Plant Biotechnology

Teacher

Dr Rukmini Mishra

Category

Core Courses

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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.

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

https://www.youtube.com/watch?v=a-EscAd0e9E

Session 4

Embryo culture and embryo rescue, protoplast isolation, culture and plant regeneration

https://www.youtube.com/watch?v=VSmDipbahlA

https://www.youtube.com/watch?v=nptapx8faUU

Session 5

Practice 
Preparation of tissue culture media
https://www.youtube.com/watch?v=ltbdM3boWmU

https://www.youtube.com/watch?v=eMv_PMNPYMc

Session 6

Practice 

Direct Organogenesis: Shoot tip culture

https://www.youtube.com/watch?v=b5kK-5NlgdY

Session 7

Protoplast isolation and fusion

https://www.youtube.com/watch?v=8grwL8daO1Q

Session 8

Somatic embryogenesis, somaclonal variation and crop improvement

https://www.youtube.com/watch?v=3ldYQRmuqtE

https://www.youtube.com/watch?v=ltaPGuwdTVc

Session 9

Germplasm conservation

https://www.youtube.com/watch?v=XXXTY4IxHrg

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 13

Genomic DNA & plasmid DNA isolation and purification
https://www.youtube.com/watch?v=8cYvyYOjzOc

Session 14

Construction of recombinant DNA and expression cassettes

https://www.youtube.com/watch?v=WwFwDGkGPlc

Session 15

Transformation (mobilization of vectors into competent bacteria), selection and analysis of recombinant clones

https://www.youtube.com/watch?v=7Ul9RVYG5CM

Session 16

Genomic DNA and cDNA libraries

https://www.youtube.com/watch?v=fmMp6avlB6I

Session 17

Vector mediated Gene transfer

https://www.youtube.com/watch?v=wHWxqsfW8r4

Session 18

Molecular basis of crown gall and hairy root diseases
https://www.youtube.com/watch?v=L7qnY_GqytM

https://www.youtube.com/watch?v=wTO-KmpZQgQ

Session 19

Features of Ti and Ri Plasmids, mechanisim of T-DNA transfer
https://www.youtube.com/watch?v=LUNL9HXvPXY

https://www.youtube.com/watch?v=wTO-KmpZQgQ&t=41s

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

https://www.youtube.com/watch?v=1rSWOSt3wVQ&t=163s

Session 22

Practice 

Agrobacterium tumefaciens mediated transformation of tobacco leaves

https://www.youtube.com/watch?v=f1_ctQSUEaE

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

https://www.youtube.com/watch?v=1GF-T3TRcHc

https://www.youtube.com/watch?v=gGZnCKvQiVA

Session 26

Practice 

Demonstration of biolistic method of gene transfer through photographs

https://www.youtube.com/watch?v=9c8E8Vs9DwM

https://www.youtube.com/watch?v=BzyuYxQpi0o

Session 27

Transgenic plants for disease resistance, nutritional improvement

https://www.youtube.com/watch?v=jsEQTTUIIW0

Session 28

Herbicide tolerance, Long shelf life, edible vaccines.

https://www.youtube.com/watch?v=aqfC-tVOnCc

https://www.youtube.com/watch?v=vwF-cMbcPzc

Session 29

Antisense RNA, construction of antisense vectors, applications of antisense technology.

https://www.youtube.com/watch?v=Bxu-_2FpPSY

https://www.youtube.com/watch?v=z47jOVyHnIc

Session 30

Gene silencing: causes (DNA methylation, homology-dependent suppression by antisense gene)

https://www.youtube.com/watch?v=Q5lGqACCthc

Session 31

Strategies for avoiding gene silencing, methods of inducing gene silencing and its application

https://www.youtube.com/watch?v=J2Y_S4EkLy8

Session 32

Regulatory RNA molecules (si RNA and miRNA)

https://www.youtube.com/watch?v=_Bl_l9SNvjA

Session 33

Session 34

CRISPR/Cas technology and its applications in plants

https://www.youtube.com/watch?v=3AQWMG2Lqvc

Our Main Teachers

Dr Rukmini Mishra

Assistant Professor
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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 […]