indirect mode of gene tranfer
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Transgenic plants are plants that have been genetically modified to express desirable traits. Genes can be transferred from one plant species to another using Agrobacterium tumefaciens, a soil bacterium. Agrobacterium transfers DNA from its tumor-inducing (Ti) plasmid into the plant genome. The Ti plasmid contains genes that cause tumors as well as a region that can be replaced with a gene of interest. Transformed plants are regenerated and screened for the introduced trait. Common traits introduced include herbicide and insect resistance, improved nutrition, and environmental stress tolerance.
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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.
Methods of gene transfer in plant
This document provides information on various methods of gene transfer in plants, including Agrobacterium-mediated gene transfer and direct gene transfer methods. Direct methods rely on delivering large amounts of DNA to plant cells through techniques like particle bombardment, electroporation, and microinjection. Agrobacterium-mediated gene transfer utilizes the bacterium Agrobacterium, which transfers genes into plant genomes. The document discusses several direct and Agrobacterium-mediated methods in detail and provides advantages and limitations of each approach.
Haploid production
This document discusses distant hybridization and various techniques used to produce haploid plants. Distant hybridization refers to crosses between individuals of different plant species or genera. Such crosses can result in fully fertile, partially fertile, or fully sterile offspring depending on chromosomal homology. Androgenesis and gynogenesis are techniques used to induce haploid plants from male and female gametes, respectively. Androgenesis involves culturing immature anthers or isolated microspores while gynogenesis involves culturing unpollinated flower parts. Wide hybridization is also used to induce maternal haploids. Factors like genotype, developmental stage, and culture conditions influence haploid induction and regeneration.
Virus resistant transgenic plants
Viral infections in plants can be controlled through several strategies including using certified seed/plants, controlling weeds that harbor viruses, and insecticide use since most viruses are vector-borne. Transgenic virus resistance involves expressing viral genes including coat proteins, replicases, movement proteins, or antisense RNA to interfere with viral replication or movement. The papaya industry was saved through a transgenic papaya resistant to papaya ringspot virus. While virus resistance holds promise, risks like recombination or heterologous encapsidation must be monitored.
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.
Direct Gene Transfer Methods
This document discusses various methods of genetic transfer, including natural genetic transfer between organisms as well as technological methods developed to manipulate genes. It describes how donor DNA can enter a recipient cell and recombine, producing genetically distinct offspring. Several gene transfer technologies are then outlined, including microinjection, biolistics, calcium phosphate precipitation, lipofection, and electroporation. The document explains the basic mechanisms and applications of each method while also noting their limitations for different purposes like gene therapy. In the conclusion, it emphasizes that gene transfer technologies now allow relatively easy and accurate introduction of genes into target cells to potentially cure diseases.
Terminator Gene Technology
This document summarizes terminator gene technology, which genetically modifies plants to produce sterile seeds. It was developed by the seed industry to prevent seed saving. There are two types: varietal GURT (V-GURT) renders all subsequent seeds sterile, while trait GURT (T-GURT) switches traits on/off using chemical treatments. While it provides benefits to industry, it is controversial due to concerns over loss of biodiversity and impact on small farmers who rely on seed saving. Most countries have imposed a moratorium on field testing and commercialization of terminator seeds.
Insect resisance ppt
1. The seminar discusses developing transgenic plants resistant to insects through the transfer of resistance genes from microorganisms, higher plants, and animals into crop plants.
2. Major objectives of plant biotechnology are to develop plants resistant to biotic and abiotic stresses. Resistance to insects has been achieved by introducing genes encoding Bt toxins from Bacillus thuringiensis and other insecticidal proteins.
3. Useful genes have been isolated from microbes like B. thuringiensis, higher plants like beans and tobacco, and animals like mammals. These genes have been successfully used to engineer insect-resistant crops like cotton, potato, tomato, and tobacco.
Recommended
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.
