Anise is a valuable medicinal plant with the scientific name of Pimpinella anisum L from the genus Apiaceae (Umbelliferea and genus Pimpinella. About 80 to 90% of its essential oil compounds are anethole. Many studies have been done on its pharmacological properties, essential oils and oils. The aim of this study was optimization of direct regeneration of this plant in an in vitro condition. In this research, different methods for germination of plant seeds were first investigated. Then different disinfection treatments were tested to decontaminate the explants. The effect of different compounds of BA, Kin and NAA on the regeneration of hypocotyl bud explants was tested with three replications. These experiments were performed in a completely randomized design. The best germination response of 70% was obtained in 24 hours of seed hydro priming and disinfection with 70% ethanol and 1.5% sodium hypochlorite in 1.2 MS medium. Also, the best direct regenerative response in terms of petiole length is related to MS supplemented with 0.5 mg/l BA + 0.5 mg/l Kin + 0.5 mg/l NAA with 4.33 cm and in terms of leaves number was related to MS supplemented with 0.5 mg /l Kin + 1 g/l and 0.25 mg/l BA + 0.25 mg/l Kin + 1 mg/l NAA with 47 leaves were obtained. Regeneration of this plant can be a prerequisite for further studies

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