Plant regeneration protocols for adventitious shoot organogenesis from apple (Malus domestica ‘Fuji’) leaf explants were developed in the present study. The effects of dark incubation periods in the early stages of culture, pre-treatment methods, the number of explants per culture container, the type of culture containers, and the orientation of the explants on culture media were evaluated to determine the optimal shoot regeneration conditions for ‘Fuji’ apple leaf explants. Light incubation of explants produced minimal response. However, dark incubation of explants for 4 weeks during the initial culture period enhanced shoot regeneration frequency. Comparing the number of explants per container, a higher percentage of shoot regeneration was obtained with nine explants per container compared with four explants per container. Pre-treatment, before culture, by dipping explants in a liquid regeneration medium containing 40 g/L of sorbitol for 2 hours produced the highest shoot formation rate, and the time of shoot formation was accelerated. The percentage of shoot regeneration and number of shoots per regenerating explant reached a maximum of 87.5% and 4.7, respectively. The regenerated shoots were elongated and rooted on a rooting medium of 1/4 MS with 0.2 mg/L IBA. The plantlets were successfully acclimatized, and the regenerated plants produced normal phenotypes.

Keywords: Apple, Culture method, Dark incubation, &lsquo,Fuji&rsquo,, Shoot organogenesis