BASIC BOTANICAL TECHNIQUES

Course Outcomes:

The students would acquire knowledge of basic botanical laboratory techniques. Gaining of knowledge on basic microscopes and their operations. Experience the hands-on training on the preparation of microscopic slides to understand the internal features of plants (anatomical studies). Learning the preparatory methods of microbial culture media. Exposure on inoculation of microbes on various culture media.

Unit 1: Microscopy:

Principles of microscopy, types of microscopes (bright field, dark field), components of microscopes. Micrometry- Micrometers (Ocular & Stage micrometers), determination of cell size. Determination cell density- Haemocytometer; Camera Lucida.

Unit 2: Micropreparation:

Sectioning plant materials- free hand sectioning and microtome  sectioning. Staining- Single staining and double staining methods

Localization of cell wall and nucleic acids using single stain- Safranin; Giemsa staining methods.

Differentiation of bacteria using double stains- Gram’s staining method.

Introduction to Fluorescent dyes.

Unit 3: Microslide preparation:

Slide preparations- fixation, embedding and staining & mounting of specimens. Temporary slide preparations of plant root, stem and leaf.

Permanent slide preparations of plant root, stem and leaf.

Unit 4: Culture of Microorganisms:

Introduction to the culture media. Simple and complex culture media. Types of culture media- liquid and semisolid media. Composition of simple media (PDA- Potato Dextrose Agar Medium) & complex media (nutrient agar medium).

Unit 5: Inoculation of Microorganisms:

Broth culture of Escherichia coli (E. coli)- serial dilution. Culture methods on semisolid media- streak plate method, pour plate method & stab culture.

PRACTICALS: 

  1. Components of the Dissection Microscope
  2.  Components of the Compound Microscope
  3.  Determination of the Cell Size using micrometers
  4.  Determination of the cell density using Haemocyometer
  5.  Drawing of plant cells using Camera lucida
  6.  Free hand sectioning of plant specimens (Root, Stem, Leaf)
  7.  Microtome sectioning of plant specimens (Root, Stem, Leaf)(demonstration)
  8. Single staining of plant specimens using Safranin & Giemsa stains
  9. Double staining of bacteria using Gram’s staining
  10. Temporary microslide preparation
  11. Permanent microslide preparation (submission of 5 slides)
  12. Preparation of microbial culture medium: PDA medium
  13. Inoculation of microbes on to the culture media: broth culture, streak plate, pour plate and stab culture

REFERENCES:

John Jothi Prakash E. 1987. A Text Book of Plant Anatomy. Emkay Publications, Delhi.

Casida L.E. 2015. Industrial Microbiology (3rd ed.), New Age International (P) Limited Publishers, New Delhi, India.

Pelczar M.J., Chan E.C.S. and Krieg N.R. 2003. Microbiology (5th ed.). Tata McGraw-Hill Publishing Company Limited, New Delhi.

Reed G. 2004. Prescott & Dunn's industrial microbiology (4th ed.), AVI Pub. Co., USA.

Waites M.J., Morgan N.L., Rockey J.S. and Higton G. 2001. Industrial Microbiology: An Introduction (1st ed.). Blackwell Science, London, UK.

Prescott S.C., Dunn C.G., Reed G. 1982. Prescott & Dunn's Industrial Microbiology (4th ed.), AVI Pub. Co., USA.

Waites M.J. 2008. Industrial Microbiology: An Introduction (7th ed.), Blackwell Science, London, UK.

ENVIRONMENTAL STUDIES