To control the plant viral diseases, identification of the virus species is considered as a first... more To control the plant viral diseases, identification of the virus species is considered as a first step. Considering the stable structure of dsRNA in comparison to RNA molecules, dsRNA extraction is a promising tool to elucidate the viral genome sequence. Majority of the RNA viruses produce dsRNA segments as the copies of virus genome; these high molecular weight molecules can be employed in virus identification. To date, several efforts have been made to extract dsRNA from many different plant species. Developing a method charging less and completing fast has been investigated in this study.
In order to address biosafety concerns regarding the constitutive expression of foreign genes in ... more In order to address biosafety concerns regarding the constitutive expression of foreign genes in crops, we applied a strategy aimed at confining foreign gene expression in insect wounding sites of cotton. For this purpose, a plant expression construct was designed by cloning the AoPR1 promoter (pathogenesis-related protein gene isolated from Asparagus officinalis) upstream from the insecticidal gene cry1Ac. The Turkish cotton cultivar cv. STN-468 was transformed using the Agrobacterium tumefaciens strain LBA4404 containing the recombinant binary vector pRD400 harboring cry1Ac under a wound-inducible promoter. The neomycin phosphotransferase (nptII) gene was used as a selectable marker at a concentration of 100 mg/L. The primary transformants were analyzed for T-DNA integration and expression using standard molecular approaches. The efficacy of insecticidal gene control of the AoPR1 promoter was investigated using leaf bioassays with 2nd instar larvae of Helicoverpa armigera and Spod...
Sequence data from genomics projects on soybean has recently been made available and can be used ... more Sequence data from genomics projects on soybean has recently been made available and can be used for cloning genes or other genetic elements. In this study a pair of primer was designed based on the exon 3 of a maize gene coding for myo-inositol-1-phosphate synthase. This primer pair was used to amplify the homologous DNA from soybean. This fragment was cloned in a T/A vector and sequenced. There was a complete similarity between the sequence of the exon 3 of the maize gene and the genes coding for myo-inositol-1-phosphate synthase and chlorophylase enzyme in soybean. Considering the biological importance of this exon, this can now be used for knock out or overexpression of this gene in soybean.
Synthetic seeds are artificially encapsulated propagules that mimic true seeds in agriculture. Al... more Synthetic seeds are artificially encapsulated propagules that mimic true seeds in agriculture. Although a variety of plant materials, such as shoot tips, axillary buds, callus, micro cuttings, and protocorm-like bodies, are used in the production of synthetic seeds, somatic embryos are the most widely used explants in the production of these seeds. Synthetic seeds compete with traditional approaches to preserve the germplasm of threatened plant species. The resulting progenies are the true clones of the main plant, thus preserving the intactness of the genetic background. Due to poor germination and low seed amount, wild Beta species are exposed to the risk of extinction. Wild relatives of Beta have agronomically important properties such as resistance to diseases and abiotic stresses. Numerous attempts have been made to give these traits to sugar beet crop through conventional breeding methods. Despite the importance of synthetic seed for wild beets, it has not yet been investigate...
Extraction of viral double stranded RNA (dsRNA) from infected plants is helpful in identification... more Extraction of viral double stranded RNA (dsRNA) from infected plants is helpful in identification of the viruses involved in infection. To date, there have been several methods developed to isolate dsRNA; however, type of the plant and virus is determinative in extraction efficiency. In this study we extracted dsRNA from different woody and herbaceous plants through a modified method which reduces the costs and time of extraction procedure. This method is based on different affinity of nucleic acids for the cellulose CF-11 in1X STE (Sodium chloride Tris EDTA) buffer containing 16 % ethanol. There is no phenol treatment or mini columns used in the isolation procedure. Extracted dsRNAs were identified by ribonuclease treatment and RT-PCR (Reverse Transcriptase-Polymerase Chain Reaction). We have applied the procedure on five different hosts representing Amaranthaceae, Vitaceae, Fabaceae and Rosaceae infected with four different viruses representing Secoviridae and Bromoviridae.
Xiphinema index is an important grapevine pathogen nematode which also vectors Grapevine fanleaf ... more Xiphinema index is an important grapevine pathogen nematode which also vectors Grapevine fanleaf virus. The viral genes involved in transmission by the vector nematode are mapped to the C-terminal residues of RNA2-encoded polyprotein. To recognize viruliferous nematodes, there are some serological and molecular methods. In this study, we extract RNA and dsRNA of the virus, then Reverse transcription-polymerase Chain Reaction was done with virus specific primers to detect virus in its vector. The virus was detected by visualizing the desired 350 and 750 bp gene fragments in electrophoresis. This study reduces the virus detection time to only couple of hours with least imposed charges, and could be employed in transmission experiments as well.
