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Background In 2018, the European Union (EU) adopted Regulation 2018/841, which sets the accounting rules for the land use, land use change and forestry (LULUCF) sector for the period 2021–2030. This regulation is part of the EU’s commitments to comply with the Paris Agreement. According to the new regulation, emissions and removals for managed forest land are to be accounted against a projected forest reference level (FRL) that is estimated by each EU Member State based on the continuation of forest management practices of the reference period 2000–2009. The aim of this study is to assess how different modelling assumptions possible under the regulation may influence the FRL estimates. Applying the interlinked G4M and WoodCarbonMonitor modelling frameworks, we estimate potential FRLs for each individual EU Member State following a set of conceptual scenarios, each reflecting different modelling assumptions that are consistent with the regulation and the technical guidance document published by the European Commission. Results The simulations of the conceptual scenarios show that differences in the underlying modelling assumptions may have a large impact on the projected FRL. Depending on the assumptions taken, the projected annual carbon sink on managed forest land in the EU varies from −319 MtCO2 to −397 MtCO2 during the first compliance period (2021–2025) and from −296 MtCO2 to −376 MtCO2 during the second compliance period (i.e. 2026–2030). These estimates can be compared with the 2017 national GHG inventories which estimated that the forest carbon sink for managed forest land was −373 MtCO2 in 2015. On an aggregated EU level, the assumptions related to climate change and the allocation of forest management practices have the largest impacts on the FRL estimates. On the other hand, assumptions concerning the starting year of the projection, stratification of managed forest land, and timing of individual management activities are found to have relatively small impacts on the FRL estimates. Conclusions We provide a first assessment of the level of uncertainty associated with the different assumptions discussed in the technical guidance document and the LULUCF regulation, and the impact of these assumptions on the country-specific FRL. The results highlight the importance of transparent documentation by the EU Member States on how their FRL has been calculated, and on the underlying assumptions. Background