This document provides information on in vitro germplasm conservation. It discusses that germplasm conservation aims to preserve the genetic diversity of plants. There are several methods of in vitro conservation including cryopreservation, cold storage, and low pressure/low oxygen storage. Cryopreservation involves freezing plant cells and tissues at ultra-low temperatures like in liquid nitrogen to bring their metabolism to zero. It allows for long term conservation of large amounts of genetic material in a small space. Cold storage conserves germplasm at low non-freezing temperatures to slow growth. Low pressure and low oxygen storage reduce atmospheric pressure and oxygen concentration to inhibit plant tissue growth.
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
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
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
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
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.
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.
Plant exploration, germplasm collection, conservation and utilizationSyed Zahid Hasan
Sequentially given germplasm exploration, collection, conservation,evaluation and utilization sof Agroforestry plants.
Some information and pictures collected from google.
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).
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
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.
Meristem tip culture for the production of the virus free plantsArjun Rayamajhi
This presentation gives general idea on the meristem tip culture for the production of the virus free plants. The principles, methods and procedures of the meristem tip culture included. General idea on different in vitro culture techniques for virus elimination meristem tip culture viz. thermotherapy, cryotherapy,chemotherapy and electrotherapy are provided.
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.
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.
Plant exploration, germplasm collection, conservation and utilizationSyed Zahid Hasan
Sequentially given germplasm exploration, collection, conservation,evaluation and utilization sof Agroforestry plants.
Some information and pictures collected from google.
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).
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
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.
Meristem tip culture for the production of the virus free plantsArjun Rayamajhi
This presentation gives general idea on the meristem tip culture for the production of the virus free plants. The principles, methods and procedures of the meristem tip culture included. General idea on different in vitro culture techniques for virus elimination meristem tip culture viz. thermotherapy, cryotherapy,chemotherapy and electrotherapy are provided.
Genetic material of plants which is of value as a resource for present and future generations of people is referred to as plant genetic resources.
The whole library of different alleles of a species or sum total of genes in a species is known as gene pool, also called germplasm, genetic stock and genetic resources.
The term gene pool was coined by Dobzhansky in 1951.
The term germplasm was first used by Weismann in 1883.
Germplasm Conservation || Presented by Mamoona GhaffarMamoona Ghaffar
Germplasm Conservation || Presented by Mamoona Ghaffar
it's all about germplasm conservation, features, types & approaches, & limitations of germplasm storage
Feel free to ask about your queries.
According to Lokesh, the goal of pure culture preservation is to preserve genetic stability and microbiological viability by use of several methods. Important techniques include the straightforward and inexpensive short-term storage of refrigeration at 4°C on agar slants; the long-term preservation of deep freezing at -70°C to -80°C with cryoprotectants like glycerol or DMSO, which guarantees little genetic changes over years; and the freeze-drying process, or lyophilization, in which cultures are frozen and dehydrated under a vacuum to preserve viability and stability over long periods of time. Research, clinical diagnostics, and industrial applications all depend on these techniques to guarantee the availability of clean microbial cultures when required.
Preservation of industrially important microorganisms, methods of preservation, periodic transfer, storage in saline suspension, storage in sterile soil, cryopreservation
presenation only for exsitu conservation includes topic (Components of ex-situ conservation
Plant genetic resources conservation in gene banks, national gene banks and gene repositories
Preservation of genetic materials under natural conditions, Perma-frost conservation
Guidelines for sending seeds to network of active/ working collections
Orthodox and recalcitrant seeds- differences in handling
Clonal repositories
genetic stability under long term storage condition)
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
The ability to recreate computational results with minimal effort and actionable metrics provides a solid foundation for scientific research and software development. When people can replicate an analysis at the touch of a button using open-source software, open data, and methods to assess and compare proposals, it significantly eases verification of results, engagement with a diverse range of contributors, and progress. However, we have yet to fully achieve this; there are still many sociotechnical frictions.
Inspired by David Donoho's vision, this talk aims to revisit the three crucial pillars of frictionless reproducibility (data sharing, code sharing, and competitive challenges) with the perspective of deep software variability.
Our observation is that multiple layers — hardware, operating systems, third-party libraries, software versions, input data, compile-time options, and parameters — are subject to variability that exacerbates frictions but is also essential for achieving robust, generalizable results and fostering innovation. I will first review the literature, providing evidence of how the complex variability interactions across these layers affect qualitative and quantitative software properties, thereby complicating the reproduction and replication of scientific studies in various fields.
