BOTN-8101                                                              Methods in Plant Cell Tissue and Organ Culture                                3(2+1)

Plant tissue culture broadly refers to the an in vitro cultivation of plants, seeds and various parts of the plants (organs, embryos, tissues, single cells, protoplasts).With the advances made in the tissue culture technology, it is now possible to regenerate species of any plant in the laboratory.  In this process the growth medium or culture solution is very important as, it is used for growing plant tissue because it contains various plant nutrients in the form of 'jelly' known as agar and plant hormones which are necessary for the growth of plant. Plant tissue culture is used widely in the plant sciences, forestry, and in horticulture. Applications include: The commercial production of plants used as potting, landscape, and florist subjects, which uses meristem and shoot culture to produce large numbers of identical individuals.

Students will know about various tissue culture methods / techniques for somatic embryogenesis,cell suspension culture, another culture and artificial / synthetic seeds. This course elaborates the role of tissue culture methods insomatic hybrids,somaclonal variationandcryopreservation for crop improvement

Contents                                                                                            

1. Different methods of aseptic techniques and their requirements, media composition, effect of chemical and physical factors, different types of media used in plant tissueculture.

2. Explant; Types of explant, meristem culture, stages of micropropagation, advantages of micropropagation and its limitations, virus free plant production.

3. 3.Types of callus;Callus development from different sources and production of secondary metabolites,organogenesis, rhizogenesis, differentiation, development of root and shoot culture, factors effecting regeneration.

4. Direct and indirect somatic embryogenesis, distinct stages in the development of somatic embryogenesis, factors effecting somatic embryogenesis, importance of somatic embryogenesis
5. Protoplast isolation, t fusion, methods of protoplast fusion, somatic hybridization, methods of selection of somatic hybrids, cybrids, major application of cybrids.
6. Cell suspension culture. Different categories of cell suspension culture, Hairy root culture; Strategies used to optimize product yield. Production ofdifferent phytochemicals by suspension culture.  Bioreactors, mass production by bioreactors, types of bioreactors
7. Principle of anther culture(microspore or pollen culture). Advantages of haploids / anther culture / pollen culture, pollenculture and its impacts in plant breeding. Ovary culture or gynogenesis
8. Artificial/Synthetic seed, methods for synthetic seed production.
9. Mechanism causing somaclonal variation, selection of somaclonal variation, problems withsomaclonal variation, methods for accessing somaclonal variation.
10. Methods of in vitro conservation at different temperatures.

Lab work                                     

1. Study of apparatus in plant tissue culture laboratory, a septic techniques used inlaboratory preparation of stock solutions MS (Murashiage& Skoog).
2. Preparation of stock solutions of growth regulators for medium M S, preparation of 1LMS medium.
3. Study of micropropagation.
4. Study of callus induction in plants.

Recommended Texts

1. Rapley, R. &Whitehouse, D.  (2015).Molecular biology and biotechnology.London: Royal Society of Chemistry.
2. Pullaiah, T., Rao, M.V.S.&Sreedevi, E. (2016).Plant tissue culture: theory and Lab work(2nd ed.).Jodhpur India.Scientific Publishers.

Suggested Readings

1. De-Smet, I. (2016). Plant organogenesis: methods and protocol.Totowa, New Jersey:Humana Press
2. Loyola-Vargas, V.M. &Neftalí, O. (2016).Somatic embryogenesis: fundamental aspects and applications.Switzerland: Springer International Publishing.

Aims and Objectives

• To quickly produce mature plants.
• The production of multiples of plants in the absence of seeds or necessary pollinators to produce seeds.
• The regeneration of whole plants from plant cells that have been genetically modified.

Assessment criteria:

Sessional (Attendance, assignment & presentation) 10 Marks

Practical Exam           15 Marks

Mid Term Exam         30 Marks

Final Term Marks       45 Marks