Endosperm culture

Triploid plant regeneration from endosperm tissues. (A) An immature endosperm explant
isolated from immature fruit (B) green callus developed from an immature endosperm
explant (C) an immature endosperm-derived callus forming adventitious buds on a MS
medium  (D) a mature endosperm-derived callus forming buds on a MS medium (E)
Embryogenesis
Embryo culture

Figure. a Torpedo-stage zygotic embryo before culture. b Production of non-

regenerative callus on medium. c Induction of SE on medium. Arrowheads point
to somatic embryos. d, e High magnification of (c) showing direct somatic
embryo production from regular row on hypocotyl (d) and at the margin of
cotyledon (e). f Necrosis of original embryo with concomitant production of
whitish somatic embryos. g Isolated somatic embryo from medium. h Plantlet
originating from somatic embryo 
Somatic embryos
Embryo rescue
Figure . Encapsulation of somatic and zygotic embryos. A) Proembryogenic mass with somatic embryos in
different development stages. B) Zygotic embryos. C) Encapsulate of somatic embryo through micropipette
with sodium alginate. D) artificial seeds rinsed with sterile water. E) Survival of somatic embryo encapsulated
after 21 d; F) elongation of somatic embryo encapsulated in sodium alginate . G) germination of zygotic
embryo encapsulated in sodium alginate.H) conversion of encapsulated zygotic embryo to plant.
Synthetic seeds
Anther culture
The procedure to establish the anther culture of the plant is as follows:
1.Select buds of the desired plant and surface sterilize them using a disinfectant.
2.Excise anthers along with their filaments under aseptic conditions and place them on a sterilized plate.
3.Crush an anther and stain it with acetocarmine to test the pollen development stage.
4.If the correct stage is found, gently detach anthers from filaments and place them horizontally in the culture medium (in some species, attachment of
even a part of filament with anthers affects the production of plants).
5.Maintain anther cultures in alternating periods of light(12-18 h; 5000-10 000 lx m2) at 28℃ and darkness (12-6 h) at 22℃. However, species of some
genus (like Brassica) are very sensitive to light and should be maintained in dark throughout.
6.As the anthers respond, wall tissues turn brown and after 3-8 weeks, they burst open due to the pressure of developing callus or embryos.
7.The embryos will germinate on the same media they were cultured or required to be transferred to another culture media.
8.When the plantlets attain a height of 3-5 cm, transfer them to a rooting medium.
9.After well-developed rooting is observed, transfer the plants to sterilize potting mix in small pots or seed trays
Disadvantages of anther culture:
•Production of genetically different/heterogeneous plants.
•Production of a heterogeneous population.
•Asynchronous pollen development will lead to the suppression of younger grains by older grains due
to the release of toxic substances.
•Plants derived from anther culture would not be purely of gametophytic origin.
In 1972, the embryo development was induced in pollen cultures by using some nurse tissue (that
provid
Isolation of protoplast