I will then present some software engineering and AI techniques that can support the strategic exploration of variability spaces. These include the use of abstractions and models (e.g., feature models), sampling strategies (e.g., uniform, random), cost-effective measurements (e.g., incremental build of software configurations), and dimensionality reduction methods (e.g., transfer learning, feature selection, software debloating).
I will finally argue that deep variability is both the problem and solution of frictionless reproducibility, calling the software science community to develop new methods and tools to manage variability and foster reproducibility in software systems.
Exposé invité Journées Nationales du GDR GPL 2024
BREEDING METHODS FOR DISEASE RESISTANCE.pptxRASHMI M G
Plant breeding for disease resistance is a strategy to reduce crop losses caused by disease. Plants have an innate immune system that allows them to recognize pathogens and provide resistance. However, breeding for long-lasting resistance often involves combining multiple resistance genes
hematic appreciation test is a psychological assessment tool used to measure an individual's appreciation and understanding of specific themes or topics. This test helps to evaluate an individual's ability to connect different ideas and concepts within a given theme, as well as their overall comprehension and interpretation skills. The results of the test can provide valuable insights into an individual's cognitive abilities, creativity, and critical thinking skills
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.
Remote Sensing and Computational, Evolutionary, Supercomputing, and Intellige...University of Maribor
Slides from talk:
Aleš Zamuda: Remote Sensing and Computational, Evolutionary, Supercomputing, and Intelligent Systems.
11th International Conference on Electrical, Electronics and Computer Engineering (IcETRAN), Niš, 3-6 June 2024
Inter-Society Networking Panel GRSS/MTT-S/CIS Panel Session: Promoting Connection and Cooperation
https://www.etran.rs/2024/en/home-english/
ESR spectroscopy in liquid food and beverages.pptxPRIYANKA PATEL
With increasing population, people need to rely on packaged food stuffs. Packaging of food materials requires the preservation of food. There are various methods for the treatment of food to preserve them and irradiation treatment of food is one of them. It is the most common and the most harmless method for the food preservation as it does not alter the necessary micronutrients of food materials. Although irradiated food doesn’t cause any harm to the human health but still the quality assessment of food is required to provide consumers with necessary information about the food. ESR spectroscopy is the most sophisticated way to investigate the quality of the food and the free radicals induced during the processing of the food. ESR spin trapping technique is useful for the detection of highly unstable radicals in the food. The antioxidant capability of liquid food and beverages in mainly performed by spin trapping technique.
Phenomics assisted breeding in crop improvementIshaGoswami9
As the population is increasing and will reach about 9 billion upto 2050. Also due to climate change, it is difficult to meet the food requirement of such a large population. Facing the challenges presented by resource shortages, climate
change, and increasing global population, crop yield and quality need to be improved in a sustainable way over the coming decades. Genetic improvement by breeding is the best way to increase crop productivity. With the rapid progression of functional
genomics, an increasing number of crop genomes have been sequenced and dozens of genes influencing key agronomic traits have been identified. However, current genome sequence information has not been adequately exploited for understanding
the complex characteristics of multiple gene, owing to a lack of crop phenotypic data. Efficient, automatic, and accurate technologies and platforms that can capture phenotypic data that can
be linked to genomics information for crop improvement at all growth stages have become as important as genotyping. Thus,
high-throughput phenotyping has become the major bottleneck restricting crop breeding. Plant phenomics has been defined as the high-throughput, accurate acquisition and analysis of multi-dimensional phenotypes
during crop growing stages at the organism level, including the cell, tissue, organ, individual plant, plot, and field levels. With the rapid development of novel sensors, imaging technology,
and analysis methods, numerous infrastructure platforms have been developed for phenotyping.
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 use of Nauplii and metanauplii artemia in aquaculture (brine shrimp).pptxMAGOTI ERNEST
Although Artemia has been known to man for centuries, its use as a food for the culture of larval organisms apparently began only in the 1930s, when several investigators found that it made an excellent food for newly hatched fish larvae (Litvinenko et al., 2023). As aquaculture developed in the 1960s and ‘70s, the use of Artemia also became more widespread, due both to its convenience and to its nutritional value for larval organisms (Arenas-Pardo et al., 2024). The fact that Artemia dormant cysts can be stored for long periods in cans, and then used as an off-the-shelf food requiring only 24 h of incubation makes them the most convenient, least labor-intensive, live food available for aquaculture (Sorgeloos & Roubach, 2021). The nutritional value of Artemia, especially for marine organisms, is not constant, but varies both geographically and temporally. During the last decade, however, both the causes of Artemia nutritional variability and methods to improve poorquality Artemia have been identified (Loufi et al., 2024).
