An abundant supply of low-cost substrate and management of green mold disease-causing fungus Trichoderma are the major hurdles in successful mushroom production. This study aimed to identify the best Agro-based industry’s by-products as a substitute for oyster mushroom production (Pleurotus ostreatus) while managing fungal contaminants eco-friendly. Two sets of In-Vitro [containing 20% extracts, from agro-based industries, such as coffee waste powder, tea dust and Mahua oil cake] and In-Vivo experiments [four substrates such as paddy straw, wood sawdust, paddy husk and banana leaves were incorporated with coffee powder, tea dust and Mahua oil cake] were prepared separately. All the experiments were conducted using a complete randomized design with three replicates. The In-Vitro data [mycelial growth and sporulation of both fungi], In-Vivo data [mycelial mushroom run, pinhead formation and yield] were subjected to ANOVA and DMRT mean separation using SAS 9.1 statistical package at P <0.05. In-Vitro results showed that the Trichoderma mycelial growth was significantly minimum in Mahua (2.5 cM) and coffee (3.6 cM) in comparison to control, whereas, with decreasing concentration of coffee, tea, and Mahua extract P. ostreatus showed enhanced growth. Trichoderma sporulation had significantly affected coffee treatment, and even not sporulate in Mahua treated plants. The In-Vivo experiment proved that spawn run was consistent and significant among the treatments when mixed tea (20 days) and coffee (21 days), respectively, at P <0.05. Treatment wise coffee treated spawn bags took an average of 32.5 days, whereas, in tea-treated substrates, it was more than 36 days to form pinhead. Mahua treated trials showed poor spawn run in all substrates, longer days of pinhead formation, and lower yield. In contrast, the paddy straw + coffee treatment produced a significantly highest yield of 200.67g. When sawdust was the substrate, the addition of tea showed a significantly higher yield of 185.00g than coffee (145.00g). In conclusion, coffee and tea extracts have a significant effect on yield with paddy straw and sawdust while minimizing the growth of Trichoderma. Keywords: Pleurotus ostreatus, eco-friendly, plant extract, substrate, coffee, paddy straw