soil micro organism

1. Biotechnological potentials of soil micro
organism: Agrobacterium tumefaciens.
Presented by : rabia naeem khan

2. Agrobacterium tumefaciens
is a gram-negative soil rhizospheric bacteria known to
cause “crown gall” disease in dicot plants and has been
scientifically applauded for its unique natural genetic
engineering potentials.
• updated scientific name
Rhizobium radiobacter,
synonym Agrobacterium
radiobacter.

3. A.tumefaciens natural genetic engineer.
Agrobacterium tumefaciens cell contains a plasmid
known as the Ti (tumor-inducing) plasmid.
Most of the Ti plasmids have following regions:
(1) T-DNA region: responsible for tumor induction.
(2) An opine synthesis (OS) region responsible for the
synthesis of unusual amino acid or sugar derivatives,
which are collectively given the name opines.

4. (3) On either side of the T-DNA, there is a short
sequence of 24 bp called border sequence. Both the left
and right border sequences are essential for tumor
induction.
(4) Origin of replication:
responsible for replication.
(5) Vir region: responsible
for virulence region and
plays a crucial role in the
Transfer of T-DNA into the
plant nuclear genome.

5. A.Tumafeciens mediated plant cell
transformation:

6. • (1) Attachment of A. tumefaciens to the plant cells.
• (2) Sensing plant signals by A. tumefaciens and
regulation of virulence genes in bacteria following
transduction of the sensed signals.
• (3) Generation and transport of T-DNA and virulence
proteins from the bacterial cells into plant cells.
• (4) Nuclear import of T-DNA and effector proteins in
the plant cells.
• (5) T-DNA integration and expression in the plant
genome.

7. How to make a useful A.tumefaciens.
First, the oncogenes from the wild-type T-DNA are
removed to eliminate the pathogenicity of the
bacterium, and the strain becomes nonpathogenic
(disarmed) without affecting its ability to transfer T-DNA.
Second, the gene of interest and selection markers for
transgenic plants need to be introduced into the T-DNA
Third ,selection of strains to be transformed in lab

8. Applications of A.tumefaciens

9. • Transformation with Agrobacterium can be achieved
in multiple ways.
• Protoplasts or alternatively leaf-discs can be
incubated with the Agrobacterium and whole plants
regenerated using plant tissue culture. (stable)
• In agroinfiltration the Agrobacterium may be injected
directly into the leaf tissue of a plant. This method
transforms only cells in immediate contact with the
bacteria, and results in transient expression of
plasmid DNA.(trasient)

11. Agrobacterium is listed as being the vector of genetic
material that was transferred to these USA GMOs
Soybean, Cotton,
Maize, Sugar Beet, alfalfa ,Wheat.

12. Flavr savr tomato
With longer shelf life.
Golden rice with
greatet provitamin A
content.

13. https://bioone.org/journals/the-arabidopsis-
book/volume-2017/issue-
15/tab.0186/Agrobacterium-Mediated-Plant-
Transformation-Biology-and-
Applications/10.1199/tab.0186.full#bibr281
reference