In this Presentation i have collected some basic information about that how Bacteria causes diseases in plants....
crown gall disease is discussed.
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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.
Agrobacterium tumifaciens
Horizontal gene transfer
Interkingdom gene transfer
Virulence or Vir a b c d e f g genes
Crown gall disease
Regulation of vir genes
Relaxosome
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.
Agrobacterium tumifaciens
Horizontal gene transfer
Interkingdom gene transfer
Virulence or Vir a b c d e f g genes
Crown gall disease
Regulation of vir genes
Relaxosome
Introduction
Components of binary vector
Development of binary vector system
Properties of binary vector
Types of binary vector
Plant transformation using binary vector
Advantage of using binary vector
Conclusion
References
Introduction
Components of binary vector
Development of binary vector system
Properties of binary vector
Types of binary vector
Plant transformation using binary vector
Advantage of using binary vector
Conclusion
References
TRANSGENIC CROPS CHALLENGES AND PROSPECTS
Transgenic Technology : Transform gene from any source.
Eg: animals, bacteria, virus etc
Traditional Breeding : Move genes only between members of a particular genus of plants.
Take multiple growing seasons to develop and test a new variety.
Lot of man power
Limited possibility of improved traits.
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.
Dr.S.KARTHIKUMAR
Associate Professor
Department of Biotechnology
Kamaraj College of Engineering and Technology, K.Vellakulam-625701, TN, India
Email: skarthikumar@gmail.com
It is a rod-shaped, Gram-negative, Peritrichous flagella, Soil bacterium. Agrobacterium is well known for its ability to transfer DNA between itself become an important tool for genetic engineering.
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The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
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2. OUTLINES
• Introduction To Transgenic Plants
• Discovery
• Requirements
• Introduction To Agrobacterium
• Agrobacterium Species
• Taxonomic Classification
• Ti-plasmid & Its Structure
• Crown Gall Disease
• Regeneration , Selection And Detection
3. TRANSGENIC PLANTS
• TRANSFORMATION
The process of
obtaining transgenic plants.
• TRANSGENIC PLANT
A plant with a foreign gene
from another plant that is
incorporated into its chromosome.
4. DISCOVERY
• 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.
6. CONTINUED
• Most common genes (and traits)
in transgenic or biotech crops
are
Herbicide resistance
Insecticide resistance
Bt genes in field corn (maize)
Virus-resistance (protein coat)
genes
99% are herbicide traits
1% are other traits
7. AGROBACTERIUM
• Discovered by Smith and Townsend
(1907).
• A soil-born gram negative
bacterium. It is a rod shaped and
motile and belongs to the
bacterial family of Rhizobiaceae.
• It is a Phytopathogen, and it is
regarded as
• Nature’s most effective plant genetic
engineer. It is the natural expert of
inter-kingdom gene transfer.
8. TYPES OF AGROBACTERIUM
There are three kinds of Agrobacterium
species
Agrobacterium
tumefaciens
Crown Gall Diseases
Agrobacterium
rhizogenes
Hairy Root Diseases
Agrobacterium
radiobacter
Avirulent Strain
10. TI-PLASMID AND VIRULENCE GENES
• A Ti Plasmid (200kb) Is A Circular Piece Of DNA Found In
Almost All Agrobacteria.
• Three Main Regions:
1. T-DNA Region – Between Right & Left DNA Border (24
Kb Each) Oncogene Opine
2. Virulence Region
3. Opine Catabolism Region
12. CROWN GALL DISEASE
BY A.TUMEFACIENS
• Infects wounded or damaged part of
plants causing plant tumor called
crown gall.
• The entry of the bacterium is faciliated by
the release of phenolic compounds like
Acetosyringone and Hydroxysyringone.
• Crown gall occurs when the
bacterium releases its Ti-plasmid
into the plant cell cytoplasm. The
T-DNA is transferred to the host
cell via wounded section.
13. CROWN GALL
• The T-DNA carries genes that
code for proteins involved in
the biosynthesis of growth
hormones (auxin and
cytokinin) and novel plant
metabolites namely- opines
and agropines.
• Growth hormones – plant cell
proliferation
• Opines & agropines – source of C
and energy
14. TRANSFORMATION OF CELLS BY
A.tumefaciens
1. Signal induction to Agrobacterium – by the phenolic
compounds and some sugars, induces biochemical changes
that help in T-DNA transfer.
2. Attachment of Agrobacterium to plant cells –
Agrobacterium attaches to cell through polysaccarides
and cellulose fibres.
3. Production of virulence protein – signal induction causes
virulence protein to form Vir A, which induces Vir G, this
then induces the production of rest of the virulence proteins
like Vir D1/D2, Vir E, Vir B.
15. 4. Production of single stranded T-DNA – this is recognised
by vir D and thus carried forward to the host by Vir D2.
5. Transfer of T-DNA out of Agrobacterium – the DNA
strand is carried out by Vir D2. Through a channel made
by Vir B.
6. Transfer of T-DNA into plant cells and integration –
integration of the DNA into the host cell is helped by Vir
E2, which protects the DNA from degradation by host
cell restriction modification system, while Vir D2 helps to
navigate the DNA to the nucleus.
This process is called Illegimate Recombination, since it
does not depend on the sequence similarity..
16. REGENARATION,SELECTION,DETECTION
• REGENERATION:
For Shoot Organogenesis,Cytokinin (LowerAmountsOf Auxin)Are
Required.
• SELECTION:
(Two Antibiotics Are Required)
1. An Antibiotic To Kill The Agrobacterium, While Not
Affecting The Plant’s Cell Growth And Division.
2. A Second Antibiotic Allows Growth Of
Transformed Shoots But Inhibits Growth Of
Untransformed Plant Cells.
17. CONTD
• Detection of the “trait” gene:
1. PCR methods can detect the presence of the trait DNA.
2. Protein detection methods are used where a gene product is
produced that defines the trait.
3. Verification of the corporation of the trait gene into the
plant’s chromosome.
By southern hybridization
By demonstrating transfer of the trait to the original
transformant’s progeny