Callus formation from mesophyll protoplasts of Fagus sylvatica L.

Viable protoplasts of European beech (Fagus sylvatica L.) were isolated from sterilized young leaves of juvenile (3-5 years) and mature (40 years) trees. Isolation in a saline solution containing 0.5% (w/v) Pectinol and 2% (w/v) Cellulase R-10 yielded 3×10(7) protoplasts per gram fresh weight. Protoplast culture in modified Kao and Michayluk (1975) medium resulted in cell wall regeneration and sustained cell divisions with the formation of colonies and microcalli.