Blepharis maderaspatensis (L.) Heyne ex. Roth is a short-term plant which contains many important secondarycompounds with high medicinal value. Currently, most of the researches focus on chemical composition and pharmacological activity, but the source of raw materials is very limited. In this study, the first step is transferring the samples from nature into in vitro culture conditions to understand the effects of the factors related to shooting and callus morphogenesis was performed, the first node from shoots apical meristem was isolated and sterilized with 1.5% NaOCl for 20 minutes to achieve high efficiency with 86.11% sterile samples and 85.56% shoot growth rate after 2 weeks of culture on MS medium. The shoot generation from axillary shoots was continued to be investigated with the highest number of shoots formed on MS medium supplemented with BA (1 mg / l) showed 1.53 shoots/implant which the height and the number of leavesare 3.65cm and 6.67, respectively. Besides, the formation of callus from leaves of MS medium supplemented with 2.4 - D (0.25 mg / l) achieved the rate of 66.67% of cultured samples, forming good callus after 4 weeks of culture. The results of the study not only contribute importantly to understanding morphogenesis for micropropagation purposes but also serve as the scientific database for further studies at the cellular and molecular levels of this plant.
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