Pramanik et al. 2025_ Supplementary Data and Tables

The study investigates the influence of basal media composition and amino acid supplementation on the morphological development and metabolic profiling of Alocasia cuprea callus cultures. The primary objective was to evaluate how specific media and amino acids affect callus proliferation, differentiation, and bioactive metabolite production. The morphological development and metabolic changes in Alocasia cuprea callus cultures induced by basal media and amino acid supplementation are discussed. The medium composition strongly affected callus proliferation and differentiation: compact organogenic callus was induced in TB9 [Murasighe and Skoog medium (MS) with the addition of 2 mg/l thidiazuron (TDZ) and 1 mg/l 6-benzyl amino purine (BAP)], whereas IA9 [MS supplemented with 2 mg/l TDZ and 1 mg/l 2,4-D] induced embryogenic callus with a distinct morphology. Among amino acids, L-arginine, in combination with TB9, gave maximum biomass and shoot initiation, while L-cysteine supported embryogenesis on IA9 but inhibited shoot development.

Metabolic profiling through GC-MS performed significant differences in the production of bioactive compounds. Treatment IA9 induced diversity in metabolites produced in comparison to TB9. Such key compounds were squalene, which was highly produced in TB9 with L-asparagine, whereas IA9 was dominated by octadecadienoic acid. PCA and heatmap clustering highlighted the activation of targeted biosynthetic pathways by specific culture conditions, supportingthe production of desirable bioactive compounds. The heatmap also revealed the similarities in bioactive compound profiles among specific callus samples. Among the bioactive metabolites, the identification of squalene is particularly notable for its commercial applications. Future studies should explore genetic and metabolic engineering approaches to enhance the biosynthesis of high-value metabolites.