welcome

DOCTORAL SEMINAR –II (GP692)

Topic: Diploid pollen and its application

in crop improvement

PRESENTED BY
Priyanka Kumari
Ph.D Student
Registration no. AP/BAU/6043/2016

DEPARTMENT OF GENETICS AND PLANT BREEDING

BIRSA AGRICULTURAL UNIVERSITY, KANKE, RANCHI - 834006
 Diploid ??
• If the cell contain two paired set of
chromosome then its is called diploid and it is
denoted by 2n.
• Normally cells are found in paired form .
 Diploid pollen: introduction
• Diploid pollen (unreduced gamete) which is developed through the
chromosomal abberations i.e. non-disjuction of sister chromatids during
meiosis (unreduced gamete) replication of DNA but no cytokinesis which is
known as  endoreplication.
 Schematic mechanism of 2n gametes formation
through abnormal meiotic division in plants

In case of plants:
Meiosis -I reffered as first division restitution and
Meiosis –II reffered as second division restitution
Resulting in the production of abnormal gametes reffered as
unreduced gametes
 Cont…

• FDR pollen are more important because of highly heterozygous 2n

gametes formed . (Bretagnolle & Thompson, 1995)
• In SDR, chrosomosome segregation is random as normal case.
• Although FDR superiority for all agronomic characterstic could not be
confirmed.
• By using molecular technique such as AFLP, RAPD or isozyme analysis
heterozygosity transmitted through 2n gametes can be calculated.
• Ex- Solanum spp. Indicate that FDR gametes transmit roughly 70-80% of
the parental heterozygosity,whereas about 30-40% through SDR. This
value also reported in othe crops:roses,ryegrass,begonia.
(Dewitte et.al.,2010a)
 Cytological abnormalities responsible for 2n gametes production as
described in recent studies
cytological 2n 2n mode species Refrences
abnormalities pollen egg
Premeiotic
doubling + Lolium. perenne
Secale cereale
Wagenvoort et al.,
1990
Lelley et al., 1987
Synaptic + + FDR Solanum spp. Watanabe & Peloquin,
mutant and 1993
SDR

+ FDR Trifolium pratense Parrott & Smith, 1984

Adiantum Rabe & Haufler, 1992
pedatum
Lack of + Manihot Nassar, 1992
MI spindle esculenta

Omission of + FDR Arachis spp. Simpson & Davis, 1983

meiotic I
division
 Importance of diploid pollen

• It is considered as the driving force behind the formation of polyploidy in

nature. (Bretagnolle & Thompson, 1995)
• Harlan & De Wet (1975) showed that almost all plant species produce
diploid pollen in some frequencies and found the importance of diploid
pollen in the origin of polyploidy.
• The use of diploid pollen in plant breeding, very useful for crop
improvement (Ramanna & Jacobsen, 2003).
• 2n pollen an effective and efficient way to transmit genetic diversity
(allelic variation) from wild variety to cultivated forms, including both
valuable qualitative and quantitative traits (Peloquin et al., 1999).
• transmit parental heterozygosity traits to offspring in a large ratio and helps
in adaptation.
• Thus it opens new avenue for the plant breeder .
 Cont…..

 It is a generation-specific phenomenon that is expressed in F1 individuals,

but not in the parental species nor in the F2 and subsequent generations.
 The fitness of F1 individuals is greatly improved by unreduced
gamete production.. 
 If two unreduced gametes unite, a fertile polyploid hybrid may form-either
autopolyploid (fertilization within species) or allopolyploid (fertilization
between species).
 Cont…..

Unreduced gametes fundamental role:

 asexual reproduction of plants,
 a speciation of flowering plants,
 and restoration of fertility of distant hybrids used to produce
new varieties with breeding-valuable traits.
 Cont……

• Different approaches have been employed for 2n-pollen production

including :-
 interspecific hybridization,
 manipulation of environmental factors and
 treatment with nitrous oxide, trifluralin, colchicine, oryzalin and other
chemicals. These chemicals can act as a stimulus to produce viable 2n
pollen. (Younis et al., 2013).
 Diploid pollen development and its use in crop
improvent
1) Spontaneous method:-
Self fertilization – Plants can combine their own chromosome after non-
disjuction.

