The present study focused on the wheat harvest grain protein content (GPC) estimation based on wheat leaf and canopy chlorophyll parameters, SPAD and SFR, which were acquired by two hand-held instruments, SPAD and Multiplex 3. The wheat GPC estimate experiment was applied on a wheat field of the Scientific Observation and Experiment Field Station for Precision Agriculture at suburb of Beijing in 2012. The wheat leaf SPAD and canopy SFR value were measured in field for all 110 wheat sample points at five different wheat growth stages from April to June. The wheat plant sample for each point was then collected after the SPAD and SFR measurement and sent to lab for leaf nitrogen content (LNC) and canopy nitrogen density (CND) analysis. Analysis results showed that the correlation coefficients of wheat GPC with wheat CND were much higher than that from wheat tillering stage to early milking stage. They were similar at the wheat middle milking stage. While the wheat leaf SPAD value was highly correlated with wheat LNC at wheat tillering, heading and early milking stage. Wheat canopy chlorophyll parameters SFR were highly correlated with wheat CND at wheat tillering, jointing, heading and milking stage. It can be seen from the study that SFR is more sensitive to the wheat CND compared with wheat LNC. The analysis also indicated that leaf SPAD value at wheat tillering, heading and milking stage was highly correlated with wheat GPC and yield of grain protein (YGP). The wheat canopy parameters, SFR_G and SFR_R were significantly correlated with wheat GPC and YGP at wheat milking stage. Then the optimal GPC and YGP estimation model was established. The R2 of GPC estimation models established by SPAD and SFR_R are 0.426 and 0.497, and the standard errors of the estimate are 0.060% and 0.055%, respectively. The R2 of YGP estimation models established by SPAD and SFR_R are 0.366 and 0.386 and the standard errors of the estimate are 125.367 and 123.454 kg·ha-1, respectively. The study reveals that SPAD value is a good indicator of single plant activity while SFR_G and SFR_R are better indicators for the wheat group activity. Wheat leaf SPAD value and canopy chlorophyll fluorescence information SFR are all feasible and valuable for GPC estimation before wheat harvesting.