ABSTRACT
Protoplasts are recognized as being investigatory structures in their own right and are providing a means of studying a wide range of cellular, biochemical, and genetic processes. One of the most important considerations in these investigations is to maximize yields, while at the same time retaining the ability to culture protoplasts. Protoplasts have been prepared from fungi, representing all the major taxonomic groups, by enzymatic digestion of all or part of the cell wall. There are a number of commercial enzymes available that have been used to isolate protoplasts from yeasts and filamentous fungi. Partial or total digestion of the mycelial walls in filamentous fungi leads to the subdivision of the cytoplasm into a heterogeneous population of protoplasts with respect to their internal morphology, physiology, and biochemistry. The influence of culture age on protoplast isolation is better documented than medium effects and growth conditions. The choice of buffer in the incubation medium can affect rate of protoplast isolation and yield.