Manipulating the oil crops to improve the oil quality has been a field of interest for genetic en... more Manipulating the oil crops to improve the oil quality has been a field of interest for genetic engineering researchers during the last years. Increasing the oleic acid content of Brassica napus has been targeted to decrease the cardiovascular diseases meanwhile biodiesel production. fad2 is a gene coding a key enzyme involved in oleic acid biosynthetic pathway in oil corps, named fatty acyl desaturase. Introducing a partial sequence of the aforementioned gene as an RNAi cassette in crops will increase the oleic acid ratio significantly in the oil extracted. Considering the start codon of the gene, the specific primers designed and following the genomic DNA extraction, the fragment of interest amplified by Polymerase Chain Reaction (PCR). After cloning in a desired vector, transformation to E. coli competent cells carried out. During the next steps the gene fragment underwent restriction and ligation reactions to achieve the desired orientation of sense and antisense fragments. At th...
Cotton (Gossypium hirsutum L.) is one of the most important fiber and oil crops with a great econ... more Cotton (Gossypium hirsutum L.) is one of the most important fiber and oil crops with a great economic value worldwide. Recent, trends in the development of agricultural crops with agronomic traits is inclined towards development of resistance to pests, herbicides, insecticides, pathogens and fiber-quality modification using genetic transformation techniques. Different varieties of cotton have evolved by hybridization that is why every regeneration protocol is cultivar specific and the researchers have to develop new protocols for each cultivar. Moreover, cotton is highly recalcitrant plant under in vitro conditions. Therefore, regeneration protocol was optimized for two cultivars SG-125 (Turkish cultivar) and Coker-310 to study the effects of media (MS, B5 & MS salt with B5 vitamin), PGRs (BAP, Kin singly or in combination with NAA), other supplements (activated charcoal, PVP & ascorbic acid with citric acid) on shoot regeneration from shoot meristem explants. Bialaphos (1, 2, 3, 5 ...
To control the plant viral diseases, identification of the virus species is considered as a first... more To control the plant viral diseases, identification of the virus species is considered as a first step. Considering the stable structure of dsRNA in comparison to RNA molecules, dsRNA extraction is a promising tool to elucidate the viral genome sequence. Majority of the RNA viruses produce dsRNA segments as the copies of virus genome; these high molecular weight molecules can be employed in virus identification. To date, several efforts have been made to extract dsRNA from many different plant species. Developing a method charging less and completing fast has been investigated in this study.
In order to address biosafety concerns regarding the constitutive expression of foreign genes in ... more In order to address biosafety concerns regarding the constitutive expression of foreign genes in crops, we applied a strategy aimed at confining foreign gene expression in insect wounding sites of cotton. For this purpose, a plant expression construct was designed by cloning the AoPR1 promoter (pathogenesis-related protein gene isolated from Asparagus officinalis) upstream from the insecticidal gene cry1Ac. The Turkish cotton cultivar cv. STN-468 was transformed using the Agrobacterium tumefaciens strain LBA4404 containing the recombinant binary vector pRD400 harboring cry1Ac under a wound-inducible promoter. The neomycin phosphotransferase (nptII) gene was used as a selectable marker at a concentration of 100 mg/L. The primary transformants were analyzed for T-DNA integration and expression using standard molecular approaches. The efficacy of insecticidal gene control of the AoPR1 promoter was investigated using leaf bioassays with 2nd instar larvae of Helicoverpa armigera and Spod...
Sequence data from genomics projects on soybean has recently been made available and can be used ... more Sequence data from genomics projects on soybean has recently been made available and can be used for cloning genes or other genetic elements. In this study a pair of primer was designed based on the exon 3 of a maize gene coding for myo-inositol-1-phosphate synthase. This primer pair was used to amplify the homologous DNA from soybean. This fragment was cloned in a T/A vector and sequenced. There was a complete similarity between the sequence of the exon 3 of the maize gene and the genes coding for myo-inositol-1-phosphate synthase and chlorophylase enzyme in soybean. Considering the biological importance of this exon, this can now be used for knock out or overexpression of this gene in soybean.
