1
Supporting Information
2
Table S1 Number of individuals, collection location and coordinates, time, technique, depth and temperature, and mortalities during exposure
3
time (as % across incubation temperatures) for the prawn species investigated in this study. *Please note that for P. montagui all mortalities
4
occurred at 25 °C.
Species
n
Locality
Palaemon
elegans
88
Mt Batten
Palaemon
macrodactylus
28
outer Thames
Estuary
Palaemon
serratus
32
Plymouth Sound
Palaemonetes
varians
100
Lymington salt
marsh
Crangon
crangon
112
Plymouth Sound
32
outer Thames
Estuary
Pandalus
montagui
Coordinates
Time
Technique
Depth (m)
Temperature (°C)
June 2012
Hand held net
0.1-0.5
12.4
May 2012
Otter trawl
5-20
9.8
March 2012
Beam trawl
8-10
10.0
June 2012
Hand held net
0.3-0.5
22.3
March 2012
Beam trawl
3-10
10.0
May 2012
Otter trawl
5-20
9.8
Mortality (%)
50° 21’ 25.16’’ N
4° 07’ 35.20’’ W
8.8
51° 29’ 37.27’’ N
0° 36’ 29.27’’ W
0
50° 21’ 09.12’’ N
4° 07’ 56.09’’ W
50° 44’ 20.1’’ N
1° 32’ 16.2’’ W
0
0
50 21’ 49.28’’ N
4 11’ 13.19’’ W
0
51° 29’ 31.80’’ N
0° 50’ 53.90’’ W
25*
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Table S2 GenBank accession numbers of the prawn species used for phylogenetic analysis.
Species
H3
12S
16S
18S
28S
Palaemon
elegans
GU117695
DQ079729
DQ079764
DQ079807
Palaemon
macrodactylus
DQ642862
JQ042297
DQ642849
EF540847
EU920915
EU920938
EU920972
GQ487498
GQ487506
GQ487514
Palaemon
serratus
JQ042291
Palaemonetes
varians
JN674357
Crangon
crangon
EU920885
Pandalus
montagui
Procaris
ascensionis
GQ487520
GQ487495
GQ487503
GQ487511
Stenopus
hispidus
DQ079701
DQ079734
DQ079769
DQ079812
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Table S3 Mean ± SE for oxygen consumption rate (ṀO2) and Upper Thermal Limits (UTL), measured as Loss of Orientation (LO), Onset of
2
Spasms (OS) and Death (D), for the prawn species investigated in this study after 7 d exposure to one of four incubation temperatures: 10, 15, 20
3
and 25 °C. Sample size and wet weight are also reported. *nr = not recorded
Species
Palaemon
elegans
Palaemon
macrodactylus
Palaemon
serratus
Palaemonetes
varians
Crangon
crangon
Temperature
(°C)
n
ṀO2 (nmol O2 g
wet weight -1 min-1)
LO (°C)
OS (°C)
D (°C)
Wet weight
(g)
Wet weight
range (g)
10
15
20
25
10
15
20
25
10
15
20
25
10
15
20
25
10
15
20
25
18
20
17
16
5
7
7
7
8
8
8
7
18
17
16
10
27
23
20
21
90.7 ± 8.4
249.2 ± 14.1
319.3 ± 20.9
699.7 ± 76.9
65.3 ± 15.1
80.9 ± 4.9
157.5 ± 10.0
304.5 ± 21.8
46.1 ± 7.1
70.2 ± 8.4
147.4 ± 21.8
171.4 ± 7.6
80.1 ± 7.7
195.3 ± 13.8
434.6 ± 50.6
799.9 ± 56.7
48.0 ± 3.1
108.3 ± 8.4
134.4 ± 13.0
215.0 ± 16.4
29.7± 0.3
31.2 ± 0.2
33.3 ± 0.3
34.5 ± 0.4
29.7 ± 0.6
32.4 ± 0.3
33.4 ± 0.5
35.7 ± 0.4
28.2 ± 1.1
30.9 ± 0.4
32.2 ± 0.3
31.6 ± 0.6
30.1 ± 0.5
31.8 ± 0.3
32.6 ± 0.2
34.1 ± 0.2
30.7 ± 0.2
32.2 ± 0.2
34.1 ± 0.2
34.3 ± 0.2
30.6 ± 0.3
32.9 ± 0.2
34.5 ± 0.3
35.0 ± 0.4
31.8 ± 0.5
33.7 ± 0.6
35.3 ± 0.2
37.8 ± 0.5
28.6 ± 0.5
32.0 ± 0.4
33.0 ± 0.8
33.5 ± 0.8
32.3 ± 0.1
33.7 ± 0.2
34.1 ± 0.1
36.0 ± 0.1
31.7 ± 0.2
33.3 ± 0.2
35.4 ± 0.2
34.4 ± 0.5
33.8 ± 0.2
35.5 ± 0.2
37.2 ± 0.3
37.9 ± 0.2
33.6 ± 0.6
35.9 ± 0.3
37.3 ± 0.1
39.9 ± 0.2
30.9 ± 0.5
33.4 ± 0.2
36.5 ± 0.2
37.6 ± 0.1
36.1 ± 0.3
36.7 ± 0.2
35.9 ± 0.1
37.3 ± 0.1
33.7 ± 0.1
35.7 ± 0.1
36.7 ± 0.1
37.4 ± 0.1
0.3 ± 0.0
0.3 ± 0.0