• for example:-
 In cabbage tetraploid production takes pace by natural sexual mating of 2n
pollen. Which is also known as autopolyploidy.
 Artificial method

 interspecific hybridization.
 manipulation of environmental factors and
 treatment with nitrous oxide, trifluralin,
colchicine, oryzalin and other chemicals.
These chemicals can act as a stimulus to
produce viable 2n pollen. (Younis et al., 2013).
interspecific hybridization,
2. Induction method:- By treating the plant parts through chemicals
such as-: Through colchicine (C22H25O6N) treatment . Other chemicals- N20
(laughing gas), Acenaphthene, 8-hydroxyquinoline.
• Theses chemicals are used to double the chromosome artificially
 Mechanism of colchicine in DIPLOIDISATION

• Acts as inhibitor of spindle formation and causes duplication of homologus chromosomes

.
• Colchicine is readily soluble in cold water, alcohol and chloroform but less soluble in hot
water.
• Used as aqueous solution – relatively unstable. So the solution prepared fresh before
each application.
• Affects only dividing cells-applied to actively dividing meristematic cells repeatedly at
brief intervals.
• Methods of application:
• ◦ Seed treatment : Freshly prepared 0.2% solution applied for 1-10 days. ◦
• Seedling treatment : At very young stage, seedlings are inverted and only the young
shoots are dipped in colchicine solution & roots are protected;3-24hrs ◦
• Application to growing apices : 0.1-1% solution applied to shoot apex using a
dropper/cotton balls dipped in colchicine solution placed on shoot apices- repeated daily
0.5-1% colchicine +lemolin paste smeared on shoot apex-repeated 2-3 times per week.
 Cytological mechanism of 2n pollen formation

 Detailed cytological investigation revealed that the main

cytological mechanisms for production of 2n gametes are:
telophase-II failure leading to the formation of triads instead
of tetrad (one is unreduced).
Example:-triad formation at the end of telophase-II in Hyola 42
and Fornax which is variety of rapseed.(Brassica napus)
respectively.( Sheidai Masoud et.al.,2003)
 Advantages

1) The length of the 2n pollen tube was longer than that of 1n

pollen tube by about 300 μm after 8–24 h of pollination, which
is during the stage when the transmitting tissue enters the
style.
 the 2n pollen tube grew fastest at a speed of 100 μm/h to 131
μm/h, respectively whereas the pollen growth rate of n was the
slowest i.e. 17 μm/h to 31 μm/h, respectively.
 The rate of increase of 2n pollen tubes in the ovary is 33.90%
more.
 The pollen tubes of 2n pollen grow faster than those of 1n
pollens on the stigmas of Solanum tuberosum (Van Breukelen
EWM, 1982).
 Cont…..

 2n pollen indicates greater viability than that of 1n pollen.

• In most angiosperms, 2n gametes are disadvantaged against 1n
gametes in double fertilization, as is the case, for example, in
Populus tomentosa(species of deciduous flowering plants in
the family Salicaceae)
• the diameter of the natural 2n pollen was about 30% larger in
size than the 1n pollen. It is well known that pollen size
increases with increasing DNA . The pollen diameters of
various Phalaenopsis polyploids are positively correlated.
(Bretagnolle F. et.al., 1995)
 Cont…..

Fig:- 2n pollen, 2n pollen tube and size of pollen grain in 2n pollen

Cont…..
Cont…..
 Disadvantages
• while 2n pollen has a potential germination ability, their
germination process is relatively slow compared with 1n
pollen and is disadvantaged against 1n gametes when
participating in fertilization, which is the primary explanation
for the low breeding rate of triploids.
• These types of gametes have been shown to enable breeding
between genotypes with different ploidy, which is usually
unsuccessful because of uneven parental share in the
development of seeds (Kohler et al., 2010).
 How 2n pollen production takes place ??

• Unreduced gamete formation derived via abnormal meiotic

cell division which is an important approach to polyploidy
breeding.
 Cont…..

