gene. Over the course of three consecutive years, 111 whiteflies were collected from different geographical regions and different plant hosts in Jordan. The results showed that, in addition to Bemisia tabaci, another nine different whitefly species were identified, including two species that were recorded for the first time in Jordan: Africaleurodes coffeacola, and Tetraleurodes neemani. A special focus has been given to economically important plant pests like the B. tabaci species complex. Three different diagnostic techniques were used to identify B. tabaci putative species based on mtCOI gene. All the collected samples of B. tabaci species complex were identified as Middle East–Asia Minor 1 (MEAM1) putativ...
Tomato yellow leaf curl disease (TYLCD) is a major threat affecting tomato production in both tro... more Tomato yellow leaf curl disease (TYLCD) is a major threat affecting tomato production in both tropical and subtropical areas worldwide. The disease is caused by the Tomato yellow leaf curl virus and other related viruses that belong to the genus Begomovirus of the family Geminiviridae, which are transmitted by the whitefly (Bemisia tabaci). Since mechanical transmission of the disease is difficult and impractical, an efficient and easy method is needed. In this study, an agrodrench inoculation method was utilized where the suspension of Agrobacterium, harbouring a plasmid with the DNA of the virus, is drenched into the soil near the roots. This method proved to be easy and can be used to overcome several problems associated with other inoculation methods. In this study, the effectiveness of the agrodrench as an inoculation method for TYLCV was found to be influenced by the virus strain, agrobacterium strain and concentration, plant age, stem-wounding and host plant species.The metho...
The non-linear transition to adulthood of today’s youth signaled by numerous theoreticians has co... more The non-linear transition to adulthood of today’s youth signaled by numerous theoreticians has consequences on the forms of political participation favoured by young people. Nevertheless, citizenship education continues to play a significant role in shaping the political behaviour of future adults. After highlighting the characteristics of citizenship education in Europe as outlined in the most recent Eurydice study (2012) and briefly presenting the alternative forms of political and social participation embraced by the young generation, the present paper aims to determine the level of interest and involvement of Romanian teenagers in political and social activities at both national and European level. The research method employed is a quantitative one, pupils from the 11 and 12grade being the subjects of a survey conducted in six Romanian high schools. Findings reveal that although teenagers are willing to make their voice heard by various means, including alternative forms of part...
A protocol for production of virus-free Vitis vinifera using meristem culture was developed. Meri... more A protocol for production of virus-free Vitis vinifera using meristem culture was developed. Meristems (0.1-0.2 mm) of V. vinifera infected with grapevine fanleaf virus and grape leafroll associated viruses were excised from 1-year-old growing vines. Shoot tips were cultured on half-strength Murashige and Skoog (MS) medium, supplemented with 0.02 mg L-1 of benzylaminopurine (BAP) and 0.01 mg L-1 of naphthalene acetic acid (NAA). BAP and kinetin resulted in differences in the number of new shoots per explant, shoot height, and number of new leaves per explant. BAP at 0.8 mg L-1 gave the highest in vitro multiplication rate, with 5.25 shoots per explant, whereas elongation was greatest in the presence of 0.2 mg L-1 of kinetin. Root initiation was tested on an MS medium supplemented with 0.0, 0.2, 0.4, 0.6, 0.8, or 1.0 mg L-1 of IBA (indole-3-butyric-acid), IAA (indole-3-acetic-acid), or NAA. Maximum root number was achieved using 0.6 mg L-1 of IBA. A survival rate of 95% was achieved ...
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