0.4 ± 0.0
0.5 ± 0.1
0.9 ± 0.1
0.7 ± 0.1
0.8 ± 0.1
0.7 ± 0.1
1.5 ± 0.3
1.1 ± 0.1
1.0 ± 0.1
1.2 ± 0.1
0.3 ± 0.0
0.3 ± 0.0
0.3 ± 0.0
0.3 ± 0.0
0.6 ± 0.0
0.8 ± 0.1
0.8 ± 0.1
0.8 ± 0.1
0.18 – 0.62
0.14 – 0.74
0.16 – 0.86
0.16 – 1.13
0.61 – 1.10
0.39 – 1.03
0.48 – 0.97
0.43 – 1.10
0.73 – 2.91
0.87 – 1.50
0.56 – 1.56
0.88 – 1.82
0.16 – 0.36
0.18 – 0.65
0.14 – 0.49
0.19 – 0.39
0.32 – 1.29
0.26 – 1.48
0.21 – 1.55
0.32 – 1.44
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Pandalus
montagui
10
15
20
25
8
8
5
8
96.6 ± 3.1
105.9 ± 7.8
193.9 ± 4.4
nr*
23.6 ± 0.2
25.2 ± 1.0
26.0 ± 1.0
nr*
24.8 ± 0.3
27.4 ± 0.2
27.6 ± 0.7
nr*
27.1 ± 0.4
30.5 ± 0.1
30.5 ± 0.3
nr*
1.3 ± 0.1
1.2 ± 0.1
1.1 ± 0.1
nr*
0.81 – 1.85
0.87 – 1.46
0.73 – 1.48
nr*
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Table S4 Mean plasticity of oxygen consumption rate (ΔṀO2) and Upper Thermal Limits
2
(ΔUTL), measured as LO, OS and D, calculated at five temperature intervals (10-15, 15-20,
3
20-25, 10-25 and 10-20 °C) as the difference between mean values of ṀO2 and UTL either
4
between consecutive or extreme incubation temperatures (considering either 25 or 20 °C as
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upper extreme temperature). nr* = not recorded
Species
Palaemon
elegans
Palaemon
macrodactylus
Palaemon
serratus
Palaemonetes
varians
Crangon
crangon
Pandalus
montagui
Temperature
(°C)
ΔṀO2 (nmol O2 g
wet weight -1 min-1)
ΔUTL LO (°C)
ΔUTL OS (°C)
ΔUTL D (°C)
10-15
15-20
20-25
10-25
10-20
10-15
15-20
20-25
10-25
10-20
10-15
15-20
20-25
10-25
10-20
10-15
15-20
20-25
10-25
10-20
10-15
15-20
20-25
10-25
10-20
158.5
70.1
380.4
609.0
228.6
15.6
76.6
147.0
239.2
92.2
24.2
77.2
24.0
125.3
101.4
115.2
239.3
365.3
719.8
354.5
60.1
26.3
80.6
167.0
86.4
1.4
2.1
1.4
4.8
3.5
2.7
1.0
2.3
6.0
3.7
2.7
1.4
-0.7
3.4
4.1
1.7
0.8
1.5
4.0
2.5
1.5
1.9
0.2
3.7
3.4
2.3
1.6
0.5
4.4
3.9
1.8
1.6
2.5
5.9
3.4
3.4
1.0
0.4
4.9
4.4
1.5
0.4
1.9
3.8
1.8
1.6
2.1
-1.0
2.7
3.7
1.6
1.8
0.6
4.0
3.4
2.4
1.4
2.6
6.4
3.8
2.6
3.1
1.1
6.8
5.7
0.6
-0.8
1.4
1.2
-0.1
2.1
1.0
0.7
3.7
3.0
10-15
15-20
20-25
10-25
10-20
9.3
88.0
nr*
nr*
97.3
1.6
0.8
nr*
nr*
2.4
3.0
-0.2
nr*
nr*
2.8
3.4
0.0
nr*
nr*
3.4
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Figure S1 Representation of plasticity, here calculated according to the concept of Reaction
2
Norms (Pigliucci, 2001; Schlichting & Pigliucci, 1998). In order to identify both the
3
magnitude and shape of Reaction Norms (sensu Pigliucci, 2001) plasticity is calculated both
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within the whole temperature range examined in this study (P10-25) (red arrow) and within
5
smaller temperature intervals (P10-15, P15-20, P20-25) (blue arrows). In the first case, plasticity is
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calculated as the difference between mean values of either oxygen consumption rate (ṀO2) or
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Upper Thermal Limits (UTL) measured at the two extreme incubation temperatures tested,
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while in the second case it is calculated as the difference between either mean ṀO2 or mean
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UTL measured at two consecutive incubation temperatures.
