Abstract
Summary Asynchronous development of berries causes metabolic chimerism in usual samples. We thus revisited the developmental changes in the metabolome of the Vitis vinifera single berries from anthesis to over-ripening. A dataset of 9,256 ions obtained by non-targeted ultra-performance liquid chromatography coupled to high-resolution mass spectrometry was submitted to an analysis workflow combining classification and dimension reduction tools, to reveal the dynamics of metabolite composition without phenological a priori . This approach led to a metabolome-based definition of developmental stages, as well as the clustering of metabolites into 12 specific kinetic patterns. The single berry intrinsic metabolomic clock alleviates constitutive asynchronicity biases in the usual combination of phenological scales and observer clock. Such increase in temporal resolution enabled the identification of metabolite clusters annunciative of the onset of ripening since the herbaceous plateau. In particular, these clusters included transient lipidic changes and the start of ABA accumulation. We also highlighted a cluster of stilbenes that accumulate after sugar loading stops, during fruit shriveling. This non-targeted approach enables a more precise and unbiased characterization of grapevine berry development through the metabolomic clock paradigm. The discovery of new metabolic milestones of berry development paves the way towards an unbiased assessment of berry physiological stages.