This study aimed to remediate petroleum-contaminated soil using co-pyrolysis biochar derived from rice husk and cellulose. Rice husk and cellulose were mixed in various weight ratios (0:1, 1:0, 1:1, 1:3 and 3:1) and pyrolyzed under 500 °C. These biochar variants were labeled as R0C1, R1C0, R1C1, R1C3 and R3C1, respectively. Notably, the specific surface area and carbon content of the co- pyrolysis biochar increased, potentially promoting the growth and colonization of soil microorganisms. On the 60th day, the microbial control group achieved a 46.69% removal of pollutants, while the addition of R0C1, R1C0, R1C3, R1C1 and R3C1 resulted in removals of 70.56%, 67.01%, 67.62%, 68.74% and 67.30%, respectively. In contrast, the highest efficiency observed in the abiotic treatment group was only 24.12%. This suggested that the removal of petroleum pollutants was an outcome of the collaborative influence of co-pyrolysis biochar and soil microorganisms. Furthermore, the abundance of Proteobacteria, renowned for its petroleum degradation capability, obviously increased in the treatment group with the addition of co-pyrolysis biochar. This demonstrated that co-pyrolysis biochar could notably stimulate the growth of functionally associated microorganisms. This research confirmed the promising application of co-pyrolysis biochar in the remediation of petroleum-contaminated soil.