FIG. 4.

Phenotypic analysis of ramosa-1 and T312 germlings. Samples were taken after cultivation for 13 h at 24°C, followed by cultivation for 4 h at 37°C. Microscopic pictures were taken for at least 50 germlings from each strain, and representative pictures are shown. (a to d) Actin immunostaining of the ramosa-1 mutant (a and b) and T312 (c and d). Polarized actin localization is indicated by arrows, an actin collar by arrowheads, and depolarized actin patches by stars. (e to h) CFW staining of the ramosa-1 mutant (e and g) and T312 (f and h). Pictures for panels e and f were taken using an automatic exposure time, and pictures for panels g and h were taken using a 100-ms exposure time. Polarized chitin localization as visible in panel f is indicated by arrows. The increase in CFW fluorescence intensity in panel g suggests enhanced chitin levels at ramosa-1 cell walls. (i and j) Microscopic images of the ramosa-1 mutant (i) and T312 (j) stained with NBT. The presence of ROS is reflected by blue hyphal tips. Bars, 5 μm for panels a to d and 10 μm for panels e to j.