Ovule and seed culture techniques and their applications are summarized. Ovules and seeds can be isolated and cultured aseptically on nutrient media. This allows for applications like in vitro pollination and fertilization, hybridization of incompatible species, production of haploid callus, circumventing the need for host stimuli in parasitic plants, and inducing polyembryony. Seed culture is useful for propagating plants with reduced embryos, parasitic plants, orchids, and producing large numbers of uniform seedlings. However, in vitro culture still poses challenges like slow growth, low multiplication, poor rooting, and somaclonal variation.
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
A novel method for triploid plant production, Increases yield of timber and fuel, Rescuing Embryos from Incompatible Crosses, Overcoming Dormancy and Shortening Breeding Cycle
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
A novel method for triploid plant production, Increases yield of timber and fuel, Rescuing Embryos from Incompatible Crosses, Overcoming Dormancy and Shortening Breeding Cycle
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.
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.
Somaclonal Variation in Plant tissue culture - Variation in somaclones (somatic cells of plants)
Somaclonal variation # Basis of somaclonal variation # General feature of Somaclonal variations # Types and causes of somaclonal variation # Isolation procedure of somaclones via without in-vitro method and with in-vitro method with their limitations and advantages # Detection of isolated somaclonal variation # Application (with examples respectively related to crop improvement) # Advantages and disadvantages of somaclonal variations.
https://www.youtube.com/watch?v=IZwrkgADM3I
Also watch, Gametoclonal variation slides to understand, how to changes occur in gametoclones of plants.
https://www.slideshare.net/SharmasClasses/gametoclonal-variation
WHAT IS ARTIFICIAL SEED..?
Artificial seed can be defined as artificial encapsulation of somatic embryos, shoot bud or aggregates of cell of any tissues which has the ability to form a plant in in-vitro or ex-vivo condition.
Artificial seed have also been often referred to as synthetic seed.
HISTORY
Artificial seeds were first introduced in 1970’s as a novel analogue to the plant seeds.
The production of artificial seeds is useful for plants which do not produce viable seeds. It represents a method to propagate these plants.
Artificial seeds are small sized and these provides further advantages in storage, handling and shipping.
The term, “EMBLING” is used for the plants originated from synthetic seed.
• The use of synthetic varieties for commercial cultivation was first suggested in Maize (Hays & Garber, 1919).
OVARY CULTURE:-
"the in-vitro culturing of ovaries in an aseptic condition from the pollinated or un-pollinated flowers, in an appropriate nutrient medium and under optimal conditions." And
OVULE CULTURE:-
"Ovule culture is an experimental system by which ovules are aseptically isolated from the ovary and are grown aseptically on chemically defined nutrient medium under controlled conditions."
Artificial Seed - Definition, Types & Production ANUGYA JAISWAL
Somatic embryogenesis is expected to be the only clonal propagation system economically viable for crops currently propagated by seeds However, it would require mechanical planting of somatic embryogenesis. Although suggestions have been made to use naked embryos for large scale planting, it would be desirable to convert them into 'synthetic seeds' or 'synseeds' by encapsulating in a protective covering.
Kitto and Janick (1982, 1985a,b) selected polyoxyethylene (Polyox r) which is readily soluble in water and dries to form a thin film, does not support growth of microorganism and is non-toxic to the embryos.
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.
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.
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.
Somaclonal Variation in Plant tissue culture - Variation in somaclones (somatic cells of plants)
Somaclonal variation # Basis of somaclonal variation # General feature of Somaclonal variations # Types and causes of somaclonal variation # Isolation procedure of somaclones via without in-vitro method and with in-vitro method with their limitations and advantages # Detection of isolated somaclonal variation # Application (with examples respectively related to crop improvement) # Advantages and disadvantages of somaclonal variations.
https://www.youtube.com/watch?v=IZwrkgADM3I
Also watch, Gametoclonal variation slides to understand, how to changes occur in gametoclones of plants.
https://www.slideshare.net/SharmasClasses/gametoclonal-variation
WHAT IS ARTIFICIAL SEED..?
