Abstract
Purpose: To compare the hyoscine contents of Datura innoxia plant grown in the wild and that grown in vitro.Methods: For callus induction from leaf explants, 15 combinations of 2,4-dichlorophenoxy acetic acid, gibberellic acid, benzyl amino purine and kinetin were used. For regeneration of shoots, 8 combinations of BAP, kinetin and indole butyric acid (IBA) were tested. Different concentrations of IBA were used for rooting. The hyoscine contents of the wild and in vitro samples was initially identified by TLC and subsequently quantified by high performance liquid chromatography using hyoscine N-butyl bromide as standard.Results: For callus induction, 2,4-D at 2 mg/L produced maximum calli (96 %). Maximum shoots regeneration was obtained with 2 mg/L BAP and 1 mg/L kinetin. IBA at 0.5 mg/L induced good rooting. Substantial amount of hyoscine was detected in the extracts of in vitro grown plants through TLC and HPLC. Wild root, stem and leaves exhibited higher amounts (approx. 2 μg/mL) of hyoscine than the parts of in vitro grown plants. Green callus grown in vitro contained a maximum concentration of hyoscine (1.01 μg/mL) followed by leaf (0.82 μg/mL) and brown callus (0.432 μg/mL).Conclusion: The callus cultures of D. innoxia are capable of yielding good amounts of hyoscine, and therefore can be exploited to increase hyoscine production using chemical/hormone treatments on a large scale. Thus, this study provides a sustainable and efficient way for the sustainable production of a natural hyoscine product via in vitro-grown D. innoxia cultures.Keywords: Datura innoxia, Hyoscine, Callus, Hyoscine, Explant, Gibberellic acid, Benzyl amino purine, Kinetin
Highlights
A number of plant-derived natural products are isolated from plants which are used to cure several ailments and infections in humans
Datura innoxia is an attractive medicinal plant [1] which is a rich source of a variety of natural products and secondary metabolites
Shoot formation from calli was successfully achieved by BAP alone as well as in combination with kinetin and indole butyric acid (IBA)
Summary
A number of plant-derived natural products are isolated from plants which are used to cure several ailments and infections in humans. Datura innoxia is an attractive medicinal plant [1] which is a rich source of a variety of natural products and secondary metabolites. Among these tropane alkaloids, hyoscyamine and hyoscine are clinically more significant [2,3,4,5]. In order to explore further opportunities for sustainable hyoscine production, in vitro culturing techniques are proposed as a good alternate These techniques can be employed to produce multiple plant nurseries for field propagation and in vitro grown callus cultures could be a direct and sustainable source of hyoscine. D. metel or D. stramonium, callogenesis [3,10,11] and organogenesis [12,13,14,15] trials have been reported
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