Abstract
Enzyme conditions for efficiently isolating protoplasts from mulberry mesophylls were examined, using cotyledons and cultured leaves as materials. The yield of protoplasts from the cotyledons was much higher than that from the cultured leaves. As for the time for enzyme treatment, 4 hours was generally adequate, although it was varied slightly, depending on the difference in enzyme solutions. Protoplast isolation efficiency was increased by adding macerozyme R10 and meicelase P1 to the enzyme solutions containing cellulase onozuka RS and pectolyase Y23. The ES5 enzyme solution, which was composed of 3% cellulase onozula RS, 0.1% pectolyase Y23, 1% macerozyme R10 and 1% meicelase P1, gave the highest yield of protoplasts, 6.2×107, when cotyledons were treated for 4 hours.
