ABSTRACT
Mycelial age is a very important factor in protoplasts release. Normally, fungal protoplasts are prepared from young mycelium which is more sensitive to the lytic action of digestive enzymes than old mycelium. Certain conditions required for preparing vacuoles from fungal protoplasts have been established. When a 2 ml volume of Fusarium culmorum protoplasts in 0.8 M mannitol was diluted with 5 ml of distilled water, disruption of most of the protoplasts occurred with the release of the cytoplasmic contents. Little work has been done on the ability of fungal protoplasts to synthesize constitutive and inducible enzymes but there is no doubt that protoplasts can make proteins and various other compounds as rapidly as intact living cells. The release of protoplasts irrespective of the lytic enzymes was similar in all filamentous fungi. In many cases, a combination of enzymes was more effective for maximum release of protoplasts than a single enzyme.
