The document discusses micropropagation, which is a method of vegetative propagation used to rapidly produce multiple genetically identical copies of plants through tissue culture techniques. It describes the five main stages of micropropagation as preparatory, initiation of culture, multiplication, rooting of shoots, and transplantation. The multiplication stage involves approaches like callus formation, adventitious bud formation, and enhanced axillary branching to produce many new shoots from an explant. Micropropagation offers advantages like producing large numbers of disease-free clones from a single plant in a relatively short time and small space.
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
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
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).
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
Introduction
Reason for cryopreservation
Selection of part of plant for cryopreservation
Technique of cryopreservation
Application
Limitation
Conclusion
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
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
Haploid Production - Techniques, Application & Problem ANUGYA JAISWAL
Haploid is applied to any plant originating from a sporophyte (2n) and containing (n) number of chromosomes.
Artificial production of haploids was attempted through distant hybridization, delayed pollination, application of irradiated pollen, hormone treatment and temperature shock.
The artificial production of haploids until 1964 was attempted through:
1. Distant hybridization
2. Delayed pollination
3. Application of irradiated pollen
4. Hormone treatments
5. Temperature shocks
The development of numerous pollen plantlets in anther cultures of Datura innoxia, first reported by two Indian scientists (Guha and Maheshwari, 1964, 1966), was a major breakthrough in haploid breeding of higher plants.
The technique of haploid production through anther culture ('anther - androgenesis') has been extended successfully to numerous plant species, including many economically important plants, such as cereals and vegetable, oil and tree crops.
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.
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,
These slides represent a comprehensive view of history of using natural products caused to appearance of pharmacognosy as a science and show several aspects of pharmacognosy and natural products use and final their importance in discovering new drugs.
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).
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
Introduction
Reason for cryopreservation
Selection of part of plant for cryopreservation
Technique of cryopreservation
Application
Limitation
Conclusion
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
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
Haploid Production - Techniques, Application & Problem ANUGYA JAISWAL
Haploid is applied to any plant originating from a sporophyte (2n) and containing (n) number of chromosomes.
Artificial production of haploids was attempted through distant hybridization, delayed pollination, application of irradiated pollen, hormone treatment and temperature shock.
The artificial production of haploids until 1964 was attempted through:
1. Distant hybridization
2. Delayed pollination
3. Application of irradiated pollen
4. Hormone treatments
5. Temperature shocks
The development of numerous pollen plantlets in anther cultures of Datura innoxia, first reported by two Indian scientists (Guha and Maheshwari, 1964, 1966), was a major breakthrough in haploid breeding of higher plants.
The technique of haploid production through anther culture ('anther - androgenesis') has been extended successfully to numerous plant species, including many economically important plants, such as cereals and vegetable, oil and tree crops.
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.
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,
These slides represent a comprehensive view of history of using natural products caused to appearance of pharmacognosy as a science and show several aspects of pharmacognosy and natural products use and final their importance in discovering new drugs.
Plants are a tremendous source for the discovery of new products of medicinal value for drug development.
Today several distinct chemicals derived from plants are important drugs currently used in one or more countries in the world. During the past decade, traditional systems of medicine have become a topic of global importance. Current estimates suggest that, in many developing countries, a large proportion of the population relies heavily on traditional practitioners and medicinal plants to meet primary health care needs.
Although modern medicine may be available in these countries, herbal medicines (phytomedicines) have often maintained popularity for historical and cultural reasons. Concurrently, many people in developed countries have begun to turn to alternative or complementary therapies, including medicinal herbs.
Clonal Propagation: Introduction, Techniques, Factors, Applications and Disadvantages
Multiplication of Apical or Axillary bud, Shoot tip or meristem culture
Production of Disease free plants by Micropropagation techniques: their Advantages and Disadvantages
Micropropagation (tissue culture or invitro culture) refers to the multiplication of plants, in an aseptic condition and in artificial growth medium from plant parts like meristem tip, callus, embryos anthers, axillary buds etc. It is a method by which a true to type and disease free entire plant can be regenerated from a miniature piece of plant in aseptic condition in artificial growing medium rapidly throughout the year.
PPT on Tissue Culture Class 10 CBSE Text Book NCERT.One Time Forever
This is a PPT Based on Class 10 Chapter How Do Organisms Reproduce, on a Small Topic of it That is Tissue Culture with easy and detailed explanation of each topic of tissue culture along with some pictures and some examples. Hopefully it Would Be Helpful To You. Thank You.
Definition of hairy root culture ,multiple shoot culture ,Production of hairy root and multiple shoot , advantages an disadvantages of hairy root and multiple shoot culture, Sterilization and sterilizing agents wit concentration and exposure time
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
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
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.
Richard's entangled aventures in wonderlandRichard 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.
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.