Artificial seed can be defined as artificial encapsulation of somatic embryos, shoot bud or aggregates of cell of any tissues which has the ability to form a plant in in-vitro or ex-vivo condition.
Artificial seed have also been often referred to as synthetic seed.
HISTORY
Artificial seeds were first introduced in 1970’s as a novel analogue to the plant seeds.
The production of artificial seeds is useful for plants which do not produce viable seeds. It represents a method to propagate these plants.
Artificial seeds are small sized and these provides further advantages in storage, handling and shipping.
The term, “EMBLING” is used for the plants originated from synthetic seed.
• The use of synthetic varieties for commercial cultivation was first suggested in Maize (Hays & Garber, 1919).
OVARY CULTURE:-
"the in-vitro culturing of ovaries in an aseptic condition from the pollinated or un-pollinated flowers, in an appropriate nutrient medium and under optimal conditions." And
OVULE CULTURE:-
"Ovule culture is an experimental system by which ovules are aseptically isolated from the ovary and are grown aseptically on chemically defined nutrient medium under controlled conditions."
Artificial Seed - Definition, Types & Production ANUGYA JAISWAL
Somatic embryogenesis is expected to be the only clonal propagation system economically viable for crops currently propagated by seeds However, it would require mechanical planting of somatic embryogenesis. Although suggestions have been made to use naked embryos for large scale planting, it would be desirable to convert them into 'synthetic seeds' or 'synseeds' by encapsulating in a protective covering.
Kitto and Janick (1982, 1985a,b) selected polyoxyethylene (Polyox r) which is readily soluble in water and dries to form a thin film, does not support growth of microorganism and is non-toxic to the embryos.
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 technique used in plant propagation and plant breeding, particularly for species that may have difficulty germinating or growing under normal conditions. It involves isolating and growing plant embryos, which are the early developmental stages of plants, in a controlled environment, typically on a nutrient medium.
• Seed culture is an important technique when explants are taken from in vitro-derived plants and in propagation of orchids.
• Embryo culture represents the earliest technique to obtain viable offspring following interspecific and intergeneric hybridizations where routine fertilization failed to produce a well-defined and full-term embryo.
• Embryo rescue holds great promise not only for effecting wide crosses, but also for obtaining haploid plants as well as for shortening the breeding cycle.
this slide is all about the different cultures in plant tissue cultures such as seed culture, root culture, cell suspension culture, anther culture etc
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.
INVITRO CULTURE: TECHNIQUES, APPLICATIOSNS & ACHIEVEMENTS.
INVITRO TECHNIQUES AND BIOTECHNOLOGY USE IN AGRICULTURE AND CROP IMPROVEMENT. APPLICATIONS OF VARIOUS BIOTECHNOLOGICAL TECHNIQUES AND METHODS. TISSUE CULTURE, MICROPROPAGATION, EMBRYO CULTURE, ANTHER CULTURE, POLLEN CULTURE, ENDOSPERM CULTURE, OVULE CULTURE, OVARY CULTURE, ETC.
Richard's aventures in two entangled wonderlandsRichard Gill
Since the loophole-free Bell experiments of 2020 and the Nobel prizes in physics of 2022, critics of Bell's work have retreated to the fortress of super-determinism. Now, super-determinism is a derogatory word - it just means "determinism". Palmer, Hance and Hossenfelder argue that quantum mechanics and determinism are not incompatible, using a sophisticated mathematical construction based on a subtle thinning of allowed states and measurements in quantum mechanics, such that what is left appears to make Bell's argument fail, without altering the empirical predictions of quantum mechanics. I think however that it is a smoke screen, and the slogan "lost in math" comes to my mind. I will discuss some other recent disproofs of Bell's theorem using the language of causality based on causal graphs. Causal thinking is also central to law and justice. I will mention surprising connections to my work on serial killer nurse cases, in particular the Dutch case of Lucia de Berk and the current UK case of Lucy Letby.
A brief information about the SCOP protein database used in bioinformatics.
