Lettershttps://doi.org/10.1038/s41477-020-0604-8

A light-dependent molecular link between competition cues and defence responses in plantsGuadalupe L. Fernández-Milmanda   1, Carlos D. Crocco1, Michael Reichelt   2, Carlos A. Mazza1, Tobias G. Köllner   2, Tong Zhang   3,6, Miriam D. Cargnel1, Micaela Z. Lichy   1, Anne-Sophie Fiorucci   4, Christian Fankhauser   4, Abraham J. Koo3, Amy T. Austin   1, Jonathan Gershenzon2 and Carlos L. Ballaré   1,5 ✉

1IFEVA, Consejo Nacional de Investigaciones Científicas y Técnicas–Universidad de Buenos Aires, Buenos Aires, Argentina. 2Max Planck Institute for Chemical Ecology, Jena, Germany. 3Department of Biochemistry, University of Missouri, Columbia, MO, USA. 4Centre for Integrative Genomics, Faculty of Biology and Medicine, Génopode Building, University of Lausanne, Lausanne, Switzerland. 5IIBIO, Consejo Nacional de Investigaciones Científicas y Técnicas–Universidad Nacional de San Martín, Buenos Aires, Argentina. 6Present address: College of Agriculture, South China Agricultural University, Guangdong, China. ✉e-mail: ballare@ifeva.edu.ar

SUPPLEMENTARY INFORMATION

In the format provided by the authors and unedited.

NATuRe PLANTS | www.nature.com/natureplants

1

Supplementary Material:

A light-dependent molecular link between competition cues and

defense responses in plants

Guadalupe L. Fernández-Milmanda1, Carlos D. Crocco1, Michael Reichelt2, Carlos A. Mazza1, Tobias G.

Köllner2, Tong Zhang3,6, Miriam D. Cargnel1, Micaela Z. Lichy1, Anne-Sophie Fiorucci4, Christian

Fankhauser4, Abraham J. Koo3, Amy T. Austin1, Jonathan Gershenzon2 and Carlos L. Ballaré1,5*

1IFEVA, Consejo Nacional de Investigaciones Científicas y Técnicas – Universidad de Buenos Aires, Ave. San Martín

4453, C1417DSE, Buenos Aires, Argentina

2Max Planck Institute for Chemical Ecology, Hans-Knöll-Str. 8, 07745 Jena, Germany

3Department of Biochemistry, University of Missouri, Columbia 65211, USA

4Centre for Integrative Genomics, Faculty of Biology and Medicine, Génopode Building, University of Lausanne, CH-

1015 Lausanne, Switzerland

5IIBIO, Consejo Nacional de Investigaciones Científicas y Técnicas–Universidad Nacional de San Martín, B1650HMP

Buenos Aires, Argentina

6Current address: College of Agriculture, South China Agricultural University, Guangdong, China

* email: ballare@ifeva.edu.ar

This PDF file includes:

Supplementary Methods

Supplementary Figures 1 to 10

Supplementary Tables 1 and 2

See attached Supplementary Data Files 1 to 3

2

Supplementary Methods

Plant material and growth conditions. Seeds were directly placed in 7 x 7 x 8-cm pots containing a standard substrate mix (80 % Fruhstorfer Nullerde pH = 6.0-6.5, 10 % vermiculite and 10 % sand). Soil was fertilized with 1 g Triabon 3-4 M (Mehrnährstoffdünger 16+8+12)/L soil, 1 g Osmocote Exact Mini 3-4 M (16:8:11)/L soil and watered with a suspension of Steinernema feltiae (Katz Biotech AG, Germany). The potas containing the seeds were stratified in the dark for 3-4 days at 4°C and then placed in a growth chamber.

DNA extraction and genotyping. To test the NASC lines leaves from individual plants were collected and used for DNA extraction. Leaf tissue was homogenized using EDM buffer and centrifuged at full speed for 5 min, to pellet cellular debris. A fraction of 300 μL of the supernatant was mixed with equal volume of isopropanol and centrifuged at maximum speed for DNA precipitation. The pellet was dried and resuspended in 100 μL of MilliQ water. PCR was performed using Pfu polymerase (PB-L, Argentina) according to the manufacturer’s instructions, with 1 μL of DNA solution and primers to a final concentration of 1 μM. Primers were designed to amplify regions adjacent to the T-DNA insertion and their sequences are listed on Supplementary Table 1. Chip-qPCR. Seedlings were crosslinked in 1% formaldehyde under vacuum for 16 min, and crosslinking was stopped by adding 2M glycine to a final concentration of 0.125 M and incubating for 5 min under vacuum. Seedlings were frozen in liquid nitrogen and chromatin was extracted on 1.1 g of frozen powder as previously described, except for the sonication step which was done for 6 x 8 cycles (30-s on/30-s off, on high intensity) on a Bioruptor (Diagenode). Glucosinolate analysis by HPLC-UV. GS were extracted with 1 mL of 80 % methanol solution containing

