Saffron,derived from the stigma of theCrocus sativusflower, is a highly valued culinary and medicinal product. Its productivity is closely linked to flower formation, which is regulated by temperature. Suboptimal temperatures can lead to flower atrophy, resulting in reduced or absent flowering. To investigate the influence of temperature on saffron flowering, we conducted morphological, transcriptomic, and metabolic analyses on meristem-enriched tissues from corms grown under non-inductive (8 °C) and inductive (25 °C) conditions during the flowering stage. Morphological and histochemical observations indicated that higher temperatures promote floral induction, whereas lower temperatures suppress it. Integrated transcriptomic and metabolomic analyses revealed a key role for sugar metabolism in temperature-mediated floral regulation. Genes and metabolites associated with starch, sucrose, trehalose and raffinose metabolism were differentially regulated between the two temperature regimes. At the inductive(25 °C) temperature, increased starch degradation and accumulation of soluble sugars—particularly sucrose and trehalose-6-phosphate—were observed. In contrast, raffinose levels were elevated under non-inductive (8 °C) conditions. These findings suggest that sucrose and trehalose-6-phosphate act as positive regulators of floral induction, whereas raffinose functions as a repressor. Exogenous application of sugars further supported these observations: sucrose enhanced, while raffinose inhibited, floral induction in saffron. Functional validation through ectopic expression and complementation studies in Arabidopsis, along with Virus-Induced Gene Silencing (VIGS) in saffron, demonstrated that CsatTPS2 and NAC14 positively regulates, while CsatTPP3 and CsGLOS1 and LHY negatively regulates flowering. Together, this study uncovers a temperature-dependent regulatory mechanism in saffron, in which the balance of sugar metabolites and their signalling pathways governs floral induction.
