Background & Aim: Withania genus has more than 200 species, of which two important species (W. somnifera) and (W. coagulans) have high medicinal value and their rapid and extensive reproduction with new methods is very important to expand the area of cultivation. Propagation of crops in vitro is an accessible method for mass propagation. Therefore, the selection of a suitable culture medium is necessary to motivate shoot regeneration from explants and proliferation. The main purpose of this study was to present a rapid and efficient method for mass propagation of virus-free and disease-free plants under in vitro culture conditions.Experimental: The experiment was designed in a completely randomized design (CRD) with three replicatesin. The seeds were sterilized and placed on MS medium after being collected from their natural habitat in Sistan and Baluchestan province. It was then cultured on MS medium containing benzyl amino purine growth regulator at four levels of 0, 0.5, 1 and 1.5 mg/l for branching.Results: The results showed that in coagulans specie in MS medium containing 0.5 mg/l of benzyl amino purine (BAP) with an average proliferation of 3.66 of shoots and a length of 2.85 cm, the highest branching was obtained. Also, the highest propagation coefficient and number of leaves were obtained in benzyl amino purine (BAP) treatment with a concentration of 0.5 mg/l. The lowest propagation rate was reported in control (without hormones). In this experiment, application of benzyl amino purine at concentrations of 0.5 and 1 mg/l improved seedling quality which was more effective in coagulans and contained higher quality and fresher seedlings. Recommended applications/industries: In general, for the production of seedlings with high branching percentages, the use of benzyl amino purine at a concentration of 0.5 mg/l is recommended for optimal branching. پرونده مقاله