Supplemental Figure 1. Yeast two-hybrid assay showing FIL-JAZ3 interaction in

the presence of 3-aminotriazole.

(A) Yeast two-hybrid assay using JAZ fused to the GAL4 Binding Domain (BD-

JAZ) and FIL/YAB fused to the GAL4 Activation Domain (AD-FIL/YAB). Yeast cells

cotransformed with the indicated combinations were selected in medium lacking

Leu and Trp (-2) and grown for 5 days in medium lacking Ade, His, Leu and Trp

without or with 5 mM 3-aminotriazole (-4, -4AT) to select for interactions. BD

represents the empty pGBKT7 vector. Numbers above the panel represent JAZs

proteins, from JAZ1 (1) to JAZ12 (12).

(B) Yeast two-hybrid assay using JAZ3 fused to the GAL4 Binding Domain (BD-

JAZ3) and FIL/YAB fused to the GAL4 Activation Domain (AD-FIL/YAB). Yeast

cells cotransformed with the indicated combinations were selected in medium

lacking Leu and Trp (-2) and grown for 5 days in medium lacking Ade, His, Leu

and Trp without or with 3-aminotriazole (-4, -4AT) to select for interactions. BD

represents the empty pGBKT7 vector.

B

AD

-FIL

AD

-YA

B2

AD

-YA

B3

AD

-YA

B5

BD

BD-JAZ3 -2

-4 3AT

BD

BD-JAZ3

BD-JAZ3

-4

A

-2

1 2 3 4 5 6 7 8 9 10 11 12

AD-FIL

AD-YAB2

AD-YAB3

AD-YAB5

AD-FIL

AD-YAB3

-4 3AT

BD-JAZ

Supplemental Data. Boter et al. (2015). Plant Cell 10.1105/tpc.15.00220

1

0

5

10

15

20

25

Control COR DEX DEX+COR

A53

0/g

FW

Col0 35s:FIL-GR

Supplemental Figure 2. Anthocyanin content of 7-day-old 35S:FIL-GR transgenic

Arabidopsis seedlings grown on MS medium supplemented with 10 µM

Dexamethasone (DEX) plus/minus 1 µM Coronatine (COR; a mimic of JA-Ile).

EtOH was the solvent used for Cor stock and was added in control treatments.

DMSO is the solvent used for DEX stock and was added in control treatment.

Error bars represent SD. Asterisks indicate statistically significant differences

between 35S:FIL-GR and Col0 in each condition (p-value<0.01, t-test).

*  

*  

Supplemental Data. Boter et al. (2015). Plant Cell 10.1105/tpc.15.00220

2

A

B

0,0

0,1

0,2

0,3

0,4

0,5

0,6

Ler fil fil/+ yab3

fil yab3 Coler yab235 fil yab235

Hyp

ocot

yl le

ngth

(cm

)

Control 50 µM JA

0

5

10

15

20

25

30

% In

hibi

tion

0,0

0,1

0,2

0,3

0,4

0,5

0,6

0,7

Dex Dex+JA

Hyp

ocot

yl le

ntgh

(cm

)

Col0

Supplemental Figure 3. Hypocotyl growth of 35s:FIL-GR and yab mutants in the presence/absence of

JA.

(A) Hypocotyl growth inhibition by JA of 10-day-old wild-type wt Col0 plants versus 35S:FIL-GR grown

in MS medium for 5 days and supplemented with 20 µM Dex plus/minus 50 µM JA for 5 days more (left

panel). Right panel represents the same data as % hypocotyl growth inhibition of JA treated plants

compared with Dex treatment. For hypocotyl growth assays seedlings were grown under continuous

white light conditions. Error bars represent standard deviations.

(B) Hypocotyl growth inhibition by JA of wild-type (Ler or Coler) plants versus yab mutants grown in MS

medium plus/minus 50 µM JA under continuous light conditions presented as in (A). Error bars

represent SD. Asterisks indicate statistically significant differences between the corresponding samples

and Ler (p-value<0.01, t-test).

 

*  *  

Supplemental Data. Boter et al. (2015). Plant Cell 10.1105/tpc.15.00220

3

0,00

0,50

1,00

1,50

2,00

2,50

3,00

3,50

Ler fil fil/+ yab3 fil yab3 Coler yab235 filyab235

Roo

t len

gth

(cm

)

control 50 µM JA

Supplemental Figure 4. Root growth inhibition of 7 days-old wild-type and yab

mutants. Seedlings were grown in MS medium plus/minus 50 µM JA under

continous light conditions. fil/+ indicate heterozygous for fil mutation. Error bars

represent standard deviations.  

Supplemental Data. Boter et al. (2015). Plant Cell 10.1105/tpc.15.00220

4

Col0

35S:FIL-GR

coi1-30

Coler

fil/+ yab235

Supplemental Figure 5. Disease symptoms in leaves of wild-type, yab mutants

and 35S:FIL-GR lines. Photographs of detached leaves were taken 4 days after

inoculation.

