Preparation and Regeneration of Mycelial Protoplasts of Alternaria eichhomiae

Preparation and regeneration of mycelial protoplasts from Aliernaria eichhorniae were examined. A commercially available muralytic enzyme, Novozym 234, was used for isolation of protoplasts. The mycelial age and the pH of the stabilized buffer affected the formation of protoplasts. The maximum production of protoplasts (3,9 X 10*/gfresh weight mycelia) was obtained from 24- h-old mycelia digested with Novozym 234 (20 mg/m!) in a stabilized buffer of pH 6.4 and incubated in the dark at 30"C on a rotary shaker (90 r.p.m.) for 6h. Morphological characteristics of the protoplasts varied and depended on the age of the mycelia used in protoplast production. Moreover, mycelial age had a highly significant influence (P = 0.0001) on the frequency of protoplast regeneration