Abstract
The aim of this study was to develop an efficient micropropagation protocol for Pueraria tuberosa, a medicinally valuable woody liana species. The nodal explants derived from in vitro-grown seedlings were cultured on Murashige and Skoog (MS) medium containing various combinations of 6-benzylaminopurine (BAP), thidiazuron (TDZ), indole-3-acetic acid (IAA), and indole-3-butyric acid (IBA). These combinations led to the substantial development of multiple shoots. The combination of TDZ (0.57 µM/L) and IBA (0.12 µM/L) resulted in the highest regeneration rate (85%), maximum number of shoots (5.52 ± 0.67), and best shoot length (4.96 ± 0.33 cm). The most effective in vitro rooting treatment consisted of half-strength MS medium accompanied by 2.46 µM/L IBA and a 10-minute pulse treatment with the same concentration of IBA. This treatment resulted in the highest root formation (100%) and maximum roots per shoot (16.72 ± 1.81), with the highest root length (3.58 ± 0.54 cm) among all treatments tested. After four weeks under field conditions, hardened in vitro plantlets showed an 88.2% survival rate. The genetic fidelity of regenerated plants was assessed using inter-simple sequence repeat (ISSR) and random amplified polymorphic DNA (RAPD) markers, which produced monomorphic bands, demonstrating the genetic homogeneity of regeneration and reliability of the in vitro propagation method.
Published Version
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