Factors Affecting Fusion Frequency of Pleurotus Protoplasts

Factors influencing the fusion frequency of protoplasts were investigated with auxotrophic mutants of Pleurotus florida and Pleurotus ostreatus. Immediately after the polyethylene glycol (PEG) solution was added, the protoplasts adhered firmly and shrank. During the subsequent dilution with 0.6 M sucrose, the protoplasts regained their normal size and larger bodies were observed. Interspecific heterokaryons were obtained by fusion of the nutritionally complementing protoplasts. Hyphae of the heterokaryotic fusants formed true clamp connections. The optimum conditions were a total of 1 to 15 million protoplasts per ml, 30% polyethylene glycol 8000 solution with adjustment to pH 8.0 and 0.6 M sucrose stabilized regeneration medium. Other parameters such as , glycine, exposure time and temperature influenced mainly the viability of the protoplasts.