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1. ITM UNIVERSITY GWALIOR
PRESENTED BY:
HIYA DASHORA
MHVN1AG23003
M.Sc Horticulture {Vegetable Science}
BREEDING FOR SEEDLESSNESS IN
HORTICULTURE CROPS
SCHOOL OF AGRICULTURE
GUIDED BY:
Prof. (Dr.) C.K. Sharma
Dr. Shubham Singh Rathore
Department of Fruit science
Master’s Seminar

2. Content
 Introduction
 Seedlessness trait and factors affecting it
 Characteristics of seedless fruit
 Parthenocarpy
 Hormonal Regulation of Fruit Set
 Breeding methods
 Genome editing technology--- CRISPR- Cas
 Genetic Engineering Strategies for Seedlessness
Breeding
 Recent Research
 Conclusion
 References.

3. Introduction
A plant is considered to be seedless when its fruits are completely devoid of seeds, contain
a much-reduced number of seeds or present aborted seeds. Seedless are obtained through
parthenocarpy if the fruits develop without fertilization and by stenospermocarpy if the
seeds abort after fertilization
Factors affecting Seedlessness or Factors
inducing Seedlessness include certain
environmental conditions, such as low or
high temperatures, chemical treatments,
chromosomal aberrations, and genetic
factors.
Seedless fruit can be obtained
through parthenocarpy if the fruits
develop without fertilization and by
stenospermocarpy if the seeds abort
after fertilization

4. A. Seedless fruits are generally smaller in size
compared to their seeded ones.
B. In seedless fruits, other forms like reduced
seed number, soft seed, hollow seed (Ex.
grapes).
C. Seedless fruits may result in thickening of pedicels in
some cases (EX. Pear)
D. Fruit surface is normally very smooth in case of
seedless fruits compared to seeded ones except in case
of some where the surface is warty . (EX. Guava)
E. Seedless fruits in general mature later than their
seeded ones.
Characteristics of Seedless Fruit
The Pharma Innovation Journal

5. Parthenocarpy Greek word, literally means “Parthenos : virgin karpos : fruit” is the
natural, artificially induced, or genetically modified production of fruit without
fertilization. In the absence of pollination, parthenocarpic plants will set seedless fruit.
(Gustafson, 1942) and According to Winkler (1907),
Parthenocarpy : Primary Requirement to
Produce Seedless Fruit
.
In artificial it is made
between vegetative
parthenocarpy, which
allows fruit set without
pollination, and
stimulatative
parthenocarpy, in which
fruit set follows
pollination, but
subsequent fertilization
is prevented, resulting in
the production of
seedless fruit.
In genetic/natural distinction
is often made between
obligatory parthenocarpy
(Ivy gourd), which always
results in seedless fruit, and
facultative parthenocarpy,
which results in seedless fruit
only when pollination is
prevented.
(Eg : Tomato, brinjal and
cucumber)
Seedlessness is
appreciated by
consumers both in
fruits for fresh
consumption (e.g.,
grape, citrus, and
banana) as well as in
conserved or
processed fruits
(e.g., frozen
eggplant, tomato
sauce

6.  Seedlessness can contribute to increase the quality of
the fruits when seeds are hard or have a bad taste.
 In the case of eggplant, the absence of seeds prevents
browning and texture reduction of the pulp.
Furthermore, seeds can produce substances that
accelerate the deterioration of the fruit, as in watermelon
and eggplant.
In this regard, the absence of seeds can increase the
shelf life of the fruits, allowing a better conservation.
Continue
http://www.jstor.org/stable/4353278
The mean temperature during the pre- flowering, flowering and fruit-setting period
is the primary factor responsible for the production of seedless fruits.

