Straw removal for bioelectricity, cellulosic ethanol, or other bio-products has become a common practice in Brazilian sugarcane areas, but long-term effects on soil organic carbon (SOC) and yield are unknown. Our objective was to quantify tillage intensity and removal levels on SOC stocks and sugarcane yield on contrasting edaphoclimatic conditions in Brazil. Conventional tillage-CT and reduced tillage-RT coupled with three straw removal levels (none-NR; moderate-MR; total-TR) were combined to create six treatment combinations: CTNR, CTMR, CTTR, RTNR, RTMR, and RTTR, respectively. Sugarcane yield was measured annually and soil samples to a depth of 40 cm were collected and analyzed before and after 5 years of treatments. Neither tillage practices nor straw removal significantly affected sugarcane yield on sandy loam soil, but CTMR, CTTR, and RTTR showed substantial yield reductions on clayey soil. SOC stocks for RTNR increased by as much as 1.64 Mg ha −1 year −1 on clayey soil, while CTMR and CTTR favored depletion of SOC stocks on both soils. Overall, each Mg of straw returned to the soil increased SOC stock by 95 kg ha −1 for clayey soil under both tillage practices and 55 kg ha −1 for sandy loam under the CT system. Furthermore, straw removal decreased microbial biomass C and b-glucosidase activity. These findings suggest that the adoption of RT attenuates the adverse impacts of straw removal on SOC stocks while ensuring sugarcane yields for a more sustainable bioenergy production in Brazil.