Figure 5.

Stable and transient transformation assays to characterize the effect of BTH application on Agrobacterium infectivity. A, BTH treatment of N. benthamiana plants did not significantly increase free SA. SA levels in the BTH-treated wild-type and TRV:GFP-inoculated plants were determined at 0 and 72 h post BTH treatment. The bars indicate the ses of the means for three biological replicates. B and C, In planta tumor assay was performed as described (Anand et al., 2007b) on the wild-type N. benthamiana and tomato plants mock or BTH treated (0.1–1 mm). Three days posttreatment, shoots were inoculated with the strain A. tumefaciens A348 and were photographed 6 weeks postinfection. D to G, Quantification of stable and transient transformation. Leaf discs derived from mock- or BTH-treated wild-type, TRV:GFP-inoculated, and ICS-silenced N. benthamiana plants were inoculated with either the strain A. tumefaciens A348 or GV2260 (carrying the binary vector pBISN1) and were incubated on hormone-free Murashige and Skoog medium or CIM, respectively. Four weeks after inoculation, fresh and dry weights of A348-infected leaves were measured (D and E). GUS activity was measured as described (Anand et al., 2007b) in GV2260-infected leaf discs at 2 and 5 dpi (F and G). These experiments were repeated at least three times with a minimum of 100 leaf discs for each plant and the data presented are the mean with se values. Letters indicate significant difference using Fisher's lsd test at P < 0.05.