Agrobactirium mediated gene transfer
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This document summarizes Agrobacterium-mediated gene transfer. It begins with an introduction to gene transformation and describes Agrobacterium as a soil bacterium that causes crown gall disease in plants by transferring genes. It then discusses the classification of Agrobacterium, the history of using Agrobacterium for gene transfer, and components involved like the T-DNA, virulence genes, and chromosomal genes. The document outlines the process of T-DNA transfer from Agrobacterium to plant cells and concludes with some common methods for Agrobacterium-mediated gene transfer.
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Agrobacterium mediated gene transfer
Agrobacterium tumefaciens is commonly used to genetically modify dicot plants through its ability to transfer DNA to plant cells. It causes crown gall disease by transferring oncogenic T-DNA from its tumor-inducing plasmid into wounded plant cells. The T-DNA encodes genes that cause tumor formation and the production of opines, which the bacteria can use as nutrients. The binary vector system was developed to overcome challenges with manipulating large Ti plasmids, allowing foreign genes to be stably introduced between the T-DNA borders and transferred to plant cells. Agrobacterium-mediated transformation is now widely used to produce transgenic plants.
Agrobacterium mediated gene transformation
This document provides an overview of Agrobacterium-mediated gene transformation. It begins with an introduction to genetic transformation methods, including direct and indirect techniques. It then discusses Agrobacterium, including its classification, the history of using it for gene transformation, and features of its T-DNA and virulence genes. The document outlines the process of T-DNA transfer from Agrobacterium to plant cells. Finally, it describes some common methods for Agrobacterium-mediated gene transfer, such as infection through wounds, leaf disk, and co-cultivation techniques.
Chloroplast transformation
Chloroplast transformation allows for the integration of foreign genes into the chloroplast genome. This is beneficial as it provides high levels of transgene expression without epigenetic effects or position effects. Chloroplast transformation requires a chloroplast specific expression vector, a method for DNA delivery such as biolistics, and an efficient selection marker such as spectinomycin resistance. Successful transformation is confirmed by PCR and Southern blot analysis showing integration of the transgene into the chloroplast genome. Applications of chloroplast transformation include production of biopharmaceuticals, vaccines, industrial enzymes, and biomaterials as well as phytoremediation.
Agrobacterium mediated gene transfer
1) Agrobacterium tumefaciens is a soil bacterium that was discovered to transfer genes between itself and plants, enabling the development of genetic engineering methods for plants.
2) The most common transgenic traits in crops include herbicide and insect resistance, with field corn often containing Bt genes for insect resistance.
3) Agrobacterium-mediated transformation is the most widely used method for producing transgenic plants. It involves using disarmed Agrobacterium strains to transfer desired gene sequences into plant cells.
Vector mediated gene transfer methods for transgenesis in Plants.
Presentation include Vector mediated gene transfer methods for trans-genesis in Plants. Only Vector-based methods are covered. Vectors includes Bacteria, Viruses, transposable genetic elements. Other possible vectors for transgenesis are also covered.
Agrobacterium mediated transformation
This document summarizes Agrobacterium-mediated plant transformation. It describes how the soil bacterium Agrobacterium tumefaciens causes crown gall disease in plants by transferring oncogenic T-DNA from its Ti plasmid into the plant genome. Scientists have exploited this natural process to develop transformation systems where they insert new genes between the border sequences of disarmed Ti plasmids, allowing transfer of the recombinant T-DNA into plant cells. While effective in dicots, transformation of monocots proved more difficult due to their limited regeneration ability, though biolistic methods using microprojectile bombardment have succeeded in some important crop species.
Agrobacterium mediated gene transfer
Agrobacterium is a soil-borne bacteria that can transfer DNA fragments called T-DNA from its tumor-inducing (Ti) plasmid into plant cells. This process allows for efficient gene transfer. The T-DNA contains oncogenes that cause tumor formation in plants as well as genes for opine synthesis. Virulence genes on the Ti plasmid regulate T-DNA transfer through a type IV secretion system. There are two main strategies for using Agrobacterium for plant genetic engineering - co-integration vectors, where the gene of interest is inserted into the Ti plasmid, and binary vectors, where the T-DNA and virulence genes are on separate replicons that complement each other in Agrobacterium.
Molecular farming
Molecular Farming, green biotechnology, milestones, general strategy, plant expression hosts, advantage & limitation, plant-based expression system, biosafety concern
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Agrobacterium mediated gene transfer
Agrobacterium tumefaciens is commonly used to genetically modify dicot plants through its ability to transfer DNA to plant cells. It causes crown gall disease by transferring oncogenic T-DNA from its tumor-inducing plasmid into wounded plant cells. The T-DNA encodes genes that cause tumor formation and the production of opines, which the bacteria can use as nutrients. The binary vector system was developed to overcome challenges with manipulating large Ti plasmids, allowing foreign genes to be stably introduced between the T-DNA borders and transferred to plant cells. Agrobacterium-mediated transformation is now widely used to produce transgenic plants.
Agrobacterium mediated gene transformation
This document provides an overview of Agrobacterium-mediated gene transformation. It begins with an introduction to genetic transformation methods, including direct and indirect techniques. It then discusses Agrobacterium, including its classification, the history of using it for gene transformation, and features of its T-DNA and virulence genes. The document outlines the process of T-DNA transfer from Agrobacterium to plant cells. Finally, it describes some common methods for Agrobacterium-mediated gene transfer, such as infection through wounds, leaf disk, and co-cultivation techniques.
