Phosphorus (P) deficiency is one of the main reasons limiting plant production of Brassica napus L. Exploring the dynamics of leaf intracellular substances and the correlations with photosynthesis and growth helps to understand the response mechanisms of B. napus L. to P deficiency. This study conducted experiments on B. napus L. plants by measuring the leaf electrophysiological parameters, leaf structure, elastic modulus (E_m), photosynthesis, and growth indices under different P treatment conditions. The dynamics of leaf intracellular water and nutrients of B. napus L. were calculated and analyzed by using the electrophysiological parameters, and the plant tolerance threshold to low-P stress was discovered. The results indicated that the status of the leaf intracellular water and nutrients remained stable when the P concentration was not lower than 0.250 mmol·L⁻¹, but maximized the photosynthesis and growth at a P level of 0.250 mmol·L⁻¹. The 0.125 mmol·L⁻¹ P concentration significantly decreased the mesophyll cell volume, and the palisade–sponge ratio and tightness degree of leaf tissue structure were remarkably increased. This led to an increase in cell elastic modulus, and significantly improved the water retention capacity of leaf cells. At the same time, the intracellular water use efficiency and total nutrient transport capacity of leaves remained stable. As a result, the photosynthesis and growth of plants were maintained at the same level as that of the control group. However, photosynthesis and growth were clearly inhibited with a further decrease in P concentration. Therefore, 0.125 mmol·L⁻¹ was the tolerance threshold of B. napus L. to low P. With the help of electrophysiological information, the effects of the dynamics of intracellular substances on photosynthesis and growth of B. napus L. under low-P stress can be investigated, and the plant’s adaptive response can be revealed. However, the findings of the current hydroponic study are not directly applicable to field conditions with naturally P-deficient soils.