Combined with the natural flowering process of Lycoris radiata (L'Hér.) Herb., the regulatory effects of drought stress on its physiological characteristics and the expression of flowering-related genes and proline metabolism genes were systematically analyzed. The results show that during the natural flowering process, the contents of proline and soluble protein in L. radiata continuously increase, the soluble sugar content and superoxide dismutase activity generally increase first and then decrease, and the peroxidase and catalase activities first decrease and then increase. The flowering-promoting genes LrFCA(Unigene33609), LrFCA(Unigene48784), LrVIN3(CL157.Contig2), LrGI(CL7790.Contig5), and LrGI(CL7790.Contig11) are upregulated before the full blooming stage, the floral repressor gene LrSVP(CL9409.Contig10) is continuously downregulated, and the key enzyme genes LrP5CS1, LrP5CS2, LrP5CR, and LrOAT in the proline synthesis pathway are continuously upregulated. Drought stress delays the flowering of L. radiata, and the flowering time is delayed with the increase of stress levels. Compared with the control group, the malondialdehyde content and the above physiological indices of L. radiata are significantly increase (P<0.05) in general under drought stress, among which, the proline content increases with the prolongation of stress time and the increase of stress levels. Overall, with the prolongation of drought stress time, the expression of flowering-promoting genes LrFCA(Unigene33609) and LrGI(CL7790.Contig11) continuously decrease, while the expression of the floral repressor gene LrSVP(CL9409.Contig10) and the key enzyme genes LrP5CS1 and LrP5CR in the proline synthesis pathway continuously increase. The correlation analysis result shows that during the natural flowering process of L. radiata, the proline content shows an extremely significant negative correlation (P<0.01) with the relative expression of LrSVP(CL9409.Contig10); while under drought stress, they show an extremely significant positive correlation. In conclusion, proline exhibits significant context-dependent effects in the regulation of L. radiata flowering: during normal development, its gradual accumulation is an intrinsic requirement for the flowering process; while under drought stress, its explosive accumulation occurs synchronously with the activation of floral repressor genes, suggesting its involvement in the drought stress-induced flowering delay response. It is speculated that proline not only acts as an osmotic regulator but may also serve as a key physiological hub integrating internal developmental timing and external environmental stress signals, achieving a trade-off between survival and reproduction by reprogramming the downstream flowering gene network.