Chloroplast transformation
Chloroplast transformation allows for the integration of foreign genes into the chloroplast genome. This is beneficial as it provides high levels of transgene expression without epigenetic effects or position effects. Chloroplast transformation requires a chloroplast specific expression vector, a method for DNA delivery such as biolistics, and an efficient selection marker such as spectinomycin resistance. Successful transformation is confirmed by PCR and Southern blot analysis showing integration of the transgene into the chloroplast genome. Applications of chloroplast transformation include production of biopharmaceuticals, vaccines, industrial enzymes, and biomaterials as well as phytoremediation.
Agrobacterium mediated gene transfer
1) Agrobacterium tumefaciens is a soil bacterium that was discovered to transfer genes between itself and plants, enabling the development of genetic engineering methods for plants.
2) The most common transgenic traits in crops include herbicide and insect resistance, with field corn often containing Bt genes for insect resistance.
3) Agrobacterium-mediated transformation is the most widely used method for producing transgenic plants. It involves using disarmed Agrobacterium strains to transfer desired gene sequences into plant cells.
Vector mediated gene transfer methods for transgenesis in Plants.
Presentation include Vector mediated gene transfer methods for trans-genesis in Plants. Only Vector-based methods are covered. Vectors includes Bacteria, Viruses, transposable genetic elements. Other possible vectors for transgenesis are also covered.
Agrobacterium mediated transformation
This document summarizes Agrobacterium-mediated plant transformation. It describes how the soil bacterium Agrobacterium tumefaciens causes crown gall disease in plants by transferring oncogenic T-DNA from its Ti plasmid into the plant genome. Scientists have exploited this natural process to develop transformation systems where they insert new genes between the border sequences of disarmed Ti plasmids, allowing transfer of the recombinant T-DNA into plant cells. While effective in dicots, transformation of monocots proved more difficult due to their limited regeneration ability, though biolistic methods using microprojectile bombardment have succeeded in some important crop species.
Agrobacterium mediated gene transfer
Agrobacterium is a soil-borne bacteria that can transfer DNA fragments called T-DNA from its tumor-inducing (Ti) plasmid into plant cells. This process allows for efficient gene transfer. The T-DNA contains oncogenes that cause tumor formation in plants as well as genes for opine synthesis. Virulence genes on the Ti plasmid regulate T-DNA transfer through a type IV secretion system. There are two main strategies for using Agrobacterium for plant genetic engineering - co-integration vectors, where the gene of interest is inserted into the Ti plasmid, and binary vectors, where the T-DNA and virulence genes are on separate replicons that complement each other in Agrobacterium.
Molecular farming
Molecular Farming, green biotechnology, milestones, general strategy, plant expression hosts, advantage & limitation, plant-based expression system, biosafety concern
Ti Plasmid
Agrobacterium tumefaciens is a soil bacterium that can transfer DNA fragments called T-DNA from its tumor-inducing (Ti) plasmid into the genome of plant cells. Scientists have harnessed this natural ability of A. tumefaciens by modifying its Ti plasmid to replace the tumor-causing genes with a gene of interest. This disarmed Ti plasmid, along with helper plasmids containing virulence genes, allows for the stable integration and expression of the new gene in plant cells. Common transformation methods using A. tumefaciens involve culturing plant explants like leaf disks with Agrobacteria containing the modified Ti plasmid, then regenerating transgenic plants from the transformed cells
T dna & transposone tagging 1 (2)
Gene tagging uses recognizable DNA fragments like T-DNA or transposons to disrupt gene function and identify genes responsible for mutant phenotypes. T-DNA tagging in plants involves random integration of Agrobacterium T-DNA that can disrupt genes and create mutants. Transposon tagging relies on the ability of transposons to move within genomes and disrupt gene function. Both techniques have been used successfully to isolate numerous plant genes involved in traits like color and development.
Gene stacking
Gene stacking refers to combining two or more transgenes into a single plant genome. It provides broader crop traits than single genes alone. Various techniques for gene stacking include sexual hybridization, retransformation, and cotransformation. Cotransformation allows integrating multiple transgenes together, making it more efficient than other methods. Coordinated expression of stacked genes requires addressing issues like promoter homology and position effects. Future work includes refining techniques for stable multigene expression and identifying new gene combinations to improve crop traits.
Transgenic plants with biotic stress resistance
This document discusses transgenic plants with resistance to biotic stress. It begins by defining biotic stress as damage caused by living organisms like bacteria, viruses, fungi and insects. Developing transgenic plants is presented as a way to improve crop yields by making plants resistant to these stresses. Specific examples of transgenic plants containing genes from Bacillus thuringiensis (Bt) that code for insecticidal proteins are described. Bt genes have been introduced into crops like corn, cotton and potatoes to resist pests like rootworms and Colorado potato beetles. While Bt crops have increased yields, there is a risk of pests developing resistance over time if not managed properly.
herbicide resistance in plants
Herbicide tolerance plants are developed to control weeds that compete with crops for resources and can reduce crop yields by 20% annually. Weeds are typically controlled mechanically or through herbicides. Glyphosate is a commonly used broad-spectrum herbicide that acts by inhibiting the shikimate pathway enzyme EPSP synthase, preventing aromatic amino acid production. Transgenic plants are made tolerant to glyphosate by introducing the aroA gene to allow continued shikimate pathway function. Phosphinothricin resistance is conferred by the bar gene, which allows detoxification of the glutamine synthase inhibitor phosphinothricin. Bromoxynil tolerance results from the bxn gene introducing nitril
Pest and herbicide resistance
Genetic engineering has led to pest and herbicide resistance in plants. The document discusses how the Bt gene from Bacillus thuringiensis was introduced into plants like cotton to make them resistant to lepidopteran insect pests. It also describes how Roundup Ready soybeans were developed to be resistant to the herbicide glyphosate by expressing a modified version of the EPSPS enzyme. The mechanisms of action of Bt toxins and glyphosate resistance are explained at the molecular level. Overall, the genetic engineering of pest and herbicide resistance traits in crops provides environmental and economic benefits over traditional pesticide and herbicide use.