7. Seedless okra production by indole 3-acetic acid
micro syringe injection on flower bud, ovary
and shoot xylem and its vitamin and mineral
content development
Scientia Horticulturae
Pusa Bedana:It is seedless watermelon
developed at IARI, by crossing Tetra 2
(tetraploid) x Pusa Rasal
Cucumis sativus (Cucumber). Noll observed
that the varieties Gurke and Grosse produced
seedless fruits when pollination was prevented.
In some the ovules made a little growth but no
embryos were observed
Luffa cylindrica. Scientist found that when
treated the pistils of flowers that had just
opened with a 1.0%o indole acetic acid-
lanolin paste he was able to produce seedless
fruits that were nearly as large as the seeded
fruits

8. Hormonal Regulation of Fruit Set
•A widespread agriculture practice for the production of seedless parthenocarpic
fruit consists in treating flowers with phytohormones before pollination.
• Auxin,
• Gibberellins
• Cytokines
• Mixtures of these hormones have all been proven to be effective in inducing
fruit development in the absence of fertilization in several crop species, for
example vegetables like tomato and eggplants.
Brinjal: Muthukrishnan and Srinivasan, 1980
Cucumber : Choudury and Phatak, 1960;Kim et al.,
1994
In crops such brinjal and
cucumber, there is evidence
that parthenocarpic fruit set is
due high levels of endogenous
auxin and GAs in the ovary.
To improve fruit set citrus
cultivars Auxin levels peak
sharply 10 d after anthesis, and
exogenous GA is essential for fruit
development and the prevention of
fruit abscission.

9. Fig: (a) different steps in fruit development, (b) t steps in seed development and (c) examples of seedless fruit.
Varoquaux et al., 2000

10. •Breeding efforts for high-quality seedless citrus fruit have increased during
the past few decades and few cultivars developed are :
•Satsuma mandarin (Citrus unshiu Marc.)
•Tahiti lime (Citrus latifolia)
• Washington Navel orange (C. sinensis)
 Seedlessness in Parthenocarpic Citrus Cultivars
Fig: Satsuma mandarin Fig: Tahiti lime
Another approach for seedlessness involves the coupling of parthenocarpy with
male sterility.
 Ploidy and Cytoplasmic Male Sterility
Washington Navel orange

11. Ploidy breeding

12. 1. Triploid in Watermelon: Seedless watermelons are gaining popularity as the mature
seedless melons become more available in the general market.
2. Triploid breeding in grape Production of large seedless berries with high quality is one of the
most important objectives in table grape breeding programs
Ploidy manipulation using crosses between diploids and tetraploids have
yielded several valuable triploid cultivars in many of crops.
Kihara, 1951
a.Marvel seedless
b.Muscat alexandria
c.Black king from
champbell

13. 5. Mutation Breeding:
• Irradiation is a valuable tool to obtain
seedless cultivars from seeded ones.
•Mutation affects pollen viability. Female and
male sterility seem to be directly related, In
Citrus spp with a lower seed number present
a lower pollen viability, and chromosomes
aberration during meiosis caused by
irradiation are responsible for sterility.
3. climacteric seedlessness is good among fruits and have good traits like high protein
content, high moisture content
4. Endosperm culture It is widely
known as the fusion of three haploid
nuclei. This is a good potential area to
isolate triploid plants directly from the
embryonic stage and cut short the
lengthy process of growing and crossing
which could develop into a possible
seedless plant.
Influence of Gamma Irradiation on Seedless

14. FIG: Visual scheme of gaining the seedlessness trait using (CRISPR/Cas) strategy
Enormous way
and endless
possiblities
Success rate is
very high
Open up new

15. In Many citrus cultivars the first
approach is male sterility. could
potentially set seedless fruit in the
absence of pollination
However, this approach is restricted to
certain conditions under which male
sterile citrus plants are isolated from
male fertile plants.
The second approach is seed coat
destruction to obtain
stenospermocarpy through the
expression of targeted suicide genes
during early seed coat development.
In this case, parthenocarpy is not
required for seedlessness.
The third approach is female sterility.
Destruction of the ovule or the stigma
by suicide genes could trigger seedless
fruit set as fertilization would be
impossible. However, this approach is
limited to cultivars with strong
parthenocarpic traits.
The fourth and last approach is
increased expression or increased
sensitivity to hormonal in the
ovary or ovule. This approach, via
genetic engineering of auxin
expression or sensitivity, has been
used successfully in several field
crops
Source: DOI: https://doi.org/10.21273/JASHS.133.1.117

