Localization studies of BcPde1 and BcPde2.

<p><b>A</b>: Germinated conidia of strain Δ<i>bcpde1</i>+<i>gfp</i>-<i>bcpde1</i> were microscopically examined in sugar-inducing conditions on object slides. GFP-BcPde1 fluorescence was observed over the whole hyphae. A very strong fluorescent signal was detected in so far unknown mobile spots (white arrows), with different sizes. DIC: differential interference contrast image; CFW: Calcofluor white staining of fungal cell wall (white), merge: GFP and CFW overlay. <b>B</b>: GFP-BcPde1 fluorescence in germinated conidia (20× magnification). <b>C</b>: 40× magnification of <i>gfp</i>-<i>bcpde1</i> expressing germinated conidia. Overlay of CFW (blue) and GFP pictures. Low exposure time presents various sizes of unexplained spots (white) of GFP fluorescence in different cellular regions. <b>D</b>: Germinated conidia of strain Δ<i>bcpde2</i>+<i>gfp</i>-<i>bcpde2</i> were microscopically examined in sugar-inducing conditions on object slides. GFP-BcPde2 fluorescence was observed over the whole hyphae. A strong signal was detected at the plasma membrane and septa (white arrows). E: Conidia of strain Δ<i>bcpde2</i>+<i>gfp</i>-<i>bcpde2</i> were microscopically examined after penetration of onion epidermis cells. Left: top of the onion epidermis. Right: Hyphae growing inside the onion epidermis after penetration. Asterisks present points of penetration. Strong fluorescence was observed at the plasma membrane and the septa. The Δ<i>bcpde2</i> phenotype (swollen hyphae after penetration) was restored by expression of <i>gfp</i>-<i>bcpde2</i>. Scale bar = 10 µm.