A simple method for the isolation of plant protoplasts.

A simple protoplast isolation protocol that was designed to recover totipotent plant protoplasts with relative ease has been described· The key elements of the protocol are, tissue digestion at slightly elevated temperatures and use of protoplastreleasing enzymes that are stable and efficient at higher temperatures· Besides enzymes, the protoplast isolation cocktail consisted of an osmoticum (mannitol or MgSO4), and a protectant (CaCl2 2H2O), all dissolved in distilled water· The protocol has ensured reproducibility, higher yields and is gentle on protoplasts as the protoplasts obtained were amenable to cell wall regeneration and cell division· Plant regeneration was demonstrated for Nicotiana tabacum cv· Thompson from protoplasts isolated by this method· Wall regeneration and cell division were obtained in other species. The merits of the protocol are, simple and easy-to-handle procedure, non-requirement of preconditioning of donor plant and explants, incubation without agitation, satisfactory yields, culturability of the protoplasts isolated and applicability of the protocol to a large number of species including mucilage-containing plants.

Plant tissue culture techniques

Plant cell and tissue culture in a simple fashion refers to techniques which utilize either single plant cells, groups of unorganized cells (callus) or organized tissues or organs put in culture, under controlled sterile conditions.