• Identification of efficient methods of inducing 2n-gamete

formation will help increase pollen germination of sterile
interspecific hybrids for inter-genomic recombination and
introgression breeding to develop new polyploid cultivars and
increase heterozygosity among plant populations.

(Younis et al., 2013).

 Cont…..

• In plants, two key groupings of 2n-gamete development have

been identified:-
• first division restitution (FDR) and
• second division restitution (SDR).
• In the FDR method, pairing and split up of homologous
chromosomes will not occur (univalent formation) during
meiosis-I(Yaqiong et al. 2007).
• while during the second division two sister chromatids of
homologous chromosome that move to opposite poles will be
produced during meiosis -II (Hermsen 1984).
 Cont…..

• The FDR gamete developed will maintain all of their parental

genes except cross-over fragments.
• During SDR, homologous chromosomes pair and separate
normally (bivalent formation) during meiosis-I, whereas
division of half-bivalent centromeres occurs during meiosis-II,
but the chromatids fail to move toward the poles Therefore,
half of parental chromosomes are present in the SDR gametes,
as occurs during normal gamete formation (random
combinations); however, there are two copies of these
chromosomes (Dewitte et al. 2010).
 Cont………..

 Lu et. al., (2013) considered high-temperature exposure ideal

for the induction of 2n female gamete in Populus and found
that plant exposure to high temperature induced[80% 2n male
gamete in Populus spp. (Kang et. al., 2000), whereas Wang et.
al., (2012) observed 66.7 % frequency of induced 2n female
gamete and found a pachytene to diplotene optimal stage for
induction of 2n-megaspore under high-temperature exposure
in Populus spp.
 In Rosa spp., the formation of 2n-gamete (*24.5 %) was
observed when they were exposed to a high-temperature
gradient (Pe´crix et. al., 2011).
 APPLICATION

• The application of 2n-gamete to obtain polyploid plants

can provide a better substitute for mitotic chromosome
doubling methods used in different breeding programs.
• The presence of 2n-gamete imparts the potential to develop
cultivars with greater ploidy and provides an opportunity for
the transfer of desired genes from 2n wild-type species to
polyploid cultivated species (Carputo et al., 2000).
• The presence of 2n-gamete can also enable the problem of F1
sterility to be overcome, provide a bridge to transfer hybrid
vigor, allow inter-genomic recombination and provide triploid
genotypes.
 Cont…

Cont…

• In plant breeding, the occurrence of polyploidy is of

great importance as produced plants are of superior
quality and have greater tolerance against biotic and
abiotic stresses.
• In potato, unreduced 2n-gametes have supported the
transfer of the Erwinia resistance gene from diploid
species such as Solanum chacoense and Solanum
tarijense to tetraploid cultivated species (Capo et al.
2002).
 Cont…
Cont…

• These unreduced gametes have also been efficiently used for

the introduction of pest and disease resistance as well as to
reduce the antinutrient factor such as to lower the cyanide
contents in cassava (Ogburia et al., 2002) and crop
improvement in alfalfa (Barcaccia et al., 2003).
• For the production of Triploid plants which facilitate the
production of seedless fruits in different crops such as citrus,
grapes, watermelons, cucumbers, etc., which improves the
overall quality of produce and fetches a premium price in the
market (Aleza et al., 2010).
• New polyploid plants either autopolyploids or allopolyploids
can be developed by manipulating 2n-gamete that has more
potential of genetic diversity and heterosis (Consiglio et
al.,2004).
Recent studies of the occurrence of 2n gametes in plants

Species Refrences

Adiantum pedatum Rabe & Hauffler, 1992.

Agropyron cristatiun Ray & Tokach, 1992.

Cyphoniandra betace Pringle & Murray, 1992o.

Dactylis glomerata De Haan et al., 1992;

Dendranthema grandiflora Bino et al., 1990

Hieracium pilosella Gadella, 1988

Iponiea batatas Jones, 1990.

Lilium spp. Van Tuyl et al., 1989.

