Protoplast formation from selected species of ectomycorrhizal fungi

Conditions for the formation of protoplasts from selected species of ectomycorrhizal fungi are described. The age of the fungal culture and extent of incubation in a lytic enzyme mixture are critical factors for efficient formation of protoplasts. There is a correlation between the distribution of nuclei in hyphal fragments and protoplasts and the frequency of protoplast regeneration. Protoplasts from at least two of the species studied are formed in sufficient numbers and regenerate at suitable frequencies to be useful for development of genetic transformation and cell fusion systems. These fungi can now be considered in experiments designed for the improvement of ectomycorrhizal associations through genetic manipulation of the fungal component.