Improving photosynthesis to raise wheat yield potential has emerged as a major target for wheat physiologists. Photosynthesis-related traits, such as nitrogen per unit leaf area (N_area) and leaf dry mass per area (LMA), require laborious, destructive, laboratory-based methods, while physiological traits underpinning photosynthetic capacity, such as maximum Rubisco activity normalized to 25 °C (V_cmax25) and electron transport rate (J), require time-consuming gas exchange measurements. The aim of this study was to assess whether hyperspectral reflectance (350–2500 nm) can be used to rapidly estimate these traits on intact wheat leaves. Predictive models were constructed using gas exchange and hyperspectral reflectance data from 76 genotypes grown in glasshouses with different nitrogen levels and/or in the field under yield potential conditions. Models were developed using half of the observed data with the remainder used for validation, yielding correlation coefficients (R² values) of 0.62 for V_cmax25, 0.7 for J, 0.81 for SPAD, 0.89 for LMA, and 0.93 for N_area, with bias <0.7%. The models were tested on elite lines and landraces that had not been used to create the models. The bias varied between −2.3% and −5.5% while relative error of prediction was similar for SPAD but slightly greater for LMA and N_area.