Abstract. Forest steppes are dynamic ecosystems, highly susceptible to changes in climate and land use. Here we examine the Holocene history of the European forest steppe ecotone in the Lower Danube Plain to better understand its sensitivity to climate fluctuations and human impact, and the timing of its transition into a cultural forest steppe. We used multi-proxy analyses (pollen, n-alkane, coprophilous fungi, charcoal, and geochemistry) of a 6000-year sequence from Lake Oltina (SE Romania), combined with a REVEALS model of quantitative vegetation cover. We found the greatest tree cover, composed of xerothermic (Carpinus orientalis and Quercus) and temperate (Carpinus betulus, Tilia, Ulmus and Fraxinus) tree taxa between 6000 and 2500 cal yr BP. Maximum tree cover (~ 50 %) occurred between 4200 and 2500 cal yr BP at a time of wetter climatic conditions. Compared to other European forest steppe areas, the dominance of Carpinus orientalis represents the most distinct feature of the woodland's composition during that time. Forest loss was under way by 2500 yr BP (Iron Age) with REVEALS estimates indicating a fall to ~ 20 % tree cover from the mid-Holocene forest maximum linked to clearance for agriculture, while climate conditions remained wet. Biomass burning increased markedly at 2500 cal yr BP suggesting that fire was regularly used as a management tool until 1000 cal yr BP when woody vegetation became scarce. A sparse tree cover, with only weak signs of forest recovery, then became a permanent characteristic of the Lower Danube Plain, highlighting recurring anthropogenic pressure. The timing of anthropogenic ecosystem transformation here (2500 cal yr BP) was in between that in central eastern (between 3700 and 3000 cal yr BP) and eastern (after 2000 cal yr BP) Europe. Our study is the first quantitative land cover estimate at the forest steppe ecotone in south eastern Europe spanning 6000 years and provides critical empirical evidence that the present-day forest steppe/woodlands reflects the potential natural vegetation in this region under current climate conditions. This study also highlights the potential of n-alkane indices for vegetation reconstruction, particularly in dry regions where pollen is poorly preserved.