Climate seasonality critically influences the functioning and dynamics of ecosystems in continental areas. The ecological importance of winter temperatures on high-latitude vegetation changes has recently been argued to be largely overlooked in comparison to summer temperatures. The Oymyakon region from eastern Siberia, with its strong continentality of extremely cold winters and moderately warm summers, is ideally suited to study the response of past vegetation to seasonal temperature changes based on long ecological time-series. However, few paleorecords are available from this area. The history of regional glacial activity and potential plant refugia since Marine Isotope Stage (MIS) 3 is not well understood. Here, we present geochemical and plant DNA metabarcoding records retrieved from a sediment core from Lake Ulu in the Oymyakon region, which provides detailed information on glacier and vegetation dynamics over the last 43 cal. ka BP. Our results suggest that glacial fluctuations were primarily driven by summer insolation, and Lake Ulu was likely initiated by glacial retreat during MIS 3. The catchment experienced multiple glacial advance/retreat cycles until the Last Glacial Maximum, and the glaciers fully retreated by 20 cal. ka BP. In addition, a tundra-steppe landscape dominated by Dryas, Papaver, Saliceae, and Anthemideae occupied the catchment for most of the time and began to collapse around 19 cal. ka BP following the expansion of trees and shrubs such as Larix, Betula, Alnus, and Vaccinium. Postglacial plant assemblages in the Oymyakon region exhibit a high sensitivity to summer temperature variations, with minimal impact from winter temperatures. This can be explained by the dominance of summer insolation amplitude, extreme continentality, extended plant growing season, and plant genetic adaptation to cold. Notably, our ancient DNA record show the earliest postglacial expansion of larch in eastern Siberia (around 18.6 cal. ka BP), which is likely related to the presence of local refugia. This implies that the Oymyakon region may be one of the earliest sources for larch recolonization and that more research should be implemented to provide insights into larch expansion and migration, and to better predict the future scenarios for Siberian larch forests.