0.05 mM intact 4-hydroxybenzylglucosinolate as internal standard. Samples were shaken on a horizontal

shaker at room temperature for 10 min, and then centrifuged at 14000 rpm for 10 min. Next, extracts

were loaded onto DEAE Sephadex A 25 columns (Sigma–Aldrich) column and washed with 80 %

methanol solution, water, and 0.02M MES buffer (pH 5.2). Sulfatase solution (arylsulfatase from Sigma-

Aldrich) was applied on the column and incubated at room temperature overnight. Distilled water (500

μl) was used to elute the desulfo-GSs into 96-deep well plates for HPLC-UV analysis. The eluted desulfo-

GSs were separated using high performance liquid chromatography (Agilent 1100 HPLC system, Agilent

Technologies) on a reversed phase C-18 column (Nucleodur Sphinx RP; 250 x 4.6 mm, 5µm particle size;

Macherey-Nagel, Düren, Germany) with a water-acetonitrile gradient (1.5% acetonitrile for 1 min, 1.5 to

5% acetonitrile from 1 to 6 min, 5 to 7% acetonitrile from 6 to 8 min, 7 to 21% acetonitrile from 8 to 18

min, 21 to 29 % acetonitrile from 18 to 23 min, followed by a washing cycle; flow 1.0 mL min-1).

Detection was performed with a photodiode array detector and peaks were integrated at 229 nm. For

quantification of individual GSs, we used the following response factors: aliphatic GS = 2.0; indole GS =

0.5. The following GSs were quantified: 4-methylsulfinylbutyl GS (4MSOB); indol-3-methylsulfinylpropyl

GS (3MSOP); 5-methylsulfinylpentyl GS (5MSOP); 7-methylsulfinylheptyl GS (7MSOH); 4-methylthiobutyl

GS (4MTB); 8-methylsulfinyloctyl GS (8MSOO); Indol-3-ylmethyl GS (I3M); 4-methoxy-indol-3-ylmethyl

GS (4MOI3M); and 1-methoxy-indol-3-ylmethyl GS (1MOI3M). ‘Total glucosinolates’ in Fig. 3 refers to

the sum of these compounds.

3

Supplementary Fig. 1. PIF4-HA binds to the promoter of ST2a in vivo as evaluated by chromatin

immunoprecipitation followed by qPCR (ChIP-qPCR) on 10-day-old seedlings exposed to low R:FR for 2

h. a, Schematic representation of the ST2A gene and two other shade-induced PIF target genes (PIL1 and

HFR1), with the regions amplified by qPCR and the position of G-boxes. b, PIF4-HA binding to PIL1 and

HFR1 in known promoter regions (see Methods for details). c, PIF4-HA binding to the ST2a promoter.

Input and immunoprecipitated DNA were quantified by qPCR. PIF4-HA enrichment is presented as

IP/Input. Small open circles represent individual data points and bars show the mean from 3 technical

replicas.. PKS4-HA seedlings are used as negative control.

c

a

b

PIL1

HFR1

ST2A

4

Supplementary Fig. 2. Phylogeny of Arabidopsis sulfotransferases (SOTs). Bayesian phylogenetic

analyses on aligned full-length sequences were performed with MrBayes v. 3.1.2 setting an MCMC

algorithm. The evolutionary distances were computed using the Dayhoff matrix based method, and are

in the units of the number of amino acid substitutions per site (see Methods for details).

adu Arachis duranensisaip Arachis ipaensisaly Arabidopsis lyrataats Aegilops tauschiibna Brassica napusbrp Brassica rapacann Capsicum annuumcrb Capsella rubellaegr Eucalyptus grandisegu Elaeis guineensiseus Eutrema salsugineumfve Fragaria vescagmx Glycine maxgra Gossypium raimondiijcu Jatropha curcasmdm Malus domesticanta Nicotiana tabacumobr Oryza brachyanthaosa Oryza sativa japonicapmum Prunus mumepxb Pyrus x bretschneiderisbi Sorghum bicolorsita Setaria itálicasot Solanum tuberosumtcc Theobroma cacaothj Tarenaya hasslerianavvi Vitis viniferazma Zea mays

