Co-transformation with one Agrobacterium tumefaciens strain containing two binary plasmids as a method for producing marker-free transgenic plants.

Co-transformation was investigated as a method that would allow the use of a selectable marker during plant regeneration followed by recovery of progeny which contain the desired gene(s) but lack a marker gene. Rapeseed (Brassica napus cv `212/86') and tobacco (Nicotiana tabacum cv `Xanthi NC') were co-cultivated with a single Agrobacterium tumefaciens strain containing two binary plasmids. Genes from both plasmids were expressed in approximately 50% of the primary transformants. Progeny expressing only one of the transgenes were observed in about 50% of the co-transformed lines, indicating that the genes were inserted at different loci. This single-strain co-transformation method allowed the use of a selectable marker during plant regeneration and subsequent recovery of marker-free progeny.