HOR 602 Biotechnology of Horticultural Crops 2+1

Syllabus

Objective

To understand the principles of biotechnology and recent developments in biotechnology along with its applications in horticultural crops.

Theory

Unit 1

Basics of Plant Tissue Culture

Biotechnology - Concepts - history and scope- Harnessing bio-technology in horticultural crops. Plant tissue culture – totipotency - sterilization techniques, explants. Tissue culture media - nutritional requirements - plant growth regulators. Types of culture – shoot tip culture, meristem culture, Anther and microspore culture, Embryo rescue, embryo and ovary culture, Protoplast culture Callus culture - organogenesis and embryogenesis. Synthetic seeds. In vitro germplasm conservation –Rooting and hardening of TC plants- National certification system for TC plants.

Unit 2

Recombinant DNA technology and genetic transformation

Tools and techniques – host, vectors – plasmid DNA and phage DNA, DNA manipulation – enzymes. Construction of recombinant DNA molecules and introduction of DNA into host cells. Isolation of genes of interest. Methods of gene delivery into plants –Agrobacterium and Particle bombardment mediated methods - Selectable markers and reporter genes, promoters. Applications of transgenic in improvement of horticultural crops. Detection of GMOs, Indian regulatory guidelines, biosafety and bioethics.

Unit 3

Molecular marker technology

DNA markers – different kinds – hybridization based – PCR based markers. Comparison among different DNA markers. Application of molecular markers - characterization of plant genetic resources – fingerprinting of varieties - gene tagging - Marker assisted selection and crop improvement.

Unit 4

Biopharming and bioprospecting

Plants as biofactories – production of vaccines, therapeutic proteins, industrial enzymes and bioplastics Discovery of novel compounds from plants - secondary metabolites – bioreactors for plant cell culture.

Practical

Laboratory organization – Equipments - sterilization techniques -Preparation of stock solution for MS and Preparation of MS medium .Inoculation of explants – shoot tip, meristem tip and embryo. Anther - microspore culture- Plant genomic DNA extraction - Quantification of DNA and Electrophoresis of plant DNA- plasmid isolation- Restriction digestion of DNA, Ligation and Bacterial transformation- Agrobacterium mediated transformation- particle bombardment- PCR- extraction of secondary metabolites.

Suggested Reading

Bhojwani, S.S. and Razdan, M.K. 2006. Plant Tissue Culture Studies – Theory and Practice. Elsevier Publication. Brown, T.A. 2006. Gene cloning - An introduction. V Ed. Chapman Hill, U.K. Lehninger. 2004. Principles of Biochemistry. CBS Publications, New Delhi. Lewin, B. 2014. Genes XI. Oxford University Press, Inc., New York. Malacinski, M. and D. Friefelder. 2003. Essentials of molecular biology. IV Ed. Jones and Bartlett publishers, Boston.

Singh, B.D. 2010. Expanding horizons in Biotechnology. Kalyani Publications, New Delhi. Slater, A. 2008.Plant Biotechnology The Genetic Manipulation of Plants 2nd edition Oxford University Press, London. Tomer. 2010. Molecular Markers and Plant Biotech. New India Pub.

N. Manikanda Boopathi. 2013. Genetic mapping and marker assisted selection: Basics, Practice and Benefits. Springer India. pp: 303. DOI 10.1007/978-81-322-0958-4 (ISBN: 978-81-322-0957-7 (Print) 978-81-322-0958-4 (Online))

e-Resources

http://www.agbiotorum.org

http://www.nal.usda.gov` http:// biotech-info.net

http://www.biotech.vt.edu/outreach/biotech-times/5_98/pharming

http://www.ncbi.nlm.nih.gov. http://www.nal.usda.gov.

http://www.agbiosafety.unl.edu/