Phosphorus deficiency induces biochemical and morphological changes which affect crop yield and production. Prompt fluorescence signal characterizes the PSII activity and electron transport from PSII to PSI, while the modulated light reflection at 820 (MR 820) nm investigates the redox state of photosystem I (PSI) and plastocyanin (PC). Therefore, combining information from modulated reflection at 820 nm with chlorophyll a fluorescence can potentially provide a more complete understanding of the photosynthetic process and integrating other plant physiological measurements may help to increase the accuracy of detecting the phosphorus deficiency in wheat leaves. In our study, we combined the chlorophyllafluorescence and MR 820 signals to study the response of wheat plants to phosphorus deficiency as indirect tools for phosphorus plant status characterization. In addition, we studied the changes in chlorophyll content index, stomatal conductance (gs), root morphology, and biomass of wheat plants. The results showed an alteration in the electron transport chain as a specific response to P deficiency in the I-P phase during the reduction of the acceptor side of PSI. Furthermore, P deficiency increased parameters related to the energy fluxes per reaction centers, namely ETo/RC, REo/RC, ABS/RC, and DIo/RC. P deficiency increased the values of MRminand MRmaxand decreased νred, which implies that the reduction of PSI and PC became slower as the phosphorus decreased. The principal component analysis of the modulated reflection and chlorophyll a fluorescence parameters, with the integration of the growth parameters as supplementary variables, accounted for over 71% of the total variance in our phosphorus data using two components and provided a reliable information on PSII and PSI photochemistry under P deficiency.