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Plant tissue culture can take anywhere from 3 to 12 months to grow, depending on the type of plant and the desired outcome, such as seedlings or mature plants.
Initial Stage: Callus Formation
The initial stage of plant tissue culture involves callus formation, where a piece of plant tissue is treated with a plant growth regulator (PGR) to induce cell division. This stage typically takes 1-3 weeks, during which the tissue is maintained in a controlled environment with optimal temperature, humidity, and light. The PGRs used include auxins, such as 2,4-D and IBA, which stimulate cell division and prevent differentiation.
Subculture and Shoot Differentiation
Once callus formation is achieved, the tissue is subcultured into a fresh medium, where shoot differentiation occurs. This stage typically takes 4-6 weeks, during which the callus tissue is encouraged to form shoots using PGRs like kinetin and BA. The ideal temperature for shoot differentiation is between 20-25°C, with a photoperiod of 16 hours of light and 8 hours of darkness.
Rooting and Acclimatization
After shoot differentiation, the tissue is transferred to a rooting medium, where it develops a root system. This stage typically takes 2-4 weeks, during which the shoots are maintained in a humid environment with optimal temperature and light. Once the roots are established, the tissue is acclimatized to outdoor conditions over a period of 1-2 weeks, during which it is gradually exposed to increasing light and temperature.
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