Auto-fluorescence of cell wall of sporidia.

<p>A and B: AS recovered from 12-day-old cultures in solid-state fermentation. C and D: SS grown in liquid-state fermentation. Arrows show the sporidia hila.</p

SEM images of <i>Pseudozyma flocculosa</i> sporidia.

<p>A: aerial sporidia produced by solid-state fermentation; B: submerged sporidia from the liquid-state fermentation.</p

UV-visible spectrophotometry of melanin extracts.

<p>MAS: Melanin extracts from aerial sporidia, MSS: Melanin extracts from submerged sporidia, DOPA: synthetic melanin. ABS: absorbance.</p

SEM and TEM images of sporidia wall surface.

<p>A: aerial sporidia (AS) wall with ornament granules. B: submerged sporidia (SS) with a smooth wall surface. C: thick walls of AS, D: thin wall (white arrows) of SS with clear nucleus (N). E: thick walls of AS with an inner fibrous electron-lucent layer (SF) and an outer granular electron-dense layer (SD) showing a high affinity for the contrasting agents, F: hilum of sporogenesis (HS) of AS and G: Thin electron-lucent wall of SS.</p

SEM views of the three different populations of aerial sporidia.

<p>A: Large sporidia (P1), medium-sized sporidia (P2) with two scars (arrows) and small sporidia (P3) with one scar only. B: Flow cytometry analyses corresponding to the AS sample, revealed different morphotypic sub-populations (results were displayed on Forward Scatter Air/Forward Scatter Height (FSC-A/FSC-H) dot-plot on the basis of 40,000 events).</p

Fluorescence spectra obtained by SpectraMax M5 in the 300–440 nm wavelength range.

<p>MAS: melanin extracts from aerial sporidia, DOPA: sample containing a synthetic melanin (DOPA melanin) (0.5mg/ml), C-: reference sample without melanin. RFU: Relative fluorescence unit.</p

Differences in the cytoplasm of aerial sporidia (AS) and submerged sporidia (SS).

<p>The sporidia were colored by Nile-red fluorescent dye marking the intracellular cells lipids. A and B: AS have a dense compact nucleus (white arrows) with large lipid stocks. C and D: SS are rich in mitochondria organelle and have a clear nucleus (white arrows).</p

<i>Pseudozyma flocculosa</i> sporidia in phase contrast microscopy.

<p>A: aerial sporidia (AS) obtained by solid-state fermentation, exhibiting one or two apiculate hilum (asterisks). B: submerged sporidia (SS) obtained by liquid-state fermentation, regularly showing protrusion bulging and inclusions, Arrows point to the apiculate hilum of the AS.</p

Germination test.

<p>A series: germination of submerged sporidia. B series: Germination of aerial sporidia. Numbers correspond to test time, expressed in hours (h).</p