The accumulation and excretion of polyols was investigated under various growth conditions and at different stages of the life cycle of Aspergillus niger. Glycerol was found to be the major solute in osmotic adjustment of the hyphae. Conidiospores contain large amounts of mannitol, which are rapidly metabolized during early germination, leading to the accumulation of glycerol. Glycerol is the major polyol in young mycelium, whereas in older mycelium mannitol and erythritol predominate. In all experiments, polyols were also excreted. The mechanism and function of this process is unknown, but it might be a way to control the levels of the intracellular polyol pools. Polyols are rapidly taken up again upon starvation. In a glycerol kinase mutant the synthesis of glycerol is unaffected but the excreted level of the polyol is higher. This glycerol is taken up again upon starvation, and accumulates intracellularly as it can not be metabolized further.