The Structural Classification of Proteins (SCOP) database is a comprehensive and authoritative resource for the structural and evolutionary relationships of proteins. It provides a detailed and curated classification of protein structures, grouping them into families, superfamilies, and folds based on their structural and sequence similarities.
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
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.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
Comparing Evolved Extractive Text Summary Scores of Bidirectional Encoder Rep...University of Maribor
Slides from:
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Track: Artificial Intelligence
https://www.etran.rs/2024/en/home-english/
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
What is greenhouse gasses and how many gasses are there to affect the Earth.
Ovule and seed culture by Prerna Jain
1. Ovule and Seed culture
Technique and
Applications
Prerna jain
11615509
1628
MSC botany (H)
A06
2. Contents
Ovule culture
Principle
Protocol
Historical perspective of ovule culture
Applications and advantages with
examples
In vitro pollination and fertilisation
Ovule culture in hybridization
Production of haploid callus
3. Contents
Ovule culture of parasitic plants
Ovule culture of orchids plants
Induction of poly-embryony by ovule culture
Virus irradiation through ovule culture
Seed culture
Protocol
4. Contents
Applications and advantages of
seed culture
Seed culture of plants with reduced embryos
Seed culture of parasitic plants
Seed culture of orchid plants
Examples of seed culture
In –vitro technologies - a challenge
5. Ovule culture is an elegant experimental
system by which ovules are aseptically
isolated from the ovary and are grown
aseptically on chemically defined nutrient
medium under controlled conditions.
6. An ovule is a mega sporangium covered by integument.
Ovules are attached with placenta inside the ovary by
means of its funiculus. An ovule contains a megaspore or
an egg cell. After fertilization, a single cell zygote is
formed which ultimately leads to form a mature embryo
possessing shoot and root primordia.
Ovules can be isolated and cultured in nutrient medium. In
vitro ovule culture helps to understand the factors that
regulate the development of a zygote through organised
stages to a mature embryo. Alternatively, it may be
possible to germinate pollen in the same culture as the
excised ovule and to induce in vitro fertilisation and sub-
sequently embryo production.
7. (1) Collect the open flower (unfertilized ovules). If
fertilized ovules are desired, collect the open flower
where the anthers are dehisced and pollination has
taken place. To ensure the fertilization, collect the
flower after 48 hrs. of anther dehiscence.
(2) Remove sepals, petals, androecium etc. from
the ovaries containing either fertilized or unfertilized
ovules.
(3) Soak the ovaries in 6% NaOCl solution.
(4) Rinse the ovaries 3-4 times with sterile distilled
water.
8. (5) Using sterile techniques, ovules are gently
prodded with the help of spoon shaped sptula
by breaking the funicles at its junction with
placental tissue.
(6) The spatula with ovules is gently lowered
into the sterile solid or liquid medium as the
culture vial is slanted about 45°.
(7) Damaged or undersized ovules are rejected
when possible, during transfer.
(8) Incubate the ovule culture in either dark or
light (16 hrs. 3,000 lux) at 25°.C
9.
10. First attempt to isolate ovules and culture
them under aseptic conditions was made by
white (1932) in Antirrhinum majus. But
perfected by Maheshwari (1958).
Maheshwari (1958) and Lal (1961) raise viable
seeds of Papaver somniferum starting with
ovules excised 6 days after pollination when
they contained zygote or 2-celled proembryo
and a few endosperm nuclei.
11. Paddubnaya -arnoldi (1959,1960) grew ovules
from pollinated ovaries of several orchids on
simply 10% sucrose solution.
Nakajima (1969) cultured ovules at the
zygote or 2-celled proembryo stage in the
medium supplemented with cucumber and
watermelon juices.
12. Joshi and Johri (1972) made extensive studies
on the nutritional and hormonal factors
concerned in ovule and fibre development
after the unsuccessful attempts at the culture
of Gossypium hirsutum due to interference of
environmental conditions.
Pundir (1967) successfully produced hybrids
by using a cross between Gossypium
arboreum and G. hirsutum.
13. Ramming (1971) reported the various in vitro
conditions for culture of ovules of immature
embryo of peach.
Ramming discovered when medium
supplemented with activated charcoal produced
larger embryos.
