To embed the protoplasts in alginate
films, 2.3% sodium alginatewasmixed in a 1:1 ratio with 0.5Mmannitol
containing the protoplasts, polymerised on CaCl2 agar [0.02M
CaCl2, 0.48M mannitol, 16 g L-1 Plant Agar (Duchefa, The Netherlands)]
and cultured in 8pmC.1 or 8pmC.2 medium, respectively