16. [A] Based on large numbers of experimental studies, Li et al. (2001) concluded that
the main factors determining the success of embryo rescue are the
i) sampling time,
ii) the parental genotype,
iii) the culture medium, and conditions.
[B] The emergence of molecular markers further sped up the breeding process for
seedless grapes.
[C] Seedlessness evolution caused majorly due to spontaneous mutation in nature.
D] Genetic makeup: Seedless fruits is a highly desirable trait and is due to natural
causes and demand .
[E] Induced mutations and Isolation of natural bud
[F] Climatic conditions (Fig) , Location (Latitudinal changes- E.g.: Grapes).
Seedlessness remains a main focus of breeders

17. • Lack of availability morphological
marker for adjudging seedlings at
the early age
•In conventional breeding, only seeded
genotypes can be used as seed parents,
hence there is lot of limitations.
•When seedless genotypes are used as
male parents, the pollen quality,
viability etc. vary considerably and
hence the results are not consistent.
•The average frequency of seedless
progeny that has involve seedless parent
can only give rise to 10-15% (on avg.)
seedless progeny which is further
reduced by poor germination.
•Introgression breeding to transfer
quality and seedlessness and an
alternative approach involving different
crosses is quite challengeing
• In many population the time for evolving seedless hybrid is reduced to 5-6
years compared to 12- 15 years under the conventional system.
•Challenges faced by breeder

18. Advantages of Seedlessness
• Year-round production
• Gustatory advantage
• processing industry
• Seed cavity can be filled
with fruit tissue (seedless
pickled gherkin, which is
more crunchy, firmer and
fleshier than its seeded
variety)
• Seedless watermelons
develop a meaty texture
and become overripe
significantly later than
seeded varieties.
• Lack of stability and
uniformity in the
expression of
Parthenocarpy.
• Malformed fruits,
• Small sized fruits
• Hampers the production
of commercial seeds
Seedlessness results in
hampered seed
production and there is
no room for the
commercial seed
production
Limitation of seedless fruits

19. Recent research (December 2021)
•It is shown that, parthenocarpy occurs more in plurispermic
(multi seeded) species than monospermic (single-seeded)
(Picarella and Mazzucato, 2019)
• It is also reported that this phenomenon occurs more in
cultivated species than in wild species

21. 1. Dokoozlian, N. 1994. Crimson Seedless research update: Reducing berry set and
increasing berry size with gibberellic acid. Vine Lines (Nwsltr. Univ. Calif. Coop. Ext.
Fresno County) April. p. 1–3.
2.Li, G.; Wang, Y.; Tang, D.; Wang, X.; Luo, Q. The studies on embryo rescue techniques of
‘Thompson Seedless’ grape. Plant Cell Tissue Organ 2001, 21, 432–436
Li, J.; Wang, X.; Wang, X.; Wang, Y. Embryo rescue technique and its applications for
seedless breeding in grape. Plant Cell Tissue Organ Cult. 2015, 120, 861–880.
3. Abbate L, Panno S, Mercati F, Davino S, Fatta Del Bosco S. Citrus rootstock breeding:
response of four allotetraploid somatic hybrids to Citrus tristeza virus induced infection.
European Journal of Plant Pathology. 2019;153(3):837-847.
4.Ahmed D, Curk F, Evrard JC, Froelicher Y, Ollitrault P. Preferential disomic segregation
and C. micrantha/C. medica interspecific recombination in tetraploid ‘Giant Key’lime;
outlook for triploid lime breeding. Frontiers in plant science. 2020;11:939.
5Aleza P, Juárez J, Cuenca J, Ollitrault P, Navarro L. Extensive citrus triploid hybrid
production by 2x × 4x sexual hybridizations and parent-effect on the length of the juvenile
phase. Plant Cell Reports. 2012;31:1723- 1735.
6.Handaji Najat HB, Najat Arsalane HB. Selection of new promising seedless mandarins
triploid hybrids from crosses between monoembryonic diploid female and diploid male
parents. Research in Biotechnology. Food and Scientific Reports. 2017;1(4):25-27.
7. Hoshino Y, Miyashita T, Thomas TD: In vitro culture of endosperm and its application in
plant breeding: approaches to polyploidy breeding. Scientia Horticulture. 2011;130:1-8.
References