Mother transformer: A High-Throughput, Cost-Effective in Planta Hairy Root Transformation Method for Cannabis

Abstract Background: Hairy root (HR) transformation assays mediated by Agrobacterium rhizogenes, both in vitro and ex vitro, are essential tools in plant biotechnology and functional genomics. These assays can be significantly influenced by various factors, which ultimately can enhance the efficiency. In this study, we optimized a two-step ex vitro HR transformation method using the actual mother plant combined with the RUBY system and compared with existing methods. Results: The two-step ex vitro method proved more efficient than both the