Optimization of the in vitro culture introduction stage of Daphne altaica Pall.

Abstract

Abstract
Background and Aim. Daphne altaica Pall. is a valuable medicinal plant used in traditional medicine for the treatment of various diseases.It is endemic Kazakhstan and has a restricted distribution range, occurring only in limited areas of Eastern Kazakhstan. Its narrow distribution, low seed productivity, and slow growth hinder the natural propagation and conservation of this species. Biotechnological propagation techniques can contribute to the preservation of the natural biodiversity of wild populations and ensure an adequate supply of plant material for the pharmaceutical industry. Therefore, this study aimed to investigate the effects of different sterilization methods and concentrations of phytohormones on the efficiency of introducing Daphne altaica into in vitro culture and on microshoot formation.
Materials and Methods. Bud explants of Daphne altaica Pall. were used as the initial plant material.
Various surface sterilization regimes were tested to obtain aseptic cultures, with the most effective treatment being a 0.01% HgCl₂ solution for 20 minutes. Micropropagation was carried out on Murashige and Skoog (MS) medium with different growth regulators, and the effects of their combinations on explant growth and morphogenetic activity were evaluated.
Results. Sterilization with 0.01% HgCl₂ provided a high level of asepsis while maintaining tissue viability. Active bud sprouting and shoot formation were observed on the culture medium. The best micropropagation results were obtained on the MS-R-4 medium supplemented with 1.0 mg/L BAP and 0.1 mg/L NAA, which promoted intensive formation of morphologically complete microshoots with
well-developed leaves in vitro.
Conclusion. The developed method enables the efficient production of sterile microshoots of Daphne altaica Pall. under in vitro conditions. This approach contributes to the conservation of the species’ genetic resources, facilitates its mass propagation, and reduces anthropogenic pressure on
natural populations.