Unlocking regeneration potential: harnessing morphogenic regulators and small peptides for enhanced plant engineering.

Plant genetic transformation is essential for understanding gene functions and developing improved crop varieties. Traditional methods, often genotype-dependent, are limited by plants' recalcitrance to gene delivery and low regeneration capacity. To overcome these limitations, new approaches have emerged that greatly improve efficiency and genotype flexibility. This review summarizes key strategies recently developed for plant transformation, focusing on groundbreaking technologies enhancing explant- and genotype flexibility. It covers the use of morphogenic regulators (MRs), stem cell-based methods, and in planta transformation methods. MRs, such as maize Babyboom (BBM) with Wuschel2 (WUS2), and GROWTH-REGULATING FACTORs (GRFs) with their cofactors GRF-interacting factors (GIFs), offer great potential for transforming many monocot species, including major cereal crops. Optimizing BBM/WUS2 expression cassettes has further enabled successful transformation and gene editing using seedling leaves as starting material. This technology lowers the barriers for academic laboratories to adopt monocot transformation systems. For dicot plants, tissue culture-free or in planta transformation methods, with or without the use of MRs, are emerging as more genotype-flexible alternatives to traditional tissue culture-based transformation systems. Additionally, the discovery of the local wound signal peptide Regeneration Factor 1 (REF1) has been shown to enhance transformation efficiency by activating wound-induced regeneration pathways in both monocot and dicot plants. Future research may combine these advances to develop truly genotype-independent transformation methods.