Course
Code: BC614T
Core
Course XIV: Plant Biotechnology
The
objective of this course is to expose the students to application of modern
tools and techniques in Biology
(Credits: Theory-4, Practical-2)
THEORY
Lectures:
60
Unit 1:
Plant Tissue Culture (16
lectures)
Historical perspective;
Composition of media; Nutrient and hormone requirements (role of vitamins and
hormones); Totipotency; Organogenesis; Embryogenesis (somatic and zygotic);
Protoplast isolation, culture and fusion; Tissue culture applications (micropropagation,
androgenesis, virus elimination, secondary metabolite production, haploids,
triploids and hybrids; Cryopreservation; Germplasm Conservation).
Unit 2:
Recombinant DNA technology (12 lectures)
Restriction Endonucleases
(History, Types I-IV, biological role and application); Restriction Mapping
(Linear and Circular); Cloning Vectors: Prokaryotic (pUC 18 and pUC19, pBR322,
Ti plasmid, BAC); Lambda phage, M13 phagemid, Cosmid, Shuttle vector;
Eukaryotic Vectors (YAC).
Unit 3:
Gene Cloning (10 lectures)
Recombinant DNA, Bacterial
Transformation and selection of recombinant clones, PCR-mediated gene cloning;
Gene Construct; construction of genomic and cDNA libraries, screening DNA
libraries to obtain gene of interest by genetic selection; complementation,
colony hybridization.
Unit 4:
Methods of gene transfer (8 lectures)
Agrobacterium-mediated, Direct gene transfer by Electroporation,
Microinjection, Microprojectile
bombardment; Selection of transgenics– selectable marker and reporter genes
(Luciferase, GUS, GFP).
Unit 5:
Applications of Biotechnology (14 lectures)
Pest resistant (Bt-cotton);
herbicide resistant plants (RoundUp Ready soybean); Transgenic crops with
improved quality traits (Flavr Savr tomato, Golden rice); Improved horticultural
varieties (Moondust carnations); Role of transgenics in bioremediation
(Superbug); edible vaccines; Industrial enzymes (Aspergillase, Protease,
Lipase); Gentically Engineered Products–Human Growth Hormone; Humulin;
Biosafety concerns.
Course
Code: BC614P
Core
Course XIV - Practical: Plant Biotechnology
1. (a)
Preparation of MS medium.
(b)
Demonstration
of in vitro sterilization and
inoculation methods using leaf and nodal explants of tobacco, Datura, Brassica etc.
2.
Study of
anther, embryo and endosperm culture, micropropagation, somatic embryogenesis
& artificial seeds through photographs.
3. Isolation
of protoplasts.
4. Construction
of restriction map of circular and linear DNA from the data provided.
5.
Study of
methods of gene transfer through photographs: Agrobacterium-mediated, direct gene transfer by electroporation,
microinjection, microprojectile bombardment.
6.
Study of
steps of genetic engineering for production of Bt cotton, Golden rice, Flavr
Savr tomato through photographs.
7. Isolation
of plasmid DNA.
8. Restriction
digestion and gel electrophoresis of plasmid DNA.
Suggested
Readings
1.
Bhojwani,
S.S. and Razdan, M.K., (1996). Plant Tissue Culture: Theory and Practice.
Elsevier Science Amsterdam. The Netherlands.
2.
Glick,
B.R., Pasternak, J.J. (2003). Molecular Biotechnology- Principles and
Applications of recombinant DNA. ASM Press, Washington.
3.
Bhojwani,
S.S. and Bhatnagar, S.P. (2011). The Embryology of Angiosperms. Vikas
Publication House Pvt. Ltd., New Delhi. 5th edition.
4.
Snustad,
D.P. and Simmons, M.J. (2010). Principles of Genetics. John Wiley and Sons,
U.K. 5th edition.
5.
Stewart,
C.N. Jr. (2008). Plant Biotechnology & Genetics: Principles, Techniques and
Applications. John Wiley & Sons Inc. U.S.A.
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