This document provides an overview of plant tissue culture and its applications. It discusses the history and development of plant tissue culture techniques. The key techniques covered are micropropagation, somatic cell genetics, transgenic plants, and the major steps of tissue culture including initiation, multiplication, and root formation phases. Different types of tissue culture are also described such as cell culture, callus culture, and protoplast culture. The applications of plant tissue culture discussed include clonal propagation, secondary metabolite production, genetic variability, somatic embryogenesis, haploid plant production, somatic hybridization, transgenic plants, and germplasm conservation. The document concludes by stating that plant tissue culture can be useful for human needs but must be performed cautiously.
Callus cultures are initiated from a small part of an organ or tissue segment called the explants on a growth supporting solidified nutrient medium under sterile conditions. Any part of the plant organ or tissues may be used as the explants.Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture medium of known composition. Plant tissue culture is widely used to produce clones of a plant in a method known as micropropagation.Tissue culture commonly refers to the culture of animal cells and tissues, with the more specific term plant tissue culture being used for plants.Plant Tissue Culture products include Murashige and Skoog media, plant growth regulators, plant growth hormones, plant transformation systems,
INTRODUCTION
2. HISTORY
3. BASIC COMPONENT OF MEDIA
1. Inorganic nutrient
2. organic supplements
3. Carbon and energy source
4. Growth Regulators
5. Solidifying Agent
6. PH
4. TYPES OF MEDIA
5. MS MEDIA
6. IMPORTANCE
7. CONCLUSION
8. REFERANCE
this slide is all about the different cultures in plant tissue cultures such as seed culture, root culture, cell suspension culture, anther culture etc
Callus cultures are initiated from a small part of an organ or tissue segment called the explants on a growth supporting solidified nutrient medium under sterile conditions. Any part of the plant organ or tissues may be used as the explants.Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture medium of known composition. Plant tissue culture is widely used to produce clones of a plant in a method known as micropropagation.Tissue culture commonly refers to the culture of animal cells and tissues, with the more specific term plant tissue culture being used for plants.Plant Tissue Culture products include Murashige and Skoog media, plant growth regulators, plant growth hormones, plant transformation systems,
INTRODUCTION
2. HISTORY
3. BASIC COMPONENT OF MEDIA
1. Inorganic nutrient
2. organic supplements
3. Carbon and energy source
4. Growth Regulators
5. Solidifying Agent
6. PH
4. TYPES OF MEDIA
5. MS MEDIA
6. IMPORTANCE
7. CONCLUSION
8. REFERANCE
this slide is all about the different cultures in plant tissue cultures such as seed culture, root culture, cell suspension culture, anther culture etc
A presentation covering the process of protoplast culture including protoplast isolation, protoplast fusion, culture of protoplast, its application, factors affecting protoplast culture and the future of protoplasts.
Gametoclonal variation in Plant tissue culture - Variation in gametes clones # Origin # Production # Application of Gametoclonal Variation in plants with their examples.
Please watch the slides and don't forget to follow our channel to getting more updates.
Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture medium of known composition. It is widely used to produce clones of a plant in a method known as micropropagation
Micropropagation is an advanced vegetative propagation technology for producing a large number of transplants in a limited time and space.
STAGES
Stage 0 — Mother Plant Selection:
Stage I — Establishment of Aseptic Culture:
Stage II — Multiplication of shoots:
Stage III — In Vitro Rooting:
Stage IV — Transplantation or Hardening:
APPLICAIONS
A presentation covering the process of protoplast culture including protoplast isolation, protoplast fusion, culture of protoplast, its application, factors affecting protoplast culture and the future of protoplasts.
Gametoclonal variation in Plant tissue culture - Variation in gametes clones # Origin # Production # Application of Gametoclonal Variation in plants with their examples.
Please watch the slides and don't forget to follow our channel to getting more updates.
Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture medium of known composition. It is widely used to produce clones of a plant in a method known as micropropagation
Micropropagation is an advanced vegetative propagation technology for producing a large number of transplants in a limited time and space.
STAGES
Stage 0 — Mother Plant Selection:
Stage I — Establishment of Aseptic Culture:
Stage II — Multiplication of shoots:
Stage III — In Vitro Rooting:
Stage IV — Transplantation or Hardening:
APPLICAIONS
Much faster rates of growth can be induced in vitro than by traditional means.
Multiplication of plants which are very difficult to propagate by cuttings or other traditional methods.
