Abstract. This study investigates the effect of Pleurotus spent mushroom compost as green supplementary nutrient in tissue culture. Different concentration of Pleurotus spent mushroom compost in powder (SMC) and activated carbon (SMCAC) were added into Murashige and Skoog media (MS) with 0.1 mg/L 6-benzylaminopurine (BAP). The shoot regeneration of Clinacantus nutans plant such as number of shoots, number of leaves and length of leaves were recorded for four weeks. Collected data was analyzed using one way analysis of variance (ANOVA) and Tukey test through the SPSS Statistics 21 software. The 0.1 g/L SMC recorded the highest leaves numbers. Meanwhile, 0.2 g/L SMC showed highest performance in length of shoot. The SMCAC showed a negative response in number of leaves and shoot as well as length of shoot. In summary, the SMC showed significant performance in number of leave and shoot length but less effective on number of shoot. In contradictory, the SMCAC reported poor performance in shoot regeneration but contribute in absorption of nutrient from environment and storage of the nutrient asfunction of biochar. Therefore, the SMC has a high potential as a green supplementary nutrient for tissue culture. The application of this material has contributes into the green technology via convert waste to product, which is in-line with zero waste concept.