Influence of Microspore Development on Pollen Protoplast Isolation in Digitalis obscura

Summary Protocols for the enzymatic isolation of Digitalis obscura pollen protoplasts are described. The developmental stage of the microspores was a critical factor for successful isolation since protoplasts were obtained only from tetrads. The highest yield of viable protoplasts was achieved with an enzymatic mixture containing helicase, cellulase and pectolyase in 0.5 M sucrose. In different culture techniques employed, cell wall regeneration took place only when protoplasts were plated in agarose drops.