THE IMPORTANCE OF MARTIAN ATMOSPHERE SAMPLE RETURN.Sérgio Sacani
The return of a sample of near-surface atmosphere from Mars would facilitate answers to several first-order science questions surrounding the formation and evolution of the planet. One of the important aspects of terrestrial planet formation in general is the role that primary atmospheres played in influencing the chemistry and structure of the planets and their antecedents. Studies of the martian atmosphere can be used to investigate the role of a primary atmosphere in its history. Atmosphere samples would also inform our understanding of the near-surface chemistry of the planet, and ultimately the prospects for life. High-precision isotopic analyses of constituent gases are needed to address these questions, requiring that the analyses are made on returned samples rather than in situ.
(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.
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.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
1. 1
Micropropagation
Multiplication of genetically identical copies of a cultivar by asexual reproduction is called clonal
propagation. In nature, clonal propagation occurs by apomixis (seed development without
meiosis and fertilization) and/or vegetative propagation (regeneration of new plants from
vegetative parts). Tissue culture has become popular method for vegetative propagation of
plants. Aseptic method of clonal propagation is called as Micropropagation and it offer the
advantage of large number of true-to-type plantlets can be produced with relatively short time
and space from a single individual. It is the fact that micropropagation is the only commercially
viable method of clonal propagation of most of the horticultural crops. E.g. Orchids.
Explants used in micropropagation
Different kinds of explants were used in micropropagation. For example, in case of
orchids, shoot tip (Anacamptis pyramidalis, Aranthera, Calanthe, Dendrobium), axillary bud
(Aranda, Brassocattleya, Cattleya, Laelia), inflorescence segment (Aranda, Ascofinetia,
Neostylis, Vascostylis), lateral bud (Cattleya, Rhynocostylis gigantean), leaf base (Cattleya),
leaf tip (Cattleya, Epidendrum), shoot tip (Cymbidium, Dendrobium, Odontioda, Odontonia),
nodal segment (Dendrobium), flower stalk segment (Dendrobium, Phalaenopsis) and root tips
(Neottia, Vanilla) are being used in micropropagation.
Different explants Differentiation of shoots directly from callus
2. A. The principle organs and tissues of the body of a seed plant; B. Cross-section of
stem; C. Cross-section of root. Apical or axillary buds are good sources of explants
2
Stages in micropropagation
Micropropagation generally involves five stages. Each stage has its own requirements.
Stage 0: Preparative stage
This stage involves the preparation of mother plants to provide quality explants for better
establishment of aseptic cultures in stage 1. To reduce the contamination problem in the
subsequent stages, mother plant should be grown in a glasshouse and watered so as to avoid
overhead irrigation. This will also reduce the need for a harsh sterilization treatment. Stage 0
3. also includes exposing the stock plants to suitable light, temperature and growth regulator
treatments to improve the quality of explants. In the case of photosensitive plants it may be
possible to obtain suitable explants throughout the year by controlling photoperiod in the
glasshouse. For example, red-light treated plants of Petunia provided leaf explants which
produced up to three times as many shoots as did the explants from untreated plants.
Stage 1. Initiation of culture
1. Explant: The nature of explant to be used for in vitro propagation is governed by the
method of shoot multiplication. For enhanced axillary branching, only the explants which
carry a pre-formed vegetative bud are suitable. When the objective is to produce virus-free
plants from an infected individual it becomes necessary to start with sub-millimeter shoot
tips. If the stock is virus-tested or virus eradication is not necessary, then the most suitable
explant is nodal cuttings. Small shoot-tip explants have a low survival rate and show slow
initial growth. Meristem tip culture may also result in the loss of certain horticultural
characteristics which are controlled by the presence of virus, such as the clear-vein
character of the Geranium cv. Crocodile. Generally, the clear vein character is transmitted in
petiole-segment culture but not in shoot-tip culture.
2. Sterilization: Special precautions need to be taken when explants are derived from field-grown
materials, which is often necessary in cloning an elite tree. In such cases an ideal
approach would be to take cuttings from the selected plant and grow them in greenhouse.
Discarding the surface tissues from plant materials while preparing the explants also
minimizes the loss of cultures due to microbial contamination.
3. Browning of medium: A serious problem with the culture of some plant species is the
oxidation of phenolic compounds leached out from the cut surface of the explant. It turns the
medium dark brown and is often toxic to the tissues. This problem is common with the adult
tissues from woody species.
3
3. Stage 2. Multiplication
This is the most crucial stage since it is the point at which most of failures in
micropropagation occur. Broadly three approaches have been followed to achieve in vitro
multiplication.
