Genetic Transformation Mediated by <em>Agrobacterium tumefaciens</em> in <em>Solanum nigrum</em>

Solanum nigrumis an orphan solanaceous relative of potato and tomato, and its fresh fruit is a nutritious and delicious berry. The whole plant of S. nigrum can be used as medicine. However, the wild S. nigrum harbors some undesirable traits. Fortunately, genetic engineering techniques, especially gene-editing technologies like CRISPR/Cas9, offer valuable opportunities to rapidly improve S. nigrum by addressing these deficiencies. In most plants, plant improvement through gene-editing technology is achieved through Agrobacterium tumefaciens-mediated genetic transformation. In this article, we have detailed the genetic transformation protocols mediated by A. tumefaciens in S. nigrum. The cotyledons and hypocotyls of S. nigrum seedlings were used as explants for genetic transformation. Because S. nigrum is resistant to kanamycin, the DsRed gene was used as a visual marker to identify transgenic plants. The genetic transformation of S. nigrum can be efficiently completed according to these operation protocols. The detection of T1 transgenic plants can be directly identified by whether they emit red fluorescence, without the need for PCR identification.