Chloroplast transformation
Chloroplasts contain their own DNA and are the site of photosynthesis. Chloroplast transformation involves delivering a vector with the gene of interest and a selectable marker flanked by chloroplast DNA sequences for homologous recombination. The vector is delivered using biolistics or PEG-mediated transformation. Transformed cells are selected using antibiotic resistance and regenerated into plants. Chloroplast transformation allows high-level expression of transgenes due to high copy number and avoids gene silencing.
Agrobacterium mediated gene transfer
This document summarizes Agrobacterium-mediated gene transfer, which is a process where genes are transferred from Agrobacterium tumefaciens bacteria to plant cells. Key points include: A. tumefaciens transfers oncogenes from its Ti plasmid to plant genomes, integrating the T-DNA and causing crown gall disease; the process involves virulence genes and results in opine production in the plant; common methods are wounding, co-cultivation, and leaf disks; transformation allows foreign genes to be stably integrated and expressed in plants.
In-planta transformation: recent advances
In-planta transformation is a direct transformation method that does not require tissue culture steps. It has several advantages over traditional transformation techniques, including being more efficient, quick, and not requiring regeneration systems. Various in-planta methods have been developed, including floral dip, vacuum infiltration, and pollen-tube mediated transformation. However, transformation efficiencies tend to be low across methods and plant species. Further optimization is still needed to improve transformation frequencies and consistency of transgene expression for many crop species using in-planta transformation approaches.
Agrobacterium-mediated Gene Transfer
An overview of the Agrobacterium-mediated gene transfer process. Moreover, studied different kinds of Agrobacterium species are involved in this mechanism.
Agrobacterium is a rod-shaped, Gram-negative bacteria found mostly in the soil. It is a plant pathogen that is responsible for causing crown gall disease in them. This bacteria is also known as the natural genetic engineer because of it's the ability to integrate its plasmid Gene into the plant genome.
Agrobacterium tumefaciens transfer of their genetic material T-DNA of Ti-plasmid into the plant cell: A: Agrobacterium tumefaciens; B: Agrobacterium genome; C: Ti Plasmid : a: T-DNA , b: Vir genes , c: Replication origin , d: Opines catabolism genes; D: Plant cell
A Ti-Plasmid (tumor-inducing plasmid) is a ds, circular DNA that often, but not always. It's a piece of genetic equipment that transfers genetic material from bacterial cells means Agrobacterium tumefaciens into plant cells used to induce tumors in the plant. The Ti-plasmid is damage when Agrobacterium is grown above 28 °C. Such cured bacteria don't induce crown gall disease in the plant due to they are avirulent. The Ti-Plasmid are classified into two types on the basis of opine genes are present in T-DNA.
The Plasmid has 196 genes that code for 195 proteins. There is no one structural RNA. The plasmid is 206.479 nucleotides long. the GC content is 56% and 81% of the genetic material is coding genes.
The modification of this plasmid is a very important source in the production of transgenic plants.
The T-DNA must be cut out of the circular plasmid. A VirD1/D2 complex nicks the DNA at the left and right border sequences. The VirD2 protein is covalently attached to the 5' end. VirD2 contains a motif that leads to the nucleoprotein complex being targeted to the type IV secretion system (T4SS).
In the cytoplasm of the recipient cell, the T-DNA complex becomes coated with VirE2 proteins, which are exported through the T4SS independently from the T-DNA complex. Nuclear localization signals, or NLS, located on the VirE2 and VirD2 are recognized by the importin alpha protein, which then associates with importin beta and the nuclear pore complex to transfer the T-DNA into the nucleus. So that the T-DNA can integrate into the host genome.
We inoculate Agrobacterium containing our genes of interest, onto wounded plant tissue explants. The Agrobacterium then transfers the gene of interest into the DNA of the plant tissue.
Biotechnology:Agrobacterium mediated Transformation
Agrobacterium mediated Transformation-Mechanism of gene transfer,Virulence induction in presence of Plant secondary metabolites,Chromosomal genes and Vir genes,Agrobacterium tumefaciens – pathogen and useful tool for genetic engineering
Bt brinjal
Bt technology uses genes from Bacillus thuringiensis to produce insecticidal crystal proteins in transgenic crops. There are several biosafety concerns regarding risks to human health from toxicity or allergies, as well as risks to the environment from increased insect resistance, gene flow to weeds or soil organisms, and effects on biodiversity. Regulatory agencies in India require various levels of approval from institutional biosafety committees, the Review Committee on Genetic Manipulation, and the Genetic Engineering Approval Committee, depending on the type and scale of field trials or commercial releases of Bt crops.
Mechanism of T-DNA to transfer genes into plants
The document summarizes the mechanism of T-DNA transfer during Agrobacterium tumefaciens infection. It explains that T-DNA is a fragment of DNA transferred from the tumor-inducing (Ti) plasmid of A. tumefaciens into the host plant genome. The T-DNA is bordered by repeats and encodes genes that cause tumors in the plant. Virulence genes are expressed in response to plant signals and produce single-stranded T-DNA, which forms a complex with other proteins and is transported into the plant cell and integrated into the plant nuclear DNA, causing uncontrolled cell growth and tumor formation. The mechanism involves multiple virulence protein complexes and integration of T-DNA is directed by the
Genetic engineering and development of transgenic plants
Genetic engineering can be used to develop transgenic plants with desirable traits. The process involves introducing foreign genes into plant cells, which are then regenerated into whole plants. Genes from Bacillus thuringiensis (Bt) have been introduced to many crops like cotton, corn and potatoes to make them resistant to pests. Other genes introduced include protease inhibitors and alpha amylase inhibitors. While transgenic plants can increase yields and reduce pesticide use, some risks include increased allergenicity and the development of resistance in pest populations.
