Soil organic carbon (SOC) content is a vital indicator for soil health. The use of moldboard (traditional) plowing for many years had led to a prominent decline in the SOC and soil organic matter (SOM) in Southern Russia. Application of no-tillage (NT) is a sustainable alternative to conventional tillage (CT) as it offers an advantage for SOC store. The aim of the study was to assess soil organic carbon dynamics in response to tillage practices in the steppe zone of Southern Russia. The conservation of SOC under different tillage systems (CT and NT) was evaluated in comparison with the soils of the virgin soils (VS) in three different regions of the steppe zone of the Lower Don region (Southern of the European part of Russia). The SOC content under the conditions of CT was significantly lower than that in the VS and demonstrated an inclining trend when using NT technology. We estimate that the transition to NT over an area of 5.5 million hectares will lead to a significant reduction of carbon emissions into the atmosphere (by ~39 × 109 g C/year), thereby SOC deposition will be (~5.1 × 1012 g C) and high economic advantages will be reaped (with cost savings of up to 27%) in the Rostov region of Russia.