Synthetic seeds are artificially encapsulated propagules that mimic true seeds in agriculture. Al... more Synthetic seeds are artificially encapsulated propagules that mimic true seeds in agriculture. Although a variety of plant materials, such as shoot tips, axillary buds, callus, micro cuttings, and protocorm-like bodies, are used in the production of synthetic seeds, somatic embryos are the most widely used explants in the production of these seeds. Synthetic seeds compete with traditional approaches to preserve the germplasm of threatened plant species. The resulting progenies are the true clones of the main plant, thus preserving the intactness of the genetic background. Due to poor germination and low seed amount, wild Beta species are exposed to the risk of extinction. Wild relatives of Beta have agronomically important properties such as resistance to diseases and abiotic stresses. Numerous attempts have been made to give these traits to sugar beet crop through conventional breeding methods. Despite the importance of synthetic seed for wild beets, it has not yet been investigate...
Extraction of viral double stranded RNA (dsRNA) from infected plants is helpful in identification... more Extraction of viral double stranded RNA (dsRNA) from infected plants is helpful in identification of the viruses involved in infection. To date, there have been several methods developed to isolate dsRNA; however, type of the plant and virus is determinative in extraction efficiency. In this study we extracted dsRNA from different woody and herbaceous plants through a modified method which reduces the costs and time of extraction procedure. This method is based on different affinity of nucleic acids for the cellulose CF-11 in1X STE (Sodium chloride Tris EDTA) buffer containing 16 % ethanol. There is no phenol treatment or mini columns used in the isolation procedure. Extracted dsRNAs were identified by ribonuclease treatment and RT-PCR (Reverse Transcriptase-Polymerase Chain Reaction). We have applied the procedure on five different hosts representing Amaranthaceae, Vitaceae, Fabaceae and Rosaceae infected with four different viruses representing Secoviridae and Bromoviridae.
Xiphinema index is an important grapevine pathogen nematode which also vectors Grapevine fanleaf ... more Xiphinema index is an important grapevine pathogen nematode which also vectors Grapevine fanleaf virus. The viral genes involved in transmission by the vector nematode are mapped to the C-terminal residues of RNA2-encoded polyprotein. To recognize viruliferous nematodes, there are some serological and molecular methods. In this study, we extract RNA and dsRNA of the virus, then Reverse transcription-polymerase Chain Reaction was done with virus specific primers to detect virus in its vector. The virus was detected by visualizing the desired 350 and 750 bp gene fragments in electrophoresis. This study reduces the virus detection time to only couple of hours with least imposed charges, and could be employed in transmission experiments as well.
Manipulating the oil crops to improve the oil quality has been a field of interest for genetic en... more Manipulating the oil crops to improve the oil quality has been a field of interest for genetic engineering researchers during the last years. Increasing the oleic acid content of Brassica napus has been targeted to decrease the cardiovascular diseases meanwhile biodiesel production. fad2 is a gene coding a key enzyme involved in oleic acid biosynthetic pathway in oil corps, named fatty acyl desaturase. Introducing a partial sequence of the aforementioned gene as an RNAi cassette in crops will increase the oleic acid ratio significantly in the oil extracted. Considering the start codon of the gene, the specific primers designed and following the genomic DNA extraction, the fragment of interest amplified by Polymerase Chain Reaction (PCR). After cloning in a desired vector, transformation to E. coli competent cells carried out. During the next steps the gene fragment underwent restriction and ligation reactions to achieve the desired orientation of sense and antisense fragments. At th...
Cotton (Gossypium hirsutum L.) is one of the most important fiber and oil crops with a great econ... more Cotton (Gossypium hirsutum L.) is one of the most important fiber and oil crops with a great economic value worldwide. Recent, trends in the development of agricultural crops with agronomic traits is inclined towards development of resistance to pests, herbicides, insecticides, pathogens and fiber-quality modification using genetic transformation techniques. Different varieties of cotton have evolved by hybridization that is why every regeneration protocol is cultivar specific and the researchers have to develop new protocols for each cultivar. Moreover, cotton is highly recalcitrant plant under in vitro conditions. Therefore, regeneration protocol was optimized for two cultivars SG-125 (Turkish cultivar) and Coker-310 to study the effects of media (MS, B5 & MS salt with B5 vitamin), PGRs (BAP, Kin singly or in combination with NAA), other supplements (activated charcoal, PVP & ascorbic acid with citric acid) on shoot regeneration from shoot meristem explants. Bialaphos (1, 2, 3, 5 ...
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Papers by Saber Delpasand Khabbazi