Anther culture is a technique where anthers are excised from flower buds and cultured to produce haploid plants. The first report of haploid tissue from anther culture was in 1964-1966 in Datura pollen grains. Over 250 species have been produced through anther culture, most commonly in families like Solanaceae, Cruciferae, and Poaceae. Anthers go through various pathways during culture, such as equal division to form two daughter cells or formation of vegetative and generative cells. Anther culture is useful for eliminating lethal genes and shortening breeding time to produce superior hybrids.
history
Lampe & Mills (1933) were the first to report the proliferation of immature endosperm tissue of Maize, grown on medium containing extract of potato.
La Rue (1947) observed that in nature, in maize , the pericarp ruptured & the endosperm exhibited a white tissue mass.
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
history
Lampe & Mills (1933) were the first to report the proliferation of immature endosperm tissue of Maize, grown on medium containing extract of potato.
La Rue (1947) observed that in nature, in maize , the pericarp ruptured & the endosperm exhibited a white tissue mass.
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
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
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.
Anther and pollen culture is the production of haploid plants exploiting the totipotency of microscope and the occurrence of single set of chromosome in microscope.
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
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.
Anther and pollen culture is the production of haploid plants exploiting the totipotency of microscope and the occurrence of single set of chromosome in microscope.
Pollen or microspore culture is an in vitro technique by which the pollen grains are squeezed out aseptically from the intact anther and then cultured on nutrient medium.
the microspores, without producing male gametes, develop into haploid embryoids or callus tissue that give rise to haploid plantlets by embryogenesis or organogenesis.
this helps to acquire the whole knowledge about anther and pollen culture.
The main objective of anther culture is production of haploid plants, which are useful in rapid production of homozygous lines and also production of doubled haploids (DH). Anther culture or pollen culture provides certain advantages over conventional breeding strategies like production of homozygous lines which takes 5-7 generations in case of conventional breeding strategies.
The Yamada et al., and Guha and Ramachandra are first to produce the haploid plants through anther culture in 1936 and 1964 respectively.
N6 is the popular media for used for anther culture.
This presentation deals with procedure, applications, limitations associated with the Anther culture.
Cotton Floral biology and breeding techniques.pdfVikraman A
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From the forgoing presentation, it can be concluded that breeding characters viz., flowering period, inflorescence, time of flower opening, time of anther dehiscence, time of stigma receptivity, pollinating agent ,time of visitor of pollinating agent and fruit set (%) in tropical species are required to be studied as they are vital for any improvement and eco-environmental planning purposes. It also throws light on how species adopts itself along with the phenomenon of speciation and reproductive isolation. From these characters we can introduce new variety which is essential for further evaluation and also the identification of the interactions between biological factors, such as animal, plant species, and non-biological factors, like temperature, RH, rain and wind, helps us to elaborate management and conservation plans for the ecosystems of the planet, which have become more and more necessary due to highly increased rate of deterioration of different ecosystems during the last few decades.
Much faster rates of growth can be induced in vitro than by traditional means.
Multiplication of plants which are very difficult to propagate by cuttings or other traditional methods.
Production of large numbers of genetically identical clones in a short time
Seeds can be germinated with no risk of damping off/ predation.
Under certain conditions, plant material can be stored in vitro for considerable periods of time with little or no maintenance
Tissue culture techniques are used for virus eradication, genetic manipulation, somatic hybridization and other procedures that benefit propagation, crop improvement, and basic research.
By means of tissue culture it is possible to produce pathogen free plantlets by mass multiplication in a very limited amount of area from a very small sterile part of a mother plant. This method is also used to produce/ multiply plants that are to be transported across national border and so for their faster multiplication.But the establishment of a tissue culturing unit needs huge financial investments, skilled labors/technicians and required areas for its establishment are major constraints. Plant tissues grow and multiply in the labs only when there is an uncompetitive, growing condition with uninterrupted supply of nutrients.
Medium:
It contains all the elements that contribute the required nutrients that aid to the growth of the tissues; it is in liquid state or semi-solid in nature. The tissues are grown on the media. It consists of 95% of water, major and minor nutrients, plant growth hormones, vitamins, sugar rich compounds and chelating agents.
