This document discusses different methods of genetic transformation in plants, including Agrobacterium-mediated gene transfer, virus-mediated gene transfer, and direct gene transfer methods like microinjection and particle bombardment. It provides details on the mechanisms of Agrobacterium-mediated transformation using Ti and Ri plasmids and describes applications of transgenic plants for herbicide resistance and insect resistance by introducing genes for Bt toxins or trypsin inhibitors.
To decrease our world hunger and to make the plant more nutritious the transgenic technique was developed. This the basis of the transgenic plant and its technique
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
Plants are continually exposed to harsh environmental conditions which is life- threatening for their survival. Drought is one of the major environmental constraints that highly affect plant growth and productivity worldwide. Osmotic stress due to limited availability of water during drought lead to the inhibition of photosynthesis which ultimately affect plant growth, yield and productivity. As sessile in nature, plants cannot escape from such adverse situations. Hence, to cope up with these adverse situations, plants have developed a complex array of adaptive strategies including intricate regulation of cellular, physiological, biochemical and metabolic processes to avoid or tolerate cellular dehydration. Under limited water availability, stomata plays an essential role to check water loss due to transpiration. In addition, upon perception of stress signal, a wide range of signaling cascade has been activated which ultimately initiates the expression of stress-responsive genes in a timely and coordinated manner. Abscisic acid (ABA), the universal stress hormone, highly accumulated under stress condition, also plays an important role in stress adaptation including stomatal closure and expression of stress-responsive genes. In recent times, whole genome sequencing analysis of different plants reveals that a large family of genes is expressed under different types of abiotic stresses that are involved in defense-related pathways. These genes can be grouped into three categories, genes involving recognition of osmotic stress, signal perception, and transduction and production of stress-adaptive components for physiological responses.
To decrease our world hunger and to make the plant more nutritious the transgenic technique was developed. This the basis of the transgenic plant and its technique
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
Plants are continually exposed to harsh environmental conditions which is life- threatening for their survival. Drought is one of the major environmental constraints that highly affect plant growth and productivity worldwide. Osmotic stress due to limited availability of water during drought lead to the inhibition of photosynthesis which ultimately affect plant growth, yield and productivity. As sessile in nature, plants cannot escape from such adverse situations. Hence, to cope up with these adverse situations, plants have developed a complex array of adaptive strategies including intricate regulation of cellular, physiological, biochemical and metabolic processes to avoid or tolerate cellular dehydration. Under limited water availability, stomata plays an essential role to check water loss due to transpiration. In addition, upon perception of stress signal, a wide range of signaling cascade has been activated which ultimately initiates the expression of stress-responsive genes in a timely and coordinated manner. Abscisic acid (ABA), the universal stress hormone, highly accumulated under stress condition, also plays an important role in stress adaptation including stomatal closure and expression of stress-responsive genes. In recent times, whole genome sequencing analysis of different plants reveals that a large family of genes is expressed under different types of abiotic stresses that are involved in defense-related pathways. These genes can be grouped into three categories, genes involving recognition of osmotic stress, signal perception, and transduction and production of stress-adaptive components for physiological responses.
Plant Disease Resistant And Genetic EngineeringShweta Jhakhar
Study the adverse effects of different viruses and other fungal diseases on the plants and their growth. Discuss the methods e.g. plant disease resistant and genetic engineering to protect the plants.
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.
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.
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
in-planta transformation technology is used to transform the desired gene into the plant without using tissue culture step is called in-planta transformation.it is useful for those plants that lack the tissue culture and regeneration system.
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
Plant Disease Resistant And Genetic EngineeringShweta Jhakhar
Study the adverse effects of different viruses and other fungal diseases on the plants and their growth. Discuss the methods e.g. plant disease resistant and genetic engineering to protect the plants.
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.
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.
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
in-planta transformation technology is used to transform the desired gene into the plant without using tissue culture step is called in-planta transformation.it is useful for those plants that lack the tissue culture and regeneration system.