Doi et al, demonstrated that the unfertilized
ovule culture techniques of gentians is a
powerful tool for obtaining haploids and double
haploids because of its reproducible and reliable
nature and application to a wide range of
genotypes.
16. TEST TUBE POLLINATION AND FERTILIZATION
An important achievement of research on
ovule culture has been the development
of the technique of test tube pollination
and fertilization.
In this technique, the ovules are planted
on a suitable culture medium which
supports growth of ovules as well as
germination of pollen grain.
Pollen grains are dusted around the
ovules. The pollen germinate and affect
fertilization.
17. EXAMPLES
Normal viable seeds develop within a few days
example Argemone mexicana , Nicotiana
developed by P. Maheshwari in 1962.
Using the same method, it has been possible to
fertilize the ovules of Melandrium album with
pollen grains from other species of
caryophyllaceae and subsequently even with
pollen of Datura stramonium. Employing ovule
culture technique, the incompatibility barrier in
Petunia axillaris has been overcome.
18.
19. In-vitro pollination via ovule culture has been
used to circumvent various problems
associated with hybridization such as failure
of pollen germination on stigmas, insufficient
growth of pollen tube, or precocious
abscission of flowers and self incompatibility
or sterility.
20. Application of Ovule Culture in
Hybridization
ovule culture has been successfully employed to obtain hybrid
seedlings.
It has been observed that in several inter specific crosses; the
hybrid embryo of Abelmoschus fails to develop beyond the heart
or torpedo-shaped embryo.
By ovule culture, viable hybrids have been obtained in three out
of five interspecific crosses attempted, namely, A esculentus x A
ficuneus Aesculentus x A moschatus and A tuberculatus x A
moschatus.
Similarly, a true hybrid between Brassica chinensis and B.
pekinensis has been obtained by culturing the fertilized ovule in
vitro.
A hybrid between Loluim perenne and Festuca rubra has also
been obtained successfully by means of ovule culture .
Pundir successfully produced hybrids by using a cross between
Gossypium hirsutum and G.arboreum.
21. Advantage
In a normal process these hybrids fail to
develop due to early embryo abortion
and premature abscission of fruits. Thus
ovule culture can be used to rescue
them.
This technique omits excision of the
embryo by culturing the entire ovule.
Thus greatly facilitating the time and
effort involved.
Also, ovule culture of cotton offers an
unique method for the studies on the
effect of phytohormones on fibre and
22. Production of Haploid Callus through
Ovule Culture
• Uchimiya et al. (1971) attempted culturing
unfertilized ovules of Solanum melongena
and obtained vigorous callus formation on a
medium supplemented with IAA and kinetin. A
cytological assay revealed it to be haploid in
nature. So it is an important attempt to obtain a
haploid cell line or plant from an alternative
source rather than anther or pollen culture.
23. Advantage
• For haploid production , one can go for anther
culture but anther cultures of some plant such as
Mimulus luteus did not form haploids. So it
is an important attempt to obtain a haploid cell
line or plant from an alternative source rather
than anther or pollen culture.
24. It is generally believed that in obligate root
parasites such as Striga or Orobanke the
formation of seedlings is dependent on some
stimulus from the host root.
Studies on ovule culture of Orobanche
aegyptica and Cistanche tubulosa have
demonstrated that the formation of shoots in
vitro can be induced in any absence of any
stimulus from the host.
25. In ovule cultures of these parasites, it is
possible to substitute the host stimulus with
some known chemical like medium enriched
with cytokinin , GA, scopoletin or strigol.
Thus one can able to study hormonal
requirement or nutritional requirement of
ovules of parasitic plants.
26. OVULE CULTURE OF ORCHID
PLANTS
Poddubnaya-Arnoldi (1959, ’60) successfully
grew the fertilized ovule of orchids in vitro.
They grew the fertilized ovule of Calanthe
veitchn, Cypripedium insigne, Dendrobium
and Phalaenopsis schilleriana.
