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HAPLOID CULTURE
IN SEARCH OF IMPROVED
TRAITS
Presented by
Husna Subhan
Depatment of Biotechnology and
Allied sciences, Haldwani 1
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Haploids
Haploid condition states a single set of
chromosome number of a organism.
There are generally two types of haploids Monohaploids: Posses half the chromosome
from a diploid specie.
Polyhaploids: Posses the half the chromosomenumber from a polyploid species.
It was in 1964 Guha and Maheshwari induced
in vitro haploid calli of Dhatura Inoxa.2
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Apomixis or parthenogenesis are the method
that lead to the development of haploids.
Apomixis states a embryo development from a
unfertilized embryo.
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Methods to produce haploids
There are generally two common strategies to
develop haploids
A. androgenesis: Androgenesis refers to theproduction of haploids from the male
gametophyte.
B. gynogenesis: Refers to the use of female
gametophyte to raise the haploids.
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Androgenesis is the most commonly use
method.
Strategy involves :A. culturing the pollens
B. culturing the anthers.
The basic principal underlying the strategy isthe diversion of pollens fate which is gamete
formation ,into a mass of cell( haploid callus).
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ANTHER
Androecium is male reproductive structure composed ofmany stamens.
Each stamen contains two parts, a long filament and anther .Each anther have two lobes called anther lobes or
microsporangium.
Microsporangium posses many microspores in it that posses
haploid sets of chromosomes in it.
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ANGIOSPERM FLOWER : TRANSVERS
SECTION
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TRANSVERSE SECTION : ANTHER
SHOWINGMICROSPORES
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WHATIS ANTHER CULTURE
Anther culture involves culturing the anther ina medium in a media which lead to productionof haploid calli.
Anther is sterilized and is then plated inmedium.
Care should be taken so that the anther wall
should not get damaged.After a period of 15 days the pollen derived
haploid callli protrudes out from the antherwall.
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WHY ANTHER CULTURE
Anther culture is generally preferred over the
pollen culture.
Anther culture is preferred because:A. Pollens are small structures and hard to
handle.
B. Anther are easy to handleC. the anther wall possess positive influences
on the development of pollen.
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BASIC STRATYGEY
Anther is storehouse of large numbers of
haploid microspores.
These microspores are destined for the futurefertilization.
Anther is placed in such a media that divert
its normal pathway i.e. production of pollen
tube, fertilization, towards the callus
formation.
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Photographs from thesis
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POLLEN CULTURE
The anther is crushed and pollens so comes out from
the anther.
Care is generally taken that the pollen nucleus should
be in binucleated stage, because they are found to be
more responsive.
Pollen culture involves direct culturing of
binucleated pollens into the media.
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WHY POLLEN CULTURE
Uncontrolled effects of anther wall can be
eliminated.
Androgenesis can be traced. Microspores are idle to for the uptake
,transformation and mutagenic stratigies.
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Pretreatment ofexplant
Generally pretreatment is done so to divert the
normal biological development of anther.
There are generally above methods ofpretreatment-
A. Cold pretreatment.
B. Hot pretreatmentC.Chemical treatment- Involves use of 2-chloro
ethylphosphonic acid (ETHREL).
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These calluses are screened for a desired trait.
These calluses are placed then on regeneration media
leading to the formation of haploid plants.
Regenerated plants so produced are haploid.
These plants parts then are placed in colchicine .
Colchicine(0.4%) promotes diploidisation by
promoting chromosome duplication without itsredistribution.
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MEDIA COMPOSITION
Media composition plays crucial role in
callusing.
Media composition varies among thegenotypes
N6 media(Chu 1978) is the most commonly
and is most specifically used for anther culture.
Different growth regulators i.e 2,4d,kinetin
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APPLICATIONS OF HAPLOID
CULT
URE Rapid generation of the pure varieties. Less time expense compaired to traditional
breeding. Maximum utilization of the variations resulted
from meiosis that are otherwise lost in nature.
Development of complete homozygous linesof desired character by fixing the traits.
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Important in studying the somatic cell
genetics.
Expression of recessive traits that areotherwise are unexposed in heterozygous
condition.
Plants of various ploidy levels have been
produced.
Generation of exclusively male plants.
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Continue..
In Asparagus plants (XY) , forming
exclusively male i.e supermale i.e YY that
are high yeilding.
In cytogenetic research.
Disease resistant varieties have been released.
Example. Q21681,Zong Hua 8 and 9 have
been found tolerent against the blast resistant.
Mutations can be induced in pollens and
succesively haploids could be generated .
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In genome mapping, as the identification of
marker is very much secured and intermediate
phenotype expressions are minimized to zero.
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Problems associated with
Haploid culture
High expertisation required.
Chances of diploid tissue formation are
involved. Little chances are there when we can isolate a
haploid because they are easy outgrown by
their competating diploid cells.
Haploids not necessarily give rise to
homozygotes
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Factors effecting haploid culture
Genotype.
Physiological status of plant.
Stage of pollen. Pretreatment.
Type of sugar use in media,sucrose have been
use but Maltose had shown some goodpromises.(Thorpe,et al2000)
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INDIAAND PRESENT STATUS
OF H
APLOI
D CULT
URE China have been leading the way towards thepresent context of haploid culture.
Till now other then BT, much progress haventbeen done.(Annual report 2010)
In India no variety with the tolerent to abiotic
stress have been released.
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Wheat- Lunghua 1, Huapei 1, Delibab, Abitus,
Florin, Zing Hua 2 and 3, Non Hua 11, Aya.
Rice- Hua Yu-1, Zhong Hua 9, Zhe Keng 66,Hirohonami.
Tobacoo- Tan Yuh 1, Tanyu 3, Hai Hua 30.
(From indian journal of crop physilogy)
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Variations derived from haploid
culture
(nationmaster.com)
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THANK YOU
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