5

Supplementary Fig. 3. st2a-1 and st2a-2 are ST2a null mutants, and ST2a is required for the production

of HSO4-JA in vivo. T-DNA position, ST2a mRNA levels, and HSO4-JA concentrations in two different st2a

null mutants: st2a-1 (GABI_149G04) and st2a-2 (SALK_ 075656). a, Representative scheme of ST2a gene,

showing coordinates for T-DNA insertion. b, ST2a gene expression showed strong regulation by

mechanical wounding (4 h after treatment) in wild type (Col-0) plants, and it was nearly undetectable in

both st2a mutant lines. c, Both st2a mutant lines failed to accumulate HSO4-JA after wounding (4 h)

under ambient or FR light conditions. The significance of the treatment effects in b and c was

determined using one-way ANOVA. Bars indicate means; thin bars = 1 SE; at each time point, n = 4

biological replicates (small open circles).

Amb Wounding

0

1000

2000

3000

ST

2a

/IP

P2

A

Col-0 st2a-1

Amb Control

P<0.0001

0

1000

2000

3000

ST

2a

/UB

C

Col-0 st2a-2

P<0.0001

0

5

10

15

20

nm

ol/g

DW

Col-0 st2a-1

Col-0 basal levels

Amb Wounded

FR Wounded

P=0.006

0

10

20

30

40

50

nm

ol/g

DW

Col-0 st2a-2

Col-0 basal levels

P=0.03

T-D

NA

inse

rtio

nH

SO4-

JAm

RN

A

st2a-1 st2a-2

Chr5: 2174955

a

b

c

Chr5: 2175453

5’ UTR3’ UTR stop ATGST2a5’ UTR3’ UTR stop ATGST2a

6

Supplementary Fig. 4. FR radiation reduced the concentrations of JA and the flux through the JA-Ile

conjugation pathway in plants exposed for 5 d to insect herbivory in a ST2a-dependent manner. Bars

indicate means; thin bars = 1 SE; at each time point, the interactive effects of genotype and light were

tested using two-way ANOVA; n = 5 independent biological replicates (small open circles). Significant (P

< 0.05) terms in the factorial analysis are indicated for each panel.

0

2

4

6

8

10

JA

nm

ol/g

FW

Col-0 st2a-1

Amb FR Amb FR

GxL P=0.02

JA-Ile

0

2

4

6

nm

ol/g

FW

Col-0 st2a-1

Amb FR Amb FR

OH-JA-Ile

0

2

4

6

Col-0 st2a-1

Amb FR Amb FR

GxL P=0.06

COOH-JA-Ile

0

2

4

6

Col-0 st2a-1

Amb FR Amb FR

GxL P=0.047

a b

7

Supplementary Fig. 5. Overexpression of ST2a increases HSO4-JA, and reduces JA concentration and

the flux through the JA-Ile conjugation pathway. a, Constitutive ST2a mRNA accumulation in the leaves

of ST2aOE plants compared to Col-0. b-c, Time course of accumulation of HSO4-JA and other jasmonate

metabolites in wounded leaves of Col-0 and ST2aOE plants. In all panels, P values for significant

differences between genotype means are indicated (Student’s t test). Bars indicate genotype means;

thin bars indicate 1 SE; n = 3 biological replicates (small open circles). gFW, gram of fresh weight.

0

2

4

6

ST

2a

/AC

T8

Col-0 st2aOE

ST2a

P=0.012

0 2 6 120

5

10

Time after wounding (h)

nm

ol/g

FW

HSO4-JA

P=0.0002

P=0.02

P=0.015

0 2 6 120

2

4

6

8

10 JA P=0.047

P=0.049

0 2 6 120

2

4

6

8

10 OH-JA

P=0.0001

P=0.038

0 2 6 120

1

2

3 JA-Ile

0 2 6 120

1

2

3 OH-JA-Ile

P=0.02

0 2 6 120

1

2

3 COOH-JA-IleP=0.036

nm

ol/g

FW

Time after wounding (h)

a b

c

ST2aOE

Col-0

8

Supplementary Fig. 6. A st2b mutant has normal concentrations of HSO4-JA. a, Genotyping of a st2b

null mutant. Images show photographs of PCR products run on an agarose gel (TAE buffer, agarose

