Supplemental material
Seasonal variations of Fucus vesiculosus fertility under ocean acidification and warming in the
western Baltic Sea
Angelika Graiff, Marie Dankworth, Martin Wahl, Ulf Karsten and Inka Bartsch
DOI 10.1515/bot-2016-0081
1
Table S1: Mean, minimum, and maximum Benthocosm temperatures in each month.
Spring
2013
Summer
2013
Autumn
2013
Winter
2014
Temperature in the Kiel Fjord (°C)
Temperature in the Kiel Fjord +5°C
Mean±SD
Min
Max
Mean±SD
April
8.2±1.4
4.3
11.7
11.3±1.1
9.0
18.4
May
11.3±2.1
6.1
17.4
15.4±2.8
10.6
21.7
June
16.1±1.3
11.7
19.4
21.1±1.3
15.5
23.7
July
20.4±1.8
14.8
24.1
25.0±2.5
16.6
29.0
August
19.5±1.2
14.7
24.8
24.5±1.1
19.5
30.0
September
17.4±1.0
13.3
19.6
22.4±1.0
18.3
24.7
October
13.1±0.5
11.6
14.6
17.7±0.9
12.7
19.0
November
10.3±1.6
7.2
13.1
15.2±1.6
12.1
18.1
December
6.9±0.5
5.5
9.5
11.8±0.5
9.7
13.3
January
4.2±1.4
1.7
7
7.8±1.3
5.4
10.6
February
4.3±0.7
2.3
6.3
8.4±0.9
6.9
11.0
March
6.7±1.1
4.4
9.1
11.5±1.2
7.8
14.4
Min
Max
2
Table S2: Summary of mean monthly seawater carbonate chemistry. pCO2 (n = 3, ± SD) was calculated from total alkalinity (TA, n = 3), dissolved
organic carbon (DIC, n = 3) and pH (n = 3) measurements of seawater corresponding to each treatment (Wahl et al. 2015b, M. Böttcher and V.
Winde pers. comm.).
Spring 2013
Summer 2013
Autumn 2013
Winter 2014
April
May
June
July
August
September
October
November
December
January
February
March
Control
pH
pCO2
TA
DIC
ppm
µmol kg–1
µmol kg–1
8.73±0.11
130±57
2066±82
1731
8.27±0.40
516±413
2007±69
1818
8.26±0.39
606±390
2059±91
1917
8.20±0.19
489±228
1971±38
1927
7.96±0.11
772±254
1906±103
1849
7.91±0.12
886±284
1901±38
1890
7.97±0.09
723±159
1968±33
1950
7.97±0.08
719±163
2058±57
2054
7.87±0.05
877±71
2077±14
2033
7.96±0.07
718±113
2223±44
2208
8.02±0.08
636±85
2173±12
2070
8.08±0.14
587±219
2142±55
2201
+CO2
pH
pCO2
TA
DIC
ppm
µmol kg–1
µmol kg–1
8.49±0.16
297±192
2069±87
1991
8.06±0.39
854±663
2008±70
1883
7.95±0.43
1244±815
2069±87
2013
8.00±0.17
786±320
1972±45
1969
7.83±0.10
1070±333
1916±112
1883
7.75±0.13
1313±397
1910±39
1921
7.87±0.09
924±198
1979±39
1970
7.85±0.09
927±212
2052±56
2085
7.73±0.05
1217±142
2086±17
2071
7.82±0.07
1066±57
2197±53
2248
7.79±0.04
1087±94
2174±10
2178
7.82±0.07
1061±210
2143±54
2221
+Temp
pH
pCO2
TA
DIC
ppm
µmol kg–1
µmol kg–1
8.52±0.12
220±121
2067±85
1929
8.03±0.38
960±837
2005±71
1905
8.13±0.18
656±244
2051±93
2069
8.04±0.26
828±538
1975±36
1926
7.74±0.13
1414±531
1921±100
1876
7.73±0.09
1373±343
1914±38
1934
7.86±0.07
952±166
1979±31
1956
7.89±0.08
878±160
2053±44
2055
7.84±0.03
953±81
2081±18
2034
7.92±0.07
808±142
2205±48
2208
7.91±0.04
826±83
2166±9
2122
7.83±0.10
1074±313
2106±69
2160
+Temp +CO2
pH
pCO2
TA
DIC
ppm
µmol kg–1
µmol kg–1
8.27±0.19
503±402
2095±89
2050
7.84±0.38
1200±947
2022±79
1953
7.86±0.25
1357±640
2059±85
2119
7.91±0.25
1385±809
1964±48
1907
7.63±0.13
1725±556
1905±108
1959
7.63±0.10
1490±885
1912±42
1970
7.81±0.11
1118±290
1976±44
1978
7.81±0.08
1092±202
2065±54
2083
7.69±0.04
1384±107
2077±8
2073
7.81±0.09
1071±215
2217±40
2244
7.73±0.04
1291±147
2168±13
2164
7.66±0.07
1585±263
2111±69
2197
3
Table S3: Three-day mean, minimum, and maximum Benthocosm temperatures in the
summer experiment (4 July-17 September 2013).
