This document discusses cell suspension culture, which involves growing plant cells, tissues, or organs in liquid nutrient medium. There are two main types of cell suspension cultures: batch culture and continuous culture. Batch culture involves growing cells in a finite volume of agitated medium, while continuous culture replaces old medium continuously to maintain cells in steady growth. The document outlines different methods for each type of culture and their characteristics, advantages, and importance for studying plant cell physiology and biochemistry.
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 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.
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
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).
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 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.
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
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).
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
Invitro culture of unpollinated ovaries and ovules represents an alternative for the production of haploid plant
First successful report on the induction of gynogenic haploid was in barley by San Noeum in 1976
Haploid plants are obtained from ovary and ovule culture of rice, wheat, maize, sunflower, tobacco, poplar, mulberry etc
Whites or MS or N6 inorganic salt medium supplement with growth substances are used
The isolation, culture and fusion of protoplasts is a fascinating field in plant research. Protoplast isolation and their cultures provide millions of single cells (comparable to microbial cells) for a variety of studies.
A process where an embryo is derived from a single somatic cell or group of somatic cells. Somatic embryos (SEs) are formed from plant cells that are not normally involved in embryo formation.
Embryos formed by somatic embryogenesis are called Embryoids.
The process was discovered for the first time in Daucas carota L. (carrot) by Steward (1958), Reinert (1959).
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
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
Invitro culture of unpollinated ovaries and ovules represents an alternative for the production of haploid plant
First successful report on the induction of gynogenic haploid was in barley by San Noeum in 1976
Haploid plants are obtained from ovary and ovule culture of rice, wheat, maize, sunflower, tobacco, poplar, mulberry etc
Whites or MS or N6 inorganic salt medium supplement with growth substances are used
The isolation, culture and fusion of protoplasts is a fascinating field in plant research. Protoplast isolation and their cultures provide millions of single cells (comparable to microbial cells) for a variety of studies.
A process where an embryo is derived from a single somatic cell or group of somatic cells. Somatic embryos (SEs) are formed from plant cells that are not normally involved in embryo formation.
Embryos formed by somatic embryogenesis are called Embryoids.
The process was discovered for the first time in Daucas carota L. (carrot) by Steward (1958), Reinert (1959).
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
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.
Induction and maintenance of callus and
cell suspension cultures.Callus Culture :
● Callus represents an unorganized or undifferentiated mass of cells.
● They are generally composed of parenchymatous cells and
usually undergo division.
● When an explant is cultured in a medium supplemented with
sufficient amount of auxins, it starts producing mass of cells from
the surface of the explant.
● The concentration of auxins required for each type of explant will
be different and is mainly dependent on the physiological state of
the explant tissue.
● Callus cultures can be maintained for a very long time by
intermittent sub-culturing to a fresh medium.
● The callus cultures can be manipulated for different purposes by
changing the hormone concentrations in the media
Usage of callus Culture :
● Rregeneration of plantlets.
● Preparation of single cells
● Suspension cultures.
● Protoplasts preparation
● Genetic transformation studies.
In some instances, it is necessary to go through a callus phase prior to
regeneration via somatic embryogenesis or organogenesis.
● Callus cultures are suitable for the generation of useful somaclonal variants.
● Callus cultures can be used for in vitro selection of cells and tissue variants
Induction of callus Culture :
● Induction of cell division in the permanent tissue is highly
dependent on the high auxin content (e.g., 2, 4-D) in the
medium.
● The hormone requirement for callus induction may be auxin
alone, cytokinin alone or auxin and cytokinin in different ratios.
● The type of growth regulator requirement and its concentration
in the medium depends strongly on the genotype and
endogenous hormone content of an explant
Transformation of any kind of tissue explant into proliferating
callus mass depends highly on the plant genotype, the source
of origin of the explant and the physiological state of the tissue
at culture.
● For the initiation of callus cultures tissues from young seedling
or from juvenile part of the mature plant either grown in vivo or
in vitro are generally taken, either steri lised (when grown in
vivo) or cut aseptically (when grown in vitro) and inoculated
aseptically on a nutrient medium provided with different
combinations of exogenous growth substances
Maintenance of Callus Culture:
● After callus induction, the callus is grown further on a new
medium, which is referred to as sub-culturing.
● During subculturing the callus tissue shows sigmoid growth
pattern on a particular media.
● The time interval for sub-culturing is calculated according to
the growth pattern of the particular tissue type and genotype
of the plant.
● When the callus growth reaches to the stationary phase of
growth then it needs to be cultured on to a fresh medium.
○Maintenance of Callus Culture:
● After callus induction, the callus is grown further on a new
medium, which is referred to as sub-culturing.
● During subculturing the callus tissue shows sigmoid growth
pattern on a particular media.
● The time interval for sub-cultur
Mass multiplication procedure for tissue culture and PTC requirementDr. Deepak Sharma
This presentation include basic Micropropagation protocol: Application and advantages of mass multiplication. Beside this the requirement of tissue culture are there (Nutrient, gelling agent, energy source, vitamins and PGRs) are also included.
