Studies on formation, regeneration and fruiting body formation of Pleurotus cystidiosus protoplasts

Abstract

Conditions suitable for the release of mycelial protoplasts of Pleurotus cystidfosus and for reversion of the isolated protoplasts were examined. The study culminated in the development of a method using commercially available preparation of 2.0 % Novozym(TM) 234, 1.5 % Chitinase, 1.5 % Driselase and 2.0 % Cellulase, by which about 3.8 x 10(6)/ml protoplasts could be obtained from the young growing mycelia, within 10 hours. The 0.05 M Na-maleate buffer containing 0.6 M MgSo(,4), pH 5.8 is suitable as protoplast buffer (buffer/stabilizer). Protoplasts from P. cystidiosus mycelia degenerated into normal hyphae with frequencies about 0.19 % in glucosemalt-yeast extract medium supplemented with 1.0 % sorbitol (regeneration medium), in embedding method. The reversion frequency was shown influence by nutrient composition, especially of carbon and nitrogen sources in the regeneration medium. Mycelium derived from protoplasts of P. cystidiosus consistently developed normal fruiting bodies.