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• Sensory analysis showed roasting reduced the perceived bitterness of WRT. • Phenolics, purine alkaloids and bitter amino acids decreased after roasting. • Human bitter taste receptors, TAS2R39 and TAS2R14, were robustly activated by WRT. • Roasting reduced the activation of WRT upon TAS2R39 and TAS2R14. The influence of roasting on tea bitterness remains unclear. With Wuyi Rock tea (WRT) as an example, this study investigated the impact of roasting on WRT's bitterness, utilizing an integrated approach involving sensory evaluation, bitter compound profiling, and cell-based calcium imaging. Sensory analysis revealed that roasting effectively reduced the perceived bitterness of WRT. This reduction was supported by decreases in various bitter compounds, including 19 flavanols, 11 flavonols, 12 phenolic acids, 2 purine alkaloids, and 9 bitter amino acids, which diminished by 16%, 26%, 19%, 2%, and 70%, respectively. Furthermore, we established two heterogeneous bitter receptor expression systems: TAS2R39/Gα15-HEK293T and TAS2R14/Gα15-HEK293T cell lines. These systems quantitatively confirmed the reduction in bitterness, demonstrating 51% and 62% decreases in intracellular calcium mobilization within the transfected cells, respectively. These findings provide compelling evidence for the bitterness-ameliorating effect of roasting, expanding our knowledge of the role of roasting in shaping the flavor of tea. [ABSTRACT FROM AUTHOR]