A convenient procedure for rapid release of protoplasts from Ustilago maydis.

Biotechniques

Dept. of Botany, University of Maryland, College Park 20742.

Published: October 1988

Filtrates from cultures of Trichoderma harzianum, grown on a liquid mineral salts medium containing vitamin and trace element supplements, 0.5 g/l glucose and 2 g/l of dried residue of ethanol-extracted commercial mushrooms (Agaricus brunnescens), are a good source of enzymes for the release of protoplasts from the sporidia of Ustilago maydis. When concentrated 50-fold by (NH4)2SO4 precipitation, the enzymes liberated 95% of the protoplasts from the sporidia within 40 min.

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