Supporting Information Legends
Supplemental Figure S1. Tomato fruit ripening stages and photosynthetic pigment
composition. Representative images of tomato fruit at the mature green (MG), orange
(OR) and red ripe (RR) stages are shown in the upper panel. Graphs show representative
HPLC-determined profiles of chlorophylls and carotenoids at these stages. The values
and bars represent the mean ± SEM from n = 3 biological replicates and they are shown
relative to those in MG samples.
Supplemental Figure S2. PSY1 response to R and FR. Quantitative PCR analysis of
PSY1 transcript levels in fruits halves irradiated with R or FR-enriched light. (a) Tomato
fruits at the MG stage were cut in two halves. One of the halves was incubated in the
dark (-) for 2h, whereas the other half was irradiated (+) with R light (30 µmol m-2 s-1 PAR)
for the same time period. (b) Tomato fruits at the OR stage were cut in two halves and
each of them was illuminated either with (-) white light (25 µmol m-2 s-1 PAR, R/FR ratio of
0.05) or with (+) white light supplemented with FR (25 µmol m-2 s-1 PAR, R/FR ratio of 3)
for 4h. Data correspond to mean ± SEM from n=3 (a) or n=4 (b) fruits.
Supplemental Figure S3. Specificity of the amiRNA against PIF1a. Transcript
abundance was assessed by quantitative PCR. Data correspond to amiPIF1a line 14.
Accessions and primers used are listed in Supplemental Table S1. The values and bars
represent the mean ± SEM from n ≥ 3 biological replicates. Values are reported relative
to the non-transformed WT controls.
Supplemental Figure S4. Light spectra at different depths of the pericarp.
Measurements correspond to tissue depths of 200 and 2000 µm from the surface of MG
and OR fruit (see Figure 3 for experimental details). Data are represented relative to
blank controls with no fruit samples.
Supplemental Figure S5. R/FR ratio of sunlight filtered through the pericarp of
tomato fruit. Measurements were performed in the greenhouse (GPS coordinates:
41°49'82"N 2°10'80"E) and correspond to R/FR ratio values of direct sunlight before (-)
and after passing through fresh hand-cut pericarp sections or pigment extracts from MG,
OR or RR fruit (n ≥ 6).
Supplemental Figure S6. Setup for experiments with tomato fruit pigment extracts.
Glass plates containing extracts of photosynthetic pigments (chlorophylls and
carotenoids) isolated from tomato fruit pericarp tissue were placed on top of light-proof
boxes with an opening in the lid so that all the light coming into the box passed through
the corresponding pigment filter. Light sensors and biological samples were placed inside
the corresponding box. When comparing different fruit extracts, the same climate
controlled chamber and source of artificial white light (fluorescent tubes providing 90 µmol
m-2 s-1 PAR) was used and the pigment concentration in the extracts was adjusted so that
the filtered light showed similar PAR values (40-50 µmol m-2 s-1).
Supplemental Figure S7. Effect of light filtered through tomato fruit pigment
extracts on Arabidopsis shade-responsive gene expression. Arabidopsis wild-type
seedlings germinated and grown under continuous light for 3 days were exposed for 24 h
to light filtered through MG or RR filters (see Supplemental Figure S4). Transcript
abundance of the indicated genes was assessed by quantitative PCR. Accessions and
primers used are listed in Supplemental Table S1. The values and bars represent the
mean ± SEM from n = 4 biological replicates. Numbers above the bars show statistical P
values according to the t-test. Values are reported relative to the MG filter condition.
Supplemental Table S1. Accession IDs and primers for genes analyzed by RT-qPCR in
this study.
Supplemental Table S2. Accession IDs for protein sequences used for molecular
phylogenetic analysis.
Supplemental Table S3. Primers used in cloning approaches in this study.
Supplemental Table S4. Primers used for ChIP-qPCR analysis.