An Optimised Protocol for Faba Bean ( Vicia faba L.) Tissue Culture and Transformation

ABSTRACT Faba bean ( Vicia faba L.) is an important legume that contributes to agricultural sustainability through nitrogen fixation and offers substantial nutritional benefits. Nonetheless, its genetic enhancement has been constrained by challenges associated with tissue culture and transformation. This study presents the development and optimisation of a robust and reproducible protocol for tissue culture and genetic transformation in various V. faba genotypes. Thirty‐three genotypes were evaluated with various hormone combinations to enhance shoot and root regeneration. Among these, 1.5 mg/L indole‐3‐acetic acid (IAA) demonstrated the highest efficacy for uniform root formation across all genotypes. Genetic transformation was performed through particle bombardment, utilising excised embryos as the target tissues. A transformation efficiency of 11.7% was achieved in cultivar Tiffany, demonstrating stable transgene inheritance. This protocol mitigates significant limitations in faba bean regeneration, such as the production of phenolic compounds and variability in rooting, thereby facilitating consistent transformation across cultivars. This study presents an efficient and reproducible protocol for tissue culture and transformation in V. faba genotypes. The approach establishes a practical foundation for subsequent applications in gene editing and molecular breeding of faba bean and a potential methodological basis for optimisation in other legume crops.