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Tissue browning is a major problem in tissue culturing of woody plants, especially for Ficus religiosa which occurs by the accumulation and oxidation of phe-nolic compounds. This study aimed to determine the effect of different concentrations of sodium nitroprusside on the appearance of callus browning from leaf explants. The results indicate that callus browning was significantly reduced by supplementation of sodium nitroprusside to the MS medium and supplemented with 2.26μM of 2,4-dichlorophenoxyacetic acid and 0.22μM of 6-benzyl amino purine. The accumulation of hydrogen peroxide and phenolic compounds in the callus tissues decreased at the 50μM concentration of sodium nitroprusside. Although catalase and peroxidase activities decreased at the 50μM concentration, the activity of superoxide dismutase and polyphenol oxidases, as well as proline content, increased exponentially. Sodium nitroprusside could be useful for the formation of non-embryogenic callus with high levels of metabolic activity for the production and isolation of sec-ondary metabolites.