In Vitro Micropropagation of Galanthus plicatus M. Bieb.: An Efficient Ex Situ Conservation Strategy for a Critically Endangered Species from the Republic of Moldova

Abstract

Galanthus plicatus M. Bieb. (Amaryllidaceae), a critically endangered geophytic species from the Republic of Moldova, has high ecological, ornamental, and pharmaceutical value as a source of biologically active alkaloids. However, excessive harvesting and habitat degradation have significantly reduced natural populations, creating an urgent need for effective ex situ conservation strategies. This study aimed to develop and optimize an in vitro micropropagation protocol for the rapid multiplication and conservation of the species’ genetic resources. Bulb-derived explants were cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of cytokinins (BAP and 2iP), auxin (IBA), and sucrose (45–60 g/L). The results showed that media supplemented with BAP (1.5–2.0 mg/L) and IBA (0.5 mg/L), combined with elevated sucrose concentrations, enhanced microbulb induction and proliferation, yielding up to 24 ± 0.43 microbulbs per explant. Increased sucrose levels also promoted reserve accumulation and microbulb maturation. An optimized sterilization protocol developed without mercuric chloride ensured a high percentage of viable and contamination-free explants. These findings confirm the effectiveness of in vitro micropropagation as a sustainable ex situ conservation strategy for G. plicatus and support the reduction of pressure on natural populations.