Supplementary Data

Figure S1. Sequence of the 1254bp Solanum lycopersicum cv. Moneymaker TPS5 promoter fragment.

Figure S2. Specific interaction of SlEOT1 with the SlTPS5 promoter in yeast.

Figure S3. Interaction of SlEOT1 with the SlTPS5 promoter in yeast.

Figure S4. Nucleotide and predicted amino acid sequence of SlEOT1 (KC331910).

Figure S5. Tissue specific expression of SlEOT2.

Figure S6. Protein alignment of SlEOT1 with related proteins from other plant species.

Figure S7. Transactivation of a mutated SlTPS5 promoter fragment by SlEOT1 in N. benthamiana leaves.

Table S1. List of clones obtained from three Y1H screens with the 207bp trichome-specific SlTPS5 promoter fragment and a tomato stem trichome cDNA library.

Table S2. List of primers used.

Supplemental figure legends

Fig. S1 Sequence of the 1254bp Solanum lycopersicum cv. Moneymaker TPS5 promoter fragment

The A of the start codon ATG (marked in bold) is designated as +1; 5’UTR is highlighted. Potential CAAT and TATA boxes are italicized. Primer sequences for the full-length and 5’ promoter deletion constructs are underlined. Selected regulatory motifs in the sequence analyzed by PLACE (http://www.dna.affrc.go.jp/PLACE/) are shown in the accompanying table. YUC element and YUC-like element are shown boxed and in enlarged bold letters respectively

Fig. S2 Specific interaction of SlEOT1 with the SlTPS5 promoter

Yeast cells with (a) the 207bp SlTPS5 promoter fragment or (b) the 326bp minimal PhODO1 promoter integrated into their genome were transformed with pAD-GAL4-2.1_SlEOT1. Cultures were grown to the same OD and plated on SD medium with 0, 5 or 10mM 3-amino-1,2,4-triazole (3-AT)

Fig. S3 Interaction of SlEOT1 with the SlTPS5 promoter

Yeast cells with the full-length (FL) or shorter SlTPS5 promoter fragments A (-1254 to -1047bp), B (-1046 to -807bp), C (-806 to -613bp),
D (-612 to -409bp), E (-408 to -208bp) and F (-207 to -1bp) integrated into their genome were transformed with pAD-GAL4-2.1_SlEOT1. Cultures were grown to the same OD in selective medium without competitive inhibitor and plated on SD medium with 5mM 3-amino-1,2,4-triazole (3-AT)

Fig. S4 Nucleotide and predicted amino acid sequence of SlEOT1 (KC331910) The predicted coding sequence is shown in capital letters, introns in small letter type. 5’ and 3’ UTRs are italicized. Start and stop codons are in bold and underlined. N-terminal and C-terminal conserved domains identified by blasting at NCBI (www.ncbi.nlm.nih.gov/Structure/ cdd/wrpsb.cgi) are highlighted

Fig. S5 Tissue specific expression of SlEOT2

Relative transcript levels for SlEOT2 as determined by Q-RT-PCR in L; leaf, WS; whole stem, BS; bald stem, T; stem trichomes, R; root, Fr; fruit, Fl; flower. Mean values (+SE) of 3 biological replicas are shown, normalized for Actin expression

Fig. S6 Protein alignment of SlEOT1 with related proteins from other plant species

Putative zinc finger domain (C3HC3H RING type) and C-terminal companion domain (IGGH type) of SlEOT1 are shown highlighted, in lowercase. The alignment was generated in CLC Workbench (www.clcbio.com) using ClustalW. Putative nuclear localization (NLS) signal as well as activation domains typically found in this protein family (glutamine (Q)-rich regions and acid residues cluster; ARC) are shown

Fig. S7 Transactivation of a mutated SlTPS5 promoter fragment by SlEOT1 in N. benthamiana leaves

Normalized GUS activity after co-infiltration with A. tumefaciens harboring the 35S:SlEOT1 effector construct and either the full-length (FL), the 207bp TPS5 promoter:GUS or a mutated (m) 207bp TPS5 promoter:GUS reporter construct. The mutation was introduced in the YUC-like element (bases -110 to -104 from the start codon; Fig. S1) of the 207bp SlTPS5 promoter fragment (ACTCTAC -> ACAAAAC). The bars represent obtained mean values and the error bars the standard error (n=3). RFP; red fluorescent protein. Letters indicate significant differences (ANOVA, P < 0.05 according to Tuckey’s B posthoc test). Representative results from two experiments are shown

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