A Protocol for in Vitro Propagation of Quercus Pubescens Willd

Abstract

In this study, a protocol was developed for the in vitro propagation of Quercus pubescens Willd. Through this research, we aimed to determine how in vitro cultivation conditions, culture medium, and the sterilisation method influence in vitro regeneration response. Explants aseptically disinfected in sodium hypochlorite solution, Benzalkonium chloride, and ethanol were inoculated onto two types of medium (WPM and GD), to which different combinations of plant growth regulators were added. Sterilisation treatment consisting of benzalkonium chloride 1%, 3 minutes + Domestos 20%, 3 minutes + Ethanol 70%, 1 minute led to cultures free of microbial contamination and the highest percentage of viable explants (80%). After one month from the initiation of the cultures, measurements were made regarding microshoot height, number, and length of the leaves. The WPM medium with gibberellin 0.25 mg/l and BAP 1 mg/l positively influenced the growth of microshoots and leaf size, leading to the best results in terms of shoot height (3.27 cm) and leaf length (0.62 cm). On the GD medium, the number of leaves was slightly higher (2.82), but their length was shorter (0.52 cm). The rooted microshoots were acclimatised in the greenhouse, and the survival rate was 84.62%.