Temperature in the Kiel Fjord (°C)
Temperature in the Kiel Fjord +5°C
Days
Mean±SD
Min
Max
Mean±SD
Min
Max
3
16.4±1.1
14.8
20.6
19.0±1.9
16.6
23.2
6
19.0±1.2
16.9
22.6
23.6±1.5
19.2
25.8
9
20.5±0.6
19.4
22.1
25.6±0.7
24.5
27.9
12
19.8±0.4
19.0
20.8
24.9±0.4
24.0
26.8
15
20.4±0.9
18.9
22.1
25.5±0.9
24.1
27.2
18
21.6±0.7
20.2
23.3
26.5±0.6
24.9
28.0
21
22.5±0.7
21.1
24.1
27.4±0.7
26.1
29.0
24
21.9±0.5
21.1
23.4
26.8±0.4
26.1
28.2
27
21.1±0.8
19.0
22.6
25.9±0.8
24.1
27.6
30
20.5±1.3
18.8
24.3
25.5±1.2
23.8
28.5
33
21.0±0.7
20.2
24.8
26.0±0.5
24
30.0
36
20.3±0.7
18.9
23.7
25.2±0.5
24.1
28.5
39
19.9±0.3
19.3
20.7
24.9±0.3
24.4
25.6
42
19.2±0.3
18.6
19.9
24.3±0.3
23.7
24.8
45
19.3±1.1
17.2
23.0
24.2±0.8
22.2
27.0
48
18.5±0.5
17.3
19.5
23.5±0.5
22.3
24.5
51
19.1±0.6
18.1
20.2
24.2±0.6
23.2
26.3
54
18.9±0.6
16.9
19.9
23.9±0.6
21.8
24.8
57
18.4±1.4
14.7
19.9
23.3±1.4
19.5
25.0
60
17.9±1.4
13.3
20.5
22.9±1.3
18.3
25.4
63
17.9±0.2
17.5
18.5
23.0±0.2
22.6
23.4
66
18.6±0.5
17.6
19.6
23.6±0.5
22.7
24.7
69
17.1±0.7
16.0
18.6
22.20.7
20.9
23.6
72
16.8±0.3
16.2
17.5
21.8±0.3
21.2
22.8
75
16.6±0.6
15.4
17.8
21.7±0.5
20.4
22.7
4
Table S4: Results of repeated-measures ANOVA for effects of temperature, CO2 and time
during the course of each experiment on proportion of mature Fucus vesiculosus receptacles
during different seasons.
Source of variation
(a) Spring
Temperature
CO2
Time
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
(b) Summer
Temperature
CO2
Time
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
(c) Autumn
Temperature
CO2
Time
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
(d) Winter
Temperature
CO2
Time
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
DF
F-value
p-value
1
1
3
1
3
3
3
4.696
1.255
11.860
11.739
0.118
0.957
0.872
0.062
0.295
<0.001
0.009
0.948
0.404
0.469
-
-
-
1
1
3
1
3
3
3
3.107
1.020
1.161
0.877
1.363
0.695
1.262
0.116
0.342
0.345
0.376
0.284
0.564
0.310
1
1
1.401
1
1.401
1.401
1.401
0.085
6.903
3.574
0.197
1.533
0.167
0.913
0.778
0.030
0.075
0.669
0.252
0.772
0.394
Seasons: spring: 4 April-19 June 2013; summer: 4 July-17 September 2013; autumn: 10
October-18 December 2013; winter: 16 January-1 April 2014. During the summer experiment
the proportion of mature receptacles on the Fucus plants was too low for statistical analyses.
