The document discusses explants used for clonal propagation in biotechnology of horticultural crops. It defines an explant as a tissue taken from a mother plant and cultured under aseptic conditions on a defined medium. The choice of explant depends on the type of culture to be initiated, its purpose, and the plant species. Common explant sources include shoots, leaves, stems, and roots. Explants must be surface sterilized to remove contaminants and allow contamination-free culture initiation. Sub-culturing is needed when cells reach the stationary state of growth.
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).
Anther culture:- the in vitro culturing of anthers containing microspores or immature pollen grains on a nutrient medium for the purpose of generating haploid plantlets.
Culturing anthers for the purpose of obtaining Double Haploid is not easy with many field crop species, particularly with the cereals, cotton, and grain legumes.
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.
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).
Anther culture:- the in vitro culturing of anthers containing microspores or immature pollen grains on a nutrient medium for the purpose of generating haploid plantlets.
Culturing anthers for the purpose of obtaining Double Haploid is not easy with many field crop species, particularly with the cereals, cotton, and grain legumes.
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.
Meristem tip culture for the production of the virus free plantsArjun Rayamajhi
This presentation gives general idea on the meristem tip culture for the production of the virus free plants. The principles, methods and procedures of the meristem tip culture included. General idea on different in vitro culture techniques for virus elimination meristem tip culture viz. thermotherapy, cryotherapy,chemotherapy and electrotherapy are provided.
history
Lampe & Mills (1933) were the first to report the proliferation of immature endosperm tissue of Maize, grown on medium containing extract of potato.
La Rue (1947) observed that in nature, in maize , the pericarp ruptured & the endosperm exhibited a white tissue mass.
HYBRIDIZATION & HAPLOID PRODUCTION
Introduction
WIDE HYBRIDIZATION
INTER-SPECIFIC HYBRIDIZATION
Barriers to distant hybridization
Techniques to overcome barriers
Haploids and Doubled Haploids in Plant
Production of haploids and doubled haploids
a) Induction of maternal haploids
Wide hybridization
3. In vitro induction of maternal haploids – gynogenesis
Induction of paternal haploids – Androgenesis
Production of Homozygous Diploid Plants
Application of Haploids in Plant Breeding
Importance and Implications of Anther and Pollen Culture
Invitro culture of unpollinated ovaries and ovules represents an alternative for the production of haploid plant
First successful report on the induction of gynogenic haploid was in barley by San Noeum in 1976
Haploid plants are obtained from ovary and ovule culture of rice, wheat, maize, sunflower, tobacco, poplar, mulberry etc
Whites or MS or N6 inorganic salt medium supplement with growth substances are used
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.
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.
Meristem tip culture for the production of the virus free plantsArjun Rayamajhi
This presentation gives general idea on the meristem tip culture for the production of the virus free plants. The principles, methods and procedures of the meristem tip culture included. General idea on different in vitro culture techniques for virus elimination meristem tip culture viz. thermotherapy, cryotherapy,chemotherapy and electrotherapy are provided.
history
Lampe & Mills (1933) were the first to report the proliferation of immature endosperm tissue of Maize, grown on medium containing extract of potato.
La Rue (1947) observed that in nature, in maize , the pericarp ruptured & the endosperm exhibited a white tissue mass.
HYBRIDIZATION & HAPLOID PRODUCTION
Introduction
WIDE HYBRIDIZATION
INTER-SPECIFIC HYBRIDIZATION
Barriers to distant hybridization
Techniques to overcome barriers
Haploids and Doubled Haploids in Plant
Production of haploids and doubled haploids
a) Induction of maternal haploids
Wide hybridization
3. In vitro induction of maternal haploids – gynogenesis
Induction of paternal haploids – Androgenesis
Production of Homozygous Diploid Plants
Application of Haploids in Plant Breeding
Importance and Implications of Anther and Pollen Culture
Invitro culture of unpollinated ovaries and ovules represents an alternative for the production of haploid plant
First successful report on the induction of gynogenic haploid was in barley by San Noeum in 1976
Haploid plants are obtained from ovary and ovule culture of rice, wheat, maize, sunflower, tobacco, poplar, mulberry etc
Whites or MS or N6 inorganic salt medium supplement with growth substances are used
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.
