The use of subtropical perennial grasses in temperate grazing systems is increasingly being promoted for production and environmental benefits. This study employed a combination of elemental and stable isotope analyses to explore whether pastures sown to either kikuyu (Pennisetum clandestinum) or a combination of panic (Panicum maximum) and Rhodes grass (Chloris gayana) could increase soil organic carbon (SOC) levels in five regions across southern Australia. Carbon was sequestered under kikuyu at a rate of 0.90±0.25MgCha-1year-1 along the south coast of Western Australia. Lower but still significant sequestration rates were found for kikuyu in South Australia (0.26±0.13MgCha-1year-1). No changes in SOC were found for panic–Rhodes grass pasture systems in the northern district of Western Australia. Additionally, we found no changes in SOC when kikuyu-based pastures were established on formerly cropped paddocks in the Namoi Catchment of New South Wales. Stable isotope results corroborated these findings and suggested that, where SOC has accumulated, the gains have been dominated by SOC derived from the perennial vegetation and have been concentrated in the upper 10cm of soil.