Protoplasts isolated from sporidia of Ustilago maydis were stored by freezing for more than two months. After thawing, the survival of protoplasts was estimated using the vital stain phenosafranine. In a buffered and osmotically stabilized medium containing polyvinyl-pyrrolidone (15% (w/v), PVP) as cryoprotectant, nearly 100% of the protoplasts remained viable, if cooled slowly to − 80 °C. Cell-wall regeneration and reversion frequency of the protoplasts after storage was investigated to further prove cell viability. No significant difference was detected comparing freshly isolated and stored protoplasts. © 1992, British Mycological Society. All rights reserved.
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