Supplementary Data

Full text

H2O2 mediates the crosstalk of brassinosteroid and abscisic acid in tomato responses to
heat and oxidative stresses
J Q Yu, Jie Zhou, Jian Wang, Xin Li, Zhixiang Chen, Yanhong Zhou, Kai Shi, and Xiaojian Xia
Supplementary Data
Supplementary Figure S1. Effects of BRs and ABA levels on heat shock and photooxidative
stress tolerance in BR- and ABA-deficient plants. (a) Electrolyte leakage values of plants after
exposure to a 6 h heat shock (42oC under 800 µmol m-2 s-1) . (b) Electrolyte leakage values of
plants after exposure to 3 h of photooxidative stress (20 μM PQ). For (a) and (b), EBR (200 nM)
or ABA (50 μM) were applied at 24 h before the plants were exposed to heat shock and PQ
stresses, respectively. Twelve plants were used for each treatment, and Electrolyte leakage values
were determined with the full 4th leaf. The data are the means of twelve replicate plants (±SD).
Means denoted by the same letter do not differ significantly at P≤0.05 according to Turkey’s test.
CR, Condine Red (wild-type); d^im, BR-deficient mutant; AC, Ailsa Craig (wild-type); not,
notabilis ABA-deficient mutant.
0
10
20
30
not
d^im
AC
CR
d
d
c c
c c
c
c c
b
b ab
a
a
d
d
Electrolyte leakage (%)
Water+25
o
C
Water+42
o
C
EBR+42
o
C
ABA+42
o
C
0
10
20
30
not
d^im
AC
CR
d
d
c c
c c
c
c bc
b
b a
a
a
d
d
Electrolyte leakage (%)
Water+PQ
Water+PQ
EBR+PQ
ABA+PQ
(a)
(b)

Supplementary Figure S2. The roles of BR and ABA in regulation of PQ tolerance. (a) and (b)
Images and values of the maximum PSII quantum yield (Fv/Fm) of leaves after exposure to a 3 h
PQ stress (20 μM) at 24 h after EBR (200 nM), or ABA (50 μM) treatments with DPI or DMTU
pretreatment. DPI (50 µM) and DMTU (5 mM) were applied 12 h before the EBR or the ABA
treatment. Twelve plants were used for each treatment and the picture of one representative leaf
is shown. Bar = 1.0 cm.
(a)
(b)
0.5
0.6
0.7
0.8
c
c
b
a
b
b
b
a
ABA
EBR
Fv/Fm
Wat er
DPI
DMTU
Water
a
DMTU
DPI
Water
Water EBR ABA

Supplementary Figure S3. In situ detection of H2O2 in leaves. Leaf samples were taken at 24 h
after (200 nM), ABA (50 μM) or at 3 h after paraquat (PQ, 20 µM) or H2O2 (50 mM) treatment,
and loaded with DAB and incubated for 6 h. DPI (50 µM) and DMTU (5 mM) were applied 12
h before the EBR or the ABA treatment. The H2O2 accumulation was detected by an Olympus
motorized system microscope (BX61, Olympus Co., Tokyo, Japan). (a) & (b) observed at 2x
magnification; (c) observed at 400x magnification.
(a)
Water EBR ABA
DPI
DMTU
Water
(c)
(b)
Water EBR ABA PQ H2O2

Supplementary Table S1. Primers used for real time RT-PCR assays
Gene
Accession numbers
Forward primer
Reverse primer
Rboh1
Sl08g081690
5’-GGAGCTCCAGCACAAGATTA-3’
5’-CTTGTTGCAGCACTCATGTC-3’
WRKY1
Sl07g047960
5’- CTAGTGCAGGGTCAAGGAAA-3’
5’- ACAGGACTCTTCGTCACTCG-3’
WRKY72
Sl02g067430
5’- AAGCAGGTTCAAAGATGTGC-3’
5’- CAGTTACTTGTGGGTTTGGG-3’
MAPK1
Sl12g019460
5’-TGCACCTCCGGTCAACAA-3’
5’-GGCAGTGCTCCTCAGATAAA-3’
HSP70
NM001246851
5’- CAAGCTGAAAGAGCTCAAGG-3’
5’- CTGTCCCAGCTGCATTACTT-3’
Cu/Zn-SOD
Sl11g066390
5’- GGCCAATCTTTGACCCTTTA -3’
5’- AGTCCAGGAGCAAGTCCAGT -3’
cAPX
Sl06g005160
5’- TCTGAATTGGGATTTGCTGA -3’
5’- CGTCTAACGTAGCTGCCAAA -3’
GR1
Sl09g091840
5’- TTGGTGGAACGTGTGTTCTT -3’
5’- TCTCATTCACTTCCCATCCA -3’
CAT1
Sl12g094620
5’-TGATCGCGAGAAGATACCTG-3’
5’-CTTCCACGTTCATGGACAAC-3’
Actin
Sl11g005330
5’- TGTCCCTATTTACGAGGGTTATGC -3’
5’- CAGTTAAATCACGACCAGCAAGAT -3’