Production of large numbers of genetically identical clones in a short time
Seeds can be germinated with no risk of damping off/ predation.
Under certain conditions, plant material can be stored in vitro for considerable periods of time with little or no maintenance
Tissue culture techniques are used for virus eradication, genetic manipulation, somatic hybridization and other procedures that benefit propagation, crop improvement, and basic research.
By means of tissue culture it is possible to produce pathogen free plantlets by mass multiplication in a very limited amount of area from a very small sterile part of a mother plant. This method is also used to produce/ multiply plants that are to be transported across national border and so for their faster multiplication.But the establishment of a tissue culturing unit needs huge financial investments, skilled labors/technicians and required areas for its establishment are major constraints. Plant tissues grow and multiply in the labs only when there is an uncompetitive, growing condition with uninterrupted supply of nutrients.
Medium:
It contains all the elements that contribute the required nutrients that aid to the growth of the tissues; it is in liquid state or semi-solid in nature. The tissues are grown on the media. It consists of 95% of water, major and minor nutrients, plant growth hormones, vitamins, sugar rich compounds and chelating agents.
Totipotency:
It is the ability of a tissue or an organ of a plant to produce the whole plant, under the optional laboratory conditions and this is called as Totipotency. This is the baseline over which plant tissue culture relies upon.
Callus Culture:
When the cells divide into an undifferentiated mass it is called as callus. Any part of a plant can be used to produce the calli. It may be a stem, leaf, meristem or any other part. It is used to produce variations among the plantlets.
Suspension culture:
The callus produced from the explants are grown on nutrient solutions (that are semi solid) for a period of time and they are induced to produce plants with new traits.
Embryo Culture:
The method of culturing mature and immature embryos in media is called embryo culture. By this method, it is possible to produce plants from dormant seeds and seeds with metabolites that inhibit germination. This method is very important in crop improvement programs.
Somatic Embryogenesis:
When the plants are grown on nutrient media, calli are formed. When these calli are subjected to growth in cytokinin medium, somatic embryos are formed. They are circular, elongated,
Single cell culture
• As stated earlier, cells derived from a single cell through mitosis constitute a clone and the process of obtaining clones is called cloning (asexual progeny of a single individual make up.
MEDICINAL PLANT BIOTECHNOLOGY UNIT 2, MPG, SEM 2. NOTES Different tissue culture techniques: Organogenesis and embryogenesis, synthetic seed and monoclonal variation
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plant tissue culture and its applications. Agricultural biotech.
1. PLANT TISSUE
CULTURE AND ITS
APPLICATIONS
Hafiz Muzammil Nisar
MPhil Biotechnology
Bahaudin Zakariya University, Multan.
2. Contents:
• History
• Introduction
• Tissue culture
• Nutrient medium
• Fundamental Principles of
plant tissue culture
• Plant tissue culture Techniques
• Micropropagation
• Somatic cell Genesis
• Transgenic Plants
• Major steps of Tissue Culture
• Initiation phase
• Multiplication phase
• Root formation phase
• Types of Tissue culture
• Cell culture
• Callus culture
• Protoplast culture
• Organ culture
• Seed culture
• Embryo culture
• Meristem culture
• Anther culture
• Applications of Tissue
Culture
• Conclusion
3. History
• Haberlandt, Father of plant tissue culture.
cultured single cells on Knop’s salt solution.
• Hanning(1904) Embryo culture of selected
crucifers.
• Robbins(1922), in vitro culture of root tips.
• Muir(1953) isolation and culture of single plant cells.
• Skoog and Miller(1957) hormonal control in tissue culture.
• Reinert and Steward(1958) report somatic embryogenesis.
4. History
• E C Cocking (1960) Isolation of protoplasts by
enzymatic degradation method.
• Murashige and Skoog (1962) Development of MS
media.
• Guha and Maheshwari (1964) Production of First
haploid plant by anther culture
5. Plant Tissue Culture
• The growth or regeneration of plant cells,
tissues, organs or whole plants in artificial
medium under aseptic conditions.
• In this technique use plant parts or cells
called Explant.
INTRODUCTION:
• The most widely used artificial nutrient
medium is MS medium.
6. Fundamental Principles of Plant Tissue Culture
Plant tissue culture depends upon;
• Totipotency, ability of plant cells to regenerate
into whole new plant
• Plasticity, ability of plants to alter their
metabolism, growth and development to best
suit their environment
Plant cell
Tissue Callus
Plant organ Embryo
New plant
7. Plant Tissue Culture Techniques:
1. Micropropagation:
• Developing high- quality clonal plants.
