Anaerobic digestion (AD), as a crucial technology for organic waste resource recovery, faces the challenge of low efficiency in converting high-load organic substance into biogas. In this study, high-load AD system with food waste and excess sludge as co-substrates was constructed. The effect and mechanism of carbon quantum dots (CQD) derived from straw in promoting the performance of AD systems have been studied. The oxidation pretreatment of straw with H2O2 and acetic acid increased the yield of hydrothermal synthesis CQD to approximately 40%. The effect of different CQD on CH4 yield performance was further explored. The cumulative CH4 yield performance of the fermenter was improved after adding CQD. The CQD synthesized from pretreated straw and the nitrogen-doped CQD synthesized using Chlorella as the nitrogen source showed competitive promotion performance, increasing cumulative CH4 yield by 17.71% and 8.87%, respectively. These CQD can effectively accelerate the degradation of dissolved organic matter and thus improve the CH4 yield performance, with the most significant effect of CQD synthesized from pretreated straw. Electrochemical analysis and the correlation analysis between microorganisms and performance parameters showed that these CQD established an electron conductive network to enhance the electron transfer of the system. This well conductive conditions enriched hydrogenotrophic methanogenic (Methanosarcina), electroactive bacteria (Clostridium_sensu_stricto_1), and hydrolytic-acidifying bacteria (norank_f__Bacteroidetes_vadinHA17). This study significantly enhanced the yield of straw-derived CQD through green methods, and deeply revealed the potential promoting mechanisms of biomass-derived CQD by investigating the correlation between system performance and microorganisms in high-load AD systems