10
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Figure S2 Bayesian majority-rule consensus phylogram of the combined 12S rDNA, 16S
12
rDNA, 18S rDNA and 28S rDNA and H3 data set. Bootstrap values (1000 replicates) for
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nodes obtained by parsimony analysis are indicated by numbers placed above the nodes. Only
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values equal to or greater than 60 are shown. Types of habitat are indicated by letters placed
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next to species names: B stands for broader/more stable thermal niches, while N indicates
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species characterised by narrower/more stable thermal niches. ‘*’ indicates that, whilst P.
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macrodactylus can occupy broad thermal niches, this species actually occurs in subtidal
18
habitats. However, for the purpose of our work, this species is classed among those occupying
19
broader/more variable thermal niches.
20
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Figure S3 The effect of temperature on UTL, measured as (a) Loss of Orientation (LO) and (b)
22
Death (D), and (c) UTL plasticity in the prawn species investigated in this study: Palaemon
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elegans, Palaemon macrodactylus, Palaemon serratus, Palaemonetes varians, Crangon
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crangon and Pandalus montagui. Histograms represent mean ± SE for UTL after 7 d exposure
25
to one of four incubation temperatures: 10 (yellow), 15 (light orange), 20 (dark orange) and
6
1
25 °C (red). Significantly different mean UTL (p < 0.05) at different incubation temperatures
2
for the same species are indicated by different letters placed above the histograms, while
3
significantly different mean UTL (p < 0.05) at the same incubation temperature among
4
different species are indicated by different numbers placed inside the histograms. Finally,
5
overall significant differences in mean UTL (p < 0.05) among different species are indicated
6
by different numbers preceded by a star placed at the top of the graph above each species.
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Pairwise comparisons were conducted using the Estimated Marginal Means test with Least
8
Significant Difference test correction. Lines in (c) represent Upper Thermal Limits–
9
Temperature (UTL–T): i.e. the patterns through which species increased their UTL in
10
response to increasing incubation temperature according to the best-fit regression model. UTL
11
are measured as LO (grey) and D (black). Raw data, regression equation and relevant
12
statistics for UTL are provided in Figure S5.
13
14
Figure S4 The relationship between temperature and ṀO2 for the prawn species investigated
15
in this study. Circles represent individual prawn ṀO2 measured after 7 d exposure to one of
16
four incubation temperatures: 10 (yellow), 15 (light orange), 20 (dark orange) and 25 °C (red).
17
Full lines represent the best-fit significant regression model; regression equation and relevant
18
statistics are as follows: P. elegans: y = 28.037e0.126x, R² = 0.784, F1,69 = 249.8, p < 0.0001; P.
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macrodactylus: y = 17.024e0.112x, R² = 0.856, F2,23 = 142.7, p < 0.0001; P. serratus: y =
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16.375e0.098x, R² = 0.743, F1,29 = 83.6, p < 0.0001; P. varians: y = 16.242e0.158x, R² = 0.839,
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F1,58 = 301.9, P < 0.0001; C. crangon: y = 19.601e0.095x, R² = 0.672, F1,89 = 182.0, p < 0.0001;
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P. montagui: y = 1.574x2 - 37.499x + 314.214, R² = 0.896, F2,17 = 73.5, p < 0.0001.
