Highly Efficient Protocol for Agrobacterium-Mediated Transformation of Tomato (Solanum Lycopersicum L.)
Tomato can be transformed using Agrobacterium tumefaciens-mediated method. Efficient and reproducible procedure for transformation of commercial cultivar of tomato using vegetative leaf has been documented. Several parameters that influence T-DNA transfer and high frequency recovery of stable transgenic plants include pre-incubation of excised vegetative leaves, Agrobacterium cell count, co-cultivation technique, kanamycin selection conditions and supplements in shoot induction and root induction medium. Supplementation of maltose in SIM supplemented and sucrose in shoot elongation medium (SEM) with optimized concentration of growth regulators enhanced shoot formation. Half-strength MS medium as root induction medium (RIM) promoted root induction in 95 + 2 % transformed shoots. The overall transformation frequency was greater than 25%. This strategy of developing stable transgenics can be feasibly deployed for its reproducibility, efficiency and robustness for rapid recovery of non-chimeric, true transgenic plantlets. The protocol takes about 100 to 105 days from co-cultivation to the planting.