Supplemental Data. Boter et al. (2015). Plant Cell 10.1105/tpc.15.00220

5

-1180 ctaggttttccatcgtacacgtaaattttcatgcaagaaagcagaaatatacaaatactaacttttagatactgaaaaatgagatcagattctagtcaaattttgttaaaagtatttataaatttaaattgcaagtcctcaaaaagtacgactaaaaatgcttttcttagaaAATGATAAtaaaccggcgttttatatataagtgtttctttttctcttctgtccagaagtaAATCATTAagaaccaatatggcttttcttaaactaatctccgtgataatcaaatctttgatcattctccacacaatcccatcaacaacatcgatctcactagatgcaccaacAATGATTctaatcggcactactaactatagagatagttgtcccaaaaaaaaaaaaaaaaactaactagagagataAATCATAttcaatacatgtactatttctactatacttaagaaaatttgtataccactatcttaactcttaacactgaacatactatacactatcttaactcccaactcttgtaaaagaatatctaattttaagaaaagacttcaaatgcttgttaaatttctagtgaagatgcacattctaaaaactggtaaaatggtaagaaaaaaatatataaaaaaatagccttattaaaatttatatctcctatttctctatccaaactacacggatgaagcttattgttattcatccaccctttttctcaattctgtcctatttcttgtgcatgaaacttctccatcttgtaatcggataAATCATAcccaaattttttctttctgaaaacatatatacccgaacattaattactatcgtcctttctcctaattttgttaagaaacatgtttgtttgtttttagtactgaaaaaggatggagatacttgctagatcctatgaaccttttctctctaggacaaatcagtaaccaaacaataacttagcaaattaagcacgacagctaatacataaaatgtggatatcaaacatgcacgtcacttccttttttccgtCACGTGtttttataaattttctcacatactcacactctctataagacctccAATCATTtgtgaaaccatactatatataccctcttccttgaccaatttacttataccttttacaatttgtttatatattttacgtatctatctttgttccATG

-1

Putative FIL binding element G-box

Supplemental Figure 6. Nucleotide sequence and graphic representation of

MYB75/PAP1 promoter.

(A) Nucleotide sequence of the MYB75/PAP1 promoter fragment used in this work,

highlighting the putative FIL DNA-binding elements (Cyan) and a G-box (yellow,

used as unspecific competitor probe in EMSA experiments) .

(B) Graphic representation of MYB75/PAP1 promoter highlighting putative FIL

binding elements (FBE1 and FBE2) and the G-box used in EMSA experiments

shown in Figure 7D. Numbers refer to nucleotide position from the ATG.

A

B FBE1 FBE2 G-box

-1 -463

Supplemental Data. Boter et al. (2015). Plant Cell 10.1105/tpc.15.00220

6

Ler fil yab3

fil yab235 Coler

Supplemental Figure 7. Anthocyanin accumulation on 5 days-old Arabidopsis fil

yab3 and fil yab2 yab3 yab5 mutants compared with their respectives wild-types

(Ler, for upper panel and Coler for bottom panel) grown in MS medium with 50 µM

JA. Arrows and arrowheads point to cotyledons and hypocotyl differences,

respectively. Major differences are detected at cotyledon whereas hypocotyl

shows minor differences. Scale bar=2 mm.  

Supplemental Data. Boter et al. (2015). Plant Cell 10.1105/tpc.15.00220

7

Supplemental Table 1. Oligonucleotides used for cloning.

YAB5 fw ggggacaagtttgtacaaaaaagcaggcttcATGGCTAACTCTGTGATGGC

YAB5 rev ggggaccactttgtacaagaaagctgggtcTTAGGCTATCTTAGCTTGCTTG

YAB1 fw ggggacaagtttgtacaaaaaagcaggcttcATGTCTATGTCGTCTATGTC

YAB1 rev ggggaccactttgtacaagaaagctgggtcTTAATAAGGAGTCACACCAAC

YAB2 fw ggggacaagtttgtacaaaaaagcaggcttcATGTCTGTAGATTTCTCATCTG

YAB2 rev ggggaccactttgtacaagaaagctgggtcTTAGTAATAGCCATTAGACTTTTG

YAB3 fw ggggacaagtttgtacaaaaaagcaggcttcATGTCGAGCATGTCCATGTCG

YAB3 rev ggggaccactttgtacaagaaagctgggtcCTAGTTATGGGCCACCCCAACG

pMYB75 fw ggggacaagtttgtacaaaaaagcaggcttcctaggttttccatcgtacacg

pMYB75 rev ggggaccactttgtacaagaaagctgggtcCATggaacaaagatagatacgt

Supplemental Data. Boter et al. (2015). Plant Cell 10.1105/tpc.15.00220

8