Loliiim spp. Sala et al., 1989

 Cont…
Cont…

Species Refrences
Lotus tennis Negri & Veronesi, 1989
Malus X domestica Zhang et al., 1988
Manihot spp. Nassar, 1992.
Medicago spp. Veronesi et al., 1986; 1989, 1990
Secale cereale Lelley et al., 1987.
Solamim spp. Werner & Peloquin, 1987; 1991;
Trifolimn nigrescens Bullita & Smith, 1992.
T. pratense Parrott & Smith, 1984;
T. repens Builita & Smith, 1992.
Vaccinium spp. Shoemaker-Megalos &
Ballington,
1988;
Ortiz et al., 1992.
METHODS USED TO DETECT THE PRESENCE
AND THE FREQUENCY OF 2n GAMETES

The main methods

are:-
 Morphological screening to detect 2n pollen

• The most direct method of screening for 2n pollen involves the

examination of the range of size of pollen produced by an
individual. It is well known that cell volume increases with
increasing DNA content, which may, in turn, influence pollen
diameter (Southworth & Pfahler, 1992; Jansen & Den Nijs,
1993).
• Because of the relatively close correlation between 'giant, big'
pollen and 2n status (Maceira et al., 1992) the presence of giant
grains has frequently been used as an indication of the
production of 2n pollen (Freyre, Iwanaga & Orjeda, 1991;
Iwanaga, Freyre & Orjeda, 1991a).
 Cont…..

• Screening of 2n pollen, based on the size and shape of the

grain, is, however, only possible in species where differences
are known to occur between haploid and diploid pollen grains
(Myers, Gritton & Struckmeyer, 1984).
• In grasses, and probably wind pollinated species in general,
buoyancy is likely to be a major selection pressure limiting
pollen size. This may help explain the broad overlap, in size
distribution, of diploid and haploid pollen in wind-pollinated
species (Van Dijk &VanDelden, 1990)
• In such cases, only potential 2n pollen producers can be
identified, since a certain amount of the 2n pollen may be the
same size as haploid pollen. When the size of diploid pollen is
distinctly larger than that of the haploid pollen, its frequency
can be directly estimated (Orjeda et. al., 1990)
 Cont…..

• Based on the measurement of pollen diameters, mathematical

modelling of the frequency of occurrence of n, 2n and 'jumbo'
(possibly 4n) pollen has been carried out in species of
Medicago and Lolium (Veronesi et.al., 1988, 1989; Jansen &
Den Nijs, 1990, 1993).
 Screening of 2n pollen by flow cytometry

• The direct quantification of nuclear DNA by flow cytometry is a

particularly powerful method of discriminating between ploidy levels.
Since its introduction to plant sciences (see Galbraith et al., 1983) flow
cytometry has been used to detect 2n gamete production in several
species (Sgorbatti et al., 1989; Lumaret, Bretagnolle & Maceira, 1992;
Maceira et al., 1992).
• A major advantage of flow cytometry is that it permits a rapid
and accurate screening of the DNA content of a large number of
individuals. Furthermore, as well as being suitable for somatic
cells, this technique has been used to study the relative DNA
content of pollen from
• different plants (Zhang et al., 1992) and, thus, to screen for the
occurrence of 2n pollen (Van Tuyl et al., 1989; Bino)
 Progeny analysis of reciprocal diploid-diploid or
diploid—tetraploid crosses

• A simple count of the 4x seeds in the progenies of reciprocal

2x-4x crosses is currently the most commonly employed
method used to detect 2n pollen and/or 2n egg production and
to estimate the frequency of 2n gamete production by
individual plants.
• When no natural polyploidy are available, the polyploidy, used
as parents in the reciprocal crosses, may be obtained by
artificial methods (nitrous oxide or colchicine treatment)
(Bingham & McCoy, 1979; Iwanaga, Freyre & Orjeda,
1991a).
• The examination of 2n gamete production, in such reciprocal
2x-4x crosses, has usually been performed with species that
are self-incompatible.
 The analysis of micro- and megasporogenesis

• The analysis of micro- and megasporogenesis provides a

useful method of estimating the frequency of 2n gamete
production (Shoemaker-Megalos & Ballington, 1988;
Orjeda et al., 1990; Tavoletti, Mariani & Veronesi, 1991
a).
• The estimation of the frequency of 2n gametes usually
involves techniques to stain and clear fixed ovaries or
anthers (see Stelly et al., 1984; Tavoletti et al. 1991a; for
technical details). A similar method, suitable for large
scale screening and detailed observations on
megasporogenesis, is described by Jongedijk (1987).
 Cont…..