Chloroplast transformation
Presented by- MD JAKIR HOSSAIN
Doctoral Research Scholar
Department of Agricultural Genetic Engineering ,
Faculty of Agricultural Sciences and Technologies,
Nigde Omer Halisdemir University, Turkey
E. Mail- mjakirbotru@gmail.com
Agrobacterium tumefaciens mediated gene transfer
Agrobacterium tumefaciens is a soil bacterium that causes crown gall disease in plants. It transfers a segment of DNA called T-DNA from its Ti plasmid into the plant genome, enabling it to modify plant cell growth. The presentation discusses the bacterial characteristics, mechanism of T-DNA transfer, use of Agrobacterium for genetic engineering of plants through insertion of foreign genes in place of tumor-causing genes on the Ti plasmid, and the regeneration of transformed plants. While it is an effective tool for plant genetic engineering, some plant species are not susceptible to Agrobacterium infection.
Agrobacterium tumefaciens
Agrobacterium tumefaciens is a soil bacterium that causes crown gall disease in plants. It does this by transferring a segment of its tumor-inducing plasmid (Ti plasmid) called T-DNA into the plant's DNA. The T-DNA contains genes that cause uncontrolled cell growth. Researchers developed techniques using the Ti plasmid and Agrobacterium to genetically transform plants by replacing the tumor-causing genes with desired genes. This involves either a binary vector system with the T-DNA on a separate small plasmid, or co-integration of a new plasmid containing the gene of interest into the Ti plasmid. Transformed plants can be regenerated from infected plant cells or tissues.
Agrobacterium mediated gene transfer in plants.
This power point presentation consist of 41 slides. Attempts have been made to illustrate how Agrobacterium behaves us natural genetic engineer. How it can infect a plant through wound and a part of DNA present on Ti plasmid is Tranferred and causes disease as crown gall in the infected plant. In second part of the presentation attempts have been made to describe how Agrobacterium can be utilized for iinsertion of desired gene into the plant,what manipulation are to be made with Agrobacterium.How infection and transfer of desired gene can be made possible.What is the role of plant tissue culture etc.
Tarns-genesis and development of transgenic plant
This document provides an overview of transgenesis and the development of transgenic plants. It defines key terms like transgene and transgenic plants. It describes traditional plant breeding techniques and compares them to transgenic technology. Transgenic technology allows genes to be transferred between any organisms, while traditional breeding is limited to the same genus. Reasons for developing transgenic plants include crop improvement, disease resistance, and stress tolerance. The document outlines the process of developing transgenic plants, including vector-mediated gene transfer using Agrobacterium and biolistic methods. It provides examples of transgenic plants created for insect resistance, herbicide tolerance, drought tolerance, and more. Both advantages and disadvantages of transgenic plants are discussed.
Agrobacterium mediated gene transfer
This document summarizes the process of plant genetic transformation using Agrobacterium tumefaciens. It describes how A. tumefaciens transfers T-DNA from its Ti plasmid into plant cells, integrating the T-DNA into the plant genome and expressing genes that cause crown gall disease. The document also outlines the key steps in the process, from gene transfer to the plant cell through regeneration of a transformed whole plant and methods to detect successful transformation events. Common genes inserted into transgenic crops are also listed, including genes for herbicide and insect resistance.
Agrobacteruim_mediated_gene_transfer.ppt
This document discusses Agrobacterium tumefaciens and its use in genetic engineering. Agrobacterium tumefaciens is a bacteria that causes crown gall disease in plants by transferring T-DNA from its Ti plasmid into the plant genome. Scientists discovered that the T-DNA transfer mechanism can be used to genetically engineer plants by replacing the T-DNA genes with a gene of interest. The virulence genes and T-DNA borders are required for transfer, while the chromosomal chv genes facilitate bacterial binding to the plant. Now Agrobacterium is widely used to insert foreign genes into plants and produce transgenic crops like tobacco and rice.
Agrobacterium tumefaciens as a tool for genetic engineering in plants
Agrobacterium tumefaciens is a tool for genetic engineering in plants. It is a soil bacterium that can transfer DNA fragments called T-DNA from its tumor-inducing plasmid into the genome of plant cells. The T-DNA can be modified to contain desirable genes for traits like herbicide or pest resistance. The bacterium recognizes wounded plant cells and transfers T-DNA using virulence genes. Integrated T-DNA is then expressed stably in the plant genome. Agrobacterium-mediated transformation is widely used for genetic engineering in plants due to its simplicity, efficiency and ability to transfer large DNA segments.
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Ti Plasmid
Agrobacterium tumefaciens is a soil bacterium that can transfer DNA fragments called T-DNA from its tumor-inducing (Ti) plasmid into the genome of plant cells. Scientists have harnessed this natural ability of A. tumefaciens by modifying its Ti plasmid to replace the tumor-causing genes with a gene of interest. This disarmed Ti plasmid, along with helper plasmids containing virulence genes, allows for the stable integration and expression of the new gene in plant cells. Common transformation methods using A. tumefaciens involve culturing plant explants like leaf disks with Agrobacteria containing the modified Ti plasmid, then regenerating transgenic plants from the transformed cells
T dna & transposone tagging 1 (2)
Gene tagging uses recognizable DNA fragments like T-DNA or transposons to disrupt gene function and identify genes responsible for mutant phenotypes. T-DNA tagging in plants involves random integration of Agrobacterium T-DNA that can disrupt genes and create mutants. Transposon tagging relies on the ability of transposons to move within genomes and disrupt gene function. Both techniques have been used successfully to isolate numerous plant genes involved in traits like color and development.
