DESCRIPTION OF COURSES
PGR 602 IN VITRO CONSERVATION AND CRYOPRESERVATION (2L+2P) II
To equip students with skills in using tissue culture and cryopreservation techniques for conservation and management of PGR.
In vitro techniques in PGR management; in vitro clonal propagation for germplasm conservation; somaclonal variation and its implication in PGR conservation; in vitro collecting; meristem culture, virus indexing and elimination, in vitro exchange and its international guidelines.
Techniques of in vitro conservation of tropical and temperate crops under slow/normal growing conditions; tissue culture in circumventing crossability barriers embryo rescue technique, rescue and survival of rare and endangered species; management of large in vitro collections; concept of active and base in vitro genebanks.
History of plant cryopreservation; structural, physiological and biochemical basis of desiccation and freezing sensitivity of plant cells, tissues and organs; cryoprotectants and their mode of action, factors affecting cryoprotection.
Methods of cryopreservation – conventional slow cooling and vitrification-based methods; factors influencing success in cryopreservation importance of pre-treatments, pre-conditioning of explant donor plants, regrowth media and culture conditions; advances in cryogenic protocols; status of application of cryopreservation in PGR conservation, application in pharmaceutical industry, forest breeding and production of trangenics.
Monitoring genetic stability of in vitro-conserved and cryopreserved germplasm.
- Preparation of stock solutions; media preparations; preparation of explants and culture initiation in monocots and dicots; meristem isolation and culture establishment; subculture of shoots in monocots and dicots, hardening and field establishment of plantlets; preparation of cryoprotectant solutions and regrowth media; cryopreservation of in vitro cultures- isolation of explants and pretreatment.
- Encapsulation-dehydration technique; encapsulation-vitrificaion technique; assessing genetic stability of in vitro conserved and cryopreserved germplasm.
- Cryobanking of germplasm,effect of duration and temperature of cryoprtectant treatment on shoot tips, effect of cold hardening - isolation of shoot tips in dicot, cryopreservation-vitrification technique, effect of thawing treatment on regrowth.
- Demonstration of embryo rescue technique/ virus indexing.
- Assessing genetic stability of in vitro conserved and cryopreserved germplasm.
- Case studies related to monitoring genetic stability of in-vitro conserved/ cryopreserved germplasm.
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- Benson, E.E. (ed.) 1999. Plant Conservation Biotechnology. Taylor & Fancis Ltd. London.
- Bhojwani, S.S. and Razdan, M.K. 1983. Plant Tissue Culture: Theory and Practice. Elsevier Science Publishing Co.Inc. New York.
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- Engelmann, F. and Takagi, H. (eds) 2000. Cryopreservation of Tropical Plant Germplasm - Current Research Progress and Application, IPGRI, Rome/JIRCAS/Japan.
- Ford-Lloyd, B.V., Newburry, J.H. and Callow, J.A. (eds.) 1998. Biotechnology and Plant Genetic Resources: Conservation and Use. CABI, Welling ford.
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- Razdan, M.K. and Cocking, E.C. (eds.) 2000. Conservation of Plant Genetic Resources In vitro, Vol. 2: Applications and Limitations. Science Publishers, Inc. USA.
- Withers, L.A. and Alderson, P.G. 1986. Plant Tissue Culture and its Agricultural Applications. Butterworth, London.