Course Name: Plant Biotechnology
Plant Biotechnology
Teacher
Dr Rukmini Mishra
Category
Core Courses
Course Attendees
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Course Reviews
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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.
Course Outcome
| COs | Course outcomes | Mapping COs with POs (High-3, Medium-2, Low-1) |
| CO1 | Understand the concepts and techniques of plant biotechnology and their applications in crop plants. | PO1-3, PO2-3, PO3-2, PO4-3, PO7-2 |
| CO2 | Laboratory skills on integrative approach for biotechnological experiments specifically on basic, techniques in micro-propagation and can get job opportunities in agribusiness, private companies, industries, universities and research laboratories | PO1-3, PO2-3, PO3-3, PO4-3, PO7-2 |
| CO3 | Design of experiments for complex scientific problems towards research in the field of conservation of plant species and can pursue higher studies | PO1-3, PO2-3, PO3-3, PO4-3, PO7-3 |
| CO4 | Experimental skills to conserve plants for sustainability | PO1-3, PO2-3, PO3-3, PO4-2, PO7-3 |
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
3. Protoplast isolation and fusion
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: Micro propagation of plants
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: Agrobacterium tumefaciens mediated transformation of tobacco 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.
Practice: Demonstration of biolistic method of gene transfer through photographs
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. CRISPR/Cas technology and its application 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
History, scope, concept cellular differentiation & Totipotency
https://www.youtube.com/watch?v=yxj5nnzhBMg
Session 2
Cell culture media and sterilization techniques
https://www.youtube.com/watch?v=ykKs5icYwq0
Session 3
Callus culture, nodal and tip culture, Protoplast and Embryo culture
Session 4
Embryo culture and embryo rescue, protoplast isolation, culture and plant regeneration
Session 5
Practice
Preparation of tissue culture media
https://www.youtube.com/watch?v=ltbdM3boWmU
Session 6
Practice
Direct Organogenesis: Shoot tip culture
Session 8
Somatic embryogenesis, somaclonal variation and crop improvement
Session 10
Production of secondary metabolites through tissue culture
https://www.youtube.com/watch?v=lTHb33TMmNA
Session 11
Industrial applications
https://www.youtube.com/watch?v=x-JcIQ2RGvI&t=3s
Session 12
Practice
Micro propagation of plants
https://www.youtube.com/watch?v=LAx2h9CeaBs
https://www.youtube.com/watch?v=byydnVwzrc8
Session 13
Genomic DNA & plasmid DNA isolation and purification
https://www.youtube.com/watch?v=8cYvyYOjzOc
Session 14
Construction of recombinant DNA and expression cassettes
Session 15
Transformation (mobilization of vectors into competent bacteria), selection and analysis of recombinant clones
Session 18
Molecular basis of crown gall and hairy root diseases
https://www.youtube.com/watch?v=L7qnY_GqytM
Session 19
Features of Ti and Ri Plasmids, mechanisim of T-DNA transfer
https://www.youtube.com/watch?v=LUNL9HXvPXY
Session 20
Role of virulence genes, Vectors based on PTi & PRi, binary and co-integrate vectors
https://www.youtube.com/watch?v=x2vxmgzsaTE
https://www.youtube.com/watch?v=1rSWOSt3wVQ
Session 21
Optimized protocols for Agrobacterium-mediated genetic transformation, physical and chemical methods of gene transfer
https://www.youtube.com/watch?v=GHc7PU_jG2M
https://www.youtube.com/watch?v=HeRgUAlH5hM
https://www.youtube.com/watch?v=VqklR_8YRfA&t=29s
https://www.youtube.com/watch?v=ulA8xsVji80
Session 22
Practice
Agrobacterium tumefaciens mediated transformation of tobacco leaves
Session 23
Direct gene transfer methods (particle bombardment/ micro projectile / biolistic, electroporation, microinjection, liposome mediated, silicon carbide fibers)
https://www.youtube.com/watch?v=e8AAyM3Qn9w
https://www.youtube.com/watch?v=WIfiwoC55bg
https://www.youtube.com/watch?v=m1ID-iJRuSY
Session 24
Chemical methods (PEG - mediated, calcium phosphate co-precipitation)
https://www.youtube.com/watch?v=mJxgtoPIGsI
https://www.youtube.com/watch?v=QLBdZN9lqqk
Session 25
Transgenic monocots and dicots via direct gene transfer, in plant transformation. Integration and fate of transgene, precision of transgene integration by site-specific
https://www.youtube.com/watch?v=Qmp7tR0mEj4
Session 26
Practice
Demonstration of biolistic method of gene transfer through photographs
Session 27
Transgenic plants for disease resistance, nutritional improvement
Session 28
Herbicide tolerance, Long shelf life, edible vaccines.
Session 29
Antisense RNA, construction of antisense vectors, applications of antisense technology.
Session 30
Gene silencing: causes (DNA methylation, homology-dependent suppression by antisense gene)
Session 31
Strategies for avoiding gene silencing, methods of inducing gene silencing and its application
Session 32
Regulatory RNA molecules (si RNA and miRNA)
Session 33
RNAi technology and its applications in plants
Session 34
CRISPR/Cas technology and its applications in plants
Our Main Teachers
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 […]