High-frequency direct organogenesis, genetic homogeneity, and root ultra-structural study in Sujanapal & Sasidh: a medicinally important wild relative of ginger.

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This study describes the efficacy of aromatic and synthetic cytokinins on direct organogenesis and root anatomy of in vitro propagated plantlets of Sujanapal & Sasidh, a medicinally important crop wild relative (CWR) of the Zingiberaceae family. A one-step in vitro plant regeneration (simultaneous production of shoot and root) micropropagation protocol using rhizome buds has been developed by optimizing the type and concentration of plant growth regulators, aiming to enhance conservation efforts and support commercial cultivation. MS medium containing only -topolin (T) in a concentration of 22.05 µM was found to be the best for shoot multiplication with the production of 13.17 ± 0.82 shoots per explant as compared to 6-benzylaminopurine (BAP) and combinations of T/BAP with α-Naphthaleneacetic acid (NAA). Longest shoots (7.58 ± 0.55 cm) and highest rooting (26.67 ± 0.76 roots per explant) were observed on MS medium containing both 4.14 µM T and 2.68 µM NAA. The plants propagated on T showed greater cellular differentiation and organization in the root ultrastructure. Well-rooted in vitro raised plantlets were hardened in soilrite and field acclimatized to natural conditions with a 92% survival rate. Clonal fidelity of the micropropagated plants with that of the mother plant was confirmed using 29 Inter Simple Sequence Repeats (ISSR) markers. In summary, an efficient micropropagation system using T was developed that offers a sustainable solution for conservation of this species and also facilitates its propagation for commercial pharmacological research.

SUPPLEMENTARY INFORMATION

The online version contains supplementary material available at 10.1007/s13205-025-04344-z.