Methods of gene transfer in plant
This document provides information on various methods of gene transfer in plants, including Agrobacterium-mediated gene transfer and direct gene transfer methods. Direct methods rely on delivering large amounts of DNA to plant cells through techniques like particle bombardment, electroporation, and microinjection. Agrobacterium-mediated gene transfer utilizes the bacterium Agrobacterium, which transfers genes into plant genomes. The document discusses several direct and Agrobacterium-mediated methods in detail and provides advantages and limitations of each approach.
Haploid production
This document discusses distant hybridization and various techniques used to produce haploid plants. Distant hybridization refers to crosses between individuals of different plant species or genera. Such crosses can result in fully fertile, partially fertile, or fully sterile offspring depending on chromosomal homology. Androgenesis and gynogenesis are techniques used to induce haploid plants from male and female gametes, respectively. Androgenesis involves culturing immature anthers or isolated microspores while gynogenesis involves culturing unpollinated flower parts. Wide hybridization is also used to induce maternal haploids. Factors like genotype, developmental stage, and culture conditions influence haploid induction and regeneration.
Virus resistant transgenic plants
Viral infections in plants can be controlled through several strategies including using certified seed/plants, controlling weeds that harbor viruses, and insecticide use since most viruses are vector-borne. Transgenic virus resistance involves expressing viral genes including coat proteins, replicases, movement proteins, or antisense RNA to interfere with viral replication or movement. The papaya industry was saved through a transgenic papaya resistant to papaya ringspot virus. While virus resistance holds promise, risks like recombination or heterologous encapsidation must be monitored.
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.
Direct Gene Transfer Methods
This document discusses various methods of genetic transfer, including natural genetic transfer between organisms as well as technological methods developed to manipulate genes. It describes how donor DNA can enter a recipient cell and recombine, producing genetically distinct offspring. Several gene transfer technologies are then outlined, including microinjection, biolistics, calcium phosphate precipitation, lipofection, and electroporation. The document explains the basic mechanisms and applications of each method while also noting their limitations for different purposes like gene therapy. In the conclusion, it emphasizes that gene transfer technologies now allow relatively easy and accurate introduction of genes into target cells to potentially cure diseases.
Terminator Gene Technology
This document summarizes terminator gene technology, which genetically modifies plants to produce sterile seeds. It was developed by the seed industry to prevent seed saving. There are two types: varietal GURT (V-GURT) renders all subsequent seeds sterile, while trait GURT (T-GURT) switches traits on/off using chemical treatments. While it provides benefits to industry, it is controversial due to concerns over loss of biodiversity and impact on small farmers who rely on seed saving. Most countries have imposed a moratorium on field testing and commercialization of terminator seeds.
Insect resisance ppt
1. The seminar discusses developing transgenic plants resistant to insects through the transfer of resistance genes from microorganisms, higher plants, and animals into crop plants.
2. Major objectives of plant biotechnology are to develop plants resistant to biotic and abiotic stresses. Resistance to insects has been achieved by introducing genes encoding Bt toxins from Bacillus thuringiensis and other insecticidal proteins.
3. Useful genes have been isolated from microbes like B. thuringiensis, higher plants like beans and tobacco, and animals like mammals. These genes have been successfully used to engineer insect-resistant crops like cotton, potato, tomato, and tobacco.
somatic hybridization
Somatic hybridization is a technique used to create hybrid plants by fusing isolated plant cells called protoplasts from two different plant species or varieties. This fusion occurs under in vitro conditions and can result in symmetric hybrids that contain chromosomes from both parents, or asymmetric hybrids that lose chromosomes from one parent. Cybrids are a type of hybrid where the nucleus comes from one species but the cytoplasm, including chloroplasts and mitochondria, comes from both parental species. Somatic hybridization and cybrid production allow for novel combinations of genes that can provide agricultural benefits like stress resistance but technical challenges remain in regenerating hybrid plants.
