Embryo culture is a laboratory method for producing plant lets from a fertilized or unfertilized embryo in invitro condition. there are several advantages are associated with the embryo culture like production of haploid plants, making distant crosses successful, sometimes aborted embryos can be rescued from a unsuccessful hybridization.
Organogenesis, in plant tissue cultureKAUSHAL SAHU
Introduction
Definition
Types of organogenesis
Organogenesis through callus formation (indirect organogenesis)
Growth regulators for indirect organogenesis
Organogenesis through adventitious organ (direct organogenesis)
Growth regulators for direct organogenesis
Factor affecting the soot bud differentiation
Organogenic differentiation
Application of organogenesis
Conclusion
References
The presentation gives overview of production of secondary metabolites using callus culture as well as tissue culture techniques. Various batch and continuous culturing process are described on the basis of secondary metabolite to be synthesised.
Embryo culture is a laboratory method for producing plant lets from a fertilized or unfertilized embryo in invitro condition. there are several advantages are associated with the embryo culture like production of haploid plants, making distant crosses successful, sometimes aborted embryos can be rescued from a unsuccessful hybridization.
Organogenesis, in plant tissue cultureKAUSHAL SAHU
Introduction
Definition
Types of organogenesis
Organogenesis through callus formation (indirect organogenesis)
Growth regulators for indirect organogenesis
Organogenesis through adventitious organ (direct organogenesis)
Growth regulators for direct organogenesis
Factor affecting the soot bud differentiation
Organogenic differentiation
Application of organogenesis
Conclusion
References
The presentation gives overview of production of secondary metabolites using callus culture as well as tissue culture techniques. Various batch and continuous culturing process are described on the basis of secondary metabolite to be synthesised.
The isolation, culture and fusion of protoplasts is a fascinating field in plant research. Protoplast isolation and their cultures provide millions of single cells (comparable to microbial cells) for a variety of studies.
A process where an embryo is derived from a single somatic cell or group of somatic cells. Somatic embryos (SEs) are formed from plant cells that are not normally involved in embryo formation.
Embryos formed by somatic embryogenesis are called Embryoids.
The process was discovered for the first time in Daucas carota L. (carrot) by Steward (1958), Reinert (1959).
Somatic embryogenesis, in plant tissue culture 2KAUSHAL SAHU
Introduction
Types of somatic embryogenesis
Developmental stages
Factors affecting somatic embryogenesis
Importance
Conclusions
References
The process of regeneration of embryos from somatic cells, tissue or organs is regarded as somatic or asexual embryogenesis.
opposite of zygotic or sexual embryogenesis.
Embryo-like structures which can develop into whole plants in a way that is similar to zygotic embryos are formed from somatic cells.
Gametoclonal variation in Plant tissue culture - Variation in gametes clones # Origin # Production # Application of Gametoclonal Variation in plants with their examples.
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The isolation, culture and fusion of protoplasts is a fascinating field in plant research. Protoplast isolation and their cultures provide millions of single cells (comparable to microbial cells) for a variety of studies.
A process where an embryo is derived from a single somatic cell or group of somatic cells. Somatic embryos (SEs) are formed from plant cells that are not normally involved in embryo formation.
Embryos formed by somatic embryogenesis are called Embryoids.
The process was discovered for the first time in Daucas carota L. (carrot) by Steward (1958), Reinert (1959).
Somatic embryogenesis, in plant tissue culture 2KAUSHAL SAHU
Introduction
Types of somatic embryogenesis
Developmental stages
Factors affecting somatic embryogenesis
Importance
Conclusions
References
The process of regeneration of embryos from somatic cells, tissue or organs is regarded as somatic or asexual embryogenesis.
opposite of zygotic or sexual embryogenesis.
Embryo-like structures which can develop into whole plants in a way that is similar to zygotic embryos are formed from somatic cells.
Gametoclonal variation in Plant tissue culture - Variation in gametes clones # Origin # Production # Application of Gametoclonal Variation in plants with their examples.
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Protoplast is a naked cell (without cell wall) surrounded by a plasma membrane. It can regenerate cell wall, grow and divide.
Spheroplast cells have their cell wall only partially removed.
Is fragile but can be cultured and grow into a whole plant.
Cells can originate from any type of tissue (Mesophyll tissue - most suitable source ).
Can be applied in somatic hybridization.
Can be applied in biotechnology and microbiology.
Somatic hybridization is the development of hybrid plants through the fusion of somatic protoplasts of two different plant species/ varieties.
Somatic Hybridization was firstly introduced by Carlson in Nicotiana
glauca.
In 1960, E.C Cocking contributed to the enzymatic isolation and culture of protoplast.
Protoplasts are naked plant cells without the cell wall, but they possess plasma membrane and all other cellular components. They represent the functional plant cells but for the lack of the barrier, cell wall. Protoplasts of different species can be fused to generate a hybrid and this process is referred to as somatic hybridization (or protoplast fusion). Cybridization is the phenomenon of fusion of a normal protoplast with an enucleated (without nucleus) protoplast that results in the formation of a cybrid or cytoplast (cytoplasmic hybrids).
The main purpose of these slides is to convey information to the Professors, Lecturers, and Students. These slides contain authentic information about this topic which is mentioned in that.
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2. Development of hybrid plants
through the fusion of somatic
protoplasts of two different
plant species/varieties is called
somatic hybridization.
