Temperature influence on stable T-DNA integration in plant cells

Abstract.

Four co-cultivation temperatures (15°C, 19°C, 25°C, and 32°C) were evaluated to determine their effects on T-DNA transfer and stable integration. Tobacco leaf explants were co-cultivated with Agrobacterium tumefaciens LBA4404 containing plasmids encoding resistance to the herbicide phosphinothricin, and Bt for insect resistance. Transgenic plants were evaluated for insect and herbicide resistance as well as at the molecular level for foreign gene integration. Even though 19°C has been reported as the optimal temperature for Agrobacterium-mediated gene transfer, co-culture at 25°C led to the highest number of stable transformed plants. Although 19°C may be the best temperature for the Agrobacterium transfer machinery, co-culture at 25°C appears beneficial for plant cell susceptibility to infection and for stable T-DNA insertion into the plant chromosomes.