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Wheat grain protein content (GPC) is a key component when evaluating wheat nutrition. It is also important to determine wheat GPC before harvest for agricultural and food process enterprises in order to optimize the wheat grading process. Wheat GPC across a field is spatially variable due to the inherent variability of soil properties and position in the landscape. The objectives of this field study were: (i) to assess the spatial and temporal variability of wheat nitrogen (N) attributes related to the grain quality of winter wheat production through canopy fluorescence sensor measurements; and (ii) to examine the influence of spatial variability of soil N and moisture across different growth stages on the wheat grain quality. A geostatistical approach was used to analyze data collected from 110 georeferenced locations. In particular, Ordinary Kriging Analysis (OKA) was used to produce maps of wheat GPC, GPC yield, and wheat canopy fluorescence parameters, including simple florescence ratio and Nitrogen Balance Indices (NBI). Soil Nitrate-Nitrogen (NO₃-N) content and soil Time Domain Reflectometry (TDR) value in the study field were also interpolated through the OKA method. The fluorescence parameter maps, soil NO₃-N and soil TDR maps obtained from the OKA output were compared with the wheat GPC and GPC yield maps in order to assess their relationships. The results of this study indicate that the NBI spatial variability map in the late stage of wheat growth can be used to distinguish areas that produce higher GPC. Full article