Regeneration, Cultivation and Differentiation of Plant Protoplasts Immobilized in Ca-alginate Beads

A method for growing plant protoplasts immobilized in Ca-alginate is presented in detail. Two types of protoplasts, isolated from hypocotyls of Brassica napus cv. Niklas and leaf mesophyll tissue of Nicotiana sanderae cv. Crimson Bedder, have been used as examples to illustrate the general applicability of the method. The major advantage of the method is improved cellular protection during the first critical weeks after isolation. This is a protection both against mechanical strain and strong gradients in environmental conditions. Higher plating efficiency may thus be achieved and handling simplified, particularly when changing nutrient medium. Both regeneration and differentiation have repeatedly been achieved.