Electronic supplementary material for: ‘Population isolation rather than ecological variation explains the genetic structure of endangered myrmecophilous butterfly Phengaris (=Maculinea) arion’ by Sielezniew and Rutkowski (JICO454)
Table S1. Characterization of microsatellite polymorphisms of P. arion sampled from 13 sites in Poland. For full names of localities and other details see Tab. 1 in the article. The sample size (n), number of alleles(A), allelic richness (R), heterozygosity observed(HO), heterozygosity expected(HE), P-values for HWE exact test for heterozygote deficiency/excess (HWE),P-values of heterozygote deficiency exact test(HD), fixation index(FIS),*FIS value significant after Benjamini-Hochbergcorrection.
Location / SOS / GUG / PIA / SOW / LUZ / TRU / HORlocus / (n = 16) / (n = 14) / (n = 20) / (n = 26) / (n = 17) / (n = 7) / (n=13)
Macu8 / A / 7.0 / 3.0 / 8.0 / 9.0 / 12.0 / 4.0 / 5.0
R / 5.36 / 2.90 / 6.15 / 5.59 / 7.57 / 4.00 / 4.04
HO / 0.938 / 0.429 / 0.900 / 0.885 / 0.941 / 0.857 / 0.769
HE / 0.758 / 0.449 / 0.824 / 0.769 / 0.849 / 0.622 / 0.666
HWE / ns / ns / ns / ns / ns / ns / ns
HD / ns / ns / ns / ns / ns / ns / ns
FIS / -0.206 / 0.082 / -0.067 / -0.131 / -0.078 / -0.309 / -0.116
Macu9 / A / 11.0 / 5.0 / 15.0 / 14.0 / 16.0 / 5.0 / 4.0
R / 7.27 / 4.51 / 9.38 / 8.22 / 9.18 / 5.00 / 3.53
HO / 0.750 / 0.571 / 0.850 / 0.885 / 0.765 / 0.714 / 0.769
HE / 0.791 / 0.737 / 0.911 / 0.881 / 0.889 / 0.724 / 0.639
HWE / ns / 0.043 / ns / ns / ns / ns / ns
HD / ns / ns / ns / ns / ns / ns / ns
FIS / 0.084 / 0.260 / 0.093 / 0.015 / 0.170 / 0.091 / -0.165
Macu11 / A / 3.0 / 3.0 / 7.0 / 2.0 / 4.0 / 2.0 / 3.0
R / 3.00 / 2.90 / 4.22 / 1.86 / 3.25 / 2.00 / 2.91
HO / 0.688 / 0.357 / 0.500 / 0.154 / 0.412 / 0.143 / 0.308
HE / 0.658 / 0.449 / 0.628 / 0.204 / 0.465 / 0.133 / 0.506
HWE / ns / ns / ns / ns / ns / ns / ns
HD / ns / ns / ns / ns / ns / ns / ns
FIS / -0.012 / 0.240 / 0.228 / 0.265 / 0.145 / 0.000 / 0.425
Macu15 / A / 4.0 / 3.0 / 5.0 / 5.0 / 4.0 / 3.0 / 5.0
R / 3.83 / 2.49 / 3.28 / 4.34 / 3.20 / 3.00 / 3.60
HO / 0.688 / 0.214 / 0.450 / 0.577 / 0.471 / 0.857 / 0.385
HE / 0.619 / 0.390 / 0.511 / 0.666 / 0.431 / 0.541 / 0.479
HWE / ns / ns / ns / ns / ns / ns / ns
HD / ns / ns / ns / ns / ns / ns / ns
FIS / -0.078 / 0.480 / 0.145 / 0.153 / -0.062 / -0.532 / 0.236
Macu16 / A / 2.0 / 4.0 / 4.0 / 5.0 / 4.0 / 2.0 / 3.0
R / 1.98 / 3.84 / 3.29 / 4.01 / 3.43 / 2.00 / 2.80
HO / 0.250 / 0.643 / 0.167 / 0.500 / 0.313 / 0.429 / 0.538
HE / 0.305 / 0.656 / 0.415 / 0.681 / 0.662 / 0.337 / 0.521
HWE / ns / ns / <0.001 / ns / 0.006 / ns / ns
HD / ns / ns / 0.005 / ns / 0.001 / ns / ns
FIS / 0.211 / 0.056 / 0.617* / 0.284 / 0.551* / -0.200 / 0.006
Macu17 / A / 8.0 / 5.0 / 6.0 / 9.0 / 9.0 / 4.0 / 5.0
R / 6.66 / 4.60 / 4.40 / 5.86 / 6.15 / 4.00 / 4.73
