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Viable mesophyll protoplasts were isolated from in vitro grown leaves of grapes (Vitis labruscana cv. Kyoho and V. vinifera cv. Cabernet Sauvignon). The best yield of mesophyll protoplasts (1.8 × 107 and 2.4 × 107 • g -1 leaf fresh weight, 'Kyoho' and 'Cabernet Sauvignon', respectively) was obtained in a combined enzyme solution of 0.5% Cellulase YC, 0.2% Macerozyme R -10 and 0.1% Driselase. More than 85% of the isolated protoplasts were viable. Most protoplasts regenerated the cell wall within first 5 days of culture. Cell division occurred after 5 to 10 days of culture in Gamborg's B5 liquid medium supplemented with 5μM 2, 4 - D, 2.5μM BA and 0.6 M sorbitol. In 'Kyoho', 12% of cell division frequency was found in the protoplasts isolated with 0.5% Cellulase YC, 0.2% Macerozyme R -10, 0.1% Driselase and 0.01% Pectolyase Y -23. In 'Cabernet Sauvignon', the frequency of cell division was low (1.4%) when the same enzyme solution was used for the protoplast isolation.

Effect of Several Enzyme Solutions on the Isolation and Culture of Grape Mesophyll Protoplasts