Totipotency:
It is the ability of a tissue or an organ of a plant to produce the whole plant, under the optional laboratory conditions and this is called as Totipotency. This is the baseline over which plant tissue culture relies upon.
Callus Culture:
When the cells divide into an undifferentiated mass it is called as callus. Any part of a plant can be used to produce the calli. It may be a stem, leaf, meristem or any other part. It is used to produce variations among the plantlets.
Suspension culture:
The callus produced from the explants are grown on nutrient solutions (that are semi solid) for a period of time and they are induced to produce plants with new traits.
Embryo Culture:
The method of culturing mature and immature embryos in media is called embryo culture. By this method, it is possible to produce plants from dormant seeds and seeds with metabolites that inhibit germination. This method is very important in crop improvement programs.
Somatic Embryogenesis:
When the plants are grown on nutrient media, calli are formed. When these calli are subjected to growth in cytokinin medium, somatic embryos are formed. They are circular, elongated,
Single cell culture
• As stated earlier, cells derived from a single cell through mitosis constitute a clone and the process of obtaining clones is called cloning (asexual progeny of a single individual make up.
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Operation “Blue Star” is the only event in the history of Independent India where the state went into war with its own people. Even after about 40 years it is not clear if it was culmination of states anger over people of the region, a political game of power or start of dictatorial chapter in the democratic setup.
The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
Safalta Digital marketing institute in Noida, provide complete applications that encompass a huge range of virtual advertising and marketing additives, which includes search engine optimization, virtual communication advertising, pay-per-click on marketing, content material advertising, internet analytics, and greater. These university courses are designed for students who possess a comprehensive understanding of virtual marketing strategies and attributes.Safalta Digital Marketing Institute in Noida is a first choice for young individuals or students who are looking to start their careers in the field of digital advertising. The institute gives specialized courses designed and certification.
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http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
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2. Definition
• Anther Culture: It is an artificial technique by which the
developing anthers at a precise and critical stage are excised
aseptically from unopened flower bud and are cultured on a
nutrient medium where the microspores within the cultured
anther develop into callus tissue or embryoids that give rise
to haploid plantlets (formation of haploid plants) either
through organogenesis or embryogenesis.
• The first report of haploid tissue from anther culture was
in 1964-1966 in pollen grains of Datura by Maheshwari
and Guha.
• Production of haploids reported in about 250 species,
Solanaceae, cruciferae, gramineae/Poaceae are most
common
• Anther/pollen culture is referred as ANDROGENESIS (the
male gametophyte (microspore or immature pollen)
produces haploid plant)
4. Haploids are useful because:
1. They carry only one allele (two or more
alternative forms of a gene) of each gene. Thus
any recessive mutation or characteristic are
apparent(CLEARLY VISIBLE).
2. Plants with lethal genes are eliminated from
the gene pool.
3. One can produce homozygous diploid (When
an individual has two of the same allele)
4. Production of haploids shorten the time for
inbreeding for superior hybrid genotypes.
6. 1. Pathway I
2. Pathway II
3. Pathway III
4. Pathway IV
5. Pathway V
7. • The microspores divide
by an equal division and
two identical daughter
cells developed.
• Vegetative and
generative cells are not
distinctly formed in the
pathway.
• Example: Datura innoxia
8. • The division of uninucleate
microspores is unusual,
resulting in the formation of
vegetative and generative cell.
• The sporophyte arises through
further division in the
vegetative cell and generative
cell does not divide.
• Examples: Nicotiana tabacum,
Hordeum vulgare, Triticum
aestivum.
9. • The uninucleate
microspore undergoes
a normal division but
pollen embryos are
formed from
generative cell alone.
• The vegetative cell
does not divide.
• Examples: Hyoscyamus
niger
10. • Both generative and
vegetative cell divide further
to the development of
sporophyte.
• Examples: Datura metal,
Atropa belladona, Datura
innoxia(occasionally).
PATHWAY V:
• In Brassica napus(Cruciferae),
1st division is symmetric and
the pollen embryos develop
the vegetative cell.
