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.
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.
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.
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
Somaclonal Variation in Plant tissue culture - Variation in somaclones (somatic cells of plants)
Somaclonal variation # Basis of somaclonal variation # General feature of Somaclonal variations # Types and causes of somaclonal variation # Isolation procedure of somaclones via without in-vitro method and with in-vitro method with their limitations and advantages # Detection of isolated somaclonal variation # Application (with examples respectively related to crop improvement) # Advantages and disadvantages of somaclonal variations.
https://www.youtube.com/watch?v=IZwrkgADM3I
Also watch, Gametoclonal variation slides to understand, how to changes occur in gametoclones of plants.
https://www.slideshare.net/SharmasClasses/gametoclonal-variation
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.
1.What is plant tissue culture?
2.Production of virus free plants.
3.History.
4.Virus elimination by heat treatment.
5.Virus elimination by Meristem Tip culture.
6.Factor affecting virus eradication by Meristem Tip culture.
7.Chemotherapy.
8.Virus elimination through in vitro shoot-tip Grafting.
9.Virus Indexing.
10.Conclusion .
11.References .
Agrobacterium mediated gene transfer in plants.ICHHA PURAK
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.
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
Somaclonal Variation in Plant tissue culture - Variation in somaclones (somatic cells of plants)
Somaclonal variation # Basis of somaclonal variation # General feature of Somaclonal variations # Types and causes of somaclonal variation # Isolation procedure of somaclones via without in-vitro method and with in-vitro method with their limitations and advantages # Detection of isolated somaclonal variation # Application (with examples respectively related to crop improvement) # Advantages and disadvantages of somaclonal variations.
https://www.youtube.com/watch?v=IZwrkgADM3I
Also watch, Gametoclonal variation slides to understand, how to changes occur in gametoclones of plants.
https://www.slideshare.net/SharmasClasses/gametoclonal-variation
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.
1.What is plant tissue culture?
2.Production of virus free plants.
3.History.
4.Virus elimination by heat treatment.
5.Virus elimination by Meristem Tip culture.
6.Factor affecting virus eradication by Meristem Tip culture.
7.Chemotherapy.
8.Virus elimination through in vitro shoot-tip Grafting.
9.Virus Indexing.
10.Conclusion .
11.References .
Agrobacterium mediated gene transfer in plants.ICHHA PURAK
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.
Surface Plasmon Resonance,
Surface Plasmons:
Plasmons confined to surface (interface) and interact with light resulting in polarities.
Propagating electron density waves occurring at the interface between metal and dielectric.
RAPD markers are decamer DNA fragments.
RAPD is a type of PCR reaction.
as the name suggest it is a fast method when compared to the traditional PCR medthod.
Agrobacterium tumefaciensppt............it is a slide presentation on interki...Anwesha Banerjee
agrobacterium gene transfer is very important for all students learning biotechnology and microbiology , i have prepaed it for my college presentation class hope all my friends like it
plant Biotechnology: The application of Plant Biotechnology by use of scientific method to manipulate living cells or organisms for practical uses (manipulation and transfer of genetic material).
Agrobacterium-mediated transformation is one of the most successful biological methods of gene transfer in plants that is used for crop improvement and the production of GMOs it is a revolutionary method that allows scientists to introduce foreign DNA into plant cells, enabling the modification of plant genomes for various purposes, including crop improvement, pharmaceutical production, and basic research.
Whether you're a student, researcher, educator, or simply curious about the fascinating world of plant biotechnology, our session on Agrobacterium-mediated transformation offers valuable insights and information to help you understand this powerful technique and its implications for plant science and beyond.
Genetic Engineering in Insect Pest management Mohd Irshad
gene incorporation is gaining attention across the globe with the aim of improving plant health, crop protection, and sustainable crop production. This versatile method of Scientific cultivation should be adopted by the growers as it has been investigated and assessed by experts and environmentalists. There is not any kind of toxic effect on mammalian.
B4FA 2013 Ghana: Genetic Engineering - Chris Leaverb4fa
Introduction to genetic engineering technologies and principles at B4FA 2013 Accra media fellowship workshop
For more information please see www.b4fa.org
: Research progress on the evaluation techniques and methods for environmenta...ssuser45befb1
Transgenic plants are plants into which one or more genes from another species have been introduced into the genome, using genetic engineering processes. Techniques include the biolistic method—in which a heavy metal is coated with plasmid DNA is shot into cells—and Agrobacterium tumefaciens mediated transformation.
Plant Genetic engineering ,Basic steps ,Advantages and disadvantagesTessaRaju
plant genetic engineering,first genetically engineered crop plant,first genetically engineered foods,genome editing,uses of GE,transgenic plants,basic process of plant genetic enginering,advantages and disadvantages of genetic engineering.
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2. Transformation – the process of obtaining transgenic plants
Transgenic plant – a plant with a foreign gene (or genes) from another
plant/animal that is incorporated into its chromosome
Marc Van Montagu and Jeff Schell, discovered the gene transfer mechanism between
Agrobacterium and plants, which resulted in the development of methods to alter the bacterium into
an efficient delivery system for genetic engineering in plants.
