Agrobacterium-Mediated Genetic Transformation in Solanum torvum L. Using Leaf Explants

A particularly promising method involves the use of leaf explants cultured on Murashige and Skoog (MS) medium, supplemented with 0.5 mg/L indole-3-acetic acid (IAA) and benzylaminopurine (BAP). Following co-cultivation with a disarmed strain of Agrobacterium tumefaciens LBA 4404 harboring a plant expression binary vector, the explants were further treated with 100 mg/L kanamycin and 200 mg/L cefotaxime to facilitate selection and prevent bacterial contamination. The co-cultivation process successfully produced transgenic adventitious shoots, with an average of 2.3 ± 0.4 shoots per explant. These transgenic plants were subsequently vernalized in a greenhouse, where they grew to maturity and displayed normal morphological characteristics. Progeny analysis of the independent transgenic lines confirmed the successful integration of the gus gene into the genome, with its inheritance following a Mendelian pattern in three independent lines.