4. 1. Through callusing: The potentiality of plant cells to multiply indefinitely in cultures and their
totipotent nature permit a very rapid multiplication of several plant types. Differentiation of
plants from cultured cells may occur via shoot-root formation (organogenesis) or somatic
embryogenesis. Somatic embryogenesis is most appealing from a commercial angle. A
somatic embryogenesis system once established lends itself to better control than
organogenesis. Since somatic embryos are bipolar structures, with defined root and shoot
meristems, the rooting stage required for microshoots gets eliminated. Above all, somatic
embryos being small, uniform and bipolar are more amenable to automation at the
multiplication stage and for field planting as synthetic seeds, offering cost advantages from
labour savings, can also be stored through cold storage, cryopreservation or desiccation for
prolonged periods. These characteristics make somatic embryogenesis potentially a less
expensive and flexible system for micropropagation. The most serious objection against the
use of callus cultures for shoot multiplication is the genetic instability of their cells.
2. Adventitious bud formation: Buds arising from any place other than leaf axil or the shoot
apex are termed adventitious buds. The shoots differentiated from calli should also be
treated as adventitious buds. In many crops, vegetative propagation through adventitious
bud formation from root (blackberry, raspberry) and leaf (Begonia, Crassula) cuttings is
standard horticultural practice. In such cases the rate of adventitious bud development can
be considerably enhanced under culture conditions. For most bulbous plants (e.g. Lilley)
adventitious bud formation is the most important mode of multiplication and the best
4
5. explants are obtained from bulb scales. A serious problem may arise when this method of
propagation is applied to varieties which are genetic chimeras. Adventitious bud formation
involves the risk of splitting the chimeras leading to pure type plants. For example, in
variegated geranium cv. Mme Salleron, the chimera is perpetuated in meristem culture but
broken down in petiole culture.
3. Enhanced axillary branching: In cultures the rate of shoot multiplication by enhanced
axillary branching can be substantially enhanced by growing shoots in a medium containing
a suitable cytokinin at an appropriate concentration with or without auxin. Due to continuous
availability of cytokinin, the shoots formed by the bud, present on the explant, develops
axillary buds which may grow directly into shoots. This process may be repeated several
times and the initial explant transformed into a mass of branches.
5
4. Stage 3. Rooting of shoots
Somatic embryos carry a pre-formed radical and may develop directly into plantlet.
However, these embryos often show very poor conversion into plantlets, especially under in
vitro conditions. They require an additional step of maturation to acquire the capability for
normal germination. Adventitious and axillary shoots developed in cultures in the presence of a
cytokinin generally lack roots. To obtain full plants the shoots must be transferred to a rooting
medium which is different from the shoot multiplication medium, particularly in its hormonal and
salt compositions. For rooting, individual shoots measuring 2 cm in length are excised and
transferred to the rooting medium.
5. Stage 4. Transplantation
The ultimate success of commercial propagation depends on the ability to transfer plants
out of culture on a large scale, at low cost and with high survival rates. The plants multiplied in
vitro are exposed to a unique set of growth conditions (high levels of inorganic and organic
nutrients, growth regulators, sucrose as carbon source, high humidity, low light, poor gaseous
exchange) which may support rapid growth and multiplication but also induce structural and
physiological abnormalities in the plants, rendering them unfit for survival under in vivo
conditions. The two main deficiencies of in vitro grown plants are – poor control of water loss
and heterotrophic mode of nutrition. Therefore, gradual acclimatization is necessary for these
plants to survive transition from culture to the greenhouse or field. During acclimatization the in
6. vitro formed leaves do not recover but the plant develops normal leaves and functional roots.
While transferring out shoots/roots their lower part is gently washed to remove the medium
sticking to them. The individual shoots or plantlets are then transferred to potting mix and
irrigated with low concentration of inorganic nutrients. This probably recommissions the
photosynthetic machinery of plants, enabling them to withstand the subsequent reduction in the
ambient relative humidity and survive under field conditions. A variety of potting mixtures such
as peat, perlite, polystyrene beads, vermiculate, fine bark, coarse sand etc. or their mixtures in
different combinations are used for transplantation. For initial 10-15 days, it is essential to
maintain high humidity (90-100%) around the plants, to which they got adapted during culture.
The humidity is gradually reduced to ambient level over a period of 2-4 weeks.
6
In vitro regenerative protocol of V. reitzii
7. 7
Advantages of micropropagation
1. Clonal mass propagation - extremely large numbers of plants can be produced. Rather
than getting 10000 plants per year from an initial cutting in vegetative propagation, one
can obtain more than 1,000,000 plants per year from one initial explant through
micropropagation.
2. Culture is initialized from small parts of plants – so no need of much space: from 1 m2
space in culture room, 20000 - 100000 plants can be produced per year.
8. 3. Production of disease and virus free plantlets. This leads to simplification of international
8
exchange of plants
4. Micropropagation enables growers to increase the production of plants that normally
propagate very slowly such as Narcissus and other bulbous crops.
