Sphaeranthus indicus is an important medicinal plant, widely used in traditional medicine for its therapeutic potential, particularly its antiviral, antimicrobial, anti-inflammatory and anticancer activities. Despite its medicinal importance, there is a need for developing efficient propagation methods to ensure its large-scale cultivation and conservation. This study aims to develop a protocol direct shoot organogenesis from nodal explants of Sphaeranthus indicus. In vitro regeneration through direct shoot organogenesis offers an efficient method for the propagation of medicinally valuable plants. Explants were subjected to surface sterilization followed by culture on MS medium (Murashige and Skoog medium) supplemented with different concentrations of BAP (6-Benzylaminopurine) for shoot induction. Root initiation was achieved on MS medium with α-naphthaleneacetic acid (NAA), and regenerated plantlets were successfully acclimatized and shifted to greenhouse and open field conditions. High shoot induction frequency of 91.6% with the lowest concentration of 1 mg/L BAP highlights the efficient regeneration protocol and initiate a pathway for micropropagation and cultivation of multipotential medicinal plant, S. indicus in large-scale. During the present investigation, an efficient in vitro regeneration protocol is developed
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