Cross‐Species Reprogramming of Developmental Plasticity and Metabolic Rewiring via Banana‐Derived WUS2 Developmental Regulator

Plant regeneration is governed by intrinsic gene regulation and phytohormonal cues. WUSCHEL (WUS) gene promotes regeneration, but its broader functional role remains unexplored. Here, we demonstrate that the constitutive and inducible expression of banana-derived WUS2 (GN-WUS2) enhances regeneration in Nicotiana tabacum (tobacco) and Musa acuminata (banana) cv. Grand Naine, even in hormone-free MS medium. Constitutive (CaMV35S::GN-WUS2) expression promoted shoot formation and modulated hormonal and morphogenic gene expression, as evidenced by molecular, biochemical and histological analyses. However, it caused some pleiotropic effects. To overcome this, glucocorticoid receptor-based inducible GN-WUS2 expression enabled healthy shoot development. The upregulated expression of HMGR1, IPPI2 and SMT1-2 in transgenic tobacco lines boosted isoprenoid and phytosterol biosynthesis and correlated with increased cell division, biomass, pod size and seed yield. Proteomic analysis of seeds from transgenic tobacco lines revealed an enrichment of lipid-associated proteins and the accumulation of the novel lipid adipic acid, supported by expression profiling of NtKA and NtSA genes. Collectively, these results establish GN-WUS2 as a master regulator that integrates developmental reprogramming with novel phytosterol biosynthesis and yield enhancement, presenting its versatile role in next-generation regeneration and crop improvement.