THEORY

 

Unit I. Plant breeding-

General idea- introduction to plant breeding, objectives of plant breeding, genetic variability And its role in plant breeding, methods of reproduction and breeding- in self pollinated, cross pollinated, vegetative propagated plants, heterocyst and inbreeding depression, genetic, physiological and biochemical basis of heterocyst. Organogenesis, factors affecting organogenesis, differentiation.

Unit II. Somatic embryogenesis

Embryogenesis - induction, development and maturation, factors, applications and synthetic seeds, molecular biology of somatic embryogenesis – induction, development, synchronizing somatic embryo development, hormonal regulation, late embryogenesis abundant (LEA).

Unit III. Exploits

Anther, pollen and ovary culture for production of haploid plants and homozygous lines, triploid production- methods and applications.

In vitro pollination and fertilization – methods, factors affecting seed setting after in vitro pollinations, applications, embryo culture, embryo rescue.

Risks and ethics of biotechnology

Unit IV. Gene expression –

Genes isolated from somatic embryos, expression of ‘non-embryonic’ genes during somatic embryogenesis, somatic embryo as a genetic system.

Uint V. Introduction to cell culture-

Totipotency, tissue culture technique to produce novel plants and hybrids, preparation and composition of tissue culture media. Callus initiation and maintenacnce, suspension culture: single cell clones. Cytodifferentiation.

Unit VI. Variation

Somaclonal and gametoclonal variations origin of somaclonal variations, mechanism underlying genetic variations, assessment of somaclonal variations, isolation of variants – disease resistant lines, herbicide resistant lines, stress tolerant lines.

Protoplast isolation – culture and fusion, selection of hybrid cells and regeneration of hybrid plants, symmetric and asymmetris hybrids, cybrids.

Unit VII. Cryopreservation-

Slow growth and DNA bank for germplasm conservation.

 

Uint VIII. Plant Transformation –

Plant Transformation applications for productivity and performance, production of transgenic plants resistant to herbicides, pathogens (Virus, fungi and bacteria), insect pests (Bt. Toxin gene) and abiotic stresses (drought, salt, chilling).

Unit IX. Genetic manipulation

Genetic manipulations in plants- Flower pigments, the time of flowering, production of male sterile plants. Antisense strategy,

PRACTICALS

1.      Sterilization and preparation of media.

2.      Isolation of explants, establishment and maintenance of culture. Subculture of callus, organogenesis and transfer of plants to soil.

3.      Micropropagation by proliferation of axillary bud.

4.      Micropropagation by adventitious shoot proliferation.

5.      Initiation and establishment of cell suspension cultures.

6.      Microspore/ anther culture for haploid production.

7.      Protoplast isolation and culture.

8.      Embryogenesis and embryo culture, synthetic seeds.

9.      In vitro pollination and culture of ovary/ ovule.

10.  Density gradient centrifugation for isolation of chloroplast and mitochondria.

11.  Isolation of DNA from chloroplast.

12.  Isolation of total RNA from plant.

13.  Demonstration of southern hybridization to check the plant transformation.

14.  Isolation of DNA from plant.