January 22, 2023
GENE TRANSFER TECHNIQUES
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01. Agrobacterium Tumefaciens mediated gene
transfer
Table of Contents
01.
Agrobacterium Tumefaciens mediated gene transfer ..................... 2
01.1 Introduction
01.1 Introduction ...................................................................................... 2
01.2
Agrobacterium Tumefaciens is a soil
phytopathogen that naturally infects plant
wound sites and causes crown gall disease
via delivery of transferred (T)-DNA from
bacterial cells into host plant cells through
a bacterial type IV secretion system (T4SS).
Through the advancement and innovation
of molecular biology technology during the
past few decades, various important
bacterial and plant genes involved in
tumorigenesis were identified. With the
help of more comprehensive knowledge of
how A. tumefaciens interacts with host
cells, A. tumefaciens has become the most
popular plant transformation tool to date.
Any gene of interest can now easily be
used to replace the oncogenes in the TDNA region of various types of binary
vectors to perform plant genetic
transformation with A. tumefaciens.
Importance .................................................................................. 3
01.3 The steps in the process of gene transfer ......................................... 4
Make It Your Own ......................................................................................... 7
Customize in Almost No Time ................................................................... 7
Overview of Programs .................................................................................. 7
Program 1 ................................................................................................. 7
Program 2 ................................................................................................. 7
Program 3 ................................................................................................. 7
Getting Started ............................................................................................. 9
What to Include? ...................................................................................... 9
Focus on What You Do Best ...................................................................... 9
Don’t Forget the Mission .............................................................................. 9
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This is a great spot for a mission statement ............................................. 9
Arabidopsis, the most-studied model plant
with powerful
Genetic and genomic resources, is readily
transformable by A. tumefaciens for stable
and transient transformation in several
ecotypes tested, although variable
transformation efficiencies in different
accessions were observed.
01.2 Importance
01.3

Plant genetic transformation
heavily relies on the bacterial
pathogen Agrobacterium
tumefaciens as a powerful tool to
deliver genes of interest into a host
plant.



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Inside the plant nucleus, the
transferred DNA is capable of
integrating into the plant genome
for inheritance to the next
generation (i.e. stable
transformation).
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01.3 The steps in the process of
gene transfer
Figure 1: Major steps of the Agrobacterium tumefaciensemediated plant transformation process
01. Attachment of A. tumefaciens
to the plant cells.
02. Sensing plant signals by A.
tumefaciens and regulation of
virulence genes in bacteria
following transduction of the
sensed signals.
03. Generation and transport of TDNA and virulence proteins from
the bacterial cells into plant cells.
04. Nuclear import of T-DNA and
effector proteins in the plant cells.
05. T-DNA integration and
expression in the plant genome.
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01.4 Advantages and disadvantage
01.4.5 Advantages
Advantages of the Agrobacteriummediated transformation in rice, like on
dicotyledons, include the transfer of pieces
of DNA with defined ends with minimal
rearrangements, the transfer of relatively
large segments of DNA, the integration of
small numbers of copies of genes into plant
chromosomes, and high quality and fertility
of transgenic plants.
01.4.6 Disadvantages
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
The need for long tissue culture
periods to recover transgenic
plants,

low frequency of stably
transformed plants

the narrow range of genotypes
within a crop species that can be
transformed
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01
5
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Make It Your Own
Overview of Programs
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1
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Getting Started
Don’t Forget the Mission
What to Include?
This is a great spot for a mission statement
We know you could go on for hours about how great your business
is. (And we don’t blame you—you’re amazing!) But since you need
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“Your company is the greatest. I can’t
imagine anyone living without you.” —
Very smart customer
A picture is worth 10,000 words, but only if it’s the right one. In marketing
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statement about your business.
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Focus on What You Do Best
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