3. Somatic hybridization technique
1. isolation of protoplast
2. Fusion of the protoplasts of desired species/varieties
3. Identification and Selection of somatic hybrid cells
4. Culture of the hybrid cells
5. Regeneration of hybrid plants
6. Mechanical Method
Used for vacuolated cells like onion bulb scale, radish and beet
root tissues .
Low yield of protoplast.
Laborious and tedious process.
Low protoplast viability.
7. Enzymatic Method
Leaf sterlization, removal of epidermis
Plasmolysed
cells
Plasmolysed
cells
Pectinase +cellulase
Pectinase
Protoplasm released Release of
isolated cells
cellulase
Protoplasm
released
Isolated
Protoplasm
8. Enzymatic Method
Used for variety of tissues and organs including
leaves, petioles, fruits, roots, coleoptiles,
hypocotyls, stem, shoot apices, embryo
microspores.
Mesophyll tissue - most suitable source .
High yield of protoplast.
Easy to perform.
More protoplast viability .
9. SOMATIC HYBRIDIZATION
The technique of hybrid production through the
fusion of isolated somatic protoplast under in vitro
conditions and subsequent development of their
product (heterokaryon) to a hybrid plant is known as
somatic hybridizaton.
In somatic hybridization the nucleus and cytoplasm of
both parents are fused in the hybrid cell.
10. Sometimes; nuclear genome of only one parent but
cytoplasmic genes (plastome) from both the parents
are present in the fused hybrid, which known as cybrid
or cytoplasmic hybrid.
Somatic hybridization involves the following aspects:
1) Fusion of protoplasts .
2) Selection of hybrid cells .
3) Identification of hybrid plants.
11. Protoplast fusion
It involves mixing of two different genomes and can be achieved
by spontaneous or induced fusion method.
Spontaneous fusion : -
I. cell fusion is natural process as is observed in case of egg
fertilization.
II. During the course of enzymatic degradation of cell walls,
adjoining protoplasts may fuse to form homokaryocytes
(homocaryons).
III. These fused cells may sometimes contain high number of
nuclei (2-40).
IV. The frequency of homocaryon formation was found to be high
in protoplasts isolated from dividing cultured cells.
12. Induced fusion method :-
I. Isolated protoplasts can be fused by induction.
II. There are several fusion inducing agents which are
collectively referred to as fusogen.
III. example: sodium nitrate ,high gylcol, polyvinyl
alcohol, lysozyme, concavalina, electrofusion dextran
and dextran sulphate , fatty acid and esters.
13. TREATMENT WITH SODIUM NITRATE
Induced fusion by NaNO3 was first reported by power
et al .(1970).
Isolated protoplasts are exposed to a mixture of 5.5%
carried NaNO3 in 10% sucrose solution.
Incubation is carried out for 5 minutes at 35 ⁰C .
Centrifugation for 5 minutes at 200x g
14. High pH and high sodium Calcium ion treatment :
The different protoplast in one solution are together
treated with condition like high calcium and high ph
so that they fuse together . In some cases such extreme
conditions has proved to be toxic to certain
protoplasts.
15.
16. Poly ethylene glycol treatment
This has proved to be one of the most effective
methods for protoplast fusion .
The cells are treated with a concentration of around
30% poly ethylene glycol which binds to plasma
membrane .
This is treated with calcium solution which being
cationic binds to PEG.
During washing the PEG pulls out the plasma lemma
leading to fusion of protoplasts in close proximately.
17. Electro fusion technique
This process involves passing low voltage electric
pulses in a solution of protoplast to be fused so that
they line of for fusion .
High voltage electric current which leads alteration of
membrane the adjacent protoplast fuse.
18. SELECTION OF HYBRID CELL
The protoplasts are actually involved in the fusion.
After the fusion process, the protoplast population
consists of heterogenous mixture of unfused
chloroplast, homokaryons and heterokaryons.
It is therefore necessary to select the hybrid cells
(heterokaryons).
The commonly used methods employed for the
selection of hybrid cells are biochemical, visual and
cytometric method.
19. Hybrid cells are cultured on suitable medium
provided with the appropriate culture conditions.
Culture of the hybrid cells
20. Regeneration of hybrid plants
Plants are induced to regenerate from hybrid calli .
These hybrid plants must be at least partially fertile, in
addition to having some useful property, to be of any
use in breeding schemes.
21. Cybrids
The cytoplasmic hybrids where the nucleus is derived
from only one parent and the cytoplasm is derived
from both the parents are referred to as cybrids .
The phenomenon of formation of cybrids regarded as
cybridization .
Normally cybrids are produced when protoplast from
two pythogenetically distinct species are fused.
Genetically are hybrids only for cytoplasmic traits.
22.
23. Advantages of somatic
hybridization
Production of novel interspecific and intergenic
hybrid.
Pomato (Hybrid of potato and tomato).
Production of fertile diploids and polypoids from
sexually sterile haploids, triploids and aneuploids.
Transfer gene for disease resistance, abiotic stress
resistance, herbicide resistance and many other
quality characters.
24. Limitations of Somatic
hybridization
Poor regeneration of hybrid plants.
Non-viability of fused products.
Not successful in all plants.
Production of unfavorable hybrids.
Lack of an efficient method for selection of hybrids.
No confirmation of expression of particular trait in
somatic hybrids.