Abstract
Rauvolfia serpentina (L.) Benth. ex Kurz., popularly known as Indian Snakeroot plant, belonging to Apocynaceae family, holds immense medicinal importance, owing to its rich source of multiple secondary metabolites such as ajmaline, ajmalicine, reserpine, and serpentine. To meet the constant demands for the key secondary metabolite (reserpine) by majority of the pharmaceutical industries, the present study assessed the effects of direct and indirect regeneration system on amelioration of reserpine accumulation in shoots of R. serpentina. In vitro multiple shoot cultures were established using shoot tip explants. Best results for shoot initiation, multiplication, and biomass production were obtained in case of Murashige and Skoog medium, supplemented with 1 mg/l N6-benzyladenine. The multiple shoots were then sub-cultured on cytokinin–auxin combination media for further proliferation. Highest shoot and leaf multiplication rates and the most enhanced biomass were obtained in case of 1–1.5 mg/l Kinetin + 0.2 mg/l α-naphthalene acetic acid (NAA). Callus induction and its subsequent proliferation was obtained using 1.5 mg/l 2,4-dichlorophenoxyacetic acid. The best indirect shoot regeneration with highest shoot and leaf proliferation from calli was observed in case of 1 mg/l thidiazuron + 0.2 mg/l NAA. Reserpine content estimation via HPTLC from in vitro shoots (direct regeneration) and calli (indirect regeneration) were recorded to undergo an almost three-fold and two-fold increment (respectively) in comparison to that of the mother plant. Thus, in vitro direct regeneration system proved to be more effective and efficient in ameliorating the reserpine content.
Highlights
Rauvolfia genus comprises of different species of plants which have diverse medicinal and allied uses
For the establishment of the in vitro shoot culture, shoot apices were inoculated in Murashige and Skoog (MS) medium supplemented with the following plant growth regulators (PGRs)—N6-benzyladenine (BA), Kinetin (KIN), thidiazuron (TDZ), zeatin
Five different cytokinins viz. BA, KIN, TDZ, zeatin, and mT were tested for their multiple shoot initiation capabilities
Summary
Rauvolfia genus comprises of different species of plants which have diverse medicinal and allied uses. 1 3 Vol.:(0123456789) 294 Page 2 of 14 By and large, this plant is propagated via seeds; stem cuttings and root cuttings have been reported in some cases as well (Ghate et al 2019). Lucrative pharmacological and ethnobotanical uses (due the presence of the above-mentioned secondary metabolites) have led to the exploitation of R. serpentina natural resources, thereby pushing the species to a threatened category as per IUCN. Keeping in view these constraints that are involved with the propagation and conservation of this plant species; in vitro tissue culture technologies offer a practical and constructive solution to the preservation and maintenance of this plant. It is hypothesized that in vitro propagation technology of R. serpentina, following direct and indirect (callus-mediated) regeneration for enhancement of biomass and secondary metabolites production, would be a significant contribution to the pharmacological industry at large, since both biomass-cum-secondary metabolite production would be facilitated simultaneously
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