P. Kunc, Gregor Osterc
Abstract Micropropagation of woody plants is often limited by explant type, physiological maturity, and season of collection. This study evaluated the influence of explant type and collection date on the in vitro propagation of Prunus subhirtella ‘Autumnalis’, an ornamental cherry from a mature (~ 60-year-old) tree. Three explant types—dormant buds, breaking buds, and nodal segments—were collected at multiple dates and cultured through induction, multiplication, and rooting stages using MS-based media supplemented with auxins, gibberellins and cytokinins. Dormant buds failed to survive in vitro , whereas nodal explants showed the highest survival (46.7%), followed by breaking buds (16%). Explant collection date significantly affected culture establishment: breaking buds showed improved survival in later collection dates, while nodal explants exhibited decreasing survival across successive dates. During multiplication and rooting, no significant differences in survival or rooting success were observed among explant types, though contamination and necrosis remained challenges. The results highlight nodal explants and optimally timed bud collections as key factors for successful in vitro propagation, providing a foundation for clonal propagation and conservation of mature Prunus subhirtella ‘Autumnalis’ genotypes.