Abandonment of croplands ongoing on 220 million ha worldwide contributes strongly to soil restoration by improvement of degraded properties and medium- and long-term carbon (C) sequestration in post-agricultural ecosystems. Two interrelated processes – decomposition and stabilization of soil organic carbon (SOC) – govern SOC dynamics and affect the C source or sink functions of former croplands. We investigated how the abandonment of arable soils affects (i) accumulation of SOC, its composition, stability, and turnover during the post-agricultural restoration of soils, and (ii) microbial activity parameters. A chronosequence study was carried in two bioclimatic zones of European Russia: deciduous forest (Luvic Phaeozems, PH-chronosequence) and dry steppe (Calcic Chernozems, CH-chronosequence). Each chronosequence included an arable soil, 3–4 soils abandoned at increasing time periods (up to 35 years), and natural soil: never cropped Phaeozem and completely restored Chernozem. We combined the results of nuclear magnetic resonance (NMR), thermal analysis including Differential Scanning Calorimetry and Derivative Thermogravimetry, long-term incubation for SOC mineralization, and microbiological activity (basal respiration and microbial C content). Degraded Phaeozems with low SOC amount had much higher relative increase in SOC content (134%) during the post-agricultural restoration compared to SOC-rich Chernozems (38%). SOC gains were recorded in all organic compound classes identified by NMR and thermal analysis, but the increase of recalcitrant SOC was more pronounced in the post-agricultural Chernozems than in the Phaeozems. The post-agricultural Chernozems were characterized by higher SOC aliphaticity and aromaticity than Phaeozems. Microbial activity and biodegradable SOC increased gradually during post-agricultural restoration. Being mostly a function of climate and vegetation, the soil type was the primary factor explaining the greatest portion (54–88%) of the total variance for most soil and microbial parameters. Concluding, despite SOC content increased in both Chernozems and Phaeozems during the post-agricultural restoration, the mechanisms of C sequestration and stabilization were dependent on climate, vegetation, and on the degradation intensity during the agricultural use. The accumulation of organic compounds was specific for virgin soils dominating in deciduous forest and steppes, and had direct consequences for microbial activities, C turnover and sequestration. © 2019 Elsevier B.V.