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.
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.
Presentation on methods of plant breeding, classification of breeding methods, methods of breeding for self-pollinated,cross-pollinated and asexually propagated species, a brief account of breeding methods
PPT on Tissue Culture Class 10 CBSE Text Book NCERT.One Time Forever
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MEDICINAL PLANT BIOTECHNOLOGY UNIT 2, MPG, SEM 2. NOTES Different tissue culture techniques: Organogenesis and embryogenesis, synthetic seed and monoclonal variation
Protoplast fusion, Hairy root multiple shoot cultures and their applications.
Micro propagation of medicinal and aromatic plants.
Sterilization methods involved in tissue culture, gene transfer in plants and their applications.
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As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
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1. To execute inoculation of explants for clonal propagation
BIOTECHNOLOGY OF HORTICULTURAL CROPS
HRT 552
2. A) Explant
Explant is a tissue taken from mother plant,
cultured under aseptic condition on a defined medium
by manipulation of medium component and other
physiological parameters. The nature of the explants to
be used for in vitro propagation is, to a certain extent,
governed by the method of shoot multiplication to be
adopted.
3. Explant
• The tissue obtained from a plant to be cultured is called an explant.
• Explants are small pieces of plant parts or tissues.
• Are aseptically cut and used to initiate a culture in a nutrient
medium.
• Explants can be taken from different parts of a plant such as:
• shoots, leaves, stems, flowers, roots, and from many types of mature
cells provided they are able to de-differentiate into totipotent cells.
4. Selection of Explants:
• The correct choice of explant material can have an important effect
on the success of tissue culture. The choice of explants depends on:
1) the kind of culture to be initiated
2) the purpose of the proposed culture
3) the plant species to be used
6. Selection of Explants:
• The correct choice of explant material can have an important effect on
the success of tissue culture. The choice of explants depends on:
1) the kind of culture to be initiated
2) the purpose of the proposed culture
3) the plant species to be used
7. S.No. Explants used Needs
1. Explants with pre-
formed vegetative bud
For enhanced axillary branching.
2. Shoot tips To produce virus-free plant from an infected
individual.
3. Nodal cuttings When the stock plant is virus-tested or,
alternatively, virus eradication is not desired.
4. Runners tips For rhizomatic plants, such as strawberry and Boston
fern.
5. Explants are derived from
root, stem, leaf or nucellus
based on their natural
capacity to form
adventitious buds.
For multiplication through adventitious bud formation,
with or without callusing
6. Leaf-base and scale-
base with small basal
plate.
For monocots.
Table: Type of explants used
8.
9. Continue..
• For successful culture initiation, there is a minimum size for explant
per unit culture volume.
• In practice, minimum inoculation size of explant varies according to
the genotype of the plant being cultured and the cultural conditions.
• For commencing suspension cultures it is commonly about 1-1.5 x
104 cells/ml.
10.
11. Contamination Free
• Plants growing in the external environment are invariably
contaminated with micro-organisms and pests.
• These contaminants are mainly confined to the outer surfaces
of the plant, although, some microbes and viruses can even
grow within the root.
12. Continue….
• Most of the surface microbes do not form tight associations
with the plant tissue.
• They are easily removed from the explant by gently rinsing,
and the remaining are killed by surface sterilization.
• Placing the explant in a 70% ethyl alcohol solution prior to
treatment with another disinfectant solution.
• Conducting the sterilization process under vacuum.
• This results in the removal of air bubbles and provides a more
efficient sterilization process.
15. •When the cells in culture are reaching at stationary
state it should be sub-cultured.
•Sub-culturing may be in similar or dissimilar type of
media with respect to the requirements.