In India, one of the major mushroom producer state is Punjab. During harvesting, fruiting bodies of mushroom are removed from compost bed. The base of the stem which is embedded in the compost is removed by cutting with knife or scissor before packaging, sorting and supplying in the market. This large amount of waste generated needs to be disposed off which is a major cause of environmental pollution. Anaerobic digestion is a technology used for the treatment of various types of wastes and produces biogas. As the mushroom waste generated is biodegradable in nature, it can be effectively used for biogas production using anaerobic digestion. Keeping this in mind, the present study was carried out in biogas Laboratory, Department of Renewable Energy Engineeering, College of Agricultural Engineering and Technology, PAU, Ludhiana. Five different sets of 2L glass digesters each containing 800 g mushroom waste along with different concentration of cow dung (as inducer) and cow dung slurry (as inoculum) were analyzed for biogas production. The biogas production was measured daily by water displacement method for a period of 80 days. The maximum cumulative biogas production was found in digester (26.013 L/Kg) carrying mushroom waste and cow dung (mushroom waste and cow dung 800g and 200g respectively) followed by digester carrying mushroom waste, cow dung and cow dung slurry(25.821 L/Kg) (mushroom waste, cow dung and cow dung slurry 800g, 100g, 100g respectively) as compared to control (3.136 L/Kg) consisting of mushroom waste and water only (mushroom waste and water 800 and 200ml respectively) after 80 days. The results obtained by the study clearly indicates that mushroom waste is a suitable substrate for methane production.