1.3%). Primers were designed to amplify the T-DNA insertion (Left photograph, showing PCR product

only in st2b mutants) or DNA flanking the region of the ST2b gene where the T-DNA is inserted (Right

photograph, showing PCR product only in Col-0, as the interruption of this region by the T-DNA (~10 kb)

is too long to produce product in st2b mutant). N = negative control. M = ladder. This experiment was

performed twice with similar results. b, ST2b gene expression is not induced by wounding or FR

radiation, and it is not altered by the st2a-1 mutation (n = 3 pools of 3 individual rosettes). Rosettes of 3-

week old plants were exposed to the indicated light treatments (Amb or FR); wounding was performed

by pressing with a forceps every mature leaf. Plants were harvested before (control) or after wounding

(4 h) for gene expression analysis. c, A st2b null mutant produces wild type levels of HSO4-JA. Plants of

the indicated genotypes were exposed to the FR treatment and harvested 4 h after wounding for

phytohormone analysis Primers used of qPCR or genotyping are listed in Supplementary Table 1. For b

and c, bars indicate treatment means; thin bars = 1 SE; n = 3 (b) or 6 (c) biological replicates (pools of 3

rosettes; small open circles).

0

1

2

ST

2b

/IP

P2

a

Col-0 st2a-1

Control Wounded

Amb FR Amb FR

Control Wounded

Amb FR Amb FR0

5

10

15

nm

ol/g

FW

Col-0 st2a-1 st2b

Amb Control

FR Wounded

T-DNA insertion ST2b gene

b

a

c

st2b st2b Col-0Col-0 N NM M

HSO4-JAST2b mRNA

9

Supplementary Fig. 7. Phytochrome B inactivation reduces the accumulation of cis-OPDA in Col-0,

which is consistent with the down-regulation of LOX2 gene expression in Col-0. a, cis-OPDA

concentration in Arabidopsis rosettes 1 h after wounding. The significance of the genotype x light (GxL)

interaction term is indicated(two-way ANOVA). Different letters indicate significant ( P < 0.05)

differences between means (Tukey); bars indicate means; thin bars = 1 SE; n = 6 biological replicates

(small open circles). b, Relationship between the relative concentration of OPDA (FR/Amb) 1 h after

wounding in Col-0 and st2a-1 rosettes and the irradiance of FR received by the plants. (n = 6 biological

replicates for each genotype and FR irradiance combination). The significance of the difference between

the slopes of the fitted regression lines was determined using the two-tailed slope comparison tool in

GraphPad Prism. c, cis-OPDA concentrations (relative to Col-0) in plants of the phyB-9 mutant, which

was used as a control for phyB inactivation (FR irradiance 28 µmol s-1 m-2); bars indicate mean

concentrations; thin bars = 1 SE; n = 4 biological replicates (small open circles).

0 1 0 10

1

2

3

4cis

-OP

DA

(n

mo

l/g

FW

)

Time after wounding (h)

a

b

a a

Col-0 st2a-1

Amb

FR

GxL (1h) P=0.005

0 20 40 60

0.4

0.6

0.8

1.0

1.2

FR irradiance (umol m-2 s-1)

cis

-OP

DA

(1

h)

(FR

/Am

b)

Col-0 Y = -0.005x + 0.967

st2a-1 Y = -0.001x + 0.985

P= 0.039

Amb FR

0.4

0.6

0.8

1.0

1.2

cis

-OP

DA

(1

h)

(FR

/Am

b)

a

b c

10

Supplementary Fig. 8. Glucosinolate concentrations in induced plants. a, Glucosinolate (GS)

accumulation requires JA synthesis to respond to wounding (left panel), and it is inducible by treatment

with exogenous MeJA (right panel). In the left panel, the significance of the genotype x wounding (GxW)

interaction term was determined using two-way ANOVA and different letters indicate significant (P <

0.05) differences between means (Tukey). In the right panel, the significance of the effect of the MeJA

treatment was determined using one-way ANOVA. b, Effects of FR on accumulation of indolic (I3M,

indol-3-ylmethyl) and aliphatic (4MSOB4‐methylsulfinylbutyl) glucosinolates in wounded Col-0 and st2a-1

plants. Samples were taken 48 h after wounding or MeJA treatment (200 µM). The significance of the

genotype x light (GxL) interaction terms was determined using two-way ANOVA and different letters

indicate significant (P < 0.05) differences between means (Tukey).. In all panels, bars indicate treatment

means; thin bars = 1 SE; small open circles = biological replicates (n = 4 in a left; n = 6 in a right; n = 4 in b

left and n = 3 in b right).

