The photochemical reflectance index (PRI), based on reflectance signatures at 531 and 570 nm, and associated with xanthophyll pigment inter-conversion and related thylakoid energisation, was evaluated as an indicator of photosynthetic function in a Mediterranean holm oak (Quercus ilex L.) coppice. The chlorophyll fluorescence pulse-amplitude-modulation and the eddy correlation techniques were used to estimate the photosystem 2 photochemical efficiency of leaves and the CO2 flux over the canopy, respectively. The reflectance and fluorescence techniques yielded identical estimates of the photosynthetic activity in leaves exposed to dark-light-dark cycles or to a variable irradiance in laboratory. However, there was no such correlation between photosynthetic performance and PRI when applied to a sun-exposed canopy in field conditions. Fluorescence profiles inside the canopy and especially a helpful use of multispectral reflectance imaging highlight the limitations of such method.