Multiple Shoot Bud Induction and Plant Regeneration in Madhuca indica J.F.Gmel.: Histological, Genetic Fidelity and GC-MS Analysis

Madhuca indica J.F.Gmel. holds significant economic and industrial value due to its applications in traditional and modern medicine. Its oil is especially important for biodiesel production, owing to its high acid value and suitability as a non-edible feedstock. However, propagation is difficult due to low seed germination, seed recalcitrance, and poor rooting of stem cuttings, limiting large-scale multiplication through conventional methods. To address these limitations, a regeneration protocol using nodal explants was developed. Murashige and Skoog (MS) medium augmented with BA (5.0 µM) and NAA (0.5 µM) produced a maximum of 7.10 ± 0.11 shoots per explant with an average shoot length of 4.53 ± 0.22 cm after six weeks. Rooting was achieved on half-strength medium supplemented with IBA (1.0 µM), resulting in 4.83 ± 0.17 roots per shoot and a root length of 4.50 ± 0.20 cm. In vitro-derived plants were successfully acclimatised in Soilrite with an 82.3% survival rate. The explants were derived from aseptic seedling material, representing juvenile rather than mature elite donor sources. Direct shoot bud development was verified by histological examination. Within the resolution of the employed marker systems, no polymorphism was found utilising RAPD and ISSR markers. SEM showed similar leaf surface characteristics, and physiological and biochemical studies were carried out throughout acclimatisation. A partial overlap in metabolite composition with qualitative and relative quantitative differences between mother and in vitro-derived plants was shown by GC–MS-based profiling. Overall, the study establishes a reproducible regeneration system for M. indica, providing a basis for further optimisation and conservation-oriented applications.