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Plant Biochemistry and Biotechnology

Course No: Bot 301Nature of the Course: Theory

Year: B.Sc. III lear

Objectives: The general aim of this coursebiochernistry and biotechnologY.

Unit A: Plant Biochemistry

Full Marks: 100Pass Marks: 35Lectures: I 50

is to provide f'undamental knowledge of plant

/5

l. Introduction: a) Plant biochemistry as a science (b) Relationship beNveen plantbiochemistry 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

scale. buffer.(b) Functional groups found in bio-molecules: general concept

(c ) Ca rbohi'd ratcs: Del'i,-r ition. c lassi fi cationMonosaccharides: 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.

6 hrs

3 hrs

(2 +5+3+3 hrs)

3. Nucleic acids: (3iZ+ lr t+3+2+3hrs):15 hrsBuilding 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 roleDNA: StructureProperties 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 nucleicacids (concept of DNase/RNase. endo- and exonuclease. DNA polymerases. their typesand fbnctions)

DNA sequencing (Determination of base sequences in DNA molecule): Maxarn Gilbertsequencing, Sanger sequencing.

4. Enzymes: Properties and chemical structure, Mechanism of action, Denaturation, F'actorsaliecting enzyme action. Allosteric protein and tbedback inhibition. Classitlcation andnomenclature 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 solublevitamins (Coenzyrne A. Vitamins Bl. 82.86. C. and Il) 5 hrs

7. Introduction to Bioinformatics 5 hrsDeflnition. role of computer in bioinformatics, Branches of bioinlormatics, Aim ofbioinfbrmatics, Scope and research areas in bioinformatics, Biological data-DNAsequence, protein sequence, macromolecular structure. Databases in bioinformatics. basicbioinformatics tools- Databases search (Entrez, SRS), Blast, Fasta.

Unit B: Plant Biotechnology

1. Overview of Biotechnology

75

( 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) Cenecloning. b) Recombinant DNA technology,c) In vitrut culture technology. d)Gcneticalll,,engineered drugs, e)Diagnosis of diseases f)Biosensors g) Biofbrtilizersh) MutationBreeding, i) [inzyme technology , .i) DNA finger printing. h)Monoclonal antibody

2. 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 requirementstor grouing plants in vitro:Laborator),organization. composition and selection o1'nutrientmedia, 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.Ovulcculture, 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 ofmaterial for deep freezing, ii)Cryoprotectors, iii) Freezing programmes. iv)Storagestrategies, v)Assessment of successful cryopreservation,b. Uses of cryopreservation --i)Cryopreservation of Semen. ii) Cryopreservation of ova and embryo, iii)Maintenance olplant germplasm for long duration, iv)Organ explants, v)Zygotic and immature embryos.r i)Shoot tips. vii)Callus. viii) Cell suspensionsx)Protoplasts. c. Significance ofcn'opreservation

3. Plant- Microbe Interaction (2+3+7) 12 hrsA. 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. Nitrogenfixing organism found in nodules, Structure and function of nodule, Mechanism of Nitrogenfixation by Nodules, Rhizobial biofertilizers, Rhizobium biolertilizers in forest trees.ii) BlueGreen Algae (BGA)-Mass production and application of Blue Green Algae, Azolla -anabaena symbiosis. Asymbiotic associations. Non symbiotic rritrogen fixing bacteria. Nons;'mbiotic bacterial biofertilizer-a) Azotobactor. b) Azospirillum.c) Carricr fbrrnulation.iii). Frankia biolertilizer. iv) Mycorrhiza-Types of mycorrhiza. beneflts fiom Mycorrhizas toplants, 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 isolationand 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

Plant Biochemistryl. 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

BioinformaticsI . 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 SpringHarboor Laboratory Press, New York'

Plant BiotechnologYI . 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 andIBH

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 AcademicPubl. Dordrecht.

/

7. Dodds,.i.H. and Roberts, L.W.1995. Experiments in Plant Ti.ssue ('ulture (3rdIidition). 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.

Marcel Dekker.I I' I.al R. and Lal S. 1995. Genetic Engineering of Plonts.for Crop Improvemerl. CRC

Press.

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.15. Satyanarayan U. 2005. Biotechnologs,.l't ed, Arunabha Sen books and Allicd Ir. l.td.

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