Establishment of a Protocol for in vitro Seed Germination and Callus Formation of Ficus religiosa L., an Important Medicinal Plant

Background: Ficus religiosa is known as a pure source in traditional medicine for the treatment of diabetes, asthma, diarrhea, gastric problems, epilepsy, sexual, infectious, and inflammatory disorders. Despite the fact that many studies have authorized its traditional medicinal uses, yet these utilized raw extracts have not been yet characterized. Therefore, there is a necessity for standardizing its phytochemical features and recognizing bioactivity, guided by bioactive metabolites. Objectives: In this study, the effect of light and dark condition, different strengths of Murashige and Skoog (MS) medium, and their interactions on seed germination and the effect of plant growth regulators on callogenesis of F. religiosa via leaf, petiole, root, and internode explants were investigated. Results: The results demonstrated the highest seed germination percentage was achieved at one-tenth strength of MS medium under the light condition. Also, the highest callus fresh weight was obtained from media supplemented with 0.5 mg/L 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 0.05 mg/L 6- benzyl amino purine (BAP) in all explants. The MS medium containing indole-3-butyric acid (IBA) produced greenish and compact calli while yellowish and compact calli was achieved in a-naphthalene acetic acid (NAA) and finally 2,4-D produced yellow-brownish calli. Conclusions: To the best of the author's knowledge, this study was the first report of seed germination and callus induction through immature explants of F. religiosa. The seed germination and callogenesis system established in this study could be applied in the future for the enrichment of certain secondary metabolites as well as the production of new secondary metabolites, with the purpose of understanding and use of medicinal properties for this valuable germplasm.