Conditions for efficient isolation and regeneration of protoplasts from Fulvia fulva

Abstract

A procedure for producing high yields and high regeneration frequencies of protoplasts from Fulvia fulva is described. This procedure was devised by systematic trials of various parameters, such as culture age and osmotic support, which are known to affect protoplast yield and regeneration. Mycelium from liquid cultures of 24–48 h, incubated with Novozym 234 in buffered 1.0 M MgSO4, gave the best conditions for protoplast release. Regeneration frequencies up to 50% were obtained with a complete medium containing 0.8 M sucrose for osmotic support.