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Before the end of the Last Glacial Maximum (LGM, ∼16.5 ka ago) 1 set in motion major shifts in human culture and population structure, 2 a consistent change in lithic technology, material culture, settlement pattern, and adaptive strategies is recorded in Southern Europe at ∼18–17 ka ago. In this time frame, the landscape of Northeastern Italy changed considerably, and the retreat of glaciers allowed hunter-gatherers to gradually recolonize the Alps. 3–6 Change within this renewed cultural frame (i.e., during the Late Epigravettian phase) is currently associated with migrations favored by warmer climate linked to the Bølling-Allerød onset (14.7 ka ago), 7–11 which replaced earlier genetic lineages with ancestry found in an individual who lived ∼14 ka ago at Riparo Villabruna, Italy, and shared among different contexts (Villabruna Cluster). 9 Nevertheless, these dynamics and their chronology are still far from being disentangled due to fragmentary evidence for long-distance interactions across Europe. 12 Here, we generate new genomic data from a human mandible uncovered at Riparo Tagliente (Veneto, Italy), which we directly dated to 16,980–16,510 cal BP (2σ). This individual, affected by focal osseous dysplasia, is genetically affine to the Villabruna Cluster. Our results therefore backdate by at least 3 ka the diffusion in Southern Europe of a genetic component linked to Balkan/Anatolian refugia, previously believed to have spread during the later Bølling/Allerød event. In light of the new genetic evidence, this population replacement chronologically coincides with the very emergence of major cultural transitions in Southern and Western Europe. • Genetic replacement in Southern European hunter-gatherers started at least 17 ka ago • The shift follows the LGM and precedes the onset of the Bølling/Allerød event • This process is chronologically associated with major cultural transitions in Europe Bortolini et al. generate genomic evidence and direct dating from an individual who lived ∼17 ka ago in northeastern Italy and backdate by about 3 ka the diffusion in Southern Europe of a genetic component linked to Balkan/Anatolian refugia previously believed to have spread during later major warming shifts. [ABSTRACT FROM AUTHOR]