OVARY CULTURE:- "the in-vitro culturing of ovaries in an aseptic condition from the pollinated or un-pollinated flowers, in an appropriate nutrient medium and under optimal conditions." And OVULE CULTURE:- "Ovule culture is an experimental system by which ovules are aseptically isolated from the ovary and are grown aseptically on chemically defined nutrient medium under controlled conditions."

2. OVARY CULTURE
OVARY CULTURE:-
"the in-vitro culturing of ovaries in an aseptic condition
from the pollinated or un-pollinated flowers, in an
appropriate nutrient medium and under optimal
conditions."
Or
"Ovary culture is a technique of culture of ovaries
isolated either from pollinated or unpollinated flowers."
The process of ovary culture refers as “Gynogenesis” as
the ovary is the female reproductive part or also refers to
as “Gynoecium”.
History:-
Gynogenesis was first studied by the scientist San Noem
in 1976. For the Gynogenesis, we can select either
pollinated or un-pollinated flower.

3. (Note:-A pollinated flower is cultured into the simple
nutrient medium whereas the un-pollinated flower is
cultured on the special medium added with synthetic
auxins and sucrose.)
Diagram of ovary (stigma , style , then ovary):-
PRINCIPLE:-
1. The principle of gynogenesis is based on the
Regeneration principle, where an ovary can
regenerate into a fully differentiated plant.
2. In the ovary culture, the flowers are excised from
the plant either in pollinated or non-pollinated

4. stage, and from that pistil containing ovary is
removed.
3. The ovary comprises of ovule which is a female
reproductive part of a plant by the growth on
nutrient medium and under controlled conditions.
Protocol:-
1. First, collect the open flowers from a healthy plant,
in a sterilized zip-lock bag.
2.Wash the flowers with the distilled water.
3.Then, dip the flowers into the 5% of Teepol solution
for 10 minutes.
4.Wash the flowers again with the distilled water.
5.Bring the flowers to the laminar airflow chamber.
6.Then subject the flowers for surface-sterilization by
immersing in a 5% of sodium hypochlorite solution
for 5-7minutes. And, then wash it with the distilled
water.
7.transfer the surface sterilized flower to a sterile
Petriplate, and by using sharp scalpel dissect out the

5. calyx, petal, anther filaments etc. to separate the
ovary.
8.Then, the ovary can culture either through induction
or regeneration process. In induction, the ovaries
float over the liquid medium. And, in regeneration
culture the ovaries on the solid nutrient medium.
9.Incubate the cultures for 16hours at 25֯ C.
( 1. For the regeneration of ovary, keep the
culture plates in a daylight regime by using a fluorescent
lamp. 2. for the induction process Place the culture tubes
in a dark for the induction process.)
10 . After 2 weeks, haploid plantlets grow either
through embryogenesis or through plant regeneration
from callus.
Application:-
1.Ovary culture helps in the study of the early stages
of embryo development.

6. 2.Gynogenesis also helps in the study of fruit
development and its physiology including
maturation.
3. From the culture of the unpollinated pistil, the
effect of phytohormones on the parthenocarpic fruit
can be studied.
4.Role of floral organs can be studied which plays a
significant role in fruit development.
5. Gynogenesis also helps in inducing Polyembryony
where instead of single plantlet, many shoots also
develop.
6.Ovary culture also explains the in-vitro pollination
and seed formation method.
7. The culture of an ovary from the apomictic plant can
help us to understand the stimulus provided by
pollination.
8. As in apomistic plant, there is no fertilization, the
only pollination occurs which alone stimulates the
ovary growth and seed development.

7. 9. Ovary culture also helps to understand the
hybridization process to obtain hybrid varieties of
seedlings by crossing over of interspecific and
intergenic species.
OVULE CULTURE
OVULE CULTURE:-
"Ovule culture is an experimental system by which ovules
are aseptically isolated from the ovary and are grown
aseptically on chemically defined nutrient medium under
controlled conditions."
Diagram:-

8. Principal:-
1.In Vitro ovule culture helps to understand the factors
that regulate the development of zygote through
organised stages to a mature embryo.
2.Alternatively, it may be possible to germinate pollen
in the same culture as the excised and to induce in
vitro fertilization and subsequent embryo
production.
Protocol:-
1.Collect the open flower for unfertilized ovule. If
fertilized ovules are desired, collect the open flowers
where anthers are dehisced and pollination has
taken place. To ensure the fertilization, collect the
flower after 48 hrs of anther dehiscence.
2.Remove sepals, petals, androecium, etc from the
overies containing either fertilized or unfertilized
ovules.
3.Soak the overies in 6% NaOCL solution.
4.Rinse the overies 3-4 times with sterile distilled
water.

9. 5.Using sterile technique, ovules are gently prodded
with the help of spoon shaped statula by breaking
the funicules at its junction placental tissue.
6.The spatula with ovules is gently lowered into the
sterile solid or liquid medium as the culture vial is
slanted about 45 0 C.
7.Damaged or unorganised ovules are rejected when
possible during transfer.
8. Incubate the culture in either dark or light at 25°C.
Application
1.Test tube pollination and fertilization:-
By this technique, it may be possible to germinate pollen
in the same culture as the excised ovule and to induce in
vitro fertilization.
2. Application in hybridization:-
In many interspecific and intergeneric crosses, the F1
hybrid embryos frequently become abortive in the
developing seeds or the F1 seeds are not capable to
support the development of embryos.Ovule culture has
been successfully em- ployed to obtain hybrid seedlings.

10. 3. Production of haploid callus through:-
Uchimiya et al. (1971) attempted culturing unfertilized
ovules of Solanum melongena and obtained vigorous
callus formation on a medium supplemented with IAA
and kinetin.
4. Induction of polyemryo:-
In horticultural practices, the artificial induction of
polyembryo holds a great potential. It has been observed
that the nucellus of monoembryonic ovule of citrus can
be induced to form adventative embryos in culture.
5. Virus irradiation:-
In the varieties of Citrus which are impossible to free of
virus by other means, the ovule culture has proved
decisively advantageous to make them virus free.
6. Ovule culture of orchid plant:-
In nature, the seeds of orchid germinate only in
association with a proper fungus. Hence many seeds are
lost due to unavailbility of proper fungus. Also the seed
capsule of many orchid takes a long time to mature. To

11. overcome such problems, several attempts have been
made to culture the fertilized ovule of orchid in vitro.
WHAT IS IN-VITRO POLLINATION:-
"The process of seed formation following stigmatic
pollination of cultured pistil has been referred to as in
vitro pollination and the development of seed through in
vitro fertilization."
Types:-
1.Ovular pollination:
2.Ovarion pollination:
3.Placental pollination:
4. Stigmatic pollination:
Application:-
1.For the production of homozygous plant.
2.For the conservation of extinct plant species.
3.Hybrid production Reducing the breeding cycle.
4.Overcome the dormant period.

12. 5.Production of hybrid species by distant
hybridization.
6.Production of haploid plant.
7.Conservation of germplasm.