Practical

Paper Code: 
BOT 423A
Credits: 
4
Contact Hours: 
60.00
Max. Marks: 
100.00
Objective: 

Course Objective

This course will enable the students to –

  1. To demonstrate and develop specific technical knowledge in field of Plant tissue culture.

Course Outcomes (COs):

     Course

Course Outcome

Learning and teaching strategies

Assessment Strategies

Paper Code

Paper Title

BOT 423A

 

Practical

On completion of this course, the students will be able to;

CO105: To explain the significance and formulations of different media compositions for the purpose of tissue culture, isolation of protoplast and organogenesis.

CO106:  To explain the concept of protoplast isolation for production of novel variants.

CO107:  To demonstrate and  execute latest techniques used for genome sequencing

CO108:  To remember and execute procedures related to basic laboratory setup, handling of explant tissue, nutrient medium and establishing the culture and incubation of cultures.

CO109:  To demonstrate suitable methods for characterizing the activity, function, diversity and composition of microbial communities.

CO110:  To demonstrate the ability to identify significant biological research questions, develop research protocols and properly analyse research questions through the use of the scientific method.

Substantial laboratory-based practical component and experiments

Technology-enabled learning

Field-based learning

Project-based learning

Internship in industry, and research establishments.

Literature surveys and evaluations

 

Practical assignments and laboratory reports

Observation of practical skills

Individual and group project reports

 

Viva voce interviews are majorly adopted assessment methods

 

 
 

 

CONTENTS:

  • Media preparation, sterilization and inoculation of given material.
  • Growth characteristics of E.coli using plating and turbid metric methods.
  • Isolation of plasmid from E.coli by alkaline lysis method and its quantitation spectrophotometrically.
  • Restriction digestion of the plasmid and estimation of the size of various DNA fragments.
  • Cloning of a DNA fragment in a plasmid vector, transformation of the given bacterial population and selection recombinants.
  • Organogenesis and somatic embryogenesis using appropriate explants and preparation of artificial seeds.
  • Isolation of protoplast from various plant tissues and testing their viability.
  • Effect of physical and chemical factors protoplast yield.
  • Demonstration of protoplast fusion employing PEG.
  • Demonstration of DNA sequencing using Sanger, s dideoxy method.
  • Demonstration of micro techniques for plant cultures.
  • Co-cultivation of the plant material with Agrobacterium and study GUS activity histochemically.

 

 

 

 

Essential Readings: 
  • Annarita Leva, ‎Laura Rinaldi. Recent Advances in Plant in Vitro Culture. (2012). Croatia: IntechOpen.
  • Bohorova, N. (1999). Laboratory protocols: CIMMYT Applied genetic engineering laboratory. Mexico: CIMMYT, Applied Biotechnology Center.
  • Dodds, J. H., Dodds, J. H., Roberts, L. W. (1995). Experiments in plant tissue culture. United Kingdom: Cambridge University Press.
  • George, E. F. (1993). Plant Propagation by Tissue Culture. United Kingdom: Exegetics.
  • Pierik, R. (2012). In Vitro Culture of Higher Plants. Netherlands: Springer Netherlands.
  • Oluf Gamborg, ‎Gregory C. Phillips ·Plant Cell, Tissue and Organ Culture: Fundamental Methods. (2013). Germany: Springer Berlin Heidelberg.
  • Shri Mohan Jain, ‎Pramod K. Gupta · Protocol for Somatic Embryogenesis in Woody Plants. (2005). Netherlands: Springer Netherlands.
  • Hvoslef-Eide & A.K,. Preil ‎W (Ed.)· Liquid Culture Systems for in Vitro Plant Propagation. (2005). Netherlands: Springer Netherlands.

 

 

                                                 

                                                

 

References: 

e-Resources:

 

Academic Year: 
2022-2023
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