• Provide rapid and large scale propagation of new
genotypes.
• Steps:
• Sterilization and introduction
• Shoot production
• Root production
• Transfer to soil
8. 2. Somatic cell genetics:
• It is used mostly in terms of haploid production
and somatic hybridization.
• Isolation of protoplast
• Fusion by using chemicals
• Plating of fused protoplast
• Selection
• Steps:
9. 3. Transgenic plants:
• They are identified as a class of
genetically modified organism
(GMO).
• Transgenic plants are plants that have been
genetically engineered, uses recombinant DNA
techniques to create plants with new
characteristics.
• The complete process is
described as:
10. Major Steps of Tissue Culture:
• Initiation Phase (Stage 1)
• Multiplication Phase (Stage 2)
• The in vitro plant material is introduced in to
the medium.
• Proliferation of the tissue and the production
of multiple shoots.
• The tissue of interest is obtained, sterilized
and introduced.
11. Major Steps of Tissue Culture:
• Roots are formed.
• Here, hormones are required in order to
induce rooting such as “Auxin”.
• Consequently complete plantlets.
• Root formation (Stage 3)
13. Cell culture:
• The culture of isolated individual cells.
• Carried out in dispersion medium.
Callus culture:
• The growth of callus from explant by
providing appropriate conditions.
• Darkness and solid medium gelled by agar
stimulate callus formation.
14. Protoplast culture:
• The protoplast culture is aimed to develop
genetically transformed plant.
• The cell in which the cell wall has been removed.
• In this, isolated protoplasts are cultured on a
suitable medium under the aseptic condition.
15. Organ culture:
• Isolated plant organs (Shoot, root, leaf, and flower) are
used as explant.
• The organ culture may be organized or unorganized.
• New growth (differentiated structures) continues
given that the organ retains its:
• Physiological features,
• Provide information on patterns of growth,
• As well as development.
16. Seed culture:
• For this method, explants are obtained from an in-vitro
derived plant.
• Introduced into an artificial environment.
• Where they get to proliferate.
• Seed culture is primarily used for plants
such as orchids
17. Embryo Culture:
• For embryo culture, the ovule, seed or fruit
from which the embryo is to be obtained is
sterilized.
• Culturing them in nutrient media, help in
developing viable seedlings.
• Embryo culture is the isolation and growth of
mature or immature embryo in-vitro.
18. Meristem culture:
• A meristem is a localized group of cells, which
are actively dividing and give rise to
permanent tissue.
• The apical meristem of shoots is
cultured.
• To get the disease-free plants.
19. Anther Culture:
• Anthers of some plants are cultured on a
suitable medium to produce haploid plants,
it is called anther culture.
• Separated anthers from flowers.
• Cultured on a suitable medium.
• This technique was first used in India
to produce haploids of Datura.
21. Clonal Propagation:
• Useful tool to get a large population of plant
species having desirable traits.
• This technique is very much used in horticulture
and silviculture—orchids and many fruit plants.
• Large scale production of plants of same
genetic stock.
22. Secondary Metabolites:
• Secondary metabolites produced by plant cell
cultures are rather small in amount.
• By clonal selection the particular high yielding
clone of cells can be isolated.
• Production of many useful compounds can
be done by plant cell culture.
23. Genetic Variability:
• The chromosomal instability in the cultured
cells play an important role in polyploidization
of cells and genetically variable plants can be
raised.
• Such kind of variations may show some useful
characters such as:
• Different kinds of morphological variations in
leaf and flowering.
24. Somatic Embryogenesis and Synthetic Seed:
• Direct or indirect somatic embryogenesis may be
achieved from:
• Pro-embryonic cell of the direct explant
• The embryoids developed within the callus
tissue.
• The application of this technique is:
• the mass production of adventitious
embryos which ultimately develop into
complete plantlet.
25. Haploid Plants:
• Haploid plants can be obtained through anther or
pollen culture (androgenesis).
• These haploids produced homozygous diploid or
polyploid lines.
• Colchicine treatment within a very short period.
26. Somatic Hybrids:
• Somatic hybridization, using isolated somatic
protoplasts is a new tool to make the wide
hybridization successful.