• In this context, the flow cytometry technique discussed above

provides a very powerful method for rapidly screening a large
number of plants, e.g. Lumaret et al. (1989, 1992), Maceira et
al. (1992) and De Haan et al. (1992).
• An interesting feature of the results obtained from both
reciprocal 2x-4x crosses and 2x-2x crosses, using known 2n
gamete producers, is the relative, or total, lack of triploids
among the progeny, the so called 'triploid block' effect. It is
characterized by the massive abortion of triploid seeds related
to the partial or total failure of the endosperm tissue associated
with the embryos.
 Cont…
Cont…
• An important consequence is that the triploid block effect may
not appear in all crosses (Freyre et al., 1991; Iwanaga et al.
1991 a) because of the existence of an effective EBN ratio of
2:1.
 The frequency of 2n gamete producers and the range of 2n
gamete production in recent studies
Species Frequency of Range of References
(2n pollen) 2n gamete production
producers (%) (mean) (%)

Agropyron cristatum ---------------- 0-1-4-9 Ray & Tokach, 1992

Dactylis glomerata 60 0-14-14 Maceira et al., 1992
Lolium perenne 11 ------------- Sala et al., 1989
Lotus tenuis 3 -28 Negri, 1992
Manihot spp. 38 < 1-35-6 Hahn et al., 1990
Medicago spp 28 (5.5) Veronesi et al., 1986
Trifoltum nigrescetis 39 1.3-34 Bullita & Smith, 1992
Vaccinium spp. ----------------- 0-3-1-9 Shoemaker-Megalos &
Ballington, 1988
 Cont…
Cont…
Species Frequency of Range of References
(2n eggs) 2n gamete production
producers (%) (mean) (%)

Dactylis glomerata 47 0-11-26 De Haan et al., 1992

Medicago spp. 37 (5.5) Veronesi et al., 1986
Solatium spp 24 4.9-23 Werner & Peloquin,
1991a
Trifoltum pratens 16 0-014-0-5 Parrott et al., 1985
Vaccinium spp. --------------- 0-4-2 Shoemaker-Megalos
& Balhngton, 1988
 The frequency of plant individuals (PI) producing 2n pollen, the
frequency of plants producing 2n gametes in the plant
individual producers and the range of 2n gametes per plant in
recent studies
Species PI with 2n 2n pollen Range References
pollen (%) plant production per
(percent) plant

Ipomoea triflda 55 10.1 < 1%-20%< Orjeda et al.,

1990
Solanum spp. (3 91 20.4 ------------- Watanabe &
spp.) Peloquin, 1989

Solanum spp. 47 5.5 1 0% > Watanahe &

(38 spp.) Peloquin, 1991a

Trifolium 100 3 1-84% Parrott et al.,

pratense 1984
 Cytological abnormalities responsible for 2n gametes production
as described in recent studies
cytological 2n 2n mode species Refrences
abnormalities pollen egg
Premeiotic
doubling + Lolium. perenne
Secale cereale
Wagenvoort et al.,
1990
Lelley et al., 1987
Synaptic + + FDR Solanum spp. Watanabe & Peloquin,
mutant and 1993
SDR

+ FDR Trifolium pratense Parrott & Smith, 1984

Adiantum Rabe & Haufler, 1992
pedatum
Lack of + Manihot Nassar, 1992
MI spindle esculenta

Omission of + FDR Arachis spp. Simpson & Davis, 1983

meiotic I
division
 Cont…
Cont…
cytological 2n pollen 2n egg mode species Refrences
abnormalities

