Effect of enzyme concentrations on protoplast isolation and protoplast culture of Spathiphyllum and Anthurium

Vital protoplasts from Spathiphyllum wallisii 'Alain' and Anthurium scherzerianum '238' were isolated from both somatic embryos and leaves. The highest yields were obtained when 1.5% cellulase, 0.5% macerase and 0.5% driselase were used for Spathiphyllum wallisii leaves and 0.5% cellulase, 0.3% macerase and 0.5% driselase for Anthurium scherzerianum embryos. About 1 x10(6) protoplasts g(-1) and 1 x10(5) protoplasts g(-1) could be isolated from leaves and embryos, respectively. For protoplast fusion Spathiphyllum wallisii 'Alain' and Anthurium scherzerianum '238' were mixed in a 1:1 ratio in a fusion solution containing 1 mM CaCl2 center dot 2H(2)O, 1 mM MES and 0.5 M mannitol. Fusion was performed by protoplast alignment under 500 V cm(-1) alternating current for 60 s and subsequent generation of two pulses of 4500 V cm(-1) direct current during 50 mu s. Development until colony stage was achieved using agarose beads for protoplast culture.