Gene stacking
Gene stacking refers to combining two or more transgenes into a single plant genome. It provides broader crop traits than single genes alone. Various techniques for gene stacking include sexual hybridization, retransformation, and cotransformation. Cotransformation allows integrating multiple transgenes together, making it more efficient than other methods. Coordinated expression of stacked genes requires addressing issues like promoter homology and position effects. Future work includes refining techniques for stable multigene expression and identifying new gene combinations to improve crop traits.
Transgenic plants with biotic stress resistance
This document discusses transgenic plants with resistance to biotic stress. It begins by defining biotic stress as damage caused by living organisms like bacteria, viruses, fungi and insects. Developing transgenic plants is presented as a way to improve crop yields by making plants resistant to these stresses. Specific examples of transgenic plants containing genes from Bacillus thuringiensis (Bt) that code for insecticidal proteins are described. Bt genes have been introduced into crops like corn, cotton and potatoes to resist pests like rootworms and Colorado potato beetles. While Bt crops have increased yields, there is a risk of pests developing resistance over time if not managed properly.
herbicide resistance in plants
Herbicide tolerance plants are developed to control weeds that compete with crops for resources and can reduce crop yields by 20% annually. Weeds are typically controlled mechanically or through herbicides. Glyphosate is a commonly used broad-spectrum herbicide that acts by inhibiting the shikimate pathway enzyme EPSP synthase, preventing aromatic amino acid production. Transgenic plants are made tolerant to glyphosate by introducing the aroA gene to allow continued shikimate pathway function. Phosphinothricin resistance is conferred by the bar gene, which allows detoxification of the glutamine synthase inhibitor phosphinothricin. Bromoxynil tolerance results from the bxn gene introducing nitril
Pest and herbicide resistance
Genetic engineering has led to pest and herbicide resistance in plants. The document discusses how the Bt gene from Bacillus thuringiensis was introduced into plants like cotton to make them resistant to lepidopteran insect pests. It also describes how Roundup Ready soybeans were developed to be resistant to the herbicide glyphosate by expressing a modified version of the EPSPS enzyme. The mechanisms of action of Bt toxins and glyphosate resistance are explained at the molecular level. Overall, the genetic engineering of pest and herbicide resistance traits in crops provides environmental and economic benefits over traditional pesticide and herbicide use.
Chloroplast transformation
Chloroplasts contain their own DNA and are the site of photosynthesis. Chloroplast transformation involves delivering a vector with the gene of interest and a selectable marker flanked by chloroplast DNA sequences for homologous recombination. The vector is delivered using biolistics or PEG-mediated transformation. Transformed cells are selected using antibiotic resistance and regenerated into plants. Chloroplast transformation allows high-level expression of transgenes due to high copy number and avoids gene silencing.
Agrobacterium mediated gene transfer
This document summarizes Agrobacterium-mediated gene transfer, which is a process where genes are transferred from Agrobacterium tumefaciens bacteria to plant cells. Key points include: A. tumefaciens transfers oncogenes from its Ti plasmid to plant genomes, integrating the T-DNA and causing crown gall disease; the process involves virulence genes and results in opine production in the plant; common methods are wounding, co-cultivation, and leaf disks; transformation allows foreign genes to be stably integrated and expressed in plants.
In-planta transformation: recent advances
In-planta transformation is a direct transformation method that does not require tissue culture steps. It has several advantages over traditional transformation techniques, including being more efficient, quick, and not requiring regeneration systems. Various in-planta methods have been developed, including floral dip, vacuum infiltration, and pollen-tube mediated transformation. However, transformation efficiencies tend to be low across methods and plant species. Further optimization is still needed to improve transformation frequencies and consistency of transgene expression for many crop species using in-planta transformation approaches.
Agrobacterium-mediated Gene Transfer
An overview of the Agrobacterium-mediated gene transfer process. Moreover, studied different kinds of Agrobacterium species are involved in this mechanism.
Agrobacterium is a rod-shaped, Gram-negative bacteria found mostly in the soil. It is a plant pathogen that is responsible for causing crown gall disease in them. This bacteria is also known as the natural genetic engineer because of it's the ability to integrate its plasmid Gene into the plant genome.
Agrobacterium tumefaciens transfer of their genetic material T-DNA of Ti-plasmid into the plant cell: A: Agrobacterium tumefaciens; B: Agrobacterium genome; C: Ti Plasmid : a: T-DNA , b: Vir genes , c: Replication origin , d: Opines catabolism genes; D: Plant cell
A Ti-Plasmid (tumor-inducing plasmid) is a ds, circular DNA that often, but not always. It's a piece of genetic equipment that transfers genetic material from bacterial cells means Agrobacterium tumefaciens into plant cells used to induce tumors in the plant. The Ti-plasmid is damage when Agrobacterium is grown above 28 °C. Such cured bacteria don't induce crown gall disease in the plant due to they are avirulent. The Ti-Plasmid are classified into two types on the basis of opine genes are present in T-DNA.
The Plasmid has 196 genes that code for 195 proteins. There is no one structural RNA. The plasmid is 206.479 nucleotides long. the GC content is 56% and 81% of the genetic material is coding genes.
The modification of this plasmid is a very important source in the production of transgenic plants.
