A brief Power Point Presentation providing an overview about Genetic improvements through Biotechnology in Cereal Crops with precise and well known procedures and examples helping students about their academics on professional level in related fields. Content for this compilation (Power Point Presentation) was collected from Different sources: Internet, books, social media, research papers, websites etc. and I definitely admire and acknowledge all these sources to provide such a helpful content. This Presentation was prepared and presented during Bachelors degree in Agriculture (Session 2014-2018) with major subject of Plant Breeding and Genetics at "University College of Agriculture and Environmental Sciences, Baghdad-ul-Jadeed Campus, The Islamia University of Bahawalpur, Pakistan" as an Assignment on the topic of "Genetic Improvement Through Biotechnology in Cereal Crops". I hope, content might be helpful on academic and professional level. Best Regards Muhammad Raza Ullah Tariq

2. Presented by
Muhammad Raza Ullah Tariq
Student at:
Department Of Plant Breeding And Genetics
University College Of Agriculture And Environmental Sciences
The Islamia University Of Bahawalpur
Pakistan

3. Genetic Improvement
Through
Biotechnology

4. Biotechnology ? (Bio= life ; Technology= developing new things
Any technique that uses living organisms(or parts of living organisms) to make or
modify products or to improve plants or animals, for beneficial use.
OR
Biotechnology is the use of a living organism, or some component of a living
system, to make a useful product.

5. Processes involved in biotechnology
at genetic level
Methods used for transferring rDNA into Host cell
 Microbial Vectors
 Micro-projectile Bombardment/ Gene gun method
 Electroporation
 Microinjection
 Transformation and Transfection
Enzymes
 Lysine Enzyme
 Endonuclease
 Alkaline Phosphatase
 Ligase
 Phosphorylase
Vectors
 pBR322 (Plasmid Bolivar & Rodriguez)
 BAC(Bacterial artificial chromosome)
 YAC(Yeast artificial chromosome)
 Cosmid (Cos sities+Plasmid)
 Ti Plasmid (Tumour inducing plasmid)

6. Enzymes
Different enzymes are used for different purposes in
biotechnological method i.e. for lysing cell, isolating
DNA, making rDNA etc.
Enzymes are macromolecular biological catalysts. Enzymes
accelerate, or catalyze, chemical reactions.

7. Lysing Enzyme
These are the enzymes that are used for digesting the
cell wall & plasma membranes in different types of cells.
These are given below:
 Plant cell
 Cellulase
 Pectinase
 Protease
 Lipase
 Animal Cells
 Lipase
 Protease
 Fungal Cell
 Chitenase
 Protease
 Lipase
 Bacterial cell
 Lysozyme

8. Endonuclease
These are called molecular scissors as well and are used to cut
the DNA in the nucleus of the cell.
 Restriction Endonuclease
These restriction endonucleases are
used to cut the DNA segments at
specific sites both in Recipient and
donor cell i.e between G & A.

9. Two types of cuts
are made
 Sticky ends or COS sites
 Blunt cut or blunt ends
A palindromic sequence is a sequence made up of nucleic acids within
double helix of DNA and/or RNA that is the same when read from 5' to 3'
on one strand and 5' to 3' on the other, complementary, strand.
(Two types of bond are broken which
are Phosphodiester bonds &
Hydrogen bonds.)
(Only one type of bond is broken
which is Phosphodiester bonds.)

10. Alkaline Phosphatase

11. Ligase
The gene of interest having both end having phosphate and hydroxyl
groups. The phosphate group of gene of interest will attach to the
hydroxyl group of linear plasmid DNA. We have to apply the ligase
enzyme for the attachment of phosphate group of gene of interest to
the hydroxyl group of the linear plasmid DNA.

12. Phosphorylase
An enzyme which introduces a phosphate
group into an organic molecule so that the end
that lacks the phosphate group (during the application
of Alkaline Phosphatase) gets the phosphate group
again and attaches to the hydroxyl group of gene
of interest and converted it to the recombinant
circular plasmid that has gene of interest.

13. Vectors
It is a vehicle (e.g. plasmids) use to transfer
genetic material such as DNA sequence
from donor organism to recipient one.
 Plasmid DNA vector discovered by
William Hays & Joshua Lederberg
(1952).
 Chang & Cohen proved the use of
plasmid as gene cloning vector.

14. Features of Vector
 Must be small in size
 Must have origin of replication (ORI site)
 Must have selectable marker
 Must have restriction sites
 Must have replication operating protein
(ROP sites)

15. Different types of Vectors
 pBR322 (Plasmid Bolivar & Rodriguez)
 BAC (Bacterial artificial chromosome)
 YAC (Yeast artificial chromosome)
 Cosmid (Cos sities+Plasmid)
 Ti Plasmid (Tumor inducing plasmid)

16. Methods Used For
Transferring rDNA
Into Host Cell

17.  Substituting the gene of interest to Agrobacterium
 Integrate specific new genetic material into the cells of target plant species
 Transformed cell then is regenerated into a whole fertile plant
 All cells in the progeny also carry and may express the inserted genes
Transformation

19. Gene gun method (discovered by Klein and colleagues (1987).)
 Plant cells by “shooting” them with microscopic pellets to which
DNA had been adhered
 Coat microscopic particles of gold or tungsten with the foreign DNA
 Particles are then loaded into the gun
 Propels the DNA-coated particles out of the gun like bullets
 Particles penetrate the cell walls of plants and release the foreign
DNA, which is now part of the plant cell

21. Electroporation
 Plant protoplasts take up macromolecules from their Surrounding fluid,
facilitated by an electrical impulse.
 The cells, stripped of their protective walls, resulting in protoplasts.
 Supplying known DNA to the protoplast culture medium.
 Applying the electrical pulse temporarily destabilizes the cell membrane
 Allowing the DNA to enter the cell.
 Transformed cells can then regenerate their cell walls and grow to whole, fertile
transgenic plants.

23. Examples of Transgenic
Cereal Crops

24. Insect pest resistant maize
Transgenic Bt maze,(the Bt gene cry1Ab)
shows resistance against Ostrinia nubilalis.

25. Bio-fortified Rice
‘The Golden Rice’

28. Demo
Transformation
Of
Soybean Crop

32. Thanks for your patience