This document provides an overview of basic techniques in plant tissue culture and plant cell immobilization. It discusses that tissue culture involves growing plant cells or tissues artificially in a controlled environment. The two main techniques are static culture (callus culture) and suspension culture. Static culture uses isolated plant tissue on a nutrient medium to form an unorganized callus mass, while suspension culture grows isolated cells or small cell aggregates in liquid medium. The document also describes various methods of plant cell immobilization including adsorption, covalent attachment, and entrapment in natural or synthetic polymers, which has advantages like continuous processing but also challenges like reduced biosynthetic capacity.
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.
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).
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.
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.
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).
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.
Until two decades ago the genetic resources were getting depleted owing to the
It was imperative therefore that many of the elite, economically important and endangered species are preserved to make them available when needed.
The conventional methods of storage failed to prevent losses caused due to various reasons.
A new methodology had to be devised for long term preservation of material.
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
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.
Suspension Culture and Single Cell Cultures, Culturing methods, maintenance a...Ananya Sinha
Suspension Culture and Single Cell Cultures, Culturing methods, maintenance and application
Generally, suspension culture is a one stop technology to produce secondary metabolites on a large scale in-vitro, irrespective of the climatic condition or nutrient availability (as required in field plants).
In this presentation, we will see the importance of suspension culture, culturing methods and it's application (mostly with respect to plants) and also focus on what exactly is a single cell culture.
Until two decades ago the genetic resources were getting depleted owing to the
It was imperative therefore that many of the elite, economically important and endangered species are preserved to make them available when needed.
The conventional methods of storage failed to prevent losses caused due to various reasons.
A new methodology had to be devised for long term preservation of material.
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
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.
Suspension Culture and Single Cell Cultures, Culturing methods, maintenance a...Ananya Sinha
Suspension Culture and Single Cell Cultures, Culturing methods, maintenance and application
Generally, suspension culture is a one stop technology to produce secondary metabolites on a large scale in-vitro, irrespective of the climatic condition or nutrient availability (as required in field plants).
In this presentation, we will see the importance of suspension culture, culturing methods and it's application (mostly with respect to plants) and also focus on what exactly is a single cell culture.
The term isolation refers to the separation of a strain from a natural, mixed population of living microbes, as present in the environment. It becomes necessary to maintain the viability and purity of the microorganism by keeping the pure culture free from contamination.
A pure culture theoretically contains a single bacterial species. There are a number of procedures available for the isolation of pure cultures from mixed populations. A pure culture may be isolated by the use of special media with specific chemical or physical agents that allow the enrichment or selection of one
organism over another.
PURE CULTURE TECHNIQUE ISOLATION AND IDENTIFICATION PROCESS .pptxVishekKumar8
Pure culture technique
INTRODUCTION
PURE CULTIURE TECHNIQE
ISOLATION PROCESS
STREAK PLATE METHOD
POUR PLATE METHOD
SPREAD PLATE METHOD
IDENTIFICATION PROCESS
BIOCHEMICAL TEST
MOLECULAR METHOD
SEROGICAL TECHNIQUE
introduction to microbial growth.
different types of growth.
different types of cultivation .
m.sc microbiology, m.sc biotech
batch, fed-batch cultivation , continous cultivation, chemostat and turbidostat
synchronous growth and diauxic growth
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.
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.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
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/
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.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
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.
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.
Basic techniques of plant tissue culture
1. BASIC TECHNIQUES OF PLANT
TISSUE CULTURE AND PLANT
CELL IMMOBILIZATION
BY
NIVETHA.B
M .PHARMACY
(PHARMACOGNOSY) I YEAR1
2. WHAT IS TISSUE CULTURE ?
Tissue culture is the term used for “the
process of growing cells artificially in the
laboratory”.
Tissue culture is invitro cultivation of plant
cell or tissue under aseptic and controlled
environmental conditions,
2
3. in liquid or on semisolid well defined nutrient
medium for the production of primary and
secondary metabolites or to regenerate plant.
Tissue culture involves both plant and animal
cells.
3
4. BASIC TECHNIQUES OT PLANT
TISSUE CULTURE
The technique of plant tissue and cell culture has
evolved over decades. This technique combine
with recent advances in developmental, cellular,
molecular genetic, metabolic engineering,
genetic transformation and using conventional
plant breeding have turned plant biotechnology
into an exciting research field with a significant
impact on pharmaceutical industries, agriculture,
horticulture and forestry.
4
5. TYPES OF TECHNIQUES :
There are mainly two major techniques in
plant tissue culture. They are explained as
follows,
STATIC CULTURE
SUSPENSION CULTURE
5
6. STATIC CULTURE
• Static culture is otherwise known as callus
culture or solid culture.
• In this technique isolated piece of plant tissue
is cultured on a nutrient medium.
• Plant growth regulators such as auxins ,
cytokinins, and gibberllins are supplemented
into the medium to initiate callus formation.
6
7. • Finally, an unorganized mass of cell appears
and it is called as callus.
7
8. 8
• This callus is transferred on to different
media to regenerate plants.
• This callus culture technique is easier and
convenient for initial maintanence of cell
lines and also for carrying out the
investigation studies related to
organogenesis.
10. SUSPENSION CULTURE
• Suspension culture is also known as liquid
culture.
• It is the culture of isolated cells or very small
cell aggregates dispersed in liquid medium.
The cell suspension is obtained by agitating
pieces of callus in liquid medium on gyrating
shaker.