The T-DNA must be cut out of the circular plasmid. A VirD1/D2 complex nicks the DNA at the left and right border sequences. The VirD2 protein is covalently attached to the 5' end. VirD2 contains a motif that leads to the nucleoprotein complex being targeted to the type IV secretion system (T4SS).
In the cytoplasm of the recipient cell, the T-DNA complex becomes coated with VirE2 proteins, which are exported through the T4SS independently from the T-DNA complex. Nuclear localization signals, or NLS, located on the VirE2 and VirD2 are recognized by the importin alpha protein, which then associates with importin beta and the nuclear pore complex to transfer the T-DNA into the nucleus. So that the T-DNA can integrate into the host genome.
We inoculate Agrobacterium containing our genes of interest, onto wounded plant tissue explants. The Agrobacterium then transfers the gene of interest into the DNA of the plant tissue.
Biotechnology:Agrobacterium mediated Transformation
Agrobacterium mediated Transformation-Mechanism of gene transfer,Virulence induction in presence of Plant secondary metabolites,Chromosomal genes and Vir genes,Agrobacterium tumefaciens – pathogen and useful tool for genetic engineering
Bt brinjal
Bt technology uses genes from Bacillus thuringiensis to produce insecticidal crystal proteins in transgenic crops. There are several biosafety concerns regarding risks to human health from toxicity or allergies, as well as risks to the environment from increased insect resistance, gene flow to weeds or soil organisms, and effects on biodiversity. Regulatory agencies in India require various levels of approval from institutional biosafety committees, the Review Committee on Genetic Manipulation, and the Genetic Engineering Approval Committee, depending on the type and scale of field trials or commercial releases of Bt crops.
Mechanism of T-DNA to transfer genes into plants
The document summarizes the mechanism of T-DNA transfer during Agrobacterium tumefaciens infection. It explains that T-DNA is a fragment of DNA transferred from the tumor-inducing (Ti) plasmid of A. tumefaciens into the host plant genome. The T-DNA is bordered by repeats and encodes genes that cause tumors in the plant. Virulence genes are expressed in response to plant signals and produce single-stranded T-DNA, which forms a complex with other proteins and is transported into the plant cell and integrated into the plant nuclear DNA, causing uncontrolled cell growth and tumor formation. The mechanism involves multiple virulence protein complexes and integration of T-DNA is directed by the
Genetic engineering and development of transgenic plants
Genetic engineering can be used to develop transgenic plants with desirable traits. The process involves introducing foreign genes into plant cells, which are then regenerated into whole plants. Genes from Bacillus thuringiensis (Bt) have been introduced to many crops like cotton, corn and potatoes to make them resistant to pests. Other genes introduced include protease inhibitors and alpha amylase inhibitors. While transgenic plants can increase yields and reduce pesticide use, some risks include increased allergenicity and the development of resistance in pest populations.
Chloroplast transformation
Presented by- MD JAKIR HOSSAIN
Doctoral Research Scholar
Department of Agricultural Genetic Engineering ,
Faculty of Agricultural Sciences and Technologies,
Nigde Omer Halisdemir University, Turkey
E. Mail- mjakirbotru@gmail.com
Agrobacterium tumefaciens mediated gene transfer
Agrobacterium tumefaciens is a soil bacterium that causes crown gall disease in plants. It transfers a segment of DNA called T-DNA from its Ti plasmid into the plant genome, enabling it to modify plant cell growth. The presentation discusses the bacterial characteristics, mechanism of T-DNA transfer, use of Agrobacterium for genetic engineering of plants through insertion of foreign genes in place of tumor-causing genes on the Ti plasmid, and the regeneration of transformed plants. While it is an effective tool for plant genetic engineering, some plant species are not susceptible to Agrobacterium infection.
Agrobacterium tumefaciens
Agrobacterium tumefaciens is a soil bacterium that causes crown gall disease in plants. It does this by transferring a segment of its tumor-inducing plasmid (Ti plasmid) called T-DNA into the plant's DNA. The T-DNA contains genes that cause uncontrolled cell growth. Researchers developed techniques using the Ti plasmid and Agrobacterium to genetically transform plants by replacing the tumor-causing genes with desired genes. This involves either a binary vector system with the T-DNA on a separate small plasmid, or co-integration of a new plasmid containing the gene of interest into the Ti plasmid. Transformed plants can be regenerated from infected plant cells or tissues.
Agrobacterium mediated gene transfer in plants.
This power point presentation consist of 41 slides. Attempts have been made to illustrate how Agrobacterium behaves us natural genetic engineer. How it can infect a plant through wound and a part of DNA present on Ti plasmid is Tranferred and causes disease as crown gall in the infected plant. In second part of the presentation attempts have been made to describe how Agrobacterium can be utilized for iinsertion of desired gene into the plant,what manipulation are to be made with Agrobacterium.How infection and transfer of desired gene can be made possible.What is the role of plant tissue culture etc.
Tarns-genesis and development of transgenic plant
This document provides an overview of transgenesis and the development of transgenic plants. It defines key terms like transgene and transgenic plants. It describes traditional plant breeding techniques and compares them to transgenic technology. Transgenic technology allows genes to be transferred between any organisms, while traditional breeding is limited to the same genus. Reasons for developing transgenic plants include crop improvement, disease resistance, and stress tolerance. The document outlines the process of developing transgenic plants, including vector-mediated gene transfer using Agrobacterium and biolistic methods. It provides examples of transgenic plants created for insect resistance, herbicide tolerance, drought tolerance, and more. Both advantages and disadvantages of transgenic plants are discussed.
