The potato is one of the most cultivated crops worldwide, providing an important source of food. The quality of potato tubers relates to their size and dry matter composition and to the absence of physiological defects. It depends on the spatial and temporal coordination of growth and metabolic processes in the major tuber tissues: the cortex, flesh and pith. In the present study, variations in the biochemical traits of each of these tissues were investigated during tuber growth under optimal and water-deficit conditions. MRI relaxometry was used as a non-invasive and quantitative method to access information on cellular water status. The presence of slight but significant variations in organic compound contents quantified in the cortex and flesh revealed a tissue-dependent metabolic pattern. The T2 and relative I0 of the bi-exponential relaxation signal allowed a distinction to be made between the pith and the cortex, whereas the flesh could be differentiated from these tissues only through its relative I0. T2 values did not vary significantly during tuber development, in accordance with the typical growth pattern of tubers, but were shown to be sensitive to water stress. The interpretation of the multi-exponential transverse relaxation times is discussed and could be further developed via microscopic analysis.