Brine shrimp (Artemia spp.) are used in marine aquaculture worldwide. Annually, more than 2,000 metric tons of dry cysts are used for cultivation of fish, crustacean, and shellfish larva. Brine shrimp are important to aquaculture because newly hatched brine shrimp nauplii (larvae) provide a food source for many fish fry (Mozanzadeh et al., 2021). Culture and harvesting of brine shrimp eggs represents another aspect of the aquaculture industry. Nauplii and metanauplii of Artemia, commonly known as brine shrimp, play a crucial role in aquaculture due to their nutritional value and suitability as live feed for many aquatic species, particularly in larval stages (Sorgeloos & Roubach, 2021).
2. WHAT IS GERMPLASM ?
The sum total of all the genes present in a crop and its related
species constitutes its germplasm. It is ordinarily represented by a
collection of various strains and species.
Plant germplasm is the genetic source material used by the plant
breeders to develop new cultivars.
They may include:
Seeds
Other plant propagules (Which can be grown into mature plant) are:
Leaf
Stem
Pollen
Cultured cells
Germplasm provides the raw material (genes) which the breeder
used to develop commercial crop varieties. Therefore, germplasm is
the basic indispensable ingredient of all breeding programmes. Thus a
great emphasis is placed on collection, evaluation and conservation of
germplasm.
3. The conservation of germplasm involves the preservation of the
genetic diversity of a particular plant or genetic stock. It can be used at
any time in future.
It is important to conserve the endangered plants or else some of the
valuable genetic traits present in the existing and primitive plants will be
lost.
Some crops produce recalcitrant or short lived seeds.
Similarly, in case of clonal crops seeds are not the best material to
conserve due to their genetic heterogeneity and unknown worth. Their
genes need to be conserved.
The roots and tubers loose viability rapidly. Their storage requires large
space, low temperature and is expensive. In addition, materials
modified by genetic engineering may some, times be unstable. Such
materials are needed to be conserved intact for future .A global
organization- International Board of Plant Genetic
Resources (IBPGR) has been established for germplasm conservation
and provides necessary support for collection, conservation and
utilization of plant genetic resources through out the world.
APPLICATIONS OR SIGNIFICANCE OF
GERMPLASM CONSERVATION
4. IN VITRO METHOD FOR GERMPLASM CONSERVATION
In vitro method employing shoots, meristems and embryos are
ideally suited for the conservation of germplasm.The plant with
recalcitrant seeds and genetically engineered can also be
preserved by this in vitro approach.
There are several advantages associated with in vitro
germplasm conservation:-
Large quantities of material can be preserved in small space.
The germplasm preserved can be maintained in an
environment free from pathogens.
It can be protected against the nature’s hazards.
From the germplasm stock large number of plants can be
obtained whenever needed.
5. THERE ARE THREE MAIN APPROACHES FOR THE
IN VITRO CONSERVATION OF GERMPLASM
CRYOPRESERVATION
COLD STORAGE
LOW –PRESSURE AND LOW OXYGEN –STORAGE
Other approaches:-
Slow Growth Cultures
Desiccated Somatic Embryos (SE) and Artificial Seeds
DNA Clones
7. CRYOPRESERVATION
Cryopreservation (Greek,krayos-frost)literally means in the
frozen state. The principle involved in cryopreservation to bring
the plant cells and tissue cultures to a zero metabolism or
non-dividing state by reducing the temperature in the
presences of cryoprotectants (DMSO (dimethyl sulfoxide),
glycerol, ethylene, propylene, sucrose, mannose, glucose,
praline, acetamide etc ).
CRYOPRESERVATION broadly means the storage of germplasm at
very low temperature using :-
Over solid carbon dioxide(at 79°C)
Low temperature deep freezer(at -80°C)
Using vapour nitrogen (at- 150°C)
In liquid nitrogen( at -196°C)
8. Among these, the most commonly used cryopreservation
is by employing liquid nitrogen. At the temperature of
liquid nitrogen(at -196°C), the cell stay in a completely
inactive state and thus can be conserved for longer period.
Infact cryopreservation has been successfully applied for
germplasm conservation of some plant species e.g
rice,wheat,peanut,sugarcane,coconut.
