The Photochemical Reflectance Index (PRI), an index based on leaf reflectance at 531 nm, has been found suitable for tracking variations in photosynthetic activity from leaf to ecosystem levels. This suitability has been attributed to PRI correlation with xanthophyll interconversion and photosynthetic radiation-use efficiency. However, other pigments and factors may be involved in such relationships. We studied the relationship between PRI and xanthophylls and other carotenoids in saplings of two widely distributed evergreen species (Scots pine and Holm oak) submitted to experimentally changing light conditions in a field experiment. PRI was strongly correlated with the de-epoxidation state of xanthophylls (DEPS, an expression of the relative concentration of the three xanthophyll cycle pigments), but also with carotenoids/chlorophyll ratio and β-carotene/chlorophyll ratio in both species. However, following momentary decreases in light due to clouds, PRI changed following the DEPS changes, while the carotenoids/chlorophyll ratio remained constant. The results show that PRI was able to reveal short-term changes in de-epoxidation state, i.e. the signal of xanthophyll interconversion, but it also tracked long-term changes in carotenoids/chlorophyll. Carotenoids other than xanthophylls, e.g. β-carotene, are also related to photoprotective processes, thus also making PRI effective as a measure of changes in photosynthetic light-use efficiency in response to stress on a long-term level.