Agrobacterium mediated gene transfer
This document summarizes the process of plant genetic transformation using Agrobacterium tumefaciens. It describes how A. tumefaciens transfers T-DNA from its Ti plasmid into plant cells, integrating the T-DNA into the plant genome and expressing genes that cause crown gall disease. The document also outlines the key steps in the process, from gene transfer to the plant cell through regeneration of a transformed whole plant and methods to detect successful transformation events. Common genes inserted into transgenic crops are also listed, including genes for herbicide and insect resistance.
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Biotechnology:Agrobacterium mediated Transformation
Biotechnology:Agrobacterium mediated Transformation
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Genetic engineering and development of transgenic plants
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Agrobacteruim_mediated_gene_transfer.ppt
This document discusses Agrobacterium tumefaciens and its use in genetic engineering. Agrobacterium tumefaciens is a bacteria that causes crown gall disease in plants by transferring T-DNA from its Ti plasmid into the plant genome. Scientists discovered that the T-DNA transfer mechanism can be used to genetically engineer plants by replacing the T-DNA genes with a gene of interest. The virulence genes and T-DNA borders are required for transfer, while the chromosomal chv genes facilitate bacterial binding to the plant. Now Agrobacterium is widely used to insert foreign genes into plants and produce transgenic crops like tobacco and rice.
Agrobacterium tumefaciens as a tool for genetic engineering in plants
Agrobacterium tumefaciens is a tool for genetic engineering in plants. It is a soil bacterium that can transfer DNA fragments called T-DNA from its tumor-inducing plasmid into the genome of plant cells. The T-DNA can be modified to contain desirable genes for traits like herbicide or pest resistance. The bacterium recognizes wounded plant cells and transfers T-DNA using virulence genes. Integrated T-DNA is then expressed stably in the plant genome. Agrobacterium-mediated transformation is widely used for genetic engineering in plants due to its simplicity, efficiency and ability to transfer large DNA segments.
Plant Genetic Engineering & Agrobacterium mediated gene transfer.pptx
Agrobacterium mediated gene transfer, Ti-plasmid, cloning vectors based on Ti-plasmid, advantages disadvantages regarding cloning vectors based on Ti-plasmid are major areas covered in this Presentation.
This bacterium has a large plasmid that induces tumor, and for this reason, it was named tumor-inducing (Ti) plasmid.
This is process of altering the genetic makeup of an organism using Recombinant DNA Technology.
Agrobacterium and other methods of plant transformation including gene gun, i...
The process of transfer, integration and expression of transgene in the host cells is known as genetic transformation. A foreign gene (transgene) encoding the trait must be incorporated into plant cells, along with a "cassette" of extra genetic material to add a desirable trait to a crop. The cassette includes a sequence of DNA called a "promoter", which determines where and when the foreign gene is expressed in the host, and a "marker gene" which allows breeders to determine by screening or selection which plants contain the inserted gene. For example, marker genes may make plants resistant to antibiotics not used routinely (e.g., agrimycin, kanamycin) or tolerant of some herbicides.
Plant transformation gene transfer methods in plants
The ultimate objective of modern plant breeding is to improve a top variety in one single additional character in a predictable and precise manner without disturbing the rest of the genome. Today this is being realised through examples of successful transfer of specific traits into higher plants by gene transfer.
Techniques that open up to the plant breeder the possibility of transferring in a planned manner characters from one organism to another have been developed in microbial genetics. It should be stressed right at the outset that the expression “gene” has different meanings in agriculture and in molecular biology.
Gene Transfer Methods:
The gene transfer techniques in plant genetic transformation are broadly grouped into two categories:
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Agrobacterium MEDIATED GENE TRANSFER
This ppt contains the detailde information of Agrobacterium mediated gene transfer. it can be used by the students of UG and PG.
Agrobacterium mediated gene transfer in plants
This document discusses Agrobacterium-mediated gene transfer in plants. It describes how the Ti plasmid of Agrobacterium tumefaciens is used to transfer T-DNA containing foreign genes into plant genomes. The Ti plasmid contains genes for transferring T-DNA as well as the T-DNA itself. Upon sensing plant signals, these vir genes are activated and work together to nick and transfer a single strand of T-DNA into plant cells, where it integrates into the plant genome. This process allows stable transformation of plant cells with foreign genes and is widely used in plant genetic engineering.
Agrobacterium CM
1) Agrobacterium tumefaciens is a soil bacterium that can transfer DNA fragments (T-DNA) from its tumor-inducing plasmid into plant cells.
2) The T-DNA is flanked by left and right border sequences and encodes genes that cause plant cells to form tumors and produce nutrients for the bacteria.
3) Upon detection of wounded plant cells, genes on the bacterial plasmid and chromosome mediate T-DNA processing and transfer into the plant cell nucleus where it integrates randomly.
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Plant expression vectors
What are an expression vector? Detailed description of plant gene structure. Plant expression vector systems are generally consists of Ri and Ti plasmids.
The other vectors which are generally used are DNA and RNA viruses.
ROLE OF Agrobacterium in plant pathology
- Agrobacterium tumefaciens is a soil bacterium that causes crown gall disease in plants by transferring tumor-inducing (Ti) plasmid DNA to host cells.
- The Ti plasmid contains genes (vir genes) required for T-DNA processing and transfer to plant cells. A key vir gene is virE2, which encodes a single-stranded DNA binding protein.
- A study introduced the virE2 gene into Nicotiana benthamiana plants to test if it provides tolerance against Sri Lankan cassava mosaic virus (SLCMV), a geminivirus.
- Results showed that virE2 reduced SLCMV infection symptoms and spread in transgenic N. bent
Agrobacterium mediated gene transfer - LIKE NEVER BEFORE!!
