Development of micropropagation protocol, assessment of genetic fidelity and phytochemical analysis of Blepharispermum subsessile DC.: a conservation concerned medicinal plant of Eastern Ghats, India

Abstract

Blepharispermum subsessile DC. is well known for its ethnomedicinal values. This plant has been used traditionally for the treatment of various diseases. Due to over-exploitation, habitat destruction, and climate change B. subsessile has been included in the list as threatened plants. Hence urgent attention is needed for the protection and conservation of this endangered plant. This plant is found in ‘Gurudangar’, Odisha, India and it is declared as Medicinal Plant Conservation Areas for in situ conservation of this plant species along with few other medicinal plants. In vitro propagation is an extremely effective alternative to overcome the limitations of conventional propagation methods and to escalate the production of B. subsessile. Keeping this in view, the present study was envisaged to develop an efficient protocol for in vitro plant regeneration of B. subsessile from cotyledonary node explant. Multiple shoots were induced from the cotyledonary node on Murashige and Skoog’s (1962) (MS) medium supplemented with different types and concentrations of plant growth regulators. A combined effect of 2.0 mg/L meta-Topolin (mT) and 1.0 mg/L indole-3-acetic acid (IAA) when augmented with MS medium was evaluated as optimum for multiple in vitro shoot regeneration from cotyledonary node. Further, in vitro nodal segments were inoculated on different plant growth regulator supplemented medium for upscaling and mT (2.0 mg/L) was found to be best for such in vitro shoot proliferation. The in vitro shoots were rooted on ½ MS medium supplemented with 0.5 mg/L indole-3-butyric acid (IBA). The in vitro regenerated plants were successfully acclimatized in the small pots containing sterile garden soil and sand (1:1) followed by the transfer to larger pots containing garden soil. The genetic fidelity of in vitro regenerated plants was assessed and ascertained by ISSR markers. The phytochemical analysis and antioxidant activity of the in vitro regenerated plants vis-à-vis mother plant were also evaluated to find out the biochemical fidelity.