Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Quantitative assays of the effect of the leucine zipper motif on WRKY protein interactions. The AD fusions were transformed with the various Gal4 DB fusion vectors into yeast cells. Proteins were isolated from the yeast cells and assayed for the β-galactosidase activity using ONPG (o-nitrophenyl-β-D-galactopyranose) as substrate. Data represent means + SE for N=5.
• Supplemental Figure 2 - Genetic Complementation of the wrky18/40/60 Mutant. (A) The wrky18/40/60 triple mutant was transformed with an empty vector or a vector containing WRKY18, 40 or 60 cDNA driven by their native promoters. At least 15 Wild type and 15 T1 transformants were inoculated by infiltration of PstDC3000 (OD₆₀₀=0.001 in 10 mM MgCl₂). Samples were taken at 0 (white bars) and 3 (black bars) dpi to determine the growth of the bacterial pathogen. The means and standard errors were calculated from 10 plants for each treatment. According to Duncan’s multiple range test, P=0.05, means of colony cfu at 0 dpi do not differ significantly it they are indicated with the same small letter; mean of cfu at 3 dpi do not differ significantly if they are indicated with the same capital letter. Expression of the WRKY transgenes in the transformants was verified by northern blotting using WRKY gene-specific DNA fragments as probes. (B) At least 15 Wild type and 15 wrky18/40/60 transformants were inoculated by spraying spore suspension at a density of 2x10⁴ spores/ml and kept at high humidity. The pictures of the plants were taken four days after the inoculation. Expression of the WRKY transgenes in the transformed wrky18/40/60 mutant plants was verified by RNA gel blotting using WRKY gene-specific DNA fragments as probes. The experiments were repeated once with similar results.