0

10

20

30

m

ol/g

DW

ab

c c

GxW P=0.01

Col-0 aos

Control

Wounded

0 4818

20

22

24

26

28

Time after MeJA (h)

m

ol/g

DW

P=0.0005

I3M

Col-0 st2a-1 0.0

0.5

1.0

1.5

2.0

m

ol/g

DW

a

b

c

GxL: P=0.01

a

4MSOB

Col-0 st2a-1 0

2

4

6

8

10

a a

b

c

GxL: P=0.02

Amb

FR

b

a Total GS

11

Supplementary Fig. 9. ST2a is required for the full expression of shade avoidance responses to low

R:FR ratio. a and b, Col-0 rosettes responded to supplemental FR radiation with a reduction in the

lamina:petiole (L:P) ratio and increased leaf hyponasty; in contrast, st2a-1 rosettes had a normal

phenotype under control conditions but, when exposed to MeJA, displayed reduced expression of the

shade avoidance syndrome (SAS). The significance of the genotype x MeJA (GxMeJA) interaction term

was determined under each light condition using two-way ANOVA; different letters indicate significant

(P < 0.05) differences between means (Tukey). In all panels, bars indicate treatment means; thin bars = 1

SE; n = 8 independent biological replicates (small open circles). c, Photos of representative leaves. FRL =

14 µmol s-1 m-2; FRH = 28 µmol s-1 m-2.

0 100 2000

2

4

6

0 100 2000

2

4

6

a ab

bcb

cc

GxMeJA: P=0.004

0 100 2000

2

4

6

aababc

bc

bc

GxMeJA: P=0.004

bc

0 100 2000

20

40

60

0 100 2000

20

40

60

0 100 2000

20

40

60

a

a a ab b

GxMeJA: P=0.04

Amb FRL FRH

MeJA (μM)

L:P

rat

ioLe

af a

ngl

e (d

eg)

Col-0

st2a-1

L

P

c

a

b

FRL

100 µM MeJA(note reduced petiole length in st2a-1)

Amb FRH

200 µM MeJA(note reduced petiole length in st2a-1)

Col-0 st2a Col-0 st2a Col-0 st2a

No MeJA(full SAS in st2a-1)

12

Supplementary Fig. 10. Spectral scans of the light sources used in the experiments. Amb = “Ambient”

light (110 µmol s-1 m-2), provided by fluorescent bulbs (solid line); FR = supplement of lateral FR radiation

(dotted line) added during the course of the photoperiod to plants of the FR treatment (except indicated

otherwise, the FR irradiance was 28 µmol s-1 m-2. Photoperiod was always 10 h.

Amb FR

13

Supplementary Table 1. Sequences of primers used in genotyping, qPCR or Chip-qPCR

Primer name Sequence 5’ to 3’

