0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.20

5

10

15

20

25

30

35

R² = 0.70253008558648

Ψl (absolute values MPa)

Val (

rela

tive

met

abol

ite c

onte

nt)

0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.20

2

4

6

8

10

12

R² = 0.791667359672369

Ψl (absolute values MPa)

Leu

(rel

ative

met

abol

ite c

onte

nt)

0 0.5 1 1.5 2 2.50

0.5

1

1.5

2

2.5

3R² = 0.956472868442063

Ψl (absolute values MPa)

Pro

(rel

ative

met

abol

ite c

onte

nt)

0 0.5 1 1.5 2 2.5350

400

450

500

550

600

650

R² = 0.779639498078526

Ψl (absolute values MPa)

Tota

l osm

olite

s (m

ole/

kg) A

B D

CCs

Sh

Figure S1. Correlation of Ψl to total osmolite concentration (A) and to the relative abundance of Proline (B), Valine (C) and Leucine (D) in Shiraz (solid circles) and Cabernet Sauvignon (empty circles). Data point represent means ± SE, n=6. for all the regressions p<0.001.

4 18 26 34

Day of the experiment

Sh IRCs IRSh D Cs D

Figure S2. PCA plot (x-1st component, y-3rd component) of Cabernet Sauvignon and Shiraz grape leaf extracts of GC/MS based metabolites analyzed using TMEV. Symbols represent different sampling days of Shiraz (Sh) and Cabernet Sauvignon (Cs) irrigated (IR) and water deficit (D) treatments.

-1.0

-0.8

-0.6

-0.4

-0.2

-0.0

0.2

0.4

0.6

0.8

1.0

-0.005 -0.004 -0.003 -0.002 -0.001 0.000 0.001 0.002 0.003 0.004

p(c

orr

)[1]P

(C

orr

ela

tion)

CoeffCS[2](Shiraz at day 34) (X Effects)

S-Plot (Shiraz Irrigated = -1, Shiraz Drought = 1)

111.0098

149.0093

164.0728191.0203

195.0509195.0363

203.0838

289.0719

955.1388

439.0842

439.1033

EZinf o 2 - combined greenhouse data negativ e mode (M5: OPLS-DA) - 2012-02-08 08:55:40 (UTC+2)

Citric acid fragment

111.0098

Phenylalanine 164.0728

Citric acid 191.0203

Tryptophan 203.0838

Catechin 389.1033

Tartaric acid 149.0093

2X Quercetin-3-O-glucuronide

955.1388

Figure S3. S-plot of the OPLS-DA model from Shiraz irrigated vs. water deficit treatments of leaf sample metabolite markers analyzed in negative ESI mode on day 34 of the experiment. Metabolites with the highest correlation and covariance values account for most of the variance between the two groups.

-1.0

-0.8

-0.6

-0.4

-0.2

-0.0

0.2

0.4

0.6

0.8

1.0

-0.010 -0.009 -0.008 -0.007 -0.006 -0.005 -0.004 -0.003 -0.002 -0.001 0.000 0.001 0.002 0.003 0.004 0.005 0.006 0.007 0.008

p(co

rr)[

1]P

(C

orre

latio

n)

CoeffCS[2](Cabernet Sauvignon at day 34) (X Effects)

S-Plot (Cabernet Sauvignon_IR = -1, Cabernet Sauvignon_DR = 1)

175.0266

EZinf o 2 - combined greenhouse data negativ e mode (M5: OPLS-DA) - 2012-02-08 14:56:41 (UTC+2)

Figure S4. S-plot of the OPLS-DA model from Cabernet Sauvignon irrigated vs. water deficit treatments of leaf sample metabolite markers analyzed in negative ESI mode on day 34 of the experiment. Metabolites with the highest correlation and covariance values account for most of the variance between the two groups.

0 0.05 0.1 0.15 0.2 0.25 0.30

10

20

30

40

50

60

70

80

90f(x) = 4.06382467446655 x^-0.62444949601184R² = 0.838758667989104

gs

AB

A (

rela

tive

met

abol

ite c

onte

nt)

Cs

Sh

Figure S5. Correlation of stomatal conductance (gs) to Abscisic acid (ABA) – as measured in the sap – in Shiraz (solid circles) and Cabernet Sauvignon (Cs) (empty circles). p<0.001

Figure S6. Symmetric difference network based on correlation between irrigated treatment and metabolites. Nodes correspond to metabolites of primary (circles) and secondary (squares) metabolism, node colors correspond to compound classes as displayed in the figure legend. Edges between nodes represent significantly identified correlations at r ≥ 0.9 and q ≤ 0.01, where blue edges correspond specifically to Shiraz and red edges correspond specifically to Cabernet sauvignon. Nodes are ordered into modules corresponding to their compound classes.

Figure S7. Symmetric difference network based on correlation between water deficit treatment and metabolites. Nodes correspond to metabolites of primary (circles) and secondary (squares), node colors correspond to compound classes as displayed in the figure legend. Edges between nodes represent significantly identified correlations at r ≥ 0.9 and q ≤ 0.01, where blue edges correspond specifically to Shiraz and red edges correspond specifically to Cabernet sauvignon. Nodes are ordered into modules corresponding to their compound classes.