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.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
2. Introduction
Plant Tissue Culture (PTC) is defined as a collection
of experimental methods of growing plant cells, tissues
and organs in an artificially prepared nutrient medium
static or liquid, under aseptic conditions.
It is also referred to as micropropagation.
It was introduced by G. Haberlandt.
The basic key used in plant tissue culture is the
totipotency of plant cells, meaning that each plant cell
has the potential to regenerate into a complete plant.
With this characteristic, plant tissue culture is used to
produce genetically identical plants (clones) in the
absence of fertilization, pollination or seeds.
3. Cell suspension Culture
The cell suspension culture also called as the
plant cell culture is a system for production of
fine chemicals.
It can be defined as “The culture of tissue and
cells cultured in liquid nutrient medium, producing
a suspension of single cells and cell clumps.”
Cell suspension culture is the primary route for
studying plant cell secondary metabolism.
The cell suspension culture requires optimization
of the cell line, the cultivation media, and the
bioreactor system.
4.
5. Types Of Cell Suspension
Cultures
There are two types of cell suspension
cultures :
A. Batch culture
B. Continuous culture
Each of these cultures have its own
advantage and all types are being used in
practice.
6. A. Batch Culture
Batch culture is a type of cell suspension
where the cell material grows in a finite
volume of agitated liquid medium.
These cultures are maintained continuously
by sub culturing.
Batch cultures are most commonly
maintained in conical flasks incubated on
orbital platform shakers at the speed of 80-
120 rpm.
It is a closed system, with no additions or
removal of nutrient and waste products during
the period of incubation.
8. Types
1. Slow rotating cultures :
o In this culture, single cells and cell aggregates are
grown in a specially designed flask, the nipple
flask.
o Each nipple flask possesses eight nipple like
projections, having a capacity of 250ml.
o They are loaded in a circular manner on the large
flat disc of vertical shaker.
o When the flat disc rotates at a speed of 1-2rpm,
the cells within each nipple of the flask are
alternatively bathed in the culture medium and
exposed to air.
9.
10. 2. Shaker cultures :
It is very and effective
system.
In this method, single cells
and cell aggregates in fixed
volume of liquid medium are
placed in conical flasks.
These flasks are then
mounted with the help of
clips on a horizontal large
square plate of an orbital
platform shaker.
The square plate moves in a
circular motion at the speed
of 60-180 rpm.
11. 3. Spinning Cultures
:
In this culture system,
large bottles are used,
usually bottles with the
capacity of 10L.
Large volumes of cell
suspension is cultured
in 10L bottles, with the
bottles spinning in a
spinner at 120 rpm at an
angle of 45° .
12. 4. Stirred Culture :
o This system is used for large scale batch culture.
o In this method, the large culture vessel (round-bottom
flask) is not rotated but the cell suspension inside the
vessel is kept dispersed continuously by bubbling
sterile air through the culture medium.
o Internal magnetic stirrer is used to agitate the culture
medium safely.
o The magnetic stirrer revolves at 200-600 rpm.
13.
14. B. Continuous Culture
In continuous culture system, the old liquid
medium is replaced continuously by the fresh
liquid medium to stabilize the physiological states
of the growing cells.
In this system, nutrient depletion does not occur
due to the continuous flow of nutrients and the
cells always remain in the steady growth phase.
Continuous culture is further divided into two types
:
1. In closed type, the used medium is replaced with
the fresh medium, hence, the cells from used
medium are mechanically retrieved and added
back to the culture and thus, the cell biomass
keeps increasing.
15. 2. In open type, both the cells and used medium are
replaced with fresh medium thus maintaining culture at
constant and submaximal growth rate.
Open continuous cell suspension culture is of two
types :
i. Chemostat :
o In this system, culture vessels are usually cylindrical
or circular in shape and possess inlet and outlet pores
for aeration and the introduction and removal of cells
and medium.
o Such a system are maintained in a steady state.
o Thus in a steady state condition the density, growth
rate, chemical composition and metabolic activity of
the cells all remain constant.
o Such continuous cultures are ideal for studying growth
kinetics and the regulation of metabolic activity in
higher plants.
16.
17. ii. Turbidostats :
o A turbidostat is a continuous culturing method
where the turbidity of the culture is held constant
by manipulating the rate at which medium is fed.
o In this system, the cells are allowed to grow upto a
certain turbidity, when the predetermined volume
of culture is replaced by fresh culture.
o The turbidity is measured by the changes of
optical density of medium
o An automatic monitoring unit is connected with the
culture vessel and such unit adjusts the medium
flow in such a way as to maintain the optical
density or PH at chosen, present level.
18. Importance of cell suspension
culture
Such systems are capable of contributing significant
information about cell physiology, biochemistry,
metabolic events, etc.
It is important to build up an understanding of an
organ/embryoid formation starting from a single cell.
Mutagenesis studies maybe facilitated by cell
suspension culture to produce mutant cell clone from
which mutant plants can be raised.
19. Advantages :
The nutrients can be continually adjusted.
This system can be scaled for large scale
production of the cells.
A whole plant can be regenerated from a
single plant cell.
Disadvantages :
The productivity of suspension cultures
decreases over extended subculture periods.
Slow growth and low productivity of plant
cells.
Cells may get damaged by shear conditions.