We explored ability of reflectance vegetation indexes (VIs) related to chlorophyll fluorescence emission (R₆₈₆/R₆₃₀, R₇₄₀/R₈₀₀) and de-epoxidation state of xanthophyll cycle pigments (PRI, calculated as (R₅₃₁- R₅₇₀)/(R₅₃₁-R₅₇₀) to track changes in the CO₂ assimilation rate and Light Use Efficiency (LUE) in montane grassland and Norway spruce forest ecosystems, both at leaf and also canopy level. VIs were measured at two research plots using a ground-based high spatial/spectral resolution imaging spectroscopy technique. No significant relationship between VIs and leaf light-saturated CO₂ assimilation (A(MAX)) was detected in instantaneous measurements of grassland under steady-state irradiance conditions. Once the temporal dimension and daily irradiance variation were included into the experimental setup, statistically significant changes in VIs related to tested physiological parameters were revealed. ΔPRI and Δ(R₆₈₆/R₆₃₀) of grassland plant leaves under dark-to-full sunlight transition in the scale of minutes were significantly related to A(MAX) (R² = 0.51). In the daily course, the variation of VIs measured in one-hour intervals correlated well with the variation of Gross Primary Production (GPP), Net Ecosystem Exchange (NEE), and LUE estimated via the eddy-covariance flux tower. Statistical results were weaker in the case of the grassland ecosystem, with the strongest statistical relation of the index R₆₈₆/R₆₃₀ with NEE and GPP.