23
24
Figure S5 The relationship between temperature and UTL, measured as (a) LO, (b) OS and (c)
25
D, for the prawn species investigated in this study. Circles represent individual prawn UTL
7
1
measured after 7 d exposure to one of four incubation temperatures: 10 (yellow), 15 (light
2
orange), 20 (dark orange) and 25 °C (red). Full and dotted lines represent the best-fit
3
significant and marginally significant regression model respectively; regression equation and
4
relevant statistics are as follows: (a) P. elegans: y = 0.330x + 26.385, R² = 0.739, F1,64 = 180.8,
5
p < 0.0001; P. macrodactylus: y = 0.375x + 26.306, R² = 0.798, F1,24 = 95.0, p < 0.0001; P.
6
serratus: y = 4.095ln(x) + 19.275, R² = 0.436, F1,25 = 19.3, p < 0.0001; P. varians: y = 0.255x
7
+ 27.689, R² = 0.544, F1,53 = 63.2, p < 0.0001; C. crangon: y = 4.285ln(x) + 20.791, R² =
8
0.657, F1,84 = 160.8, p < 0.0001; P. montgui: y = 3.468ln(x) + 15.685, R² = 0.217 F1,13 = 3.6, p
9
= 0.080; (b) P. elegans: y = 4.981ln(x) + 19.286, R² = 0.655, F1,61 = 115.8, p < 0.0001; P.
10
macrodactylus: y = 0.390x + 27.775, R² = 0.905, F1,21 = 199.4, p < 0.0001; P. serratus: y =
11
5.335ln(x) + 16.775, R² = 0.595, F1,23 = 33.8, p < 0.0001; P. varians: y = 0.226x + 30.041, R²
12
= 0.808, F 1,53 = 222.9, p < 0.0001; C. crangon: y = 3.575ln(x) + 23.724, R² = 0.410, F1,77 =
13
53.6, p < 0.0001; P. montgui: y = 2.233x2 - 0.065x + 8.975, R² = 0.814, F2,15 = 32.9, p <
14
0.0001; (c) P. elegans: y = 4.564ln(x) + 23.284, R² = 0.734, F1,68 = 187.7, p < 0.0001; P.
15
macrodactylus: y = 0.406x + 29.580, R² = 0.919, F1,23 = 260.9, p < 0.0001; P. serratus: y =
16
7.730ln(x) + 12.922, R² = 0.920, F1,29 = 335.0, p < 0.0001; P. varians: y = 0.050x + 35.618,
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R² = 0.079, F1,53 = 4.5, p = 0.038; C. crangon: y = 4.100ln(x) + 24.354, R² = 0.903, F1,87 =
18
805.9, p < 0.0001; P. montgui: y = -0.069x2 + 2.415x + 9.860, R² = 0.962, F 1,18= 216.1, p <
19
0.0001.
20
21
Figure S6 The relationship between ṀO2 and UTL, measured as (a) LO and (b) D, for the
22
prawn species investigated in this study. Circles represent individual prawn ṀO2 and UTL
23
measured after 7 d exposure to one of four incubation temperatures: 10 (yellow), 15 (light
24
orange), 20 (dark orange) and 25 °C (red). Full and dotted lines represent the best-fit
25
significant and marginally significant regression models respectively; regression equation and
8
1
relevant statistics are as follows: (a) P. elegans: y = 2.243ln(x) + 19.689, R² = 0.677, F1,64 =
2
134.4, p < 0.0001; P. macrodactylus: y = 3.108ln(x) + 17.963, R² = 0.808, F1,24 = 101.2, p <
3
0.0001; P. serratus: y = 1.792ln(x) + 22.777, R² = 0.294, F1,25 = 10.4, p = 0.003; P. varians: y
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= 1.369ln(x) + 24.478, R² = 0.486, F1,53 = 50.2, p < 0.0001; C. crangon: y = 1.759ln(x) +
5
24.608, R² = 0.383, F1,84 = 52.1, p < 0.0001; P. montgui: y = 2.790ln(x) + 11.619, R² = 0.219,
6
F1,13 = 3.7, p = 0.078; (b) P. elegans: y = 1.836ln(x) + 25.908, R² = 0.640, F1,68 = 121.0, p <
7
0.0001; P. macrodactylus: y = 3.332ln(x) + 20.669, R² = 0.834, F1,23 = 115.6, p < 0.0001; P.
8
serratus: y = 3.491ln(x) + 18.839, R² = 0.641, F1,29 = 51.8, p < 0.0001; P. varians: y =
9
0.306ln(x) + 34.780, R² = 0.093, F1,53 = 5.4, p = 0.023; C. crangon: y = 1.856ln(x) + 27.194,
10
R² = 0.642, F1,87 = 156.1, p < 0.0001; P. montgui: p > 0.05.
9