Germination (% of control
120
100
Aa
A ab
A
A
b
b
80
60
40
20
c
c
0
0
50
100
150
200
NaCl concentration (mM)
Supplemental Figure S1. The effect of NaCl on germination of Arabidopsis and Eutrema. Seeds were sown on MS
medium or MS supplemented with the indicated NaCl concentrations. Germination was recorded at 7 d after stratification
and is presented as percentage compared to the respective control (0 mM NaCl) medium. Data are mean ± SD (n=3). Each
replicate plate contained ca. 100 seeds. Data are representative of similar results from three independent experiments. Bars
with different letters (upper case, Arabidopsis; lower case; Eutrema) indicate significant difference (p ≤ 0.01, student’s ttest). Each species was statistically analyzed separately between all salt levels. Dark grey bars, Arabidopsis; light grey bars,
Eutrema.
A
B
D
Number of leaves
Fresh weight (mg)
C
8
6
4
2
0
N/T
T
N/T
E
T
6
5
4
3
2
1
0
N/T
T
N/T
T
F
Supplemental Figure S2. The effect of transfer by mesh on growth and development of Arabidopsis and Eutrema. Seeds
were sown on MS medium overlaid with circles of autoclaved silk mesh and either transferred to fresh MS plates at stage
0.5 (radical emergence, Boyes et al. (2001) or grown on the same plates until harvested. A and B, Images of plates with
overlaid mesh with Arabidopsis (A) and Eutrema (B) at 12 days after stratification (DAS). C, Fresh weight of seedlings at
12 DAS. D, Number of true leaves at 12 DAS. E and F, Appearance of Arabidopsis (E) and Eutrema (F) seeds at the time of
transfer to mesh. Data are mean ± SD (n=3). Each replicate plate contained ca. 15 seedlings. Data are representative of
similar results from two independent experiments. No significant differences (p ≤ 0.01, student’s t-test) were observed in
either fresh weight or number of leaves between transferred and non-transferred plants. Dark grey bars, Arabidopsis; light
grey bars, Eutrema; T, transferred seedlings; N/T, non-transferred seedlings.
A
140
Hull area (mm2)
120
Arabidopdis
A
Eutrema
100
A
a
80
ab
60
B
b
40
c
20
0
0
B
Hull area (mm2)
1800
1600
50
100
150
A
Arabidopsis
Eutrema
1400
1200
a
1000
B
b
800
B
b
600
C c
c
200
300
400
200
0
0
50
100
Rosette leaf area (mm2)
C
1000
900
800
700
600
500
400
300
200
100
0
A
Arabidopsis
Eutrema
B
B
a
b
b
C
c
0
50
100
200
NaCl concentration (mM)
c
300
Supplemental Figure S3. Actual hull and rosette areas of in vitro- and soil-grown Arabidopsis and Eutrema in response to
salt stress. A, Hull area, in vitro-grown plants. B, Hull area soil-grown plants. C, Rosette area, soil-grown plants. Data are
mean ± SD (n=3, in vitro; n=8, soil). Each replicate in vitro plate contained ca. 15 seedlings. Only Eutrema plants could
grow on soil under 300 mM NaCl. Data are representative of similar results from two independent experiments. Bars with
different letters (upper case, Arabidopsis; lower case; Eutrema) indicate significant difference (p ≤ 0.05, student’s t-test).
Each species was statistically analyzed separately between all salt levels. Dark grey bars, Arabidopsis; light grey bars,
Eutrema.
Arabidopsis
240
0 mM NaCl
50 mM NaCl
200
100 mMNaCl
200 mM NaCl
160
Individual leaf area (mm2)
120
80
40
0
90
Eutrema
0 mM NaCl
50 mM NaCl
100 mM NaCl
200 mM NaCl
300 mM NaCl
80
70
60
50
40
30
20
10
0
с
с
1
2
3
4
5
6
7
8
9
10
Leaf number
Supplemental Figure S4. The effect of salt stress on the area of individual soil-grown Arabidopsis and Eutrema leaves. To
better visualize the overall effect of salt, all salt treatments were placed on one graph for each species. Data are mean ± SD
(n=8). Data are representative of similar results from two independent experiments. Dark grey bars, Arabidopsis; light grey
bars, Eutrema.
Arabidopsis
0 mM NaCl
50 mM NaCl
100 mM NaCl
200 mM NaCl
Eutrema
0 mM NaCl
50 mM NaCl
100 mM NaCl
200 mM NaCl
Supplemental Figure S5. Microscopic images of adaxial Arabidopsis and Eutrema leaf surfaces. Red scale bar = 20 m.