REGENERATION OF ENZYMATICALLY ISOLATED PROTOPLASTS OF THE MOSS FUNARIA-HYGROMETRICA L SIBTH

Protoplasts from protonemata of the moss Funaria hygrometrica were obtained by one-step enzymatic degradation of the cell walls. A pre-incubation with a non-ionic detergent Tween 80 and the use of a hemicellulase in combination with cellulase and Macerozyme appeared to be important for successful protoplast isolation. Protoplasts were cultured in hanging drops of a mineral nutrient medium supplied with 0.3 M mannitol and 0.5% glucose. The presence of glucose was required for survival and the subsequent division of protoplasts. Cultured protoplasts regenerated cell walls within 3 days of isolation and developed into multi-cellular protonemata after one month of culture. © 1979.