Abstract
The freezing tolerances of protoplasts isolated from the epicotyls of darkgrown seedlings of cold-hardened and non-hardened winter rye were examined. When these isolated protoplasts were frozen extracellularly (slow freezing) in the appropriate media, they exhibited freezing sensitivities similar to their tissues of origin. Over 80% of the protoplasts isolated from the hardened epicotyls survived freezing to −15°C whereas only 20% of those isolated from non-hardened tissues survived. The most suitable freezing medium was the saline medium of Y. Meyer and W.O. Abel (Planta, 125 (1975) 1). Sucrose alone was cryoprotective for the protoplasts whereas mannitol was injurious even for hardened protoplasts. The retention of differences in freezing sensitivities between protoplasts isolated from hardened and non-hardened winter rye, and the ease with which large quantities of such protoplasts could be isolated make it an ideal system to study the involvement of membranes in freezing tolerance and injury.
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