Delayed + FDR Solanum spp. Werner & Peloquin,

meiotic SDR 1991a
division

Lack of + SDR Solanum spp Werner & Peloquin,

Mil spindle 1987; 1991a
+ SDR Lolium perenn Sala et al., 1989

+ SDR Matus x Zhang et al., 1988

domestica
+ SDR Manihot Nassar, 1992
esculenta
+ SDR Agropyron Ray & Tokach, 1992
cristatum
 The genetic basis of 2n gamete production

• Meiosis is a continuous process involving several cytological

events. There is evidence that it is controlled by a large
number of genes, the majority of which are present in a
dominant state (Baker et al., 1976; Kaul & Murthy, 1985).
• Depending on the phase at which they act, these author have
classified genes as pre-meiotic (controlling meiotic initiation
events), meiotic (infiuencing the course of meiosis) and post-
meiotic (controlling post-meiotic events and gametogenesis).
• The mutation of such genes alters meiosis and, thus, affects
gametic fertility and/or may lead to the production of 2n
gametes (Baker et al., 1976; Kaul & Murthy, 1985).
 Factors influencing the expression of meiotic genes

• Meiotic gene expression is influenced by genotype,

environment and their interaction (De Wet, 1980; Koduru &
Rao, 1981; Kaul & Murthy, 1985).
• Several studies have shown the influence of seasonal and
environmental factors, such as high and low temperature on 2n
gamete production (Veilleux & Lauer, 1981; Hermsen, 19846;
Felber, 1991)
• For example, the cultivation of several 2n pollen producers of
Solanum tuberosum in a cold coastal field strongly increased
the frequency of 2n pollen (76 % and 80 % during two
successive years of cultivation).
 Cont…
Cont…
• In contrast, under warmer cultivation conditions in a
greenhouse, 2n pollen was produced at a frequency of 35%
and 27% in two successive years (McHale, 1983). Thus, some
meiotic abnormalities leading to 2n gamete production may be
due solely to environmental factors.
• The relative importance of genotype and environment effects
have recently been documented, especially the impact of
temperature on the frequency of 2n gamete production.
 THE GENETIC CONSEQUENCES OF FDR AND
SDR

• In FDR:-A 2n gamete produced by FDR will possess two

non-sister chromatids and, thus, may contain equivalent
levels of heterozygosity and the complex epistasis
combinations of its parents (Hermsen,1984;Veilleux, 1985).
• In FDR, for a given chromosome pair, all loci from the
centromere to the first chiasma (if one occurs), that are
heterozygous in the 2x parent, will be heterozygous n the
FDR gamete, and, on average, half of the loci
heterozygous beyond the first chiasma will also be
heterozygous (Hermsen,1984; Werner &
Peloquin,1991c).
 Cont…
Cont…
• Thus, in Solanum, it has been calculated that a 2n gamete
produced by FDR with crossing-over will transmit roughly
80% of the parental heterozygosity to its progeny, whereas
without crossing-over 100% of the parental heterozygosity
will be transmitted (Werner & Peloquin,1916).In all such
cases, the FDR gamete possesses a large fraction of the
epistatic interactions present in the parent.
 Cont…
Cont…
• In contrast, a SDR type gamete will possess the two sister
chromatids from the centromere to a chiasma (if any) and,
thus, the progeny will contain les heterozygosity and epistatic
combinations than the
parent(Bingham,1980;Hermsen,1984;Werner&
Peloquin,1991c).
 Case studies:
(Discussion about The production of functional unreduced gametes has been
reported in several plants)
 Analysis of the Microsporogenesis

It doesnot provide any information about the pollen viability.

Since the production of n and 2n also depends on the balanced chromosome
segregation during meiosis and further maturation steps after meiosis.
2n pollen in Potato (Solanum tuberosum) (2N=4X=48)

• The first meiotic mutants in Solanum were discovered by

Hanneman and Peloquin (1969).

