Plant Cell and Tissue Culture Techniques

The whole plants can be regenerated virtually from any plant part (referred to as explant) or cells.

Plant tissue culture techniques involve the following steps:
a) Preparation and selection of suitable nutrient media
b) Selection of explants such as shoot tip.

c) Surface sterilization of the explants by disinfectants e.g. sodium or calcium hypochorite solution 0.3- 0.6% ) followed by washing the explants with sterile distilled water.

d) Inoculation or Transfer of the explants onto the suitable nutrient medium (sterilized by autoclaving) in culture vessels under sterile conditions (using laminar flow hood).

e) Incubation or Growing the cultures in the growth chamber or plant tissue culture room at optimum physical conditions of light (16 hours of photoperiod), diurnal illumination, temperature (25+/- 20C and relative humidity (50%-60%).

f) Regeneration of plants from cultured plant tissues.

g) Hardening: it is the gradual exposure of plantlets for acclimatization to environmental conditions

h) Transfer of plants to the field conditions following the acclimatization/ hardening of the regenerated plants.

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