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CH5. Types of Culture

CH5. Types of Culture. In vitro of explant. The non-dividing, differentiated, quiescent cells of the explant when grown on a nutrient medium first undergo changes to achieve the meristematic state. Callus.

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CH5. Types of Culture

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  1. CH5. Types of Culture

  2. In vitro of explant • The non-dividing, differentiated, quiescent cells of the explant when grown on a nutrient medium first undergo changes to achieve the meristematic state.

  3. Callus • An amorphous mass of loosely arranged thin-walled parenchyma cells arising from the proliferating cells of the parent tissue cultured on agar medium.

  4. Cellular totipotency • Cytodifferentiation • Cell differentiation, mainly emphasis on vascular differentiation, tracheary element differentiation, etc. • Dedifferentiation • The phenomenon of mature cells reverting to a meristematic state and forming undifferentiatedcallus tissue. • Redifferentiation • The ability of the component cells of the callus to differentiate into a whole plant or organ.

  5. Types of culture Embryo culture Seed culture Meristem culture Cell culture Plant tissue culture Protoplast culture Organ culture Bud culture Callus culture

  6. Seed Culture • Seed culture is an important technique when explants are taken from in vitro-derived plants and in propagation of orchids. • In nature germination of orchid seedlings id dependent on a symbiotic relationship with a fungus. • In vitro, it is possible to independent on asymbiotic germination.

  7. Embryo culture • Mature embryo culture • Seed dormancy – ex. ripe seeds • Immature embryo culture / embryo rescue • To avoid embryo abortion • Ex. Hybrid embryo.

  8. Application of embryo culture • Prevention of embryo abortion in wide crosses. • Production of haploids • Overcoming seed dormancy • Shortening of breeding cycle • Prevention of embryo abortion with early ripening stone fruit • In vitro clonal propagation

  9. 1. Meristems 2. Leaf sections 3. Bulb sections 4. Embryos 5. Anthers 6. Nucellus Callus formation De-differentiation Re-differentiation Explants Callus Protoplasts Development   Suspension cells Organs (leaves, roots, shoots, flowers,...) ?

  10. Callus formation Stimuli : In vivo : wound, microorganisms, insect feeding In vitro : Phytohormones   1. Auxin 2. Cytokinin 3. Auxin and cytokinin 4. Complex natural extracts

  11. Callus formation Stimuli : In vitro : Phytohormones   1. Auxin 2. Cytokinin 3. Auxin and cytokinin 4. Complex natural extracts

  12. # Auxins : 0.01 - 10 mg / L Activity IAA IBA NAA 2,4 - D 2,4,6 - T Dicamba Picloram # Cytokinins : 0.01 - 10 mg / L TDZ BA (BAP) Kinetin 2 - ip Zeatin

  13. Callus formation Stimuli : In vitro : Phytohormones   1. Auxin

  14. Callus formation Stimuli : In vitro : Phytohormones   1. Auxin 2. Cytokinin 3. Auxin and cytokinin 4. Complex natural extracts

  15. Callus formation Stimuli : In vitro : Phytohormones   1. Auxin 2. Cytokinin 3. Auxin and cytokinin 4. Complex natural extracts Coconut endosperm ( milk / water ) Yeast extract

  16. Callus growth Growth characteristics of callus : friable cultures : lignified and hard cultures :

  17. Callus growth Growth characteristics of callus : pigmentation: yellowish, white, green, pigmented,

  18. Maintenance : Subculture : sub-division of a culture for transfer to fresh medium. ?

  19. Q : How? Q : Where? Medium

  20. Q : How? Q : Where?

  21. Q : When? growth time

  22. 3年 ? Q : What happened in long-term cultures? Physiological condition

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