The factors influencing the predominance of one of the two mineral forms of calcium oxalate (CO), the monohydrated whewellite (COM) and the di-hydrated weddellite (COD), forming the pruina of the upper cortex of lichens, have been investigated through a simple, sensitive histochemical assay: toluidine blue O (TBO), a metachromatic staining test. The differential reactivity of 43 thalli of 17 pruinose foliose species, supplemented by X-ray diffraction analysis and observations with polarizing and scanning electron microscopy, suggests that the histochemical reactivity of hyphal walls and cementing substances of the upper cortex are related to the density of anionic charges. These factors are probably due to the occurrence of polyuronic acid substances that strongly affects the mineralization of CO. Di-hydrated wedellite is always associated with TBO metachromatic reactivity, and COM with orthochromatic reactivity. When the material has an ambiguous ortho/metachromatic reactivity, COD and COM may occur together. This study presents the first experimental evidence that in lichens CO biomineralization is at least partially biologically controlled.