Gene transformation methods
Transformation is the process of altering an organism's genetic makeup by inserting new genes. Common transformation methods include Agrobacterium-mediated transformation, particle bombardment, protoplast transformation using polyethylene glycol or electroporation, and fibre-mediated DNA delivery. Agrobacterium transformation involves the bacteria transferring T-DNA from its Ti plasmid into the plant genome, while direct methods introduce naked DNA into plant cells using physical methods like particle bombardment or chemical treatments that make cell membranes permeable. Transformation allows improving crop traits like yield and stress resistance.
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 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.
Transgenic plants and their Application
Genetically modified or transgenic plants are plants that have been modified using genetic engineering techniques to introduce new traits. This document discusses the history and process of creating transgenic plants. It describes how transgenic plants are generated by transferring genes from other species into the target plant using either indirect methods like Agrobacterium-mediated transformation or direct physical methods like biolistics. The document provides details on the various applications of transgenic plants including producing herbicide resistance, insect resistance, virus resistance, and improving nutritional quality.
Herbicide resistance
Weeds reduce crop yields by 10-15% by competing for resources. Herbicides were developed to control weeds, but can also damage crops. Glyphosate is a broad-spectrum herbicide that inhibits the shikimic acid pathway in plants, blocking growth. To develop resistant crops, scientists have introduced the petunia EPSPS gene to overexpress the enzyme, used a mutant version of EPSPS that cannot bind glyphosate, and introduced bacterial genes that detoxify glyphosate. Combining these strategies provides high levels of herbicide resistance.
Gene transfer in plants
Gene Transfer and its applications; Plant tissue culture and its application for production of secondary metabolites
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.
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.
TRANSPOSON TAGGING
This document summarizes transposon tagging as a method to identify genes. Transposon tagging involves inserting a transposon near a gene of interest, which then allows the gene to be identified based on its proximity to the transposon. The document discusses different types of transposons used for tagging in plants and animals. It describes approaches for both targeted and non-targeted tagging and methods for identifying the tagged gene, including RFLP analysis and inverse PCR. As an example, it summarizes how the Cf-9 gene conferring resistance to leaf mold in tomato was identified using Ds transposon tagging.
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.
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.
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.
Gene isolation methods
There are three main methods for isolating genes:
1. Using an automated gene machine to synthesize genes from predetermined nucleotide sequences.
2. Gene cloning, which involves inserting a DNA fragment into a vector that is then transferred into a host cell to produce multiple copies.
3. Polymerase chain reaction (PCR), which amplifies a specific DNA sequence using primers that flank the target sequence.
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
Somatic embryogenesis, in plant tissue culture 2
Introduction
Types of somatic embryogenesis
Developmental stages
Factors affecting somatic embryogenesis
Importance
Conclusions
References
The process of regeneration of embryos from somatic cells, tissue or organs is regarded as somatic or asexual embryogenesis.
opposite of zygotic or sexual embryogenesis.
Embryo-like structures which can develop into whole plants in a way that is similar to zygotic embryos are formed from somatic cells.
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.
Genetic transformation
1. There are two main methods of gene transfer - direct and indirect gene transfer. Indirect transfer uses Agrobacterium-mediated transformation while direct transfer uses physical or chemical methods.
2. Agrobacterium-mediated transformation uses Agrobacterium tumefaciens to transfer T-DNA containing the gene of interest into the plant genome. The process involves co-cultivation of plant explants with Agrobacterium followed by selection and regeneration of transgenic plants.
3. Direct physical methods include biolistic transformation, microinjection, electroporation, and macroinjection. Direct chemical methods include PEG-mediated, calcium phosphate co-precipitation, and liposome-mediated transformation
Direct gene transfer
This document discusses various vectorless and direct gene transfer methods for recombinant DNA technology. It describes 10 different methods: chemical methods, electroporation, particle bombardment, lipofection, microinjection, macroinjection, pollen transformation, delivery via growing pollen tubes, laser induced transformation, and fibre mediated transformation. Each method directly introduces DNA into host cells without the use of biological vectors. The document provides details on the mechanisms and procedures for several of these direct gene transfer techniques.
