Figure 4.

In planta tumorigenesis and leaf disc transformation assays in the gene-silenced plants of N. benthamiana. A, The in planta tumor assay was performed as described (Anand et al., 2007b) on ICS-, NPR1-, and SABP2-silenced and control (TRV:GFP) N. benthamiana plants. Shoots of control and silenced plants were inoculated with the tumorigenic strain A. tumefaciens A348 and the tumors were photographed 6 weeks after inoculation. B and C, Quantification of stable transformation. Axenic leaf discs derived from control and gene-silenced plants were inoculated with A. tumefaciens A348 and were incubated on hormone-free Murashige and Skoog medium. Four weeks after inoculation, the fresh and dry weights of infected leaves were measured. D and E, Quantification of transient transformation. The leaf discs derived from gene-silenced plants were inoculated with the disarmed strain A. tumefaciens GV2260 (carrying the binary vector pBISN1) and GUS activity was determined as described (Anand et al., 2007b) at 2 and 5 dpi. The experiments were replicated three times with a minimum of 100 leaf discs for each gene-silenced plant and the data indicate the average with se values. Letters indicate significant difference using Fisher's lsd test at P < 0.05.