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rajesh bhammar, bihari group, plant tissue culture, genetics, agriculture, tissue, culture, hybridization, application of tissue culture, application, nvpas, ty genetics , vallabh vidhyanagar, anand
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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
Conti..
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THANK YOU
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