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
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.
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
Palestine last event orientationfvgnh .pptxRaedMohamed3
An EFL lesson about the current events in Palestine. It is intended to be for intermediate students who wish to increase their listening skills through a short lesson in power point.
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
Embracing GenAI - A Strategic ImperativePeter Windle
Artificial Intelligence (AI) technologies such as Generative AI, Image Generators and Large Language Models have had a dramatic impact on teaching, learning and assessment over the past 18 months. The most immediate threat AI posed was to Academic Integrity with Higher Education Institutes (HEIs) focusing their efforts on combating the use of GenAI in assessment. Guidelines were developed for staff and students, policies put in place too. Innovative educators have forged paths in the use of Generative AI for teaching, learning and assessments leading to pockets of transformation springing up across HEIs, often with little or no top-down guidance, support or direction.
This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
Biological screening of herbal drugs: Introduction and Need for
Phyto-Pharmacological Screening, New Strategies for evaluating
Natural Products, In vitro evaluation techniques for Antioxidants, Antimicrobial and Anticancer drugs. In vivo evaluation techniques
for Anti-inflammatory, Antiulcer, Anticancer, Wound healing, Antidiabetic, Hepatoprotective, Cardio protective, Diuretics and
Antifertility, Toxicity studies as per OECD guidelines
How to Make a Field invisible in Odoo 17Celine George
It is possible to hide or invisible some fields in odoo. Commonly using “invisible” attribute in the field definition to invisible the fields. This slide will show how to make a field invisible in odoo 17.
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
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2. NORMAL TOMATO E. coli
Desired gene
vector Modified
vector
Normal
tomato plant
GM PLANT
3. METHODS OF GENE TRANSFER IN
PLANTS
GENE TRANSFER
HORIZONTAL GENE
TRANSFER
VERTICAL GENE
TRANSFER
SOMATIC
HYBRIDISATION
GENETIC
TRANSFORMATION
DIRECT GENE
TRANSFER
AGROBACTERIUM
MEDIATED GENE
TRANSFER
VIRUS
MEDIATED GENE
TRANSER
4. MECHANISM OF
TRANSFORMATION
There are mainly 3 methods for genetic transformation :-
Agrobacterium mediated gene transfer
Ti plasmid
Ri plasmid
Virus mediated gene transfer
Direct gene transfer/physical method
microinjection , electroporation , chemical
transformation, particle bombardment
5. AGROBACTERIUM MEDIATED GENE
TRANSFER
Most classical example of HGT in plants is the
infection of dicot plant cells with Agrobacterium. It
has 2 species that are useful in gene transfer ,
Agrobacterium tumifaciens & Agrobacterium
rhizogenes.
These are gram negative soil bacteria responsible for
causing crown gall disease and hairy root in plants
respectively.
The plasmids carrying for these characters are known
as Ti (tumour inducing) plasmid & Ri (root inducing)
plasmid respectively.
6. Ti plasmid :- Plasmid
conjugation
Virulence
region
Right
borderLeft
border
T - DNA
Over drive
ORI
Norpaline /
octopine
catabolism
(2,00,000 nucleotide pairs)
7. Ti plasmid vector :- (onco genes deleted)
They are of 2 types based on whether the
DNA to be introduced is physically linked to
vir gene (co-integrating or cis-vectors) or
not (binary or trans vectors).
8. Ri plasmid :-
Both Ti and Ri plasmid have similar
appearance except for that, the Ri plasmid
lack the cytokinine synthesis region and
have homology only for auxin and opine
synthesis sequences.
Mechanism of transfer for Ti & Ri plasmids
are the same.
10. Virus mediated gene transfer :-
Of all the viruses cauliflower mosaic virus (CaMV)
is considered a most potential vector for plant
transformation as its genome is composed of
double stranded DNA.
