SUPPLEMENTARY DATA
Fig. S1. Phenotype of selected mutants.
(A-M) Twelve mutant candidates obtained after re-screening of M3 population exhibited a
resistance to 2,4-D. All of these mutant lines (B-M) showed an increased fluorescent signal of
PIN2:GFP and AUX1:YFP after prolonged treatment (20 h, 1 µM 2,4-D) when compared
with the control seedlings (A). (N) Besides the resistance to 2,4-D mutant lines ade1- ade3
showed a morphological phenotype. ade1, ade2 displayed smaller rosette leaves whereas ade3
line showed early flowering phenotype as compared with the control. Related to Fig. 1.
Fig. S2. Relative root length.
Five-day-old control and mutant seedlings were grown on standard 0.5 x MS or 0.5 x MS
supplemented with 20 nM NAA or 10 nM 2,4-D. The data for the mutants for each condition
(MS, NAA, 2,4-D) were normalized to the one obtained respectively for the control. The
normalized data were used to quantify the relative root length as the ratio of the normalized
root length observed on medium containing synthetic auxins versus MS medium. ade1 mutant
roots behaved and responded similarly to control, whereas ade2 and ade3 mutant lines
showed less sensitivity to auxins. Error bars represent SD. *, P<0.05 and ***, P<0.001
indicate significant differences when each mutant was compared to each other. Related to Fig.
2D.
Fig. S3. Lateral root development in ade3 mutant.
Eight-day old control and ade3 seedlings were grown on 0.5 x MS or on 0.5 x MS
supplemented with 10 nM 2,4-D. Lateral root primordia (LRP) include all stages until
emergence. Em stands for emerging lateral roots. Lateral root means fully emerged lateral
roots. The mutant line grown on 2,4-D showed a reduced number of fully emerged lateral
roots and an increased number of emerging lateral roots. Results from one representative
experiment were presented. Error bars represent SD. * indicates significant difference P ˂
0.05. Related to Fig. 2F.
Fig. S4. Vasculature phenotypes.
The cotyledon venation pattern was analysed in 12-day-old seedlings of Col-0, ade1, ade2,
ade3 mutant and control lines. The vasculature arrangement was used for categorization of the
observed pattern in seven classes based on unusually disconnected veins. The number of
cotyledons with venation defects was quantified. Related to Fig. 4
Fig. S5. Three independent replicates for the data presented in Fig. 2 and 3.
Box plots showing the distribution of the results for the three replicates of experiments shown
in Figures 2 and 3. (A) Results from measurements of the root length of control, ade1, ade2
and ade3 of five-day old seedlings grown on standard 0.5 x MS or 0.5 x MS supplemented
with 20 nM NAA or 10 nM 2,4-D. (B) Results from measurements of the gravitropic index of
control, ade1, ade2 and ade3 of five-day-old seedlings grown on standard 0.5 x MS or 0.5 x
MS supplemented with 20 nM NAA or 10 nM 2,4-D. (C) Results from measurements of the
hypocotyl lengths of control, ade1, ade2 and ade3 of five-day-old seedlings grown on
standard 0.5 x MS or 0.5 x MS supplemented with 20 nM NAA or 10 nM 2,4-D.