Germplasm and its conservation
This document discusses germplasm and its conservation. It begins by defining germplasm as a collection of genetic resources for an organism, such as a seed bank or gene bank, that contains the genetic information for a species. Germplasm conservation is important to preserve genetic diversity and provide plant breeders resources to develop new crop varieties. Methods of conservation include in situ conservation of plants in their natural habitat and ex situ conservation of seeds, tissues, cells or DNA stored outside the natural habitat. Cryopreservation in liquid nitrogen at -196°C is an effective long-term storage method that stops cellular metabolism. The document outlines the cryopreservation process and applications for conserving plant species and genetic variations.
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
Biotechnology_transgenic plants.pptx
Genetically modified crops can be created through transgenic approaches using Agrobacterium tumefaciens. A. tumefaciens contains a tumor-inducing plasmid that allows it to transfer DNA fragments into plant cells. Researchers have developed methods to modify this plasmid to transfer desirable genes instead of tumor-causing ones. The modified plasmid is introduced into A. tumefaciens along with the gene of interest, and the bacteria is used to infect plant cells and transfer the new gene. This allows crops to be engineered for traits like pest resistance, herbicide tolerance, or improved nutrition more efficiently than traditional breeding methods.
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somatic hybridization
Somatic hybridization is a technique used to create hybrid plants by fusing isolated plant cells called protoplasts from two different plant species or varieties. This fusion occurs under in vitro conditions and can result in symmetric hybrids that contain chromosomes from both parents, or asymmetric hybrids that lose chromosomes from one parent. Cybrids are a type of hybrid where the nucleus comes from one species but the cytoplasm, including chloroplasts and mitochondria, comes from both parental species. Somatic hybridization and cybrid production allow for novel combinations of genes that can provide agricultural benefits like stress resistance but technical challenges remain in regenerating hybrid plants.
Gene transformation methods
Transformation is the process of altering an organism's genetic makeup by inserting new genes. Common transformation methods include Agrobacterium-mediated transformation, particle bombardment, protoplast transformation using polyethylene glycol or electroporation, and fibre-mediated DNA delivery. Agrobacterium transformation involves the bacteria transferring T-DNA from its Ti plasmid into the plant genome, while direct methods introduce naked DNA into plant cells using physical methods like particle bombardment or chemical treatments that make cell membranes permeable. Transformation allows improving crop traits like yield and stress resistance.
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 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.
Transgenic plants and their Application
Genetically modified or transgenic plants are plants that have been modified using genetic engineering techniques to introduce new traits. This document discusses the history and process of creating transgenic plants. It describes how transgenic plants are generated by transferring genes from other species into the target plant using either indirect methods like Agrobacterium-mediated transformation or direct physical methods like biolistics. The document provides details on the various applications of transgenic plants including producing herbicide resistance, insect resistance, virus resistance, and improving nutritional quality.
Herbicide resistance
Weeds reduce crop yields by 10-15% by competing for resources. Herbicides were developed to control weeds, but can also damage crops. Glyphosate is a broad-spectrum herbicide that inhibits the shikimic acid pathway in plants, blocking growth. To develop resistant crops, scientists have introduced the petunia EPSPS gene to overexpress the enzyme, used a mutant version of EPSPS that cannot bind glyphosate, and introduced bacterial genes that detoxify glyphosate. Combining these strategies provides high levels of herbicide resistance.
Gene transfer in plants
Gene Transfer and its applications; Plant tissue culture and its application for production of secondary metabolites
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.
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.
TRANSPOSON TAGGING
This document summarizes transposon tagging as a method to identify genes. Transposon tagging involves inserting a transposon near a gene of interest, which then allows the gene to be identified based on its proximity to the transposon. The document discusses different types of transposons used for tagging in plants and animals. It describes approaches for both targeted and non-targeted tagging and methods for identifying the tagged gene, including RFLP analysis and inverse PCR. As an example, it summarizes how the Cf-9 gene conferring resistance to leaf mold in tomato was identified using Ds transposon tagging.
