With the increasing accumulation of cow manure, the increasing sewage discharge and the improper treatment of Ganoderma lucidum residues, it is urgent to utilize them as resources. This papaer studied the enhancement of anaerobic digestion (AD) of Ganoderma lucidum residue biochar. Ganoderma lucidum residue biochar prepared at different pyrolysis temperatures had different enhancement effects on AD of cow manure. Among them, the addition of Ganoderma lucidum residue biochar prepared at 450℃ has the best enhancement effect. Compared with the control group, the biogas and methane production increased by 20.7% and 20.59%, respectively. Additionally, the biogas and methane of AD is not positively correlated with the dosage of water-soluble and water-insoluble Ganoderma lucidum residue biochar pyrolyzed at 450℃. The optimal dosage of water-soluble Ganoderma lucidum residue biochar was W3 (0.015 g), which increased the biogas and methane production by 8% and 14.28%, respectively. The best dosage of insoluble Ganoderma lucidum residue biochar is R1 (0.603 g), which can increase biogas by 19.09% and methane by 26.91%, respectively. However, 1.203 g of water-insoluble Ganoderma lucidum residue biochar will inhibit the anaerobic digestion of biogas and methane from cow manure, which maybe be due to the increase in the number of the Candidatus Nitrobacteria. It will compete with the Methanobacterium for CO2, and inhibit the hydrogen methane production pathway in AD, thus reducing biogas and methane production in AD.