Monitoring autophagy in the filamentous fungus Podospora anserina.

Autophagy has been monitored in the filamentous fungus Podospora anserina using electron, light, and fluorescence microscopy. In this organism autophagy can be induced either by starvation or rapamycin treatment or by het gene incompatibility. Incompatible HET products signal a cell death reaction referred to as cell death by incompatibility. In het-R het-V strain bearing the two incompatible het-R and het-V genes, cell death is induced by a simple shift in growth temperature, as incompatibility is thermosensitive. In this strain large autophagosomes are formed as revealed by electron microscopy or using the GFP-PaATG8 marker. This strain constitutes an alternative model to study autophagy. Analysis of the three autophagy mutants, DeltaPaATG1, DeltaPaATG8, and DeltapspA, reveals that autophagy is essential for aerial hyphae and female organ differentiation and involved in spore germination. During the incompatibility reaction, autophagy might protect cells from cell death as suggested by accelerated cell death observed in autophagy mutants.