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.
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.
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.
Gene isolation methods
There are three main methods for isolating genes:
1. Using an automated gene machine to synthesize genes from predetermined nucleotide sequences.
2. Gene cloning, which involves inserting a DNA fragment into a vector that is then transferred into a host cell to produce multiple copies.
3. Polymerase chain reaction (PCR), which amplifies a specific DNA sequence using primers that flank the target sequence.
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
Somatic embryogenesis, in plant tissue culture 2
Introduction
Types of somatic embryogenesis
Developmental stages
Factors affecting somatic embryogenesis
Importance
Conclusions
References
The process of regeneration of embryos from somatic cells, tissue or organs is regarded as somatic or asexual embryogenesis.
opposite of zygotic or sexual embryogenesis.
Embryo-like structures which can develop into whole plants in a way that is similar to zygotic embryos are formed from somatic cells.
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.
Genetic transformation
1. There are two main methods of gene transfer - direct and indirect gene transfer. Indirect transfer uses Agrobacterium-mediated transformation while direct transfer uses physical or chemical methods.
2. Agrobacterium-mediated transformation uses Agrobacterium tumefaciens to transfer T-DNA containing the gene of interest into the plant genome. The process involves co-cultivation of plant explants with Agrobacterium followed by selection and regeneration of transgenic plants.
3. Direct physical methods include biolistic transformation, microinjection, electroporation, and macroinjection. Direct chemical methods include PEG-mediated, calcium phosphate co-precipitation, and liposome-mediated transformation
Direct gene transfer
This document discusses various vectorless and direct gene transfer methods for recombinant DNA technology. It describes 10 different methods: chemical methods, electroporation, particle bombardment, lipofection, microinjection, macroinjection, pollen transformation, delivery via growing pollen tubes, laser induced transformation, and fibre mediated transformation. Each method directly introduces DNA into host cells without the use of biological vectors. The document provides details on the mechanisms and procedures for several of these direct gene transfer techniques.
Germplasm and its conservation
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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
Biotechnology_transgenic plants.pptx
Genetically modified crops can be created through transgenic approaches using Agrobacterium tumefaciens. A. tumefaciens contains a tumor-inducing plasmid that allows it to transfer DNA fragments into plant cells. Researchers have developed methods to modify this plasmid to transfer desirable genes instead of tumor-causing ones. The modified plasmid is introduced into A. tumefaciens along with the gene of interest, and the bacteria is used to infect plant cells and transfer the new gene. This allows crops to be engineered for traits like pest resistance, herbicide tolerance, or improved nutrition more efficiently than traditional breeding methods.
17 agrobacterium tumefaciens
Agrobacterium tumefaciens is a soil bacterium that causes crown gall disease in plants like apples, pears, and roses. It transfers part of its tumor-inducing plasmid (Ti plasmid) DNA to plant cells, which integrates into the plant genome and causes tumor formation. The transferred T-DNA segment contains genes encoding enzymes and hormones that alter plant metabolism and induce tumor growth. Agrobacteria have been modified for plant genetic engineering by removing tumor-causing genes and adding genes of interest between the plasmid's left and right border sequences, which allows expression of the added genes in plants.
Agrobacterium mediated transformation, its mode of action and applications in...
Agrobacterium mediated transformation, its mode of action and applications in...Dr. Shobha D. Surbhaiyya
here described in detail gene transformation technique in plant by natural method for improvement of crop Agrobacterium tumefacience
Agrobacterium tumefaciens is a soil bacterium that naturally transfers DNA to plant cells and causes crown gall disease. It contains a tumor-inducing plasmid (Ti plasmid) that can be engineered to transfer foreign genes of interest into plant genomes. This process, called Agrobacterium-mediated transformation, is commonly used in genetic engineering due to its high efficiency. The Ti plasmid contains T-DNA that is transferred to plant cells, along with virulence genes that facilitate transfer without being transferred themselves. Transformed plants are regenerated from cultured plant cells or tissues containing the novel genes. Agrobacterium-mediated transformation is useful for genetic engineering but has limitations such as a narrow host range and time-consuming
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.