Brisson et al. (1984) introduced methotrexate
resistant dihydrofolate reductase (DHFR) gene of E.
coli into turnip plants using CaMV as the vector.
11. Limitation of virus mediated
transformation :-
It is not expected to bring about heritable
transformation as neither the gene gets
integrated into plant genome nor is the virus
transmitted through seeds.
As a plant transformation vector is that genes
larger than a few hundred bp (DHFR is 234 bp)
interfere with the packaging of the genome into
the virion particle.
12. Direct gene transfer :-
(i) Chemical transformation :-
Many chemicals such as polyamines (poly-L-
ornithine and poly-L-lysine) and dextran sulphate
stimulate DNA uptake into protoplasts. However,
they also highly reduce cell viability. Of the many
chemicals tested for their ability to stimulate DNA
uptake into protoplasts polyethylene glycol (PEG)
has proved most effective. Treatment of
protoplasts with MgC12 (5-25 mM)
significantly improves transformation rates.
13. (ii) Electroporation :-
A popular physical method
for introducing new genes
into protoplasts is the use
of electric field which
makes the protoplasts
temporarily permeable
to DNA.
14. (iii) microinjection :-
Direct injection of
DNA into plant
protoplast or cells
using fine tipped
pipette has also been
tried to transform plant cells.
15. (iv) Particle bombardment method:-
Particle bombardment
(or Biolistics), wherein
microscopic tungsten or
gold particles (1-3 ttm)
coated with genetically
engineered DNA are
explosively hurled into
plant cells with high
velocity.
16. Application :-
Herbicide resistant plants :-
Several classes of herbicides are quite effective for broad
spectrum weed control. However, as they act by
inactivating target proteins (mostly enzymes) essential for
vital functions in plants, such as photosynthesis, shared by
both crop and weed plants, they are either nonselective
and kill the crop plants or cause significant injury to them
at the application rates required to eliminate the weeds.
Production of herbicide resistant/tolerant crop plants can
considerably reduce these losses.
17. Three approaches have been followed to obtain herbicide
resistant transgenic plants:-
(1) modification of the plant enzyme or other sensitive
biochemical target of herbicidal action to render it
insensitive to the herbicide
(2) overproduction of the unmodified target protein
permitting normal metabolism to occur even in the presence
of the herbicide
(3) introduction of an enzyme or enzyme system to degrade
and/or detoxify the herbicide prior to its action.
18. Insect resistance :-
Insects are serious pests of agricultural products in the
field and during storage.
Insects belonging to the orders Coleoptera, Lepidoptera
and Diptera are the most serious plant pests.
Bacillus thuringiensis (Bt), employed as an
insecticide for over 20 years and its demand is
continuing to increase because of its specificity
towards lepidopteran pests and being environmentally
safe.
Insect resistant transgenic plants have also been
created by introducing trypsin inhibitor gene.
19. (i) Bt toxin gene expression:-
Sporulating Bt produces non-toxic proteinaceous
endotoxin (protoxin) of 130-140 kDa which upon
hydrolysis, under highly alkaline conditions (pH 9-12) of
the insect gut, produces smaller insecticidally active
peptides of 65 -70kDa.These polypeptides specifically
attack the brush bordermembrane of midgut epithelial
cells paralysing the insect and eventually killing it.
The first Bt gene was cloned and characterized in 1985.
The first transgenic plants showing field resistance to
insects were produced by introducing the truncated Bt
toxin gene.
20. (ii) Trypsin inhibitor gene expression :-
The occurrence of certain proteins in plants which
inhibit the common insect digestive protease have
been implicated in natural mechanism of plant
defence against insect attack.
For example, the leaves of a number of plants,
including tomato, synthesize protease inhibitors in
response to insect attack. Similarly, cowpea trypsin
inhibitor (CpTI), a 80 amino acid polypeptide, has
been identified as an insecticidal component of
cowpea seeds, which is active against a range of
economically important field and storage pests,
including bruchid beetle, a major pest of stored
cowpea.
