Abstract
Rice mesophyll protoplasts were isolated with ease and with high yield from expanded 2nd leaves, which were grown on agar plates under 23°C, 16.5 W/m2 for 25 days, by using a new cell wall lytic enzyme 1% "funcelase" together with 1% cellulase "Onozuka" R-10, although the isolation was difficult by the conventional method using cellulase and macerozyme. The yield ranged from 104-106/g of fresh weight of the 2nd rice leaves by enzyme treatment for 4 h. The survival cells just after isolation were 80-90% of the isolated total protoplasts. The yield and survival rate of the protoplasts could be elevated by shorter treatment time using cellulase "Onozuka" RS in place of cellulase "Onozuka" R-10. Treating in the enzyme solution for 4 h or less would be better when the subsequent culture of protoplasts was carefully considered. The protoplast isolation from rice mesophyll, as long as the present method is used, is no necessity for using macerozyme or pectinase. The application of 1% BSA was effective for the isolation and culture of rice mesophyll protoplasts. A satisfactory isolation in this study would be dependent considerably on the decomposition of β(1-3) glucan by laminarinase contained in funcelase.