• Obtain somatic hybrid plants between sexually
compatible and incompatible plants.
• Production of cybrid is also of immense
importance in the plant breeding program.
27. Transgenic Plants:
• The genetically modified (GM) plants, in which a
functional foreign gene has been incorporated by
biotechnological method.
• A number of transgenic plants have been
produced carrying genes for different traits.
• The direct DNA uptake through protoplast is
the most ideal method.
28. Germplasm Conservation:
• Many of the important crop species produce recalcitrant
seeds.
• Plant tissue culture may be applied for this purpose.
• Mainly the plant species which are endangered,
needed to be conserved by ex-situ method of
germplasm conservation.
29. Conclusion:
• Useful to human kind and ecosystem.
• Hope to end world hunger.
• Performed with cautions to minimize the risks.
• Researcher must be ethical.
• Use for cloning purposes and increase yield.
This is a method of biological research in which fragments of tissue from an animal or plant are transferred to an artificial environment in which they can continue to survive and function. inorganic salts (both micro and macro elements),
a carbon source (usually sucrose),
vitamins, amino acids,
growth regulators (e.g. auxins and cytokinin and gibberellins).
Fig 1: banana plantlets transfer to soil produced by micropropagation.
PEG, sodium nitrate, calcium ions.
in order to prevent any microorganism from negatively affecting the process. It is during this stage that the tissue is initiated in to culture.
Here, the medium is composed of appropriate components for growth including regulators and nutrients.
Callus is the unorganized and undifferentiated dividing mass of cells.
Darkness is required as light can encourage differentiation of the callus.
Somatic hybridization of two plant species through protoplast fusion in done by protoplast culture.
In the event that a plant material is used directly for this process, then it has to be sterilized to prevent tissue damage and ensure optimum regeneration.
In some plants, seed dormancy may be due to chemical inhibitors or mechanical resistance etc. Embryo culture may involve the use of a mature of immature embryo. Whereas mature embryos for culture are essentially obtained from ripe seeds, immature embryo (embryo rescue) involves the use of immature embryos from unripe/hybrid seeds that failed to germinate. In doing so, the embryo is ultimately able to produce a viable plant.
For embryo culture, the ovule, seed or fruit from which the embryo is to be obtained is sterilized, and therefore the embryo does not have to be sterilized again.
Silviculture is the art and science of controlling the establishment, growth, composition, and quality of forest vegetation for the full range of forest resource objectives. in the plants which have long seed dormancy, tree species,
Production of many useful compounds like alkaloids, Steroids (Diosgenin), Glycosidic compounds and many other essential oils (Jasmine), flavoring and coloring agents (saffron) can be done by plant cell culture. This aim can be achieved by selection of specific cells producing high amount of desired compounds and development of a suitable medium.
Flowering time, plant height, lodging resistance, yield, nutrient content and
Synthetic seed production, germination and field transfer of synthetic seeds of S. chirayita. (a)-encapsulated torpedo shaped somatic embryos in 100 mM CaCl 2 solution (bar = 0.5 cm); (b)-synthetic seeds produced using torpedo shaped somatic embryos (bar = 0.5 cm); (c)-germination of a synthetic seed on MS medium after 30 days of culture (bar = 1.0 cm); (d)-cluster of plantlets derived from a synthetic seed (bar = 1.2 cm); (e)-complete rooted plant from synthetic seed (bar = 1.0 cm); (f)-acclimatized plant after transfer to soil in ex vitro conditions.
Ginseng transformation by Agrobacterium tumefaciens for the transgenic adventitious root induction.
Colchicine is a toxic chemical that is often used to induce polyploidy in plants. Basically, the colchicine prevents the microtubule formation during cell division, thus the chromosomes do not pull apart like they normally do. The end result is a cell that now has double the number of chromosomes that it would normally have.
This technique has been mainly used for introgression many useful criteria from the wild genotype to cultivated crop variety. Cybrid-the fusion between two protoplasts—one partner with nucleus and another partner with cytoplasm,
like insect resistance, herbicide tolerance, delayed ripening, increased amino acid and vitamin content, improved oil quality, etc.
The direct DNA uptake through protoplast is the most ideal method for production of transgenic plants. Any gene of interest that may be of eukaryotic or prokaryotic origin can be used for this purpose but should be expressed.
Recalcitrant seeds (subsequently known as unorthodox seeds) are seeds that do not survive drying and freezing during ex-situ conservation and vice versa