Isolation and Culture of Protoplasts from Apple Callus and Cell Suspension Cultures

Abstract
Protoplasts were isolated from actively growing callus and cell suspension cultures initiated from apple leaf discs. Callus and cell suspension cultures incubated in a solution of 0.7 m mannitol, 0.01 m CaCl2, 2% cellulysin, 0.5% macerase yielded protoplasts from about 75% of the callus cells and 90% of the suspension cells. Protoplast viability was 95–98% upon isolation, and 80–85% after a 2-week culture in a modified Murashige and Skoog (MS) medium. About 60% of the protoplasts regenerated cell walls, and 5–10% of the original protoplasts exhibited cell division.

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