10
11. Growing cells in a liquid medium under
controlled conditions.
Transfer of an established callus culture into
the liquid medium in Erlenmeyer flask.
Medium with high auxin content, appropriate
concentration of auxin and yeast or auxin vs
kinetin helps to maintain suspension.
Incubated at 25˚ C in darkness or with less light
intensity fluorescent light.
11
12. It contains a uniform suspension of separate cells in a
liquid medium
callus
liquid medium
agitated continuously
finally cells separated
sub-culture the cells
This can be achieved by rotary shaker attached within
the incubator at a rate of 50-150 rpm .
12
13. TYPES OF SUSPENSION CULTURE
There are three types of suspension culture.
1.Batch suspension culture
2. Continuous culture
(I)Open type
(II)Close type
3.Semi-continuous culture
13
14. BATCH CULTURE
• These cultures are maintained continously by
propagating a small aliquot of inoculum in the
moving liquid and transferring it to fresh
medium at regular intervals.
• Generally cell suspension are grown in
flasks(100-250 ml) containing 25-27 ml of the
culture medium.
14
15. • When the cell number in suspension culture
is plotted against the time of incubation , a
growth curve is obtained.
15
16. CONTINUOUS CULTURE
• In this system , the liquid medium is
continuously replaced by the fresh liquid
medium to stabilize the physiological stage of
growing cells.
• Cell proliferation takes place under constant
conditions
16
17. • As a result ,the active growth phase of the cell
declines the depletion of nutrient.
• The cells passing through out flowing medium
are separated mechanically.
• The cells are always kept in exponential
growth phase.
17
18. MEASUREMENT OF GROWTH OF
CELL CULTURES
Methods used to determine are..
CELL NUMBER
• By counting the cell number in
haemocytometer under a microscope.
• Suspension culture is preferable.
18
20. PACKED CELL VOLUME
Cell suspension is transfer to graduated
centrifuge.
• Centrifuged at 2000 rpm for 5mints.
• Cell will form pellets called biomass volume,
expressed by ml-1
20
25. VIABLE CELL TEST
• The staining method such as fluorescein di-
acetate is used for accessing the cell viability.
• Dead cells appear as fluorescein red.
25
27. PLANT CELL IMMOBILIZATION
• Immobilization is the newest culture of plant
cell and considered has to be the most
‘natural’.
• It has been defined as a technique, which
confines to a catalytically active enzyme or to
a cell within a reactor system and prevents its
entry into mobile phase, which carries the
substrate and product.
27
28. ADVANTAGES:
• Retention of biomass enables its continuous
reutilization as a product system, a definite
advantage with slow growing plant cells
e.g.Papaver somniferum have remained stable
and active for up to six months.
• Continuous process: Immobilization allows a
continuous process ,which increase volumetric
productivity and allows the removal of
metabolic inhibitors.
28
29. • Separation of cells from medium:The
immobilization separates cells from medium
and the desired product in extra cellular,
which will simplify down stream processing
compared to extraction from tissue.
• Decoupling of growth and product formation:
Immobilization is compatible with non growth
associated product formation.
29
30. • Reduces problems such as aggegrate,growth
and foaming.
• High biomass levels.
30
31. DISADVANTAGES:
• Secretion of secondary metabolites requires
cellular transport or a artificially altered
membrane permeability’
• The efficiency of the production process
depends on the rate of release of products
rather than actual raye of biosynthesis.
31
32. • The immobiliztion process may reduce
biosynthetic capacity.
• Products must be released from the cell into
medium.
• Relese of single cells from cell aggregate may
make processing of the product more
difficult.
32
33. TYPES OF IMMOBILIZATION
• Direct intracellular binding due to natural
affinity(adsorption , adhesion and
agglutination).
• Covalent coupling
• Intracellular connection via bi or poly
functional reagent(cross-linking)
33
34. • Mixing with suitable materials,changing their
consistency with temperature(embedding).
• Physical retention within the framework of
diverse pore size and permeability
(entrapment , micro encapsulation).
34
35. METHOD OF IMMOBILIZATION
1. Adsorption
2. Covalent attachment
3. Entrapment
a) Natural polymer
Alginate
Agar
Agarose
35
37. ENTRAPMENT:
• GEL ENTRAPMENT BY POLYMERIZATION:
A monomer or a mixture of monomers is
polymerized in the presence of cell
suspension, which is entrapped inside the
lattice of the polymer.
The most common example is
polyacrylamide.
37
38. GEL ENTRAPMENT FORMATION BY
PRECIPITATION:
• Gels may be formed by precipitation of some
natural and synthetic polymers by changing
one or more parameters in the solution, such
as temperature, salinity or pH of solvent .
• Several methods can be used for entrapment.
38
39. Gel entrapment by precipitation
SPECIES GEL
Catharanthus roseus Agarose , Agrose, Agar
Silybum marianum Agar
39
40. SURFACE IMMOBILIZATION:
• Surface immobilization may occur on both
natural and other matrices.
• Eg for natural matrices are deeper callus
layers and cellulose. For synthetic steel and
nylon are used.
40
41. REFERENCE:
• Medicinal Plant Biotechnology by Ciddi
Veeresham.
• Pharmaceutical Biotechnology by S S Kori.
• Trease and evans pharmacognosy by W.C evans.
• A textbook of industrial pharmacognosy by
A.N.Kalia.
• Pharmacognosy and photochemistry-part 2 by
Vinod.D.Rangari. 41