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 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 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.
Agrobacterium presentation
This document discusses Agrobacterium tumefaciens, a soil bacterium that causes crown gall disease in plants. It transfers tumor-inducing (Ti) plasmid DNA into wounded plant cells, integrating it into the plant genome and causing tumors. The Ti plasmid contains the T-DNA region flanked by borders, which is transferred to plants. It also contains virulence genes and opine catabolism genes. The infection process involves signal induction, bacterial attachment, production of virulence proteins, T-DNA production and transfer into the plant cell nucleus where it integrates.
Plant biotechnology with having refrence from glick
Plant biotechnology can be used to genetically engineer plants for insect and disease resistance. Common methods include expressing genes from Bacillus thuringiensis (Bt) that code for insecticidal proteins or genes for plant proteins that inhibit insect growth. Other strategies prevent the development of Bt-resistant insects. Virus resistance has been achieved by expressing viral coat proteins or antiviral plant proteins. Fungus and bacteria resistance can be conferred by overexpressing plant defense genes or salicylic acid production. Transgenic plants now provide protection from major agricultural pathogens and pests.
Agrobacterium mediated gene transfer
Agrobacterium tumefaciens is a soil bacterium that can transfer DNA fragments called T-DNA into plant cells. It causes crown gall disease in susceptible plants like tomatoes and sunflowers by integrating T-DNA into the plant cell's genome. During Agrobacterium-mediated gene transfer, wounded plant cells produce signal molecules that activate virulence genes in Agrobacterium, causing it to synthesize single-stranded T-DNA. The T-DNA is then transferred into the plant cell nucleus where it integrates and expresses genes that promote tumor formation. This natural process allows for stable transformation of plant cells and regeneration of whole transgenic plants.
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:
I. Vector-mediated gene transfer
II. Direct or vector less DNA transfer
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
Ti plasmid vector
An agrobacterium based vector to introduce plant genomic fragments., very much used to produce genetic modified plants Pla
Agrobacterium mediated gene transfer in plants
1) Agrobacterium tumefaciens is a soil bacterium that causes crown gall disease in dicot plants by transferring oncogenic DNA (T-DNA) from its Ti plasmid into the plant genome.
2) The T-DNA contains genes that produce hormones causing uncontrolled cell division and the formation of tumors.
3) By removing the oncogenic genes and inserting foreign DNA between the border repeats, the Ti plasmid can be used as a vector to stably introduce foreign genes into plant genomes through the natural plant-Agrobacterium interaction and T-DNA transfer process.
Agrobacterium mediated transformation
In this Presentation i have collected some basic information about that how Bacteria causes diseases in plants....
crown gall disease is discussed.
Contact 0311 9469029
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.
agrobacterim vector
The document summarizes how Agrobacterium tumefaciens is used for plant genetic engineering. It describes the key components of the Ti plasmid, including the T-DNA, vir genes, and opine catabolism genes. It explains that the Ti plasmid is modified by deleting tumor-causing genes and inserting a gene of interest, along with selectable markers. The modified plasmid is transferred to A. tumefaciens via triparental mating and then used to transform plant cells. Transformed cells are selected using antibiotics and regenerated into transgenic plants.
Ti plasmid and ca mv
This document discusses genetic engineering techniques using Agrobacterium tumefaciens and its tumor-inducing (Ti) plasmid. The Ti plasmid contains a segment of transferred DNA (T-DNA) that is integrated into the host plant genome and causes tumor formation. Genes of interest can be inserted into the T-DNA, which is then transferred to the plant cell. However, wild-type Ti plasmids also contain oncogenes, so alternative vectors like plant viruses have been developed that can introduce foreign DNA into plant cells without causing tumors. Cauliflower mosaic virus is one such virus vector that has been used, as its small, 8kb double-stranded DNA genome can be easily manipulated.