Genotyping

st2a-1 Fw AAAGTTCTTGATCGACTTGT

st2a-1 Rv AACATTTCCAATCCCTCG

st2a-2 Fw CACAAGTGGAAAGATTGTCAG

st2a-2 Rv TAATCATTGTGGTTCAGTCTC

st2b Fw ATTGCTCAACAACCCCCTC

st2b Rv ATTCGGTCGAGAATCCCAG

Gene cloning

ST2a_ORF_1 GCTCTAGAATGGCTACCTCAAGCATGAAG

ST2a_ORF_2 GCGTCGACTTAGCTCAACCTGAAAGTG

qPCR

IPP2a Fw ATGGTTCAGATTGGTGGTGGAC

IPP2a RV AAAGATGTTCAGAGTTTGTGGATGG

UBC Fw CTGCGACTCAGGGAATCTTCTA

UBC Rv TTGTGCCATTGAATTGAACCC

ST2a Fw CTGAGGGCCTACTATATACG

ST2a Rv CGACAAACTTCGGTGTTGAC

MYC2 Fw CCGAAAACCCGAATCTGGAT

MYC2 Rv GGGTCTGAGAATGAACCGGAC

LOX2 Fw AAGACTGACCAGCGGATTACG

LOX2 Rv CAGGCATCTCAAACTCGCAC

VSP2 Fw TGACCGTTGGAAGTTGTGGA

VSP2 Rv CGAACCATTAGGCTTCAATATGAG

IAR3 Fw GATGCACTTGCTATGCAGGA

IAR3 Rv ACACTCCAGCCTCCACAATC

ILL6 Fw AGGCATTGTATCCCGTGAAG

ILL6 Rv CGGGTATATCGCATTCTGCT

JOX2 Fw CGGCGAAGAGCTAGTGAAGC

JOX2 Rv TGGTCATACCGCCAGGATCG

JOX4 Fw GAGGAGGCGACAAAGTCGGA

JOX4 Rv CATCACACACGATGGACCTGA

CYP94B3 Fw TGGCTTACACGAAGGCTTGTC

CYP94B3 Rv AGTCCCACGAAACTGGAGGAT

JAR1 Fw TCACGCTTTTAGAACCTTTGAACAG

JAR1 Rv GGACCGATGGGACAGTAATACG

CYP94C1 Fw GGCCCGGATTACGAAGAGTTT

CYP94C1 Rv GGCAACTTACCTTCGTT

ST2b Fw GATCCAGAACTATGAGAACCGG

ST2b Rv CTGAAAGTGAGACCAGATCCAG

Chip-qPCR

PIL1_peak_1 GAATCACGCGGCATTCAC PIL1_peak_2 ACCTTCACGCCATTATTAAGAC PIL1_control_1 GGGATGAACAATGCACCACCACAA

14

PIL1_control_2 AAACACACGAAGGCACCACGAATG

HFR1_peak_1 ACGTGATGCCCTCGTGATGGAC

HFR1_peak_2 GTCGCTCGCTAAGACACCAAC

HFR1_control_1 ACGCAACAAACGAACCACAC

HFR1_control_2 AGAGCGATCGGATCAGATAG

ST2a_region1_1 TGTGTGGAAGTGAACGTGGT

ST2a_region1_2 GGCTTCAAAGCACACTCACA

ST2a_region2_1 TATTCGCACACGCCGTTTAT

ST2a_region2_2 TTCGAGATGAAGTTGGGTGTTT

ST2a_region3_1 GATCTGGTCTCACTTTCAGGTTG

ST2a_region3_2 TCGACAAACTTCGGTGTTGA

ST2a_control_1 TGGATACAATGCCAACCAACT

ST2a_control_2 CGAAATGATTTGTTGGTGATGC

15

Supplementary Table 2. Details of analysis of phytohormones by LC-MS/MS [HPLC 1260 (Agilent

Technologies)-QTRAP6500 (SCIEX)] in negative ionization mode

Q1 Q3 RT (min) Compound

Internal std

RF DP EP CE CXP

136.93 93 3.3 SA D4-SA 1.0 -20 -8 -24 -7

263 153.2 3.4 ABA D6-ABA 1.0 -20 -12 -22 -2

209.07 59 3.6 JA D6-JA 1.0 -20 -9 -24 -2

322.19 130.1 3.9 JA-Ile D6-JA-Ile 1.0 -50 -4.5 -30 -4

290.9 165.1 4.6 OPDA D6-JA 1.0 -20 -12 -24 -2

263 165 4.2 dinor-OPDA D6-JA 0.7 -20 -10 -20 -10

338.1 130.1 3 OH-JA-Ile D6-JA-Ile 1.0 -50 -4.5 -30 -4

225.1 59 2.6 OH-JA D6-JA 1.0 -20 -9 -24 -2

352.1 130.1 3 COOH-JA-Ile D6-JA-Ile 1.0 -50 -4.5 -30 -4

305 97 2.4 SulfoJA D6-JA 6.0 -20 -10 -60 -10

387.1 207 2.4 JA-Gluc D6-JA 3.7 -50 -10 -28 -21

140.93 97 3.3 D4-SA -20 -8 -24 -7

269 159.2 3.4 D6-ABA -20 -12 -22 -2

215 59 3.6 D6-JA -20 -9 -24 -2

214 59 3.6 D5-JA -20 -9 -24 -2

328.19 130.1 3.9 D6-JA-Ile -50 -4.5 -30 -4

327.19 130.1 3.9 D5-JA-Ile -50 -4.5 -30 -4

++++++++++++

Supplementary Data File 1. Overrepresented GO categories in RNAseq data reported on Fig 3b (Excel

File).

Supplementary Data File 2. Identification codes for sulfotransferase sequences (Excel File).

Supplementary Data File 3. List of genes included in the heat map of Supplementary Fig. 1a (Excel

File).

++++++++++++