Overseeding legumes in natural grasslands coupled with phosphorous fertilization are management practices oriented to increase forage production and quality, and to restore nutrient losses generated by livestock. Several studies show increases in forage due to this practice, but less is known about impacts on soil fertility and carbon sequestration. The objective of this study was to evaluate under real farm conditions changes in root C and N stocks and soil organic carbon (SOC) and nitrogen (SON) stocks in two different soil pools, the particulate organic matter (POM) and the mineral associated organic matter (MAOM), after the introduction in natural grasslands of a legume species, Lotus subbiflorus cv. "El Rincón", accompanied with phosphorous fertilization. We also evaluated changes in the natural abundance of 15N and 13C in soils and roots to understand changes in N fixation and species composition. We selected 12 adjacent paddocks of natural grasslands (NG) and natural grasslands overseeded with legumes and fertilized with phosphorous (NGLP) located in commercial farms in Uruguay. We found that overseeding legumes increased root C and N stocks and SOC and SON stocks in some farms but decreased them in others. On average, no significant differences arose between NGLP and NG paddocks in total stocks of 0-30 cm depth. However, higher C stocks were observed in POM of NGLP paddocks in 0-5 cm layer and lower contents in 5-10 cm layer indicating a change in the vertical distribution of C in POM. Changes in δ15N suggest that atmospheric N is being fixed by legumes in NGLP paddocks, but not translated into more N or C stocks in the MAOM fraction, probably due to high N losses promoted by cattle grazing. Our work suggests that carbon sequestration can be achieved after legumes introduction in grazed natural grasslands but will depend on grazing management practices.