Meta-topolin mediated improved micropropagation, phytochemical profiling, and assessment of genetic fidelity in Chlorophytum borivilianum Sant. et Fernand., an endangered medicinal herb

Abstract

A highly effective method has been developed to examine the impact of different combination of meta-topolin and various cytokinins on Chlorophytum borivilianum Sant. et Fernand. using apical bud explants. The effects of benzyl adenine purine (BAP), kinetin (Kin), meta-topolin (mT), and thidiazuron (TDZ) were explored separately and in different concentrations (ranging from 0.5 to 3.0 mg L−1) on shoot proliferation, as well as the influence of auxins like Indole -3- acetic acid (IAA), α-Naphthalene acetic acid (NAA) and Indole-3-butyric acid (IBA) on rooting. When grown on MS medium supplemented with mT, highest growth rates were observed followed by BAP. Within the permutations tested, 2.0 mg L−1 of mT and 1.0 mg L−1 of BAP produced the best results with 96.3% success rate and average of 19.2 ± 1.92 shoots measuring 6.72 ± 0.49 cm in length after 8 weeks of culture. Among the various auxins used for in vitro root formation, 95.5% of multiple shoots effectively established roots on MS medium supplemented with 1.0 mg L−1 of IBA resulting in an average of 14.0 ± 1.39 root numbers and 3.36 ± 0.27 cm in length after 4 weeks of culture. Furthmore, 94% of the acclimated plants not only survived but also thrived when grown in a mixture of loam, sand and fym (in 1:2:1 ratio) with an average weight of 16 g. High-performance thin layer chromatography (HPTLC) analysis revealed the presence of diosgenin and stigmasterol in both in vitro regenerants and mother plant, although there were variations in their content. To ensure genetic fidelity among regenerants, ISSR and SCOT markers were used, but no somatic differences were detected. Therefore, the established procedure seems to be easy and consistent for large scale development of genetically identical clones with a greater content of bioactive compounds.