The basic technique of plant tissue culture involves the following steps:
(i) Selection of suitable explants like shoot tip, leaf, cotyledon and hypocotyls.
(ii) Surface sterilization of the explants by disinfectants (e.g. sodium hypochlorite) and then washing the explants with sterile distilled water.
(iii) Inoculation (transfer) of the explants onto the suitable nutrient medium (which is sterilized by autoclaving or filter-sterilized to avoid microbial contamination)in culture vessels under sterile conditions (i.e., in laminar flow cabinet).
(iv) Growing the cultures in the growth chamber or plant tissue culture room, having the appropriate physical conditions [i.e., artificial light (16 h photoperiod), temperature (~26 C) and relative humidity (50-60%)].
(v) Regeneration of roots and shoots from cultured plant tissues and their elongation.
(vi) Transfer to the green house and then to fields.
- Applications of Plant cell culture-
- Micropropagation
- Producing virus free plants
- Producing artificial seeds
- Embryo rescue in interspecific & intergeneric hybrids
- Generating haploids & triploids
- Somatic hybrids & Cybrids
- In vitro germplasm conservation
- Somaclonal variations
- Production of secondary metabolites
