Agrobacterium-mediated Transformation of Nicotiana tabacum cv. Xanthi Leaf Explants

Abstract

Plant transformation has become an important technology for the large-scale production of a number of goods, ranging from vaccines and pharmaceutical products to biofuel. A plant that is frequently used for this purpose is Nicotiana tabacum due to several advantages: it can be easily regenerated, its genome is completely sequenced and genetic transformation is straightforward. We describe here the most common method for tobacco genetic transformation, which involves Agrobacterium tumefaciens. Many plasmid vectors are compatible with this bacterium, allowing the transformation of a wide range of plant species, as well as the insertion of more than two genes of interest in the plant genome. Furthermore, the capability to transfer a single copy DNA fragment to its host reduces the probability to switch off the gene or genes of interest. Here, we present a simple and reliable methodology to transform tobacco leaf explants using Agrobacterium tumefaciens to insert two genes of interest.