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Agrobact transfer.pptx
Agrobacteria are soil bacteria that can transfer genes to plant genomes. They contain Ti plasmids which transfer a segment of DNA (T-DNA) into the host plant genome. This causes tumors and induces the plant to produce unique amino acids called opines that the bacteria can use as nutrients. The Ti plasmid and bacterial chromosome work together with virulence genes on the plasmid producing proteins needed for T-DNA transfer. The T-DNA is processed and transferred via a T-strand into the plant cell nucleus where it integrates randomly. This process allows genetic engineering of plants through Agrobacterium.
agrobacterium-mediatedgenetransfer-200528100800 (1).pptx
Agrobacterium tumefaciens is a soil bacterium that can naturally infect plant wounds and transfer DNA (T-DNA) from its tumor-inducing (Ti) plasmid into the plant genome. This leads to crown gall disease in infected plants. The T-DNA contains genes that produce phytohormones causing tumors and opines that the bacteria can use. By replacing the T-DNA with a gene of interest, A. tumefaciens can be used to genetically transform plants. The process involves sensing wounded plant signals, transferring T-DNA and virulence proteins into plant cells, and integrating the T-DNA into the plant genome. A. tumefaciens-mediated transformation is
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Ti plasmid ss
1. Agrobacterium tumefaciens is a soil bacteria that causes crown gall disease in plants by transferring a segment of DNA (T-DNA) from its tumor-inducing plasmid (Ti plasmid) into the plant genome.
2. The T-DNA contains oncogenes that cause tumor formation as well as genes for producing nutrients called opines that the bacteria can use.
3. Ti plasmids are modified to create disarmed vectors for plant transformation by deleting the oncogenes from the T-DNA. Binary vectors further separate the T-DNA and virulence genes into two plasmids.
Agrobacterium -MGT.pptx
This document discusses Agrobacterium-mediated gene transfer. Agrobacterium is a genus of bacteria that can transfer DNA between itself and plants, causing tumors. The most commonly studied species is A. tumefaciens, which causes crown gall disease in plants. A. tumefaciens contains a Ti plasmid that can transfer a segment of DNA (T-DNA) into the host plant genome. The T-DNA contains oncogenes that cause tumor formation and opine synthesis genes. Virulence genes on the Ti plasmid (Vir genes) are required for T-DNA transfer.
Agrobactreium nature’s genetic engineer
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Agrobacterium tumefaciens as a tool for genetic engineering in plants
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Agrobactirium mediated gene transfer
- 1. AGROBATERIUM MEDIATED GENE TRANSFER BY M.sobika III B.sc Biotechnology Ayya Nadar Janaki Ammal College ,Sivakasi.
- 2. CONTENTS Introduction to gene transformation. Agrobacterium general introduction. Classification of agrobacterium. History of agrobacterium mediated gene transformation. T-DNA . Virulence genes . Chromosomal genes . Agrobacterium tumefaciens T-DNA transfer process. Method of agrobacterium tumefaciens . References.
- 3. GENE TRANSFER INTRODUCTION Gene transfer is to transfer a gene from one DNA molecule to another DNA molecule . The direct desirable gene transfer from one organism to another and the subsequent stable integration & expression of foreign gene into the genome is referred as genetic transformation . Transient transformation occur when DNA is not integrated into host genome
- 4. Agrobacterium mediated gene transfer Agrobacterium: Soil borne ,gram negative ,rod shape motile found in rizosphere . Causative agents of “crown gall "disease of dicotyledons . Have ability transfer bacterial genes to plant genome . Attracted to wound site via chemotaxis in response to chemical (sugar and phenolic molecules :acetosyringone )released from damaged plant cells . Contain Ti plasmid which can transfer its T-DNA region into genome of host plants
- 7. CLASSIFICATION OF AGROBACTERIUM Scientific classification Kingdom :Bacteria Phylum :Proteobacteria Class : Alproteobacthaperia Order :Rhizobiales Family :Rhizobiaceae Genus :Agrobacterium Species :A. tunefaciens
- 8. HISTORY OF AGROBACTERIUM MEDIATED GENE TRANSFORMATION: Smith & Townsend ( 1907 ) –said bacteria caused crown gall disease . Brown & Stonier ( 1958 ) –proposed that not whole bacteria but some part of it cause disease . Zenen et.al (1974 ) –noted virulent stain Agrobacterium tunefaciens . Chilton et.al ( 1977 ) –reported Ti & Ri plasmid transfer to plant causing disease .
- 10. T-DNA The transfer DNA ( T-DNA ) is the transferred DNA . The T-DNA initiates at right border and terminates at left border . Its about 24000 base pairs . Contains genes for enzymes synthesizing opiens and phytohormones . It also contains a vir .
- 12. AGROBACTERIUM TUNEFACIENS T-DNA TRANSFER PROCESS
- 13. VIRULENCE GENES Virulence region consists of 24 genes in total Virulence genes are located in 8 operons from virA-virH Vir A vir F and vir G are monocistronic operons, where as vir B,C,D,E,H are polycistronic Vir A senses Acitosyringone Vir G transcriptional activator of vir box Vir B conjugational pores between plant cell and bacteria Vir D1 essential for cleavage of super coiled stranded substrate Vir E responsible for gene transfer protein Vir C helps in DNA transfer Vir B11 ATPase activity –provides energy for DNA
- 17. AGROBACTERIUM TUNEFACIENS T-DNA TRANSFER PROCESS Steps of gene transfer in plants I. Bacterial clonization II. Induction of virulence system III. Generation of T-DNA transfer complex IV. T-DNA transfer V. Integration of T-DNA into plants
- 18. METHODS OF AGROBACTERIUM TUMEFACIENS GENE TRANSFER Infection through wound Leaf disk method Co –cultivation