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indirect mode of gene tranfer
- 1. Transformation :- Transfer of desirable gene from one plant species to another Transgene : Transferred gene Transgenic plant : Transformed plant Reasons for developing transgenic plants :- To improve the agriculture , horticulture or ornamental value of plants To study the action of gene in plants during development & various biological process
- 2. To develop plant bioreactors for inexpensive manufacture of commercially important products e.g:- proteins , medicine , pharmaceutical compound Genetic traits introduced into plants :- • Resistance to herbicides • Protection against viral infections • Insecticidal activity • Improved nutritional quality • Altered flower pigmentation • Tolerance to environmental stresses
- 3. The gene transfer technique in plant genetic transformation is broadly divided in two categories :- 1. Vector mediated / indirect gene transfer 2. Vector less mediated / direct gene transfer
- 4. Vector mediated :- Carried by agrobacterium or by use of plant virus as vectors Most vectors carried marker genes which allow recognition of transformed cells – selectable markers E.g:- npt is a common selectable marker providing kanamycin resistance Common features of vectors :- 1. Multiple unique restriction sites 2. Ori
- 5. Agrobacterium ti plasmid is preferred all over other vectors because of wide range & capacity to transfer genes due to presence of T DNA border sequence Agrobacterium mediated gene transfer :- soil born gram negative bacterium Rod shaped Family – rhizobiaceae A natural genetic eng capable of integrating its DNA into plant genome
- 6. Species of agrobacterium :- A. tumefaciens - crown gall disease A. rhizogenes - hairy root disease Characteristics of ti plasmid :- 200 kb plasmid Varying length of 12 – 24 kb 4 major regions • T DNA region • An ori
- 7. T DNA region is a part of Ti plasmid it can be integrated in to the plant genome . T DNA has right & left borders Virulence region genes helpful for T DNA transfer in to plant genome . Mutations in this region abolishes virulence The opine catabolism region synthesis opine Ori is responsible for replication
- 8. General structure of T DNA :- T DNA consists of 1. Onc (oncogenisity region) :- Onc region consists of tms 1 & tms 2 , tmr These are responsible for biosynthesis of 2 phytohormones namely IAA , isopentyl adenosine 5ˡ monophosphate ( cytokine ) These genes causes crown gall disease 2.Nos region :- Responsible for the synthesis of aa or sugar derivatives called opines
- 9. Depending upon opine, the bacteria is octopine type or nopaline type
- 10. Virulence region :- Organized in to 6 operons ( a,b,c,d,e,g ) A,B,D,G – required for virulence C,E – tumor formation A - located on the inner membrane of bacteria . Chemoreceptor for plant hormones B - form a pore between bacteria & plant cell D - cutting of T DNA E - preventing degradation of T DNA , covers T DNA
- 11. production of acetosyrengone binding of bacteria to plant cell vir genes activated by phenolic compounds vir A & vir G & all vir genes are activated vir D cleaves T DNA tube formed between bacteria & plant cell DNA is transferred in to plant cell
- 13. In T DNA the genes b/w the L&R border are removed Any fragment can be transformed in to plant genome by replacing T DNA with our gene of interest Prerequisites • L&R borders • Vir genes • Chromosomal genes
- 17. A.rhizogenes infect plants results in hairy root . These plasmids are referred as Ri plasmids Some of these Ri plasmids posses genes that are homologous to Ti plasmid e.g:- auxin biosynthetic genes Instead of virulence genes Ri plasmid contains a series of open reading frames on TDNA Hairy root system are used in the production of secondary metabolites particularly pharmaceutical products
- 18. Advantages :- Natural method of gene transfer Agrobacterium can conveniently infect any explant Large fragment of DNA can be effectively transferred Transformed plant can be effectively regenerated Limitations:- Extensively utilized for dicotyledonous species monocotyledons could not be successfully utilized for agrobacterium gene transfer(failure of gene response in
- 19. Cells that regenerate more efficiently are often difficult to transformed Deep layers are not easy target