Download pce12214-sup-0001-si

Suplementary Figure:
Peptide/ Antibody
PIP2
PIP2PH
Pept PIP2
1.21
0.11
Pept PIP2PH
0.09
1.02
BSA
0.08
0.07
Table S1 Specificity of antibodies to recognize their corresponding peptides. The table shows the
specificity of each antibody to link at different aquaporin proteins and BSA (bovine serum albumin).
We have antibodies against the C- terminal peptide PIP2 and one antibody against phosphorylated
PIP2 at the C-terminal, like PIP2PH (Ser 280). These results were obtained using ELISA technique.
Compound
Transition
225>151
225>133
227>151
d2-Methyl jasmonate
227>133
Methyl jasmonate
Dwell
(ms)
80
80
80
80
DP (V)
EP (V)
CEP (V)
CE (V)
26
26
26
26
8
8
8
8
15
15
15
15
18
21
19
21
Table S2. Compounds dependent parameters for methyl jasmonate.
Untreated
MeJA
7000
[MeJA] pmoles g-1
*
6000
*
5000
4000
3000
2000
1000
0
1Hour
24 Hours
Time of exposition
Fig. S1 MeJA concentration (pmol g-1 root FW) of roots of P. vulgaris plants after 1 or 24 h of exposure to 100
µM MeJA. Bars represent mean ± SE (n = 4). Asterisks mean significant differences (p < 0.05) after t-student
test.
Untreated
0,1 mM MeJA
0,5 mM MeJA
1 mM MeJA
L( gH2O g-1 root DW MPa-1 h-1)
40
a
35
30
a
25
20
a
a
a
15
10
b
b
b
5
0
1 Hour
24 Hours
Time of exposition
Fig. S2 Root hydraulic conductivity (L) of P. vulgaris plants subjected to 0 (untreated), 0.1, 0.5 and 1 mM
MeJA during 1 and 24 h. L was measured by the free exudation method. Bars represent mean ± SE .
Within each time period different letters mean significant differences (p<0.05) after ANOVA and LSD test
(n =8).
Untreated
MeJA
L (gH2O g-1 root DW MPa-1 h-1)
70
*
60
50
*
40
30
20
10
0
1 Hour
24 Hours
Fig. S3 Root hydraulic conductivity (L) measured with a pressure chamber in roots of P. vulgaris plants after 1
or 24 h of exposure to 100 µM MeJA. Bars represent mean ± SE (n = 8). Asterisks mean significant
differences (p < 0.05) after t-student test.
L (gH2O g-1 root DW MPa-1 h-1)
L (gH2O g-1 root DW MPa-1 h-1)
(a)
Untreated
MeJA
8
7
a
6
a
a
5
4
A
3
2
B
1
C
0
30
60
Time of exudation (min)
(b)
90
16
a
14
12
10
8
b
6
b
4
2
A
0
30
B
B
60
90
Time of exudation (min)
Fig. S4 Root hydraulic conductivity (L) of P. vulgaris plants, untreated and treated with 100 µM MeJA
during 1 (a) or 24 h (b) . L was measured each 30 min during 90 min by the free exudation method . Bars
represent mean ± SE . Within each treatment different letters mean significant differences of L along the
time (p<0.05) after ANOVA and LSD test (n =8).
WT
def-1
def-1
1.6
Gene expression(r.u)
1.4
1.2
*
1
0.8
0.6
0.4
0.2
0
AOS 2
MC
Fig. S5 Relative expression of SlAOS2 and SlMC genes of S. lycopersicum in WT and def-1 roots. Bars
represent mean ± SE (n = 6). Asterisks mean significant differences (p < 0.05) after t-student test.
Fig. S6 Phylogenetic tree of P. vulgaris and Arabidopsis thaliana PIP aquaporins using neighbour-joining
method. The accession numbers of the proteins used are: PvPIP1;1 (AAB72149), PvPIP1;2 (AAY22204),
PvPIP1;3 (ABH85411), PvPIP2;1 (AAY22203), PvPIP2;2 (ABU94630), PvPIP2;3 (ABU94631), AtPIP1;1
(CAB71073), AtPIP1;2 (AAC28529), AtPIP1;3 (AAF81320), AtPIP1;4AAF02782), AtPIP1;5 (CAA20461), AtPIP2;1
(CAB67649), AtPIP2;2 (AAD18142), AtPIP2;3 (AAD18141), AtPIP2;4 (BAB09839), AtPIP2;5 (CAB41102),
AtPIP2;6 (AAC79629), AtPIP2;7 (CAA17774), AtPIP2;8 (AAC64216).
Ca2+ concentration (µmol ml-1h-1g-1)
25
b
20
a
15
10
5
0
Untreated
EGTA
Fig. S7 Apoplastic calcium concentration of untreated plants and plants treated with (1mM) EGTA
(apoplastic calcium chelator) during 24 h. Scholander chamber at 0.25 MPa was used to get the exudates
(n =4)(t-test, P≤0.05).