BOT-356

BOT-356: PRINCIPLES OF BIOTECHNOLOGY

Credits: 3(2+1)

THEORY:
Concept of plant biotechnology, History of plant tissue culture and plant genetic engineering, Scope and importance of plant biotechnology in crop improvement, Totipotency and morphogenesis, Nutritional requirements of plant tissue culture, Techniques of in vitro culture, Test tube fertilization, Endosperm culture, Factors affecting on test tube fertilization and endosperm culture, Applications and achievements and types of somaclonal variation, Somatic embryogenesis and synthetic seed production, Protoplast: Isolation and culture, Protoplast manipulation and fusion, Somatic hybrids and cybrids and their applications in crop improvement. Concept of genetic engineering, Restriction enzymes and their uses, Concept of vectors and their types for gene transfer, Gene cloning, Direct method of gene transfer, Indirect method of gene transfer, Transgenic plant and their applications, Different type of blotting techniques, DNA fingerprinting, DNA based markers: RFLP, RAPD, SSR and DNA probes, Mapping of QTL and future prospects, MAS and its application in crop improvements.

PRACTICAL:
Plant tissue culture laboratory specifications and organization of equipments and their use, Different Sterilization techniques and aseptic manipulations, Preparation of tissue culture media and their composition, Preparation of explants (Banana / Sugarcane) establishment and maintenance of callus cultures from different explants, sub culture of callus, Regulation of morphogenesis from different explants, roots, stem, leaf, bud, Micropropagation with shoot apex culture in different plants (Banana / Sugarcane), Meristem culture, Anther and pollen culture, Embryo and endosperm culture, Somatic embryogenesis and artificial seed production, Isolation and culturing of protoplast, Isolation of genomic DNA, Gene transfer method: direct method, Gene transfer method: indirect method, Gel electrophoresis technique, Confirmation of genetic transformation.

Practical (Experiments):
1) Plant tissue culture laboratory specifications and organization of equipments and their use.
2) Different Sterilization techniques and aseptic manipulations.
3) Preparation of tissue culture media and their composition.
4) Preparation of explants (Banana/Sugarcane) establishment and maintenance of callus cultures from
different explants, sub culture of callus.
5) Regulation of marphogenesis from different explants, roots, stem, leaf, bud.
6) Micropropagation with shoot apex culture in different plants (Banana / Sugarcane).
7) Meristem culture.
8) Anther and pollen culture.
9) Embryo and endosperm culture.
10) Somatic embryogenesis and artificial seed production.
11) Isolation and culturing of protoplast.
12) Isolation of genomic DNA.
13) Gene transfer method: direct method.
14) Gene transfer method: indirect method.
15) Gel electrophoresis technique.
16) Confirmation of genetic transformation.