BIOCHEM-231

BIOCHEM-231: FUNDAMENTALS OF PLANT BIOCHEMISTRY AND BIOTECHNOLOGY

Credit: 3(2+1)

THEORY:

Importance of Biochemistry.Plant cell structure and function of cell orgenells.Properties of Water and significance of weak interactions and biomolecules.Bioenergetics and significance of redox reactions. Carbohydrate: Structure, classification, properties and functions. Lipid: Structures and properties of fatty acids. Structure, classification, properties and functions. Amino acids and Proteins: Structure, classification, properties and functions. Structural organization of proteins. Enzymes: General properties; Classification; Mechanism of action and enzyme kinetics and inhibition. Enzyme regulation. Nucleic acids: Importance and classification; Structure of Nucleotides, DNA structure and forms of RNA and function.Mitochondrial electron transport and oxidative phosphorylation. Metabolism of carbohydrates, lipids and nucleic acids.Biochemistry of nitrate assimilation and photosynthesis. Introduction to recombinant DNA technology: PCR techniques and its applications.

Concepts and applications of plant biotechnology: Scope, organ culture, embryo culture, cell suspension culture, callus culture, anther  culture, pollen culture and ovule culture and their applications;  Micro-propagation methods; organogenesis and embryogenesis, Synthetic seeds and their significance;  Embryo rescue and its significance; somatic hybridization and cybrids;  Somaclonal variation and its use in crop improvement; cryo-preservation;  Introduction to recombinant DNA methods:  physical (Gene gun method),  chemical (PEG mediated)  and Agrobacterium mediated gene transfer methods; Transgenics and its importance in crop improvement;  PCR techniques and its applications; RFLP, RAPD, SSR; Marker Assisted Breeding in crop improvement; Biotechnology regulations.

PRACTICALS:

Preparation of solution, pH & buffers, Qualitative tests of carbohydrates and amino acids.Quantitative estimation of soluble sugars and amino acids. Estimation of starch, total carbohydrate and  soluble proteins . Determination of crude fat and qualitative tests of fats and oils. Enzyme assay: Alpha amylase , Nitrate reductase, lipase and protease. Paper chromatography/ TLC demonstration for separation of amino acids/ Monosaccharides.Demonstration on isolation of DNA. Demonstration of gel electrophoresis techniques and DNA finger printing.

Practicals (Experiments):

1) Preparation of solution, pH & buffers

2 & 3) Qualitative tests for carbohydrates and amino acids

4) Estimation of reducing sugars by Nelson-Somogyi method

5) Estimation of starch by Anthrone method

6) Determination of soluble protein by folin-lowry method

7) Estimation of free amino acids by Ninhydrin method

8) Determination of total crude fat/oil by Soxhlet method

9) Qualitative tests for oil

10) Determination of alpha amylase activity from germinating seed

11) Determination of invivo nitrate reductase activity from leaf tissue

12) Paper chromatography/ TLC demonstration for separation of amino acids

13) TLC for separation of sugars

14 & 15) Isolation of genomic DNA from plant. Purification, Quantification and quality determination

16) Amplification of genomic DNA using different primers and resolution of PCR products on agarose gel