Studies of hyphal morphology and the effects of various chemicals on growth are often difficult to perform on filamentous fungi because of the difficulty of observing the protoplasm through the rigid hyphae wall and because most activity occurs in a limited region near the hyphal tip. As an alternative, hyphae can be reacted with certain cell wall degrading enzymes to remove the cell wall to produce protoplasts. Procedures for producing fungal protoplasts have been developed for a number of fungi, including Lentinulus edodes, but there have been few efforts to develop similar protocals for producing protoplasts from common wood decay fungi. This paper describes methodology for producing protoplasts from hyphae of Phanerochaete chrysosporium, Postia placenta, Gloeophyllum trabeum, and Trametes versicolor using commercially prepared cell wall degrading enzymes (Novozyme 234). The effect of culture age, osmotic stabilizers, pH, and exposure time are explored in relation to the number of protoplasts produced and the number of protoplasts which can be regenerated. Protoplasts can be used for a number of studies including chemical sensitivity and enzyme production.

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