Rapid and efficient stem segment regeneration propagation system for Euonymus bungeanus

Euonymus bungeanus Maxim. is a tree species with high ornamental, industrial, and medicinal value. Establishing a method for rapid and efficient regeneration of E. bungeanus is essential to achieve industrial-scale production. The aim of this study was to establish a rapid tissue propagation technique for E. bungeanus and provide a foundation for the industrial production of tissue-cultured seedlings. Using stem segments of E. bungeanus as explants, we investigated effects of explant collection time, sterilization method, various culture media, and ratios of plant growth regulators on the initiation, subculture, and rooting stages of the tissue culture process for E. bungeanus. The optimal explant collection time was mid-April; a combination of 75% ethanol for 20 s, followed by 0.1% HgCl₂ for 7 min was suitable for disinfection, yielding a survival rate of 55.00% for the explants. Initiation culture using the woody plant medium (WPM) supplemented with 1.0 mg L^-1 of 6-benzylaminopurine (6-BA) and 0.2 mg L^-1 of α-naphthalene acetic acid (NAA) achieved an induction rate of 87.22% for explants. Proliferation culture on ¼ WPM medium containing 2.0 mg L^-1 of 6-BA and 0.1 mg L^-1 of NAA resulted in a propagation coefficient of 4.98. Rooting culture on ½ Murashige and Skoog (MS) medium supplemented with 0.2 mg L^-1 of indole-3-butyric acid achieved a rooting rate of 78.33%. However, the rooting rate was low, indicating the need for further optimization of rooting and acclimatization. This study is the first to develop a rapid and efficient regeneration system for E. bungeanus using stem segments.