Optimum conditions for the release and regeneration of protoplasts of Alternaria alternata

Best release of Alternaria alternata protoplasts was obtained when 20 h old mycelia were incubated in a hydrolytic enzyme mixture of Novozym 234, Driselase, and β -glucuronidase. Numbers of nuclei/protoplast varied but generally decreased with increased time of incubation. While salts, were better osmotic stabilizers for protoplast release, sucrose and sugar alcohols were better for regeneration.