The technique of freeze preservation is based on the
transfer of water present in the cells from a liquid to
solid state. Due to the presence of salts and organic
molecules in the cells,the cell water requires much more
lower temperature to freeze(even up to -68°C) compared to
the freezing point of pure water(around 0°C). When stored
at low temperature, the metabolic processes and
biological deteriorations in the cells/tissues almost come
to standstill.
MECHANISM OF CRYOPRESERVATION
9.
10. TECHNIQUE OF CRYOPRESERVATION
The cryopreservation of plant cell culture followed the
regeneration of plants broadly involves the following
stages.
Development of sterile tissue culture.
Addition of cryoprotectant and pretreatment.
Freezing
Storage
Thawing
Reculture
Measurement of survival/viability
Plant regeneration
11. 1.DEVELOPMENT OF STERILE TISSUE CULTURE
The selection of plant species and the tissue with particular
references to the morphological and physiological
characters largely influences the ability of the explants to
survive in cryopreservation.Any tissue from a plant can be
used for cryopreservation
e.g.meristems,embryos,endosperm,ovules,seeds,culture
plants.
2.ADDITION OF CRYOPROTECTANT
Cryoprotectant are the compound that can prevent the
damage caused to cells by freezing or thawing.There
are several cryoprotectant which include:
(DMSO, GLYCEROL, ETHYLENE, PROPYLENE,
SUCROSE, MANNOSE, GLUCOSE…..)
12. 3.FREEZING
The sensitivity of the cells to low temperature is visible and
largely depends on the plant species.Four different types of
freezing are used.
Slow freezing method
Rapid freezing method
Stepwise freezing method
Dry freezing method
4.STORAGE
Maintenance of the frozen cultures at the specific temperature is
as important as freezing.In general,the frozen cells/tissues are
kept for storage at temperature in the range of -72 to-196°C.
Storage is ideally done in liquid nitrogen refrigerator at -
150°C in the vapour phase,or at -196°C in the liquid phase.
The ultimate objective of storage is to stop all the cellular
metabolic activities and maintain their viability .For long term
storage temperatue at -196°C in liquid nitrogen is ideal.
14. 5.THAWING
Thawing is usually carried out by plunging the frozen
sample in ampoules into the warm water (temp 35-
45°C) bath with vigorous swirling. By this approach,rapid
thawing(at the rate of 500-750°Cmin-1)occurs, and this
protects the cell from the damaging effects ice crystal
formation.
As the thawing occurs (ice completely melts) the ampoules
are quickly transferred to a water bath at temperature 20-
25°C. This transfer is necessary since the cells get
damaged if left for long in warm(35-45°C) water bath.
6.RECULTURE
In general thawed germplasm is washed several times to
remove cryoprotectant. The material is then cultured in a
fresh media.
15. 7.PLANT REGENERATION
The ultimate purpose of cryopreservation of
germplasm is to regenerate the desired plant. For
appropriate plant growth and regeneration,the
cryopreserved cell/tissue have to be carefully
nursed and grown. Addition of certain growth
promoting substances ,besides maintenance of
appropriate environmental conditionis often
necessary for successful plant regeneration.
16. COLD STORAGE
Cold storage is a slow growth germplasm conservation
method. It conserves the germplasm at a low and non-
freezing temperature (1- 9°C). The growth of the plant
material is slowed down in cold storage in contrast to
complete stoppage in cryopreservation. Thus it prevents
cryogenic injuries. Long term cold storage is simple, cost
effective. It yields germplasm with good survival rate. Virus
free strawberry plants could be preserved at 10°C for about
6 years. Several grape plants have been stored for over 15
years by using a cold storage at temperature around 9°C
and transferring them in the fresh medium every year.
17. LOW PRESSURE AND LOW OXYGEN STORAGE
In low- pressure storage, the atmospheric pressure
surrounding the plant material is reduced. In the low
oxygen storage, the oxygen concentration is reduced. The
lowered partial pressure reduces the in vitro growth of
plants. In the low-oxygen storage, the oxygen
concentration is reduced and the partial pressure of
oxygen below 50 mmHg reduces plant tissue growth.
Due to the reduced availability of 02, and reduced
production of CO2, the photosynthetic activity is reduced. It
inhibits the plant tissue growth and dimension. This method
has also helped in increasing the shelf life of many fruits,
vegetables and flowers.
18. SLOW GROWTH CULTURES
Slow-growth of plantlets in-vitro provides an attractive
alternative to freeze preservation of germplasm as it is
simpler, cheaper and very effective. Slow growth may be
achieved by maintaining the plantlets either at a low
temperature (4-9°C or Ca. I5°C) or on a medium having
high osmotic concentration (e.g., 20% sorbitol or
sucrose) or both.
