Plant Biotechnology


Dr Rukmini Mishra


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Course Name: Plant Biotechnology

Code(Credit) : CUTM1439(3-1-0)

Course Objectives

•To understand the basics principles of plant sciences and molecular biology and their integration towards trait improvement in plants.
•To have a thorough knowledge of laboratory techniques used in plant biotechnology.
•To understand the industrial applications of biotechnology in developing new products.
•To undertake research in plant biotechnology.

Learning Outcomes

•The students will understand the concepts and techniques of plant biotechnology and their applications in crop plants.
•They can get job opportunities in agribusiness, private companies, industries, universities and research laboratories.
•They can pursue higher studies and research in the field of plant biotechnology.

Course Syllabus

Module I: Basics of Tissue Culture

History, scope, concept of cellular differentiation and Totipotency, Cell culture media and sterilization techniques, Callus culture, nodal and tip culture, Protoplast and Embryo culture, Embryo culture and embryo rescue, protoplast isolation, culture and plant regeneration

Practice: 1. Preparation of tissue culture media
2. Direct Organogenesis: Shoot tip culture

Module II: Applications of Plant Tissue Culture

Somatic embryogenesis, Somaclonal variation and crop improvement, Germplasm conservation, Production of Secondary metabolites through Tissue culture, Industrial applications.

Practice: Micropogation techniques

Module III: Recombinant DNA technology

Genomic DNA & plasmid DNA isolation and purification, construction of
recombinant DNA and expression cassettes, Transformation (mobilization of vectors into competent bacteria), selection and analysis of recombinant clones, genomic DNA and cDNA libraries.

Practice: DNA isolation and purification

Module IV: Genetic Engineering in Plants

Vector mediated Gene transfer, Molecular basis of crown gall and hairy root diseases, features of Ti and Ri plasmids, mechanism of T-DNA transfer, role of virulence genes, vectors based on PTi & PRi, binary and co-integrate vectors, optimized protocols for Agrobacterium-mediated genetic transformation, physical and chemical methods of gene transfer.

Practice: 1. Callus induction
2. Agrobacterium mediated gene transfer in plants

Module V: Methods of Gene transfer in plants

Direct gene transfer methods (particle bombardment/ micro projectile / biolistic, electroporation, microinjection, liposome mediated, silicon carbide fibers), chemical methods (PEG - mediated, calcium phosphate co-precipitation), transgenic monocots and dicots via direct gene transfer, in plant transformation. Integration and fate of transgene, precision of transgene integration by site-specific.

Module VI: Applications of Genetic Engineering

Transgenic plants for disease resistance, nutritional improvement, herbicide tolerance,
Long shelf life, edible vaccines.

Module VII: Gene silencing in plants

Antisense RNA technology: Antisense RNA, construction of antisense vectors, applications of antisense technology. Gene silencing: causes (DNA methylation, homology-dependent suppression by antisense gene), strategies for avoiding gene silencing, methods of inducing gene silencing and its application. Regulatory RNA molecules (si RNA and miRNA), RNAi technology and its applications in plants.

Text books:
Glick, B. R. and Pasternak (2003). Molecular Biotechnology: Principles and Applications of Recombinant DNA. ASM Press, Washington, D. C., USA.
Kyte, L. and Kleyn, J. (1996). Plants from Test Tube to: an Introduction to Micro propagation, 3rd Ed. Timber press, Port land, USA.
Reference Book
Pollard, W. J. and Walker (1990).Plant Cell and Tissue Culture Vol VI. Humana press Clifton, USA.

Session Plan

Session 1

Cellular differentiation & Totipotency

Session 2

Tissue culture media and sterilization techniques

Session 3

Session 4

Session 5

Session 6

Embryo culture and embryo rescue

Session 8

Protoplast isolarion and plant regeneration

Session 9

Session 10

Session 11

Somaclonal variation and crop improvement

Session 12

Germplasm conservation, Cryopreservation

Session 13

Production of secondary metabolites through tissue culture

Session 14

Industrial applications of tissue culture

Session 15

Plasmids, vectors and enzymes used in genetic engineering

Session 16

Vector construction, transformation and selection of expression cassettes

Session 17

Genomic DNA and plasmid DNA isolation and purification

Session 19

Genomic library and cDNA library, screening of libraries.

Session 21

Session 22

Ti and Ri Plasmids, vir genes, mechanisim of T-DNA transfer

Session 23

vectors based on PTi & PRi, binary and co-integrate vectors

Session 24

Agrobacterium mediated gene transfer in plants

Session 25

Chemical methods (PEG - mediated, calcium phosphate co-precipitation)

Session 26

Session 27

Session 28

Integration and fate of transgenic plants

Session 29

Session 30

Session 31

Applications of genetic engineering in industry

Session 32

Applications of genetic engineering in enviromental waste mangement

Session 33

Gene Silencing in Plants-Introduction

Session 34

Antisense RNA technology, antisense vectors, applications

Session 36

T-DNA Tagging, Transposon Tagging, Applications of RNAi technology in plants

Case Studies

Case Studies

Our Main Teachers

Dr Rukmini Mishra

Assistant Professor

Dr Rukmini Mishra is presently working as an Assistant Professor (Botany), School of Applied Science, Centurion University of Technology and Management. She has a PhD in Agricultural Biotechnology from ICAR-National Rice Research Institute, India. After PhD, she received the prestigious Young Scientist Award from the Dept. of Science & Technology, SERB, Govt. of India to […]