Course of Study of Biochemistry and Biotechnology (B.Sc. 3rd Year)
Course No: Bot 301
Nature of the Course: Theory
Year: B.Sc. III lear
Objectives: The general aim of this course biochernistry and biotechnologY.
Unit A: Plant Biochemistry
Full Marks: 100
Pass Marks: 35
Lectures: I 50
is to provide f'undamental knowledge of plant /5
l. Introduction: a) Plant biochemistry as a science (b) Relationship beNveen plant biochemistry and plant sciences (genetics. cell biology, molecular biology. plant physiology and other branches of bio sciences) 2 hrs
2. Bio-molecules 37 hrs (a) Water: motecular structure, properties, water as a solvent; lonization of water. pH
(b) Functional groups found in bio-molecules: general concept (c ) Ca rbohi'd ratcs: Del'i,-r ition. c lassi fi cation
6 hrs 3 hrs
Monosaccharides: classification (based on lunctional group: based on no of C atoms). Properties ( Esteri fi cation, ox idation. red uction. osazone lormat ion. cyanohyd ri n reaction. furfural lbrmation. enolization): dcrivativcs ol'monosaccharides (dc-oxy clerivatives. acids. alcohols. amino sugars. sialic acid): tliological impo(ancc: Disaccharides. Polysaccharides and their biological importance ( I +6+3 hrs)
(d) Lipids: Definition, structure of glycerol and fatty acids
Properties of fatty acids, kinds of lipids (Clycero lipids; Iipids without glycerol; complex lipids), properties of lipids, Biological roles of lipids (l+4hrs)
(e) Proteins: Dellnition. Amino acids and their classillcation (essential. semi essential and 1on-essential: polar and non-polar: acidic. basic or neutral)
properties ol'Amino Acids: Amino acids as zwitterions. Amino acids as electrolytes" Reactions of arnino aci,Js (ninhycJrirt reactiott: Sanget"s reaction: [:drtran"s reaction" Darrsyl Ch Ioride reaction ; Phosgene reaction : clecarboxylation : [:steri tlcation : Acylation): Peptide bonds and Polypeptide: Structure of polypeptides primary/secondary/tertiary and quaternary structures;
Classification of proteins. Functions of proteins.
(2 +5+3+3 hrs)
3. Nucleic acids: (3iZ+ lr t+3+2+3rs):15 hrs Building components of nucleic acids: N2 bases and their derivatives. sugars, phosphates. modifled bases.
Biological roles of nucleotides
'fypes of nucleic acids (DNA. RNA and their types)and their role
Properties of Nucleic acids: Denaturation/ renaturation. hydrolysis. non-enzymatic transfbrmation of nucleotides and nucleic acids (deamination. depurination. alkytation. formation of'pyrimidine dimers and photoproduct); F.nzymatic transformation of nucleic acids (concept of DNase/RNase. endo- and exonuclease. DNA polymerases. their types and fbnctions)
DNA sequencing (Determination of base sequences in DNA molecule): Maxarn Gilbert sequencing, Sanger sequencing.
4. Enzymes: Properties and chemical structure, Mechanism of action, Denaturation, F'actors aliecting enzyme action. Allosteric protein and tbedback inhibition. Classitlcation and nomenclature 7 hrs
5. Plant pigments: Structure and lunctions of chlorophylls. carotenoids. anthocyanins, phycobi I I ins 4 hrs
6. \/itamins and their role in plants: Fat soluble vitamins (A. tr and K): Watcr soluble vitamins (Coenzyrne A. Vitamins Bl. 82.86. C. and Il) 5 hrs
7. Introduction to Bioinformatics 5 hrs Deflnition. role of computer in bioinformatics, Branches of bioinlormatics, Aim of bioinfbrmatics, Scope and research areas in bioinformatics, Biological data-DNA sequence, protein sequence, macromolecular structure. Databases in bioinformatics. basic bioinformatics tools- Databases search (Entrez, SRS), Blast, Fasta.
Unit B: Plant Biotechnology 1. Overview of Biotechnology
( l+3+ l0 hrs): l4 hrs
Introduction : A)Origin and History of biotechnology.l)) Scope and irnportance ol' biotechnolog),: a) Iliotechnology in Medicinc. b) Blotechnologl, in t'oocl industr.r,. c)lliotechnology in agriculturc. d) Biotechnology in lrermentation technologl,, e) Biotechnology in environmental engineering: C)Achievements ol'biotechnology: a) Cene cloning. b) Recombinant DNA technology,c) In vitrut culture technology. d)Gcneticalll,, engineered drugs, e)Diagnosis of diseases f)Biosensors g) Biofbrtilizersh) Mutation Breeding, i) [inzyme technology , .i) DNA finger printing. h)Monoclonal antibody2. Plant tissue culture technique and application (2-r5i-25-r3) :35 hrs A. Introduction to plant tissue culture.
B.Basic principles and tcchniques oflz vitro culture: 1-otipotenclr. Basic requirements tor grouing plants in vitro:Laborator),organization. composition and selection o1'nutrient media, steril ization, culture room
C.Types of plant tissue culture and their applications: a.Seed culture. b.organ culture (Meristem culture, root culture" shoot culture). c.['.mbryo culture. d.Anthcr Culturee.Ovulc culture, f .Endosperm culture.g.Callus cultureh. Protoplast culturc. i. Cell suspcnsion culture. j Shortening of breeding c,r"cle.
