Ranunculus asiaticus L. is a quantitative long-day geophyte, grown in a cold greenhouse for cut flowers and potted plants. Flowering in ranunculus is a complex process, strongly steered by temperature and photoperiodism. Vernalization of rehydrated tuberous roots anticipate sprouting and leaf rosette formation and flowering. It is known that the time for flowering and the sensitivity to cold treatment, in terms of flowering anticipation, varies in numerous hybrids, while no information seems to be available on the influence of hybrids and on the vernalization on the photosynthetic process and primary metabolite profiling. We investigated the influence of two ranunculus hybrids, MDR and MBO, and two preparation procedures of tuberous roots, only rehydration (Control, C) and rehydration followed by vernalization (V), on the photosynthesis and photochemistry of plants grown in a climatic chamber, under a controlled environment. In addition, in MBO plants, in which the vernalization showed the main effects, carbohydrate, amino acid and protein levels were also investigated. In control plants, the response of leaf photosynthesis, to increasing white light, revealed higher photosynthetic activity in MDR than in MBO. The quantum yield of PSII (ϕPSII), electron transport rate (ETR) and non-photochemical quenching (NPQ) did not differ between the two hybrids. The maximal photochemical efficiency (Fv/Fm) was higher in MBO than in MDR and showed a decrease in both hybrids after vernalization. The preparation treatment of propagation material affected the light response of photosynthesis in the two hybrids differently, which increased in plants from vernalized tuberous roots, compared to those from only rehydrated in MBO and decreased in MDR, in accordance to the effects of vernalization observed in leaf photosynthetic pigments. In MBO vernalized tuberous roots, starch was rapidly degraded, and the carbon skeletons used to synthesize amino acids. Control plants of MBO, developed more leaves than those of MDR and a consequent larger plant leaf area. Compared to only rehydration, vernalization of rehydrated tuberous roots increased the plant leaf area in both the hybrids. Compared to the control, vernalized tuberous roots of MBO showed higher concentrations of sucrose and free amino acids, which could act as a long-distance signal promoting floral transition in young leaf primordia.