Sutini Sutini, Nurul Jadid, Ilham Misbakudin Al Zamzami, Umang Wildan Pratama
The production of high-value flavanol metabolites in Camellia sinensis L. is often inconsistent due to environmental fluctuations, developmental variability, and limitations inherent in conventional field cultivation. In vitro culture offers a controlled and reproducible platform to optimize secondary metabolite biosynthesis while supporting rapid and uniform plant propagation. This study aimed to develop an integrated in vitro system capable of simultaneously enhancing flavanol production and inducing pro-embryo formation. Leaf tip explants were cultured on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP), 2,4-D, and manganese (Mn2+) as an abiotic elicitor. Callus biomass was evaluated through wet and dry weight analysis, and flavanol content was quantified using high-performance liquid chromatography (HPLC). The results indicated that Mn2+ elicitation was associated with increased callus biomass, and HPLC profiling confirmed the presence of flavanol-related peaks by retention-time matching with a reference standard. Embryogenic calli were subsequently transferred to a benzylaminopurine-naphthalene acetic acid (BAP–NAA) liquid medium, where pro-embryonic structures were observed under microscopic examination. Because quantitative flavanol determination and embryogenesis efficiency metrics were not assessed in this study, the findings are presented as an integrated qualitative workflow linking elicitation, metabolite profiling, and early embryogenic initiation. Overall, this study provides the first integrated evidence that Mn2+ elicitation and hormone-regulated suspension culture can concurrently enhance metabolite production and somatic embryogenic initiation in C. sinensis. The optimized system represents a promising biotechnological strategy for sustainable industrial-scale flavanol production and the mass propagation of elite tea genotypes. © 2026 The authors.
Department of Agrotechnology, Faculty of Agriculture, UPN Veteran East Java, Surabaya, 60294, Indonesia; Department of Biology, Faculty of Mathematical and Natural Sciences, Sepuluh Nopember Institute of Technology, Surabaya, 60115, Indonesia; Coastal and Marine Research Center, University of Brawijaya, Malang, 65145, Indonesia; Department of Economics, Faculty of Economics and Business, Universitas Pembangunan Nasional Veteran Yogyakarta, Yogyakarta, 55283, Indonesia