28. ADVANTAGE
In nature, the seeds of orchid germinate only
in association with a proper fungus. As a
result numerous seeds are lost due to
unavailability of proper fungus. Secondly ,
the seed capsule of many orchid takes a long
time to mature. thirdly ,minute size of seeds.
To overcome such problems, several
attempts have been made to culture the
fertilized ovule of orchid in vitro.(asymbiotic
germination).
29. Ovule culture induces polyembryony in
various parts of the plant.
It has been observed that the nucellus of
mono-embryonic ovule of citrus can be
induced to form adventive embryos in
culture.
30. In horticultural practices, the artificial
induction of polyembryo holds a great
potential. Therefore, such achievement is
very significant.
Polyembryony associated with polyploids
helps them to counteract the disadvantage of
being sexually sterile and thus help them in
their perpetuation generation after
generation.
31. Virus Irradiation through
Ovule Culture
In the varieties of Citrus which are
impossible to free of virus by other
means, the ovule culture has proved
decisively advantageous to make
them virus free.
32. • Thus , due to all these advantages
and applications, ovule culture is a
boon for the plant breeders in
obtaining seedlings from crosses
which are normally unsuccessful
because of abortive embryos.
33. Germination of seeds in vitro conditions
involves the proper inoculation of seed in the
medium. It is better to inoculate the seeds
without seed coats as it reduce the
germination time and increases the
germination potential.
34. Surface sterilize the seeds with teepol and
then 0.1% Hgcl2 for about 5 min.
Wash 3-4 times thoroughly with deionized
water and then inoculate the seeds in basal
MS media under laminar air flow.
36. Plants such as Eranthis (ranunculaceae)
characterized by the presence of embryos
that lack differentiation into various
embryonic organs, namely radicle, plumule
and cotyledons.
Thus in such a case the culture of seeds to
raise sterile seedlings is the best method.
38. Seed culture of parasitic
plants
In obligate root parasites such as
Orobanche and Striga , the seeds
germinate close to host roots ,and it is
only after the parasite establishes
contact with the host –roots that it
develops shoot .
Thus ,through seed cultures of these
parasites it is possible to substitute the
host stimulus with some known
chemicals.
39. Example of seed culture of
parasitic plants
The seeds of Orobanche germinate in
vitro in the absence of a natural host if
the nutrient medium is enriched with a
cytokinin ,GA, scopoletin or strigol
The seeds of Scurrula pulverulenta a
stem parasite can also germinate on
plain white’s basal medium also
seedlings grow better when casein
hydrolysate is added to the medium.
40. Seed culture of orchid
plants
There are several plant species in which
seed sizes are minute and do not germinate
well. orchid is one of them.
Thus , to meet requirements and to conserve
natural resources in-vitro seed germination
has been utilized to produce large quantities
of uniform seedlings.
In vitro seed culturing of such plants is
dependent on the growth medium
supplemented and not required fungus/orchid
symbiotic relationship dependent germination
and thus known as asymbiotic germination.
42. Seed culture is widely used for large, rapid
propagation and large-scale seedling
production. For producing large-scale
seedlings of Capparis spinosa sterilized
the seeds with 0.1% HgCl2 (12 min) and
utilized optimum MS medium
supplemented with activated carbon.
Several examples of seed cultures are
given in following table-
43. Examples of seed culture
Plant description method Effective media
Cephalanthera falcata Asymbiotic
germination
Kano medium and ND
medium
Malaxis khasiana Rapid in-vitro
propagation
Ms medium 2 %
sucrose + casein
hydrolysate +BAP
Dendrobium
hookerianum
Asymbiotic seed
germination
MS medium
Coelogyne nervosa Asymbiotic seed
germination
Ms medium + 30%
coconut water
Cymbidium mastersii Mass propagation Ms medium + BAP
+NAA
44. In –vitro technology –a challenge
• In-vitro culture technologies are still a challenge
because of the slow growth of plantlets, low
multiplication rate, poor rooting, and
somaclonal variation. In this regard,
micropropagation through protocorm-like
bodies obtained from germinating embryos and
somatic tissues is an important strategy in
obtaining genetically stable plants and the
improvement of quality.