Elicitor-mediated enhancement of tropane alkaloid production in callus cultures of Datura stramonium L.

Abstract Callus cultures provide an alternative in vitro platform for the production of secondary metabolites, although their potential remains underexploited in many plant species. This study evaluated the capacity of elicited callus cultures of Datura stramonium L. to produce the tropane alkaloids hyoscyamine and scopolamine. Callus cultures were induced from root, stem, and leaf explants on media supplemented with several combinations of auxins and cytokinins. Callus biomass gain was assessed as fresh weight over a 16-week cultivation period. Several elicitors significantly promoted callus biomass accumulation, as reflected by increased fresh weight. For alkaloid production and elicitation experiments, roots-derived callus cultures were used. Untreated calli produced relatively low levels of hyoscyamine and scopolamine. However, specific elicitors substantially enhanced alkaloid accumulation. Yeast extract at 300 mg/L was the most effective, increasing hyoscyamine content up to 24.7 mg/g dry weight (DW), representing a 38.8-fold increase compared over the untreated control. A plant extract containing cucumber mosaic virus (CMV) also exhibited strong elicitation potential. Both yeast extract (300 mg/L) and the CMV-containing extract increased scopolamine levels by more than 30-fold. These findings demonstrate that root-derived callus cultures of D. stramonium offer a promising in vitro system for tropane alkaloid production and highlight the critical role of optimizing plant growth regulator combinations and elicitor treatments to enhance secondary metabolite biosynthesis.