Cytological, genetical and phytochemically stable meta-Topolin (mT) - induced mass propagation of underutilized Physalis minima L. for production of withaferin A

Abstract

Abstract Physalis minima L. is commonly known as wild Gooseberry. Traditionally this plant has been used in different ethnopharmacological and modern medicinal practices. No systematic evaluation is done from an underutilized alternative resource of withaferin A content from P. minima. Withaferin A is a steroidal lactone withanolide mainly found in the plant Withania somnifera and other few members of family Solanaceae. The present investigation evaluated withaferin A content from 30 chemotypes collected from different geographical regions in India. Among 30 different chemotypes, an elite clone (17.13 ± 0.31 mg/g dry weight (DW) withaferin A) was selected for in vitro shoot multiplication. Well-organized micropropagation has been studied using five different cytokinins in Murashige and Skoog (MS) medium. Among them, aromatic cytokinin “meta-Topolin” (mT) at 1.0 mg/l triggered the highest number (83.26 ± 1.02) of multiple shoots and shoot length (4.87 ± 0.69) after 28 days of explant implantation. Maximum root induction was showed in ½ MS medium with 0.5 mg/l indole-3-butyric acid (IBA) to trigger 75.65 ± 1.03 roots and root length 4.56 ± 0.25 cm after 14 days of culture. In vitro well-rooted plantlets were excised for hardening and show; 99.02% survival rate. Genetic stability of micropropagated plants were performed with cytological and molecular marker-based selection. Simultaneously, phytochemical stability was performed with HPLC (High performance liquid chromatography) analysis. In our experiment no significant difference showed between the mother plant and micropropagated plants. Thus, our protocol can be used for large-scale commercial cultivation of elite chemotype of this medicinal plant.