Tobacco plants were transformed by Agrobacterium rhizogenes infection during their evolution.

We discovered that the origin of cT-DNA in the genome of wild-type Nicotiana glauca is the T-DNA of the mikimopine-type Ri plasmid (pRi) harbored in Agrobacterium rhizogenes. The cT-DNA was inserted into the genomic DNA of N. glauca from the position corresponding to the right border of mikimopine-type pRi. The cT-DNA contained two mikimopine synthase gene (mis) homologs, NgmisL and NgmisR, both of which were transcribed at low level in all N. glauca organs. NgMisR protein expressed in Escherichia coli has preserved Mis activity, which converts l-histidine and alpha-ketoglutaric acid to mikimopine. The mis homolog was also found in the genome of three other Nicotiana species: N. tomentosa, N. tomentosiformis, and N. tabacum; however, the site of insertion differed from that in N. glauca, suggesting that A. rhizogenes harboring mikimopine-type pRi independently infected the ancestors of some Nicotiana plants. This is the first clear evidence of a host-parasite relationship during the early evolution of Nicotiana plants. We propose that a new phylogenetic approach using opine type cT-DNA is applicable for presuming divergence in the genus Nicotiana.