Accurate sensor-based prediction of crop yield and grain quality in-season would enable growers to adjust nitrogen (N) fertilizer management for optimized production. This study assessed the feasibility (and compared the accuracy) of wheat (Triticum aestivum L.) yield, grain N uptake, and protein content prediction with in-season crop spectral reflectance measurements (Normalized Difference Vegetative Index, NDVI) obtained with a handheld GreenSeeker (GS) sensor and an Unmanned Aerial Vehicle (UAV)-mounted sensor. A strong positive correlation was observed between GS NDVI and UAV NDVI at Feekes 5 (R² = 0.78) and Feekes 10 (R² = 0.70). At Feekes 5, GS NDVI and UAV NDVI explained 42% and 43% of wheat yield, respectively. The correlation was weaker at Feekes 10 (R² of 0.34 and 0.25 for GS NDVI and UAV NDVI, respectively). The accuracy of wheat grain N uptake prediction was comparable to that of yield: the R² values for GS NDVI and UAV NDVI were 0.53 and 0.37 at Feekes 5 and 0.13 and 0.20 at Feekes 10. We found that neither GS NDVI nor UAV NDVI in-season data were useful in prediction of wheat grain protein content. In conclusion, wheat yield and grain N uptake can be estimated at Feekes 5 using either handheld or aerial based NDVI with comparable accuracy.