Plant Tissue Culture rajesh bhammar

Rajesh Bhammar

PLANT TISSUE CULTUREprinciple and applications

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PLANT TISSUE CULTURE PRINCIPLE A. INTRODUCTION B. PLASTICITY C. TOTIPOTENCY D. CYTO-DIFFERENTIATION E. GENETICS F. PROCESS OF PLANT TISSUE CULTURE APPLICATIONS IN… A. STUDY B. AGRICULTURE C. MEDICINE D. PHARMA INDUSTRIES E. GENETICS F. R-DNA TECHNOLOGY

Index

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It is a technique of growing and maintaining the plant cell ,tissue or organ in a suitable medium in in vitro under aseptic condition.

Plant tissue culture

Introduction

Principles of plant tissue culture

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Plants, due to their long life span, have developed a greater ability to endure extreme conditions .

Many of the processes involved in plant growth and development adapt to environmental conditions.

This plasticity allows plants to alter their metabolism, growth and development to best suit their environment.

Plasticity

Particularly important aspects of this adaptation, as far as plant tissue culture and regeneration are concerned, are the abilities to initiate cell division from almost any tissue of the plant

And to regenerate lost organs or undergo different developmental pathways in response to particular stimuli.

When plant cells and tissues are cultured in vitro they generally exhibit a very high degree of plasticity, which allows one type of tissue or organ to be initiated from another type.

In this way, whole plants can be subsequently regenerated.

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Conti…

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The ability of an individual cell to develop into a whole plant is referred as to as cellular totipotency.

The inherent characteristic feature of plant cell namely dedifferentiation and redifferentiation are responsible for the phenomenon of totipotency.

Totipotency

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The phenomenon of mature cell (differentiated) reverting to meristematic state to produce callus is known as dedifferentiation.

Dedifferentiation

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The ability of callus cells to differentiate into a plant organ or a whole plant is regarded as redifferentiation.

Redifferentiation

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In plant tissue culture , during growth and maturation of the callus tissue or free cells in suspension culture, few dedifferentiation cells undergoes cytoquiescence and cytosenescence.

These two changes are mainly associated with the process of cyto-differentiation which will ultimately form the vascular tissue , particularly xylem elements.

This phenomenon is known as cytodifferentiation

Cyto-differentiation

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SUMMARY

As per principle

Procedure of plant tissue culture

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Explant: Any piece of the differentiated or undifferentiated tissue

taken from the plant to start plant tissue culture. laboratory:

1. washing area

2. transfer area

3. culture room

4. green house Nutrient medium: Aseptic condition 1. incubator

2. autoclave

3 .lamina air flow

Pre-requisites

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cultures

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EXPLANT: For callus culture may be the differentiated

tissue from any part of the plant.

Due to their genetic totipotency.

E.g. meristematic cell, leaf measophyll cell, phloem cell, cambium Cells etc..

Callus culture

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Callus culture

Cell suspension culture

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Undifferentiated and unorganized mass of plant cells.

Parenchymatous in nature although not truly homogenous.

Some quantity of differentiated tissue besides the bulk of non-differentiated tissue.

Callus

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Explant and its genotypeComposition of mediumPhysical factorsAge of the plantLocation of ExplantGrowth regulators

FACTORS AFFECTING CALLUS CULTURE

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Callus is Slow growth plant culture system , this enables to conduct several studies like…

1. Nutritional requirement of plants2. Cell and organ differentiation3. Development of suspension and protoplast

culture4. Somaclonal variation5. Product of secondary metabolites and their

regulation

Application of callus culture

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Cell culture technique1. Isolation of single cell2. Callus culture3. Suspension culture growth and subculturing4. Synchronization of suspension culture

Cell suspension culture

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1. To study pathways of cellular metabolism2. Serve as good target for mutation and

selection of desirable mutant3. Production of secondary metabolites of

commercial interest4. Good potential for crop improvement

Application of cell culture

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Callus culture

Cell suspension culture

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Hardening

Introduction to the benefits

Applications of plant tissue

cultures

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Embryo culture

Ovary and ovule culture

Control of fertilization

Endosperm culture

Applications in agriculture

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Anther culture

Somatic hybridization & genetic modification

Propagation through though tissue culture

Production of disease free plants

Synthetic seed production

Tissue culture as a novel tool

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Protoplast from different species can be fused to generate a hybrid and this process is known as somatic hybridization

New possibilities for the in vitro genetic manipulation of plant to improve the crops

Applications 1. Disease resistance2. Environmental tolerance3. Quality characters4. Cytoplasmic male sterility5. Study Cytoplasmic gene and their function

Somatic hybridization

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The genetic variations found in the in vitro cultured cell are collectively referred as Somaclonal variation.