Bold type indicates p-values < 0.05.
5
Table S5: Results of repeated-measures ANOVA for effects of temperature, CO2 and time
during the course of each experiment on proportion of vegetative Fucus vesiculosus apices
during different seasons.
Source of variation
(a) Spring
Temperature
CO2
Time
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
(b) Summer
Temperature
CO2
Time
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
(c) Autumn
Temperature
CO2
Time period
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
(d) Winter
Temperature
CO2
Time
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
DF
F-value
p-value
1
1
3
1
3
3
3
0.908
1.013
38.249
12.274
1.702
0.272
0.386
0.368
0.344
<0.001
0.008
0.193
0.845
0.764
1
1
1.164
1
1.164
1.164
1.164
283.953
3.953
162.821
9.899
109.597
0.818
0.433
<0.001
0.082
<0.001
0.014
<0.001
0.407
0.556
1
1
1.554
1
1.554
1.554
1.554
1.398
1.329
28.786
0.786
2.405
0.389
0.320
0.271
0.282
<0.001
0.401
0.138
0.635
0.679
1
1
1.114
1
1.114
1.114
1.114
0.817
7.750
1.183
1.988
0.862
0.058
0.337
0.392
0.024
0.313
0.196
0.390
0.841
0.600
Seasons: spring: 4 April-19 June 2013; summer: 4 July-17 September 2013; autumn: 10
October-18 December 2013; winter: 16 January-1 April 2014. Bold type indicates p-values <
0.05.
6
Table S6: Results of repeated-measures ANOVA for effects of temperature, CO2 and time
during the course of each experiment on proportion of immature Fucus vesiculosus
receptacles during different seasons.
Source of variation
(a) Spring
Temperature
CO2
Time
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
(b) Summer
Temperature
CO2
Time
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
(c) Autumn
Temperature
CO2
Time
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
(d) Winter
Temperature
CO2
Time
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
DF
F-value
p-value
1
1
1.11
1
1.11
1.11
1.11
0.406
1.988
9.116
1.776
2.744
1.034
4.156
0.542
0.196
0.013
0.219
0.131
0.346
0.070
-
-
-
1
1
1.978
1
1.978
1.978
1.978
0.499
1.349
33.298
0.551
1.685
1.004
0.268
0.500
0.279
<0.001
0.479
0.217
0.388
0.766
1
1
1.878
1
1.878
1.878
1.878
0.168
0.002
5.198
2.622
0.162
0.312
1.049
0.693
0.968
0.021
0.144
0.839
0.724
0.370
Seasons: spring: 4 April-19 June 2013; summer: 4 July-17 September 2013; autumn: 10
October-18 December 2013; winter: 16 January-1 April 2014. During the summer experiment
the proportion of immature receptacles on the Fucus plants was too low for statistical
analyses. Bold type indicates p-values < 0.05.
7
Table S7: Results of repeated-measures ANOVA for effects of temperature, CO2 and time
during the course of each experiment on proportion of decayed Fucus vesiculosus receptacles
during the spring experiment (4 April-19 June 2013).
Source of variation
(a) Spring
Temperature
CO2
Time
Temp x CO2
Temp x Time
CO2 x Time
Temp x CO2 x Time
DF
F-value
1
1
1.326
1
1.326
1.326
1.326
15.030
0.100
18.522
3.311
25.934
9.274
6.465
p-value
0.005
0.760
0.001
0.106
<0.001
0.008
0.022
During summer, autumn and winter experiments the proportion of decaying receptacles on the
Fucus plants was too low for statistical analyses. Bold type indicates p-values < 0.05.
8
Figure S1: (a) pH in the control (blue) and +CO2 (red) experimental units of a tank with
ambient temperature and in Kiel Fjord (in situ, black). (b) pH in the +Temp (blue) and +Temp
+CO2 (red) experimental units of a tank with increased temperature and in Kiel Fjord (in situ,
black). Note that Kiel Fjord pH measurements prior to 30 May were taken at a shallower
depth than the inlet of the flow-through. After that date pH in Kiel Fjord was measured close
to the inlet (modified according to Wahl et al. 2015).
9