1. Describe various steps involved in tissue culture.
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Describe the various steps involved in tissue culture.
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.
5) Incubation:
- After inoculation, the cultures are incubated so as to provide proper conditions for their growth & 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 & shoots & eventually the complete plant.
7) Sub-culturing:
- The cultured cells or tissues are transferred regularly to new nutrient medium called subculturing.
8) Regeneration:
- 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 & this transfer to fields is known as hardening.
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