There is a strong trend toward reforestation of abandoned grasslands in alpine regions which may impact the carbon balance of alpine ecosystems. Here, we studied the effects of afforestation with Norway spruce (Picea abies L.) on an extensively grazed subalpine pasture in Switzerland on soil organic carbon (SOC) cycling and storage. Along a 120-year long chronosequence with spruce stands of 25, 30, 40, 45, and >120 years and adjacent pastures, we measured tree biomass, SOC stocks down to the bedrock, natural 13C abundances, and litter quality. To unravel controls on SOC cycling, we have monitored microclimatic conditions and quantified SOC decomposability under standardized conditions as well as soil respiration in situ. Stocks of SOC were only moderately affected by the afforestation: in the mineral soil, SOC stocks transiently decreased after tree establishment, reaching a minimum 40–45 years after afforestation (−25 %) and increased thereafter. Soils of the mature spruce forest stored the largest amount of SOC, 13 % more than the pasture soils, mainly due to the accumulation of an organic layer (23 t C ha−1). By comparison, C accumulated in the tree biomass exceeded the SOC pool by a factor of three in the old forest. In contrast to the small impact on C storage, afforestation strongly influenced the composition and quality of the soil organic matter (SOM). With increasing stand age, δ13C values of the SOM became consistently more positive, which can be interpreted as a gradual replacement of grass- by spruce-derived C. Fine roots of spruce were enriched in 13C, in lignin and had a higher C/N ratio in comparison to grass roots. As a consequence, SOM quality as indicated by the lower fraction of readily decomposable (labile) SOM and higher C:N ratios declined after the land-use change. Furthermore, spruce plantation induced a less favorable microclimate for microbial activity with the average soil temperature during the growing season being 5 °C lower in the spruce stands than in the pasture. In situ soil respiration was approximately 50 % lower after the land use conversion, which we primarily attribute to the colder conditions and the lower SOM quality, but also to drier soils (−25 %) and to a decreased fine root biomass (−40 %). In summary, afforestation on subalpine pastures only moderately affected SOC storage as compared to the large C sink in tree biomass. In contrast, SOC cycling rates strongly decreased as a result of a less favorable microclimate for decomposition of SOM, a lower C input by roots, and a lower litter quality.