Most common genes (and traits) in transgenic or biotech crops
herbicide resistance
Insecticide resistance
Bt genes in field corn (maize)
virus-resistance (coat-protein) genes
Transgenic plants – An introduction
3. Plant Transformation Methods
Physical Chemical Biological
Microinjection
Pressure
Biolistics - gene gun/
particle bombardment
Electroporation
Silica/carbon fibers
Lazer mediated
SAT
PEG
DEAE-dextran
Calcium phosphate
Artificial lipids
Proteins
Dendrimers
A. Tumefaciens
A. Rhizogenes
Virus-mediated
4. 4
Introduction to Agrobacteria…
A rod shaped, Gram
negative soil bacterium
A natural genetic engineer
2 species
A.tumefaciens (produces
a gall)
A. rhizogenes (produces
roots)
Oncogenes (for auxin and
cytokinin synthesis) +
Opines
• Disease producing
Agrobacterium are higly
pathogenic and do not
benefit the plant
• In the presence of
exudates (e.g.
acetosyringone) from
wounded plants, Virulence
(VIR) genes are activated
and cause the t-DNA to be
transferred to plants.
Everything between the
left and right border is
transferred.
5. Scientific classification
• Kingdom :Bacteria
• Phylum :Proteobacteria
• Class :Alproteobacthaperia
• Order :Rhizobiales
• Family :Rhizobiaceae
• Genus :Agrobacterium
• Species :A. tumefaciens
6. Ti plasmid and virulence genes
• A Ti plasmid is a circular piece of DNA found
in almost all Agrobacteria
• Three main regions
T-DNA region(Between right and left T-DNA
border)
Oncogene
Opine
Virulence region
Opine catabolism region
7. Cellular process of Agrobacterium–host interaction
Tzvi Tzfira and Vitaly Citovsky, 2002, Trends in Cell Biol. 12(3), 121-129
13. Production of transgenic plants
Isolate and clone gene of interest
Add DNA segments to initiate or enhance gene
expression
Add selectable markers
Introduce gene construct into plant cells
(transformation)
Select transformed cells or tissues
Regenerate whole plants
14. Procedure for plant transformation
IMAGE: Mol bio of the cell by Albert (pg no:599)
15. Regeneration, Selection And Detection...
Regeneration: for shoot organogenesis, cytokinin (lower
amounts of auxin) are required
Selection: two antibiotics are required
• an antibiotic to kill the Agrobacterium, while not affecting the
plant's cell growth and division
• a second antibiotic allows growth of transformed shoots
(w/selectable marker) but inhibits growth of untranformed plant
cells.
Detection of the "trait" gene
PCR methods can detect the presence of the "trait" DNA
protein detection methods are used where a gene product is produced
that defines the trait
verification of the incorporation of the trait gene into the plant's
chromosome
• by Southern hybridization
• by demonstrating transfer of the trait to the original transformant's
progeny
17. Types of genetic modification
>99% of all
transgenic crops
are either herbicide
or insect resistant
<1% have other
traits
0
5
10
15
20
25
Herbicide
Insect
resistance
Others
Millionsofhectares
19. Ø Scientists can insert any gene they want into the plasmid in place of the tumor
causing genes and subsequently into the plant cell genome
Ø By varying experimental materials, culture conditions, bacterial strains, etc.
scientists have successfully used A. tumefaciens Gene Transfer to produce BT Corn
Ø This method of gene transfer enables large DNA strands to be transferred into
the plant cell without risk of rearrangement whereas other methods like the Gene Gun
have trouble doing this
Ø The vast majority of approved genetically engineered agriculture has been
transformed by means of Agrobacterium tumefaciens Mediated Gene Transfer
Ø Original problems existed in that Agrobacterium tumefaciens only affects
dicotyledonous plants
Ø Monocotyledon plants are not very susceptible to the bacterial infection
Benefits and Problems with
Agrabacteria….
20. Transformed plants will help the world to meet the human demands for food,
energy, medicine, shelter, clothing, and a cleaner environment.
Gene transfer technology is being used to produce plants capable of hyper
accumulating toxic metals from soil and water, thus contributing to a cleaner
environment
Increases in plant production are needed to meet growing demands for food.
Transformed Plants offers significant advantages over traditional plant breeding
in developing improved crops.
CoNClusioN…..
21. References
Introduction to plant biotechnology by H.S Chawla
Molecular biology of the cell by Albert et al
Online References
http://www.nepadbiosafety.net/for-regulators/resources/subjects/biotechnology/
plant-transformation-agro
http://www.bio.davidson.edu/people/kabernd/seminar/2002/method/dsmeth/ds.htm
http://arabidopsis.info/students/agrobacterium/uses.html
http://microbewiki.kenyon.edu/index.php/Agrobacterium_tumefacien0073
http://www.apsnet.org/edcenter/intropp/lessons/prokaryotes/Pages/CrownGall.aspx
http://www.ag.ndsu.edu/pubs/plantsci/crops/a1219w.htm