• The unusually high seedset obtained from 4x×2x crosses, as

well as the tetraploid chromosome number of the resulting
progeny strongly suggested that a meiotic mutation
systematically leading to 2n pollen formation was present in
the diploid parents.
 Cont…
Cont…
• They make the potato the best organism in which to
manipulate all sets of chromosomes for breeding purposes.
• They allow breeders to broaden the genetic diversity,
introducing both new genes for the improvement of traits of
interest and allelic diversity to maximize heterozygosity in
tetraploid varieties.
Variation for 2n Pollen Production and Male Fertility in wild Solanum
Germplasm Resistant to Phytophthora infestans (Mont.) de Bary (US-8 )
• Wild potato species possess genetic variability for valuable
traits including resistance to Phytophthora infestans, the
causal agent for potato late blight disease.

• Resistance to P. infestans was found to be independent of 2n

pollen production; therefore, identifying genotypes combining
2n pollen production, fertility, and resistance to P. infestans
was possible. (ZLESAK and THILL, 2002)
 2n pollen role in hybridization and breeding of
multiploids in Rosa hybrida.
• The ploidy inheritable characteristic of ‘Orange Fire’ × ‘Old Blush’ were
analyzed.
• The results of the cytological observations indicated that 2n pollen
developed from the defeated cytoplasmic division or nuclear division in
the meiosis metaphase II of PMC (pollen mother cell) in ‘Old Blush’.
• the results suggested that the germination rate of 2n pollen on the stigma
was not superior to that of 1n pollen, but that the proportion of 2n pollen
increased to 30.90 and 37.20%, respectively, while it traversed the stigma
and entered into style. The length of the 2n pollen tube was longer than
that of 1n pollen tube by about 300 μm (Gao et al.,2015)
 First division restitution in hybrids of Langdon durum
disomic substitution lines with rye and Aegilops
squarrosa
• In order to determine the genetic control of FDR a complete set of 14
Langdon durum D-genome disomic substitution lines (LDNDS) was
crossed with “Gazelle” rye and one accession (RL4175)of A. squarrosa
The microsporogenesis and fertility of the hybrids were studied.
• These results suggest that the reduced or absent FDR in such hybrids might
be related to the substitution of chromosomes with an FDR gene and poor
compensation ability of the D genome chromosomes for their
homoeologous A or B genome chromosomes.
• Cytological analysis suggested that chromosome 3A in LDNDS most
likely carries a gene for high frequency of FDR in hybrids. (Xu and Joppa
1995)
 Brassica U triangle
• U’s Triangle includes three diploid species with
genome complements AA, BB and CC (B. rapa, B.
nigra and B. oleracea respectively) and three
allotetraploid species AABB, AACC and BBCC (B.
juncea, B. napus and B. carinata respectively).
• Interspecific trigenomic hybrids between the
allotetraploid species (B. juncea × B. napus, AABC;
B. juncea × B. carinata, BBAC; and B. napus × B.
carinata, CCAB) may easily be created and the
hybrids often flower and produce viable gametes.
 2n pollen in cabbage
• Chinese cabbage (2n=2×=20) which naturally produce 2n pollen.
The diploid non-heading Chinese cabbage (2n=2×=20) can be
hybridized with tetraploid male sterile line (2n=4×=40) to
produce tetraploid plants, and therefore, it was used to study the
cytological mechanism of unreduced gamete (n=2×=20)
formation.
•  In the meiotic study of microspore mother cells (MMCs), it was
found that the first meiotic division was normal, but at the
second meiotic division, parallel and tripolar spindle appeared
and dyads and triads were found at the tretrad stage, which lead
to the formation of 2n pollen. Therefore, the formation of 2n
gamete in non-heading Chinese cabbage is due to the first
division restitution. It plays important role in the sexual
polyploidization breeding of non-heading Chinese cabbage. (Yu-
Hong-Xu et. al.,2012)
 CONCLUSION
 The exploitation of 2n gametes creates a new opportunities for practical
breeding
 2n pollen may transmit a high level of heterozygosity (genetic variation) to
the progeny.
 it helps in the creation of new species.
 It helps to overcome the incomaptibility between the different species .
 Helps in the production of biotic and abiotic resistant crops.
 Evolution of new species takes place.
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