In addition, the nutritional status of the medium may be
lowered to restrict the growth of plantlets. Under the
conditions of slow-growth, cultures may be attended to only
once in several months. Its subculture may, be necessary
only after long periods, once every 236 months.
19. The slow-growth approach is being utilized for germplasm
conservation of specified root, tuber and tree species by the
NBPGR, New Delhi. A National Facility for Plant Tissue
Culture Repository has been created for this purpose. It has
so far developed the slow-growth protocols for ginger,
garlic, banana, sweet potato, etc
The techniques for desiccation of SE s and for production
of desiccated, artificial seeds are now becoming available.
The desiccated SEs and artificial seeds can be stored at
low (4°C) or ultralow (-20°C) temperatures for prolonged
periods in a manner similar to zygotic seeds..
Desiccated Somatic Embryos (SE) and
Artificial Seeds
20. DNA CLONES
Germplasm can also be conserved in form of DNA
segments cloned in a suitable vector, e.g., cosmids,
phasmids or YACs. The technique is highly
sophisticated, technically demanding and expensive.
It is likely to be used for conservation of valuable .genes
or DNA segments from threatened species. It can also be
used for the conservation of the entire genomes of
various germplasm lines of different species.
21. LIMITATION OF IN-VITRO GERMPLASM
CONSERVATION
The expensive equipment needed to provide controlled
and varible rates of cooling/warming temperatures can
however be a limitation in the application of in vitro
technology for large scale germplasm conservation.
Formation of ice crystal inside the cell should be
prevented as they cause injury to the cell.
Sometimes certain solutes from the cell leak out during
freezing.
Cryoprotectant also effect the viability of cells.
22. STORAGE STABILITY USING CRYOPRESERVATION: A CASE STUDY IN
PAPAYA [2011]
Ashmore, S.E.Drew, R.A.Kaity, A.
Abstract
Ex situ conservation of Carica papaya and its crop wild relatives has been the subject of ten
years of research in our laboratory. This paper summarises the achievements and key
findings of this work. Both clonal and seed materials have been investigated to allow storage
of elite clonal material as well as maximum genetic diversity. Clonal material from several
genotypes has been successfully stored using micropropagation followed by shoot tip
cryopreservation, with >70% regeneration after 12 months storage in LN. The
regeneration and acclimatisation of cryopreserved plants has been achieved for field testing
and observations of morphological characteristics, including flowering and fruiting, have not
detected any changes associated with cryopreservation. Genetic stability following shoot tip
cryopreservation has also been monitored using both RAF (Random Amplified DNA
Fingerprinting) and AMP (Amplified DNA Methylation Polymorphism) techniques. Whilst both
RAF and AMP changes were observed following cryopreservation, these were not associated
with cryopreservation per se and did not correlate with any modification in plant morphology.
Investigations of papaya seed storage indicate that seed is essentially orthodox, but
dormancy breaking treatments were required for germination. However, germination post
storage for 12 months was >68% at ultra-low temperatures (LN), but <5% at
conventional seed storage temperatures (-20°C). This is in line with recent evidence
that cryopreservation enhances storage stability and longevity, even in orthodox seeds,
when compared with standard seed bank approaches. This highlights the importance of
cryopreservation for the long-term management of genetic resources of both clonal and seed
genetic resources of papaya.
23. CRYOPRESERVATION OF ORCHID GENETIC RESOURCES BY DESICCATION:
A CASE STUDY OF BLETILLA FORMOSANA
Rung‐Yi Wu, Shao‐Yu Chang, Ting‐Fang Hsieh, Keng‐Chang Chuang,IeTing, Yen‐Hsu Lai
and Yu‐Sen Chang
Abstract
Many native orchid populations declined yearly due to climate change.
This resulted in some wild orchids being threatened. In order to maintain
the orchid genetic resources, development of proper methods for the
long-term preservation is urgent. Low temperature or dry storage
methods for the preservation of orchid genetic resources have been
implemented but are not effective in maintaining high viability of certain
orchids for long periods. Cryopreservation is one of the most acceptable
methods for long-term conservation of plant germplasm. Orchid seeds
and pollens are ideal materials for long-term preservation (seed
banking) in liquid nitrogen (LN) as the seeds and pollens are
minute, enabling the storage of many hundreds & thousands of
seeds or pollens in a small vial, and as most species germinate
readily, making the technique very economical. This article describes
cryopreservation of orchid genetic resources by desiccation and a case
study of Bletilla formosana.