Supplemental Figure 1

Supplemental Figure 1

Supplemental Figure 2

Supplemental Figure 3

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Supplemental Figure 7

Supplemental Figure 8

Supplemental Figure 9

Supplemental Figure 1. Nucleotide sequence of the NtPMT1a gene promoter from -201 to +67.

Supplemental Figure 2. Functional analysis of the GAG motifs from four N. tabacumPMT genes.

Supplemental Figure 3. Transactivation of gene expression by tobacco NtERFs mediated by binding the GAG motif in yeast one-hybrid assays.

Supplemental Figure 4. Phylogenetic relationships of Group IX NtERFs inferred from maximum likelihood tree using DNA plus MEME sequences.

Supplemental Figure 5. Characterization of the overexpression of NtERF32 in transgenic BY-2 cells.

Supplemental Figure 6. Expression levels of four group IX NtERFs in WT and NtERF32-overexpression lines before and after MeJA treatment.

Supplemental Figure 7. Expression levels of four group IX NtERFs in WT and NtERF32 RNAi knockdown lines.

Supplemental Figure 8. Expression levels of NtERF32 in NtMYC2 RNAi knockdown lines.

Supplemental Figure 9. Expression levels of NtERF32 in Nicotiana tabacum cv Burley 21 and nic1, nic2, and nic1nic2 tobacco plants.

Supplemental Table 1. Expression levels of tobacco Group IX ERF TF genes in MeJA treated BY-2 cells.Given in the table are the fold-gene expression changes in the various members of the tobacco Group IX ERF family (Rushton et al., 2008b) as determined by microarray analysis. Underlined gene names indicate ERF family members up-regulated in BY-2 cells in response to MeJA treatment. Gray shaded boxes indicate the time point of maximum induction.

Supplemental Table 2Ct values and fold inducibilities for the ERF genes NtERF10, NtERF32, NtERF121,NtERF210 and NtERF221 during a timecourse of BY-2 cell treatment with MeJA. Fold inducibilities are calculated by the 2DCt method. Two different sets of primers were used for both NtERF32 and NtERF121.

Supplemental Table 3 Alkaloid levels in the WT and transgenic BY2-cell lines 72h after addition of 100 µM MeJA. Alkaloid levels in the un-induced WT and transgenic BY2-cell lines were not detectable by GCMS.

Supplemental Table 4 Gene specific PCR primers used for qRT- PCR for determination of the MeJA-inducibility of tobacco group IX ERF genes in BY-2 cells identified in microarray experiment.

Supplemental Table 5 Gene specific PCR primers used for semi-quantitative RT- PCR in gene expression analysis.

Supplemental Table 6 Gene specific PCR primers used for quantitative RT- PCR in gene expression analysis.