Isolation of protoplasts from Aspergillus nidulans conidiospores

Canadian Journal of Microbiology
C J Bos, S M Slakhorst

Abstract

Protoplasts were prepared from conidiospores of Aspergillus nidulans. The mononucleated conidia gave protoplasts of a uniform size, approximately 5-micron diameter, depending on the strain and the stabilizing medium used. Conidia were preincubated with 2-deoxy-D-glucose in a minimal medium at 37 degrees 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 degrees 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 also successful for Aspergillus niger.

Citations

Jan 1, 1988·Mutation Research·C J BosJ H van der Veen
Aug 1, 1987·Molecular & General Genetics : MGG·K WernarsH W van den Broek

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