Question

Explain briefly the steps involved in tissue culture.

Answer 1

Tissue culture involves the following steps:

Small amounts of parent tissue or a number of cells are taken.

The plant material is transferred to plates containing sterile nutrient agar jelly.

Plant hormones are added to stimulate the cells to divide.

Answer 2

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.