phiona scabra ( family Astraceae), is a medicinal endangered wild plant growing in Sinai Peninsula. It is rich in flavonoids and used in traditional medicine as an antispasmodic drug. Flavonoids have various biological activities like antiinflammatory, antioxidant, Alzheimer’s disease suppression and anti-cancer. The aim of this study was to use tissue culture technique for callus production of I. scarba and enhancing productivity of callus biomass and total flavonoids content (TFC) in solid culture medium. Shoot tips have been cultured on Murashige and Skoog (MS) medium fortified with 4-dichlorophenoxyacetic acid (2.4-D) at concentrations 0.25, 0.50 and 1.00 mg/l with kinetin (Kin) at 0.10 mg/l for the initiation of callus. For mass production of callus it was transferred to MS supplemented with thidiazuron (TDZ) 0.25, 0.50 and 0.75 mg/l combined with 0.5mg/l 1-naphthaleneacetic acid (NAA). Callus biomass was transferred to MS medium with different concentrations of either yeast extract as an elicitor or phenylalanine as a precursor at concentrations of 0.0, 25, 50, 100 and 200 mg/l. The best concentration that gave the highest accumulation of TFC was MS medium supplemented with 25 mg/l yeast extract. It gave 5.44 fold increase in callus biomass on the fourth week. The best treatment was MS medium fortified with 25 mg/l phenylalanine on the sixth week. That gave the highest fresh weight of callus 7.93 g/jar and accumulation of TFC 5.33 fold. Such potential method biotechnological strategies ensure better homogenous and stable production of sustainable flavonoids throughout the year under controlled environmental conditions.