Aralia spinosa L. is a promising industrial crop for the biosynthesis of triterpenoid saponins and phenylpropanoid derivatives, however scalable production of genetically and phytochemically uniform plant material remains limited. This study describes an efficient multi-route somatic embryogenesis (SE) system established with genetic and phytochemical fidelity evaluation. Primary indirect SE was induced from leaf-derived callus, and regeneration efficiency reached 31.65 plantlets g −1 callus under the optimized protocol. To intensify propagation, cotyledonary somatic embryos were subjected to cyclic secondary direct SE induction using medium with 70 g L −1 sucrose that yielded ∼300 plantlets per flask. Regenerants acclimatized readily, with 73.3 – 93.3% survival after eight weeks under ex vitro conditions. Genetic fidelity evaluated with combined Start Codon Targeted polymorphism (SCoT) and Inter Simple Sequence Repeat (ISSR) markers (213 loci) revealed 92.02% monomorphic bands and 98.12% retention of donor loci, while analysis of molecular variance (AMOVA) attributed only 5.5% variance to differences among SE-derived groups. UHPLC-DAD-ESI-IT-MS 3 profiling detected 72 secondary metabolites. Among them 17 phenolic acids, 6 flavonoid glycosides, and 36 saponins were tentatively identified, while no qualitative gains or losses were observed across SE pathways or generations. Targeted quantification indicated comparable or moderately higher levels of key metabolites (e.g., calenduloside E, chikusetsusaponin IVa and ginsenoside Ro in shoots, and araloside A and chlorogenic acid in roots). Overall, the developed SE protocol allows the combination of high propagation rates with genetic and phytochemical stability, which provides the basis for large-scale biotechnological production of standardized A. spinosa biomass. • Efficient regeneration of Aralia spinosa L. via primary somatic embryogenesis. • Cyclic secondary somatic embryogenesis markedly increased embryo proliferation. • High acclimatization success and normal morphology of regenerated plantlets. • ISSR and SCoT markers revealed high genetic fidelity among regenerated plantlets. • UHPLC-DAD-ESI-IT-MS 3 fingerprinting confirmed phytochemical profile stability.
Kiełkiewicz et al. (Fri,) studied this question.