Understanding the impacts of straw removal on quantity and quality of soil organic carbon (SOC) is crucial for sustaining or improving soil functions and producing economically viable sugarcane yields. Field experiments were carried out on commercial farms to quantify effects of straw removal on sugarcane yields, SOC stocks and the degree of soil organic matter (SOM) humification under three diverse edaphoclimatic conditions (Quirinopolis-GO, Chapadao do Ceu-GO and Quata-SP) in Brazil, while considering the effects of cover crop on sugarcane yields during two harvest seasons. Three straw removal rates (0, 50, and 100\%) were arranged in a randomized block design with four replications within two paired areas, one seeded with Crotalaria spectabilis (cover crop-CC) and the other kept under bare fallow (BF) during the sugarcane-replanting period. Sugarcane yields were measured annually using an instrumented truck equipped with load cells, and soil samples were collected to a depth of 40-cm two years after establishing the trials. Straw removal for two years did not significantly influence cane yields in Quata-SP, but in Chapadao do Ceu-GO and Quirindpolis-GO, complete straw removal resulted in cumulative yield losses of up to 28 and 62 Mg ha(-1) respectively. The inclusion of Crotalaria spectabilis within sugarcane cropping cycle increased two-year cumulative yields by 25 and 27 Mg ha-1 in Chapadao do Ceu-GO and Quirinopolis-GO respectively. SOC response to straw removal was highly site-specific after two years. In fine-textured soils, straw removal rates did not significantly affect SOC stocks in Chapadao do Ceu-GO, but in Quirinopolis-GO complete straw removal favored the depletion of SOC stock relative to partial or no straw removal in both soil layers (0-10 and 0-40 cm) of the BF area, while within the CC area SOC depletion was observed in the 0-40 cm layer. For sandy soil (Quata-SP), complete straw removal decreased SOC stock only in the top 10 cm of the CC area. Laser-Induced Fluorescence Spectroscopy showed high degree of SOM humification in soils with depleted SOC stock, indicating that excessive straw removal was degrading soil quality by reducing the amount of labile C in the SOM. Conclusions drawn from this study indicate that, even on a short-term basis, complete straw removal already began to modify the quantity and quality of SOC, while both moderate or complete removals adversely affected sugarcane yields.