D. Cryopreservation : a. Principles, Method of cryopreservation-i)Preparation of material for deep freezing, ii)Cryoprotectors, iii) Freezing programmes. iv)Storage strategies, v)Assessment of successful cryopreservation,b. Uses of cryopreservation -- i)Cryopreservation of Semen. ii) Cryopreservation of ova and embryo, iii)Maintenance ol plant germplasm for long duration, iv)Organ explants, v)Zygotic and immature embryos. r i)Shoot tips. vii)Callus. viii) Cell suspensionsx)Protoplasts. c. Significance of cn'opreservation
3. Plant- Microbe Interaction (2+3+7) 12 hrs A. Biological Nitrogen fixation: mechanism and its impclrtance
B. Symbiotic and asymbiotic organisms for soil fertility and crop improvement.
C. Biofertilizers (Symbiotic associations): i) Rhizobium- F'ormation ol'nodulc. Nitrogen fixing organism found in nodules, Structure and function of nodule, Mechanism of Nitrogen fixation by Nodules, Rhizobial biofertilizers, Rhizobium biolertilizers in forest trees.ii) Blue Green Algae (BGA)-Mass production and application of Blue Green Algae, Azolla - anabaena symbiosis. Asymbiotic associations. Non symbiotic rritrogen fixing bacteria. Non s;'mbiotic bacterial biofertilizer-a) Azotobactor. b) Azospirillum.c) Carricr fbrrnulation. iii). Frankia biolertilizer. iv) Mycorrhiza-Types of mycorrhiza. beneflts fiom Mycorrhizas to plants, establislrment of Mycorrhiza associations in vitro
4. Gene transfer in plants: ( l0+4 hrs;-.14 lrrt
A) Concept of gcne cloning: Basic requirements fbr gene cloning in plants: sene isolation and cloning; Concept ol'vcctors; marker and reporler genes and thcir rolcs in plant
transformation; identification and analysis of cloned genes (colony hybridization' immunological detection, PCR, blotting)
B. Gene transfer techniques in plants: ilgene transfer methods: (Direct/vectorless and indirect/ vector mediated); transformation of mitochondria and chloroplasts iii)CMcrops: applications and limitations iv) General concept of molecular farrning fiom transgenic plants. v) ethical issues in plant genetic engineering
Text and Referencc books
l. Bhattarai, T. 2005. Experiments on Plant Biochemistry and Plant Biotechnology. BhundiPuran Prakashan, Kathmandu
2. Bhattarai, T. 2OO7 . Plant Physiology. Bhundipuran Prakashan, Kathmandu' 3. Jain, J.L. 2004. Fundamentals of Biochemistry. S Chand and Company Ltd' New
4.|:llirr.r, A.1.., Netson, D.L. and cox, M.2004. Principles of Biochemistrv.4'r' edition. McMillan Limited, USA (lndian Reprint )
5. Rastogi. S.C. 1993. Biochemistry. Tata McGraw-Hill Publishing Company Ltd. Delhi
I . Attwood, T.K. and Parry-Smith. D. J. 2001 . lntroduction to Bioinfbrmatics. Prentice Hall Inc.
2. Ghosh, Z. and Malllick. B. 2008. Llioinlbrmatics -principle and applications. Oxtbrd University Press, lndia'
3. Mount. D. W.2001. Bioinfbrmatics Sequence and Cenome Analysis. Cold Spring Harboor Laboratory Press, New York'
I . Altman, A. and Hasegawa, P. 2012. Agricultural Biotechnology. Academic Press' 2. Bhojwani S. S. I 990. Planr Tissue Culture: Applications and Limitation's. Elsevier Science Publishers.
3. Bhojrvani S.S. and Razdan, M. K. 1996. Plant Tissue Culture: T-heorv and Practice' Elsevier Science Publishers.
4. Chawala Il.S. 2009. InrroCuction to P!ont 7'i.;sue Cu!tttre. 'Ihird Edition. Oxford and
5. Crispeels, M.J. and Sadava, D.E. 2006. Plants. Genes and crop lmprovement' American Society of Plant Biologists- l'JSA'
6- Debergh, P.C. and Zimmerman. R.H. 1990. Micropropagation. Kluwer Academic Publ. Dordrecht.
7. Dodds,.i.H. and Roberts, L.W.1995. Experiments in Plant Ti.ssue ('ulture (3rd
Iidition). Cambridge University press. Cambriclge. UK.
8. Dubey R.C. 2009. A text Book of Biotechnology S Chand and company Limited.
9. Gamborg O.L. and Phillips C.C. 1995. Plant Cell. 7-i.s.yue ancl Organ Culture -
F-undamental Methods (Lab. Manual). Springer-Verlag.
10. Greene J'-l' and Rao V.B. 1998. Recontbinant Dlr,lA Principle.s antl llfethocktlogia.s.
I I' I.al R. and Lal S. 1995. Genetic Engineering of Plonts.for Crop Improvemerl. CRC
12. Pierik R.L.M. 1997.ln vitroCulture o/'Higher Plant.s. Kluwer Academic publisher. Netherlands.
13. Punia M'S. 1999. Plant BiotechnoloEy and Molecular Biology: A Laborotort, Manual. Scientific Publishers. India.
14. Razdan M. K' 2003. tntroduction to Plant Tissue C'ulture.Agritech publications.