Two spatially localized phases of glycogen accumulation were detected by electron microscopy after cytological staining of thin sections of Streptomyces coelicolor A3(2) colonies. In phase I, glycogen granules were present in hyphae in the air-agar interface region of colonies that were undergoing aerial mycelium formation, though absent from aerial hyphae themselves. With one exception (a bldF mutant, which contained abundant glycogen), the absence of aerial mycelium caused by various developmental mutations (bldA, bldB, bldC, bldD, bldG and bldH mutations) was associated with a virtual absence of detectable glycogen. Mutations that allow aerial hyphae to form but prevent or interfere with the septation needed for spore formation (whiA, whiB, whiG, whiH and whiI mutations) did not impair phase I deposition. In phase II, abundant glycogen granules were present in aerial hyphal tips during intermediate stages of sporulation, but disappeared as spores matured. Phase II glycogen accumulation was observed with bldA, bldC, bldD and bldG mutants grown with mannitol as carbon source - conditions that allowed normal aerial mycelium development and sporulation; but phase I deposition was still at a very low level in these colonies. Glycogen was also deposited in the coiling tips of aerial hyphae of whiA, whiB, whiH and whiI mutants, and sporadic clusters of granules were present throughout whiG colonies. Significantly, glycogen was deposited in spore chains that developed ectopically in the normally spore-and glycogen-free substrate mycelium when multiple copies of whiG were present. Overall, the two phases of glycogen synthesis (and degradation) appear to be under separate developmental control rather than being mainly responsive to external growth conditions. Phase II glycogen levels were particularly high in a whiE mutant defective in spore pigment biosynthesis, and particularly low when hyper-pigmentation was induced by additional copies of the whiE genes. Spore pigment may therefore be a major sink for carbon stored as glycogen during sporulation. The possibility is discussed that, in addition to supplying carbon and energy at particular locations, glycogen synthesis and degradation may also play a part in morphogenesis by influencing turgor pressure.