Biochar-amended soils influence the degradation of herbicides depending on the pyrolysis temperature, application rate, and feedstock used. The objective of this study was to evaluate the influence of sugarcane straw biochar (BC) produced at different pyrolysis temperatures (350 °C, 550 °C, and 750 °C) and application rates in soil (0, 0.1, 0.5, 1, 1.5, 5, and 10%w/w) on metribuzin degradation and soil microbiota. Detection analysis of metribuzin in the soil to find time for 50% and 90% metribuzin degradation (DT50and DT90) was performed using high-performance liquid chromatography (HPLC). Soil microbiota was analyzed by respiration rate (C-CO2), microbial biomass carbon (MBC), and metabolic quotient (qCO2). BC350 °C-amended soil at 10% increased the DT50of metribuzin from 7.35 days to 17.32 days compared to the unamended soil. Lower application rates (0.1% to 1.5%) of BC550 °C and BC750 °C decreased the DT50of metribuzin to ~4.05 and ~5.41 days, respectively. BC350 °C-amended soil at high application rates (5% and 10%) provided high C-CO2, low MBC fixation, and highqCO2. The addition of low application rates (0.1% to 1.5%) of sugarcane straw biochar produced at high temperatures (BC550 °C and BC750 °C) resulted in increased metribuzin degradation and may influence the residual effect of the herbicide and weed control efficiency.