ABSTRACT. Accurate estimation of aerial net primary production (ANPP) using remotely acquired data is one of the main challenges in both environmental monitoring and precision agriculture. Reflectance-based techniques have been widely used for decades, but detection of fluorescence emission by chlorophyll has emerged as a promising alternative in recent years. Although passive sun-induced fluorescence (SIF) monitoring has shown interesting results, the information it provides is limited to few wavelengths (Fraunhofer and telluric lines). On the other hand, active measurements of steady-state fluorescence and its spectral distribution cover the full-emission spectrum but have not been fully explored due to obvious experimental limitations. In this work we develop a novel active fluorescence measurement procedure, based on lamps and sensors mounted on a field tractor. This technique allowed the detection of the full spectrum of fluorescence emission of a plant crop for the first time in the literature. The main objective of this work was to analyze how the information based on reflectance and fluorescence, recorded by the new proposed methodology, tracks the differences caused by different irrigation treatments in the ANPP of three soybean varieties. We observed that reflectance-based vegetation indices showed limited sensitivity to these cumulative differences, as only EVI2, NDWI and SRWI were able to distinguish between rainfed and irrigation treatments in some few cases. Passive, irradiance-normalised SIF showed this same trend, but active fluorescence peak ratio (FRed/FFar-red) revealed statistically significant differences for the three cultivars studied. In addition, the latter showed a significant correlation with ANPP for two soybean varieties after correction for light re-absorption and scattering (p 0.5), which was observed for only EVI and foliar water status VIs among passive indicators. Active fluorescence measurements at leaf level by PAM fluorometry did not show differences between treatments in the upper part of the canopy but revealed a biomass-dependent decrease in PSII yield along the vertical axis. Our study demonstrated that fluorescence emission spectrum holds highly valuable information that might allow monitoring ANPP changes upon irrigation from remote sensing applications, and therefore should be carefully studied. Lastly, it highlights the potential of SIF retrieval at both O2-A and O2-B lines. © 2021 Elsevier Inc.