ABSTRACT
This chapter describes the protoplast culture, protoplast fusion techniques available for the improvement of crops, and utilization of protoplasts for the integration of useful genes into plant. Protoplast isolation can be carried out by means of many protocols designed for specific types of materials. The mechanical method is dependent on preliminary plasmolysis of cell within tissues and the subsequent dissection of the tissue and deplasmolysis to release the preformed protoplasts. The isolation of protoplasts depends on the concentration of the enzyme osmoticum, and incubation period. Enzymes or enzyme mixtures used vary with species and material being employed as a source of protoplasts. These enzymes singly or in mixtures degrade cell components, primarily cellulose, hemicellulose, and pectin. The osmoticum for a protoplast culture medium may be metabolically active or inert. Active substances are metabolized, thereby gradually lowering the osmotic strength of the medium.
