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The berries of Embelia ribes Burm f. are a rich source of embelin, a compound known for its anthelmintic, antidiabetic, and anticancer activities. Due to over-exploitation, the natural habitat of Embelia ribes is now considered vulnerable. Traditional propagation methods are insufficient to meet current demands, necessitating alternative production approaches. This study was designed to explore in vitro culture as a viable alternative for the production of embelin. Although numerous studies have focused on extracting embelin from callus cultures, research on enhancing the embelin content in both organogenic and embryogenic callus cultures remains limited. The objective of this study was to enhance embelin production in callus cultures by using both biotic and abiotic elicitors. Additionally, efforts were made to optimize rapid in vitro shoot induction. We achieved the best response rate for the induction of organogenic and embryogenic calli using MS basal medium supplemented with BAP and TDZ in combination with 2-4-D. Among the various elicitors tested, chitosan at 200 mg/L was the most effective, yielding the highest embelin content, at 6.44% in embryogenic calli and 5.72% in organogenic calli. Subsequent subculturing enabled successful differentiation of callus cultures into shoot buds on MS medium supplemented with a combination of BAP (6-benzylaminopurine) and IAA (indole-3-acetic acid) at concentrations of 2.0 mg/L and 0.1 mg/L, respectively. An effective protocol has been developed for obtaining the highest embelin content from embryogenic and organogenic callus cultures, coupled with a high frequency of shoot multiplication. The protocol can be instrumental for large-scale embelin production, ex-situ conservation, sustainable utilization, and industrial applications.