ABSTRACT
A plant protoplast is a cell without a wall. Due to the absence of a cell wall and consequent exposure of the plasma membrane, the protoplast becomes a very fragile structure. It is disrupted easily, unless it is maintained in an osmoticum. The key operation in the release of a plant protoplast is the removal of cell wall in a way that it does not damage the protoplast and impair its ability to regenerate into a cell. The pioneering attempt to isolate protoplasts from a higher plant was by a mechanical method. The mechanical method involved a preplasmolysis of tissue followed by its random sectioning which resulted in the release of a few undamaged protoplasts along with several broken cells. Large-scale isolation of protoplasts was possible when enzymatic digestion of cell wall was demonstrated from root cells of tomato. To digest the cell wall the enzyme source was a culture filtrate of the fungus Myrothecium verucaria.
