Isolation of protoplasts from Aspergillus nidulans conidiospores

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Abstract
Protoplasts were prepared from conidiospores of A. nidulans. The mononucleated conidia gave protoplasts of a uniform size, .apprx. 5-.mu.m diameter, depending on the strain and the stabilizing medium used. Conidia were preincubated with 2-deoxy-D-glucose in a minimal medium at 37.degree. C for 3 h. The swollen conidia were collected, resuspended in a buffer containing 0.4 M (NH4)2SO4 as stabilizer and incubated with Oerskovia lytic enzymes at 30.degree. C for 3 or 4 H. Approximately 80% of the conidia were converted into protoplasts. The protoplasts were separated from cell wall fragments and intact conidia by centrifugation over 30% sucrose. This isolation procedure gives a suspension of mononucleated or binucleated protoplasts suitable for recombination experiments and other studies for which a homogenous protoplast suspension is required. The procedure was successful for A. niger.