Cytological comparison of early stages of wall regeneration of Cercospora nicotianae and Neurospora crassa protoplasts

Freshly isolated protoplasts of Cercospora nicotianae and Neurospora crassa are equally sensitive to the toxin, cercosporin. After a 12-h regeneration period C. nicotianae cells are resistant, but N. crassa cells remain sensitive. Production of cell wall material by both C. nicotianae and N. crassa was monitored by transmission electron microscopy and fluorescence microscopy. Freshly isolated protoplasts lacked cell wall material as shown by observation with electron microscopy and inability to bind the fluorescent brightener Tinopal 5BM. After a 12-h incubation, electron micrographs of regenerating protoplasts showed well-developed cell walls for N. crassa, whereas C. nicotianae displayed variations in wall structure. Ability to bind Tinopal 5BM was acquired very early by regenerating cells of both fungi. Percentages of cells that could bind Concanavalin A did not differ between the two fungi at any time after protoplast isolation. Ability to bind wheat germ agglutinin and Bandeiraea simplicifolia agglutinin II was detected earlier in C. nicotianae than in N. crassa. These data demonstrate the presence of cell wall materials in both C. nicotianae and N. crassa at the time that differential sensitivity to cercosporin is observed. These results suggest that components in the C. nicotianae cell wall may play a role in cercosporin resistance.