Calliclonesprotoclones Applications 1. Production of agronomically important useful

plants.2. Resistance to diseases3. Resistance to abiotic stresses4. Resistance to herbicides5. Improve seed quality i.e. lathyrus seed.

Somaclonal variation

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CROP CHARACTERS

Rice Flowering period; plant height; leaf length; shape and color; frequency of fertile seed

Wheat Plant height; seed storage protein; grain color

Maize Reduced pollen fertility and male sterility

Sugarcane High sugar yield; increase stalk length; diameter ;weight and density

Tomato Early flowering; orange fruit color

Carrot Higher carotene content

potato Higher yield; growth habit; maturity and morphology

Somaclonal variants

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High rate of plant propagation

Production of diseased free plant

Cost effective process

Automated micropropagation

Micropropagation

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Prevention of embryo abortion

Overcoming seed dormancy

Shortening of breeding cycle

Production of haploids

Overcoming of seed sterility

Clonal propagation

Embryo culture

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Possessing only a single set of chromosomes in the sporophyte.

Applications:1. Development of homozygous lines2. Generation of exclusive male plant3. Induction of mutation4. Production of disease resistant plants5. Production of insect resistant plant6. Production of salt tolerance plant7. Cytogenetic research8. Induction of genetic variability9. Evolutionary studies

Haploids

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Increase in human population , pressure on the forest and land resources have increased which result in decline in the population of medicinal and economically important plant species.

Threatened speciesConserve the Germplasm by preserving the

genetic material,1 . In situ preservation

2 . Ex situ preservation

Germplasm preservation

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Most of the natural products are compound derived from primary metabolites such as amino acid, carbohydrate and fatty acids.

Secondary metabolites are considered as the end product of primary metabolites.

Plant tissue culture gives the excellent tool for study the various metabolic activity of the cell.

From this study it is easy to produce medically important compounds (as secondary metabolite).

E.g. Morphine, Codeine etc..

Application in medicine

Ajmalicine Digoxin papain reserpine

Atropine Ephdrine Papaverine Scorpolanine

Caffeine Hyoscyamine Psedoephedrine

Sennosides

Codein L-Dopa Quinidine Vincaleukoblasti-ne

Colchicine Morphine Quinine

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Plant derived medicines used in western medicine.

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PhytochemicalsPrimary metabolitesSecondary metabolitesApplication of secondary metabolites in…1. Pharmaceuticals 2. Flavors3. Perfumes4. Agrochemicals5. Insecticides6. Raw material for industries

Production of secondary metabolites

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Production under control conditions as per demand

Culture system are independent of1. environmental factors2. seasonal variation3. Pest and microbial diseases4. Geographical constrainsCell growth can be controlled to

facilitate improved product formationQuality of the product will be consistent

as its product from specific cell line

Advantages by plant tissue culture

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Recovery of product will be easyEasy to provide desired environment than in

the fieldMutant cell line can be developedBiotransformation reactions can be carried

out with certain culture cellsThe production time is less and labour cost

are minimal

Conti…

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The conversion of one chemical into another by using biological system (cell suspension) .

Conversion of less important compound to valuable medicinal or commercially important product.

Bioconversion may involve many type of reaction e.g. Reduction etc…

It is important to select such cell line that possess the enzyme for catalysing the desired reaction

Biotransformation

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Substrate Product

Digitoxin Digoxin

Codeinone codeine

carvoxin cavaxone

vanilin Vanillin-D-glucuside

steviol Steviocide;steviobiocide

Examples…

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Expression and regulation of gene

Site directed mutagenesis

rDNA technology

Function of gene product

In genetics…

Conclusion

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U.Satyanarayana,2007.callus culture, cell culture, secondary metabolites, somatic hybridization, production of haploid plant, Somaclonal variation, micropropagation, synthetic seeds, embryo culture, Germplasm conservation, 497-516,523-537, 538-545, 546-551, 552-564, 565-566, BIOTECHNOLOGY, Book and allied (p) Ltd.

K.G.Ramawat, G.Shaily,2010.application of plant tissue culture in agriculture, medicine, horticulture, forestry, Germplasm preservation and industry.325-340,molecular biology and biotechnology, S.chand

Bibliography

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Kalyan Kumar de,1997,cytodifferentiation, totipotency, 55-58, 59-66, an introduction to plant tissue culture, new central book agency(p) Ltd.

Important URL For videos:http://www.youtube.com/watch?v=4EWju4JNcx

c&p=7AEB2041844CA224&playnext=1&index=6

http://www.youtube.com/watch?v=oYkpu41LVgk

http://sciencehack.com/videos/view/jKMwLrbYyJ0

http://www.youtube.com/watch?v=D7Glb3l9mbY

http://www.youtube.com/watch?v=Jew0j1RGgvM&feature=related 

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THANK YOU

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