5. Introduction of disease free new cultivars is possible through micropropagation
6. Vegetative propagation of sterile hybrids can be used as parent plants for seed
production. Eg. Cabbage
7. One of the rapid methods for cloning of disease free trees.
8. In vitro cultures can be stored for long time through cryopreservation.
9. Breeding cycle can be shortened.
Disadvantages of micropropagation
1. Expensive laboratory equipment and service
2. No possibility of using mechanization
3. Plants are not autotrophic
4. Poor Acclimatization to the field is a common problem (hyperhydricity)
5. Risk of genetic changes if 'de novo' regeneration is used
6. Mass propagation cannot be done with all crops to date. In cereals much less success is
achieved
7. Regeneration is often not possible, especially with adult woody plant material.
8. More problems in inducing rooting
9. May not get uniform growth of original plant from tissue culture. Each explant has
different in vitro growth rates and maturation. Thus cannot be used for floriculture crop
production where uniformity is critical.
Horticultural uses for plant tissue culture
1. Clonal mass propagation. The important point here is that extremely large numbers of
plants can be produced. Rather than getting 10000 plants per year from an initial cutting, one
can obtain upwards of 1,000,000 plants per year from one initial explant.
2. Difficult or slow to propagate plants. Micropropagation enables growers to increase the
production of plants that normally propagate very slowly such as narcissus and other bulbous
crops.
9. 3. Introduction of new cultivars eg. Dutch iris. Get 5 daughter bulbs annually. Takes 10 years
for commercial quantities of new cultivars to be built up. Can get 100-1000 bulbs per stem
section.
4. Vegetative propagation of sterile hybrids used as parent plants for seed production. Eg.
cabbage.
5. Pathology - Eliminate viruses, bacteria, fungi etc. Use heat treatment and meristem
culture. Used routinely for potatoes, carnation, mum, geranium, garlic, gypsophila
6. Storage of germplasm
Generally the only successful method to date is keeping them in refrigerator. Slows down, but
does not eliminate, alterations in genotype.
9
10. Questions
1. Multiplication of genetically identical copies of a cultivar by asexual reproduction is called
……………….
b)Regeneration of new plants from
vegetative parts
b)Regeneration of new plants from
vegetative parts
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a) Clonal propagation b) Apomixis
c) Vegetative propagation d) None of the above
2. In nature, clonal propagation occurs by ……………….
a) Vegetative propagation b) Apomixis
c) Both a & b d) None of the above
3. Apomixis is by ……………….
a) Seed development without meiosis
and fertilization
c) Both a & b d) None of the above
4. Vegetative propagation is by ……………….
a) Seed development without meiosis and
fertilization
c) Both a & b d) None of the above
5. Advantage of tissue culture is/are ……………….
a) Production of large number of true-to-type
plantlets from a single plant
b) Less time requirement
c) Less space requirement d) All the above
6. Total number of stages in micro propagation is ……………….
a) 3 b) 4
c) 5 d) 6
7. ………. stage involves the preparation of mother plants
a) Stage 0 b) Stage 1
c) Stage 2 d) Stage 3
8. ………. stage involves the preparation of mother plants
11. 11
a) Stage 0 b) Stage 1
c) Stage 2 d) Stage 3
9. Stage 0 in tissue culture is
a) Preparative stage b) Initiation of culture
c) Multiplication d) Rooting of shoots
10. Stage 1 in tissue culture is
a) Preparative stage b) Initiation of culture
c) Multiplication d) Rooting of shoots
11. Stage 2 in tissue culture is
a) Preparative stage b) Initiation of culture
c) Multiplication d) Rooting of shoots
12. Stage 3 in tissue culture is
a) Preparative stage b) Initiation of culture
c) Multiplication d) Rooting of shoots
13. Stage 3 in tissue culture is
a) Preparative stage b) Initiation of culture
c) Multiplication d) Transplanting
14. Stage 3 in tissue culture is
a) Preparative stage b) Initiation of culture
c) Multiplication d) Transplanting
15. The suitable explant for producing virus free plant is …………..
a) Shoot tip b) Leaf bit
c) Stem bit d) None of the above
16. The oxidation of phenolic compounds leached out from the cut surface of the explant in
tissue culture leads to …………..
a) Browning of the medium b) Blackening of the medium
12. c) Whitening of the medium d) None of the above
17. The oxidation of phenolic compounds leached out from the cut surface of the explant in
tissue culture leads to …………..
12
a) Browning of the medium b) Blackening of the medium
c) Whitening of the medium d) None of the above
18. Browning of medium is a common problem in …………..
a) Adult tissues from woody species b) Juvenile tissues from woody species
c) Both a and b d) None of the above
19. The crucial stage in tissue culture is
a) Preparative stage b) Initiation of culture
c) Multiplication d) Rooting of shoots
20. In tissue culture, the multiplication is through
a) Callusing b) Adventitious bud formation
c) Enhanced axillary branching d) All the above