12. Protocol (Nicotiana tabacum)
1. Collect the flower buds of Nicotiana tabacum at the onset
of flowering. Select the flower bud of 17-22 mm in length
when the length of the sepals equals that of the petals.
Reject all flower buds which are beginning to open.
2. Transfer the selected flower buds under the laminar airflow.
Each flower bud contains five anther and these are normally
surface sterile in closed buds. The flower buds are surface
sterilized by immersion in 70% ethanol for 10 seconds
followed immediately by 10 minutes in 20% sodium
hypochlorite. They are washed three times with sterile
distilled water. Finally transfer the buds to sterile Petridish.
3. To remove the anthers, slit the side of the bud with a sharp
scalpel and remove them, with a pair of forceps, place the
five anthers with the filaments to another Petridish. The
filaments are cut gently. Damaged anthers should be
discarded.
13. 4. Anthers are placed on agar solidified basal MS or White or Nitsch
and Nitsch medium.
5. The culture is kept initially in dark. After 3-4 weeks, the anthers
normally undergo pollen embryogenesis and haploid plantlets
appear from the cultured anther. In some cases, anther may
undergo proliferation to form callus tissue which can be induced
to differentiate into haploid plants.
6. At this stage the cultures are incubated at 24-28°C in a 14 hrs day
light regime at about 2000 lux.
7. Approximately 50mm tall plantlets are freed from agar by gently
washing with running tap water and then transferred to small pots
containing autoclaved potting compost. Cover each plant with
glass beaker to prevent desiccation and maintain in a well-lit-
humid green house. After some week, remove the glass beaker
and transfer the plant to larger pots when the plants will mature
and finally flower.
14.
15. A) Swollen anther (after 2 weeks in culture).
B) Cristal, after 6 weeks in culture, producing callus masses at both sides.
C)After 6 weeks in culture, producing both callus masses and embryos (arrowhead).
D) After 6 weeks in culture, producing only embryos (arrowheads).
E) Regenerating in vitro plantlet
F) fully acclimated plant
G) Shifted to a pot
16. 1) Genotype of donor plant :
• Determine the frequency of pollen plant
production.
• Eg. In Hordeum vulgare each genotype differs
with respect to androgenic response in anther
culture.
• High responsive anthers should be taken.
17. 2) Anther wall factor :
• Act as conditioning factors and promote culture
growth.
• Report: glutamine alone or in combination with
serine and myoinositol could replace the anther wall
factor.
3) Stage of pollen:
• Development stage of pollen varies with species.
• Before/after 1st pollen mitosis- Datura, tobacco,etc.
4) Physiological status of donor plant:
a) Grown under best environmental conditions with
good anthers.
b) Flowers obtained at the beginning of flowering
season are highly responsive.
18. 5) Pretreatment of anthers :
• Appropriate treatment required for good success
of haploid production(depend on donor plant
species).
6) Temperature influence:
a) Induction of androgenesis is better if stored at
low temperature prior to culture, e.g. maize , rye.
b) Pre-treatment of anthers at higher temperature
stimulates androgenesis
19. Cold Treatment (3 to 5° C) Enhances
Symmetric Division of Microspores or
Division of Vegetative Nuclei
20. 6) Effect of light:
• Pre-treatment of anthers at elevated temperatures(
3°C) stimulate androgenesis in some Brassica and
Capsicum.
7) Culture medium:
• It vary with the genotype and age of the anther.
• Culture maintained on an auxin medium for longer
period develop a friable callus.
• A compound related to auxin namely 2,3,5-
triiodobenzoic acid (TIBA) gives +ve result at low
concentration.
• Incorporation of activated charcoal/2-chloroethyl-
phosphate stimulates androgenesis in some
systems.
21. Applications
• Utility of Anther and Pollen Culture for Basic
Research
• Simple
• Less time consuming
• Responsive
• Mutation Study
• Use of Haploids for Cryogenic Study
• For Plant Breeding and Crop Improvement
• Application of Haploid Culture for
Horticultural Plants
• For study of Secondary metabolites Content
22. Requires skill to remove anthers without
causing damage.
Not much successful in case of cereal crop.
Risk of chimera and callus formation from
anther wall.