Studies of the structure and functional organization of foreign DNA integrated into the genome of Nicotiana tabacum

Abstract

In transgenic plants obtained either by Agrobacterium tumefaciens-mediated transformation or by direct DNA transfer, the structure of integrated chimeric donor DNA remains stable during vegetative proliferation, during sexual transmission, and under various selection conditions. We correlate the level of expression of the introduced gene in independent transformants and their offspring with the particular arrangement and modification of their integrated DNAs. Genetic analysis of transgenic plants shows that the chimeric gene is transmitted in a Mendelian fashion to the F1 and F2 progeny as a single dominant trait. Deviations from the expected segregation pattern are discussed with respect to different levels of gene activity. We compare the gene activity in heterozygotes versus homozygotes, and show variation in activity between plants regenerated independently from the same transformed callus. Cotransformation studies with two physically unlinked and partly homologous plasmids carrying two different marker genes indicate that they are physically linked after integration into the host genome.