Responses of Enzymically Isolated Aleurone Cells of Oat to Gibberellin A3

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Abstract
Oat (Avena sativa L.) aleurone layer cells (spheroplasts) were isolated by maceration of the aleurone layer with a mixture of commerically available cellulase and pectinase. About 20% of the cells present in intact layers were released as spheroplasts and 79 .+-. 9% of the spheroplast population was viable as judged by methylene blue staining. The spheroplasts became disorganized in solutions containing less than 0.4 M D-mannitol. When the spheroplasts were incubated for 48 h, total activities of acid p-nitrophenyl phosphatase and acid proteinase increased and protein levels decreased. These changes were not effected by GA3. Isolated aleurone layers incubated under the same conditions as the spheroplasts showed reduced responses to GA3. It is concluded that the necessary presence of an osmoticum limits the value of spheroplasts as a system for studying the mechanism of action of GA3 in the aleurone cell.