describe the various steps involved in tissue culture
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Answers
Explanation:
tissue culture :
tissue culture is the growth of tissue or cells in an artificial medium separate from the organisms
steps in tissue culture technique :
- selection of mother plant
- initiation of culture
- multiplication
- root formation
- transfer of soil
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It involves following steps:
1. selection of plant - The plant which has to be cultured in vitro is selected.
2. Isolation of explant- Explant is any part of plant excised out for tissue culture.
3. Sterilization of explant- Explant is surface sterilized so as to avoid contamination.
4. Inoculation of explant- The explant is inoculated on the nutrient medium. For plant tissue culture, the most common medium used is MS media.
5. Incubation: After inoculation, the cultures are incubated so as to provide proper conditions for their growth and regeneration like temperature, moisture etc.
6. Initiation of callus: Callus is a mass of undifferentiated cells formed by the dedifferentiation of plant cells or the explant. Callus further regenerates to form roots and shoots and eventually the complete plant.
7. Sub-culturing: The cultured cells or tissues are transferred regularly to new nutrient medium and this is called subculturing.
8. Regenerstion: Regeneration is the formation of organized structures like roots, shoots, flower buds etc from the cultured cells
9. Hardening: The plantlets are removed from the cultures and prepared for soil transfer and this transfer to fields is known as hardening.