HO / 0.625 / 0.786 / 0.600 / 0.808 / 0.529 / 0.714 / 0.769
HE / 0.844 / 0.727 / 0.636 / 0.814 / 0.779 / 0.602 / 0.754
HWE / ns / ns / ns / ns / ns / ns / ns
HD / 0.006 / ns / ns / ns / <0.001 / ns / ns
FIS / 0.289 / -0.044 / 0.082 / 0.028 / 0.347 / -0.111 / 0.020
Mean
/ A / 5.8 / 3.8 / 7.5 / 7.3 / 8.2 / 3.3 / 4.2R / 4.68 / 3.54 / 5.12 / 4.98 / 5.46 / 3.33 / 3.60
Overall / HO / 0.656 / 0.500 / 0.578 / 0.635 / 0.572 / 0.619 / 0.590
HE / 0.662 / 0.568 / 0.654 / 0.669 / 0.679 / 0.493 / 0.594
HWE / ns / ns / 0.001 / ns / 0.01 / ns / ns
HD / ns / ns / 0.008 / ns / <0.001 / ns / ns
FIS / 0.042 / 0.156 / 0.142* / 0.071 / 0.188* / -0.182 / 0.047
Table S1. continued
Location / ORC / SUK / HUT / BAB / KLU / SRO / Totallocus / (n = 18) / (n = 26) / (n = 20) / (n = 16) / (n = 22) / (n = 18) / (n = 233)
Macu8 / A / 8.0 / 10.0 / 11.0 / 7.0 / 8.0 / 8.0 / 26.0
R / 5.93 / 6.38 / 7.80 / 5.61 / 5.37 / 5.13 / -
HO / 0.889 / 0.885 / 0.950 / 0.813 / 0.727 / 0.667 / 0.828
HE / 0.793 / 0.810 / 0.874 / 0.781 / 0.749 / 0.705 / 0.827
HWE / ns / ns / ns / ns / ns / ns / ns
HD / ns / ns / ns / ns / ns / ns / ns
FIS / -0.092 / -0.073 / -0.062 / -0.008 / 0.052 / 0.083 / 0.001
Macu9 / A / 13.0 / 17.0 / 15.0 / 11.0 / 10.0 / 14.0 / 29.0
R / 7.85 / 9.88 / 9.15 / 8.10 / 6.41 / 9.31 / -
HO / 0.889 / 0.962 / 0.950 / 0.813 / 0.955 / 1.000 / 0.858
HE / 0.846 / 0.926 / 0.905 / 0.877 / 0.824 / 0.907 / 0.931
HWE / ns / ns / ns / 0.02 / ns / ns / < 0.001
HD / ns / ns / ns / ns / ns / ns / < 0.001
FIS / -0.023 / -0.019 / -0.024 / 0.106 / -0.135 / -0.074 / 0.080*
Macu11 / A / 3.0 / 5.0 / 7.0 / 4.0 / 4.0 / 6.0 / 13.0
R / 2.27 / 2.91 / 4.45 / 2.57 / 3.45 / 3.80 / -
HO / 0.167 / 0.346 / 0.500 / 0.250 / 0.591 / 0.444 / 0.382
HE / 0.245 / 0.358 / 0.651 / 0.229 / 0.563 / 0.593 / 0.487
HWE / ns / ns / ns / ns / ns / 0.035 / < 0.001
HD / ns / ns / ns / ns / ns / ns / ns
FIS / 0.346 / 0.053 / 0.256 / -0.062 / -0.026 / 0.277 / 0.219*
Macu15 / A / 5.0 / 8.0 / 8.0 / 6.0 / 5.0 / 7.0 / 14.0
R / 4.04 / 5.52 / 5.78 / 5.13 / 4.21 / 4.57 / -
HO / 0.556 / 0.769 / 0.900 / 0.938 / 0.818 / 0.556 / 0.635
HE / 0.622 / 0.786 / 0.800 / 0.740 / 0.737 / 0.630 / 0.707
HWE / 0.003 / ns / ns / ns / 0.021 / ns / 0.013
HD / ns / ns / ns / ns / ns / ns / < 0.001
FIS / 0.135 / 0.040 / -0.100 / -0.236 / -0.088 / 0.146 / 0.104*
Macu16 / A / 3.0 / 3.0 / 5.0 / 3.0 / 3.0 / 4.0 / 8.0
R / 2.92 / 2.61 / 3.51 / 2.44 / 1.86 / 3.62 / -
HO / 0.133 / 0.308 / 0.300 / 0.188 / 0.045 / 0.167 / 0.295
HE / 0.498 / 0.329 / 0.419 / 0.518 / 0.129 / 0.662 / 0.558
HWE / <0.001 / ns / ns / 0.002 / 0.023 / <0.001 / < 0.001
HD / < 0.001 / ns / ns / 0.003 / 0.02 / <0.001 / < 0.001
FIS / 0.748* / 0.085 / 0.307 / 0.656 / 0.661 / 0.761* / 0.473*
Macu17 / A / 9.0 / 12.0 / 12.0 / 9.0 / 9.0 / 10.0 / 25.0
R / 5.70 / 7.69 / 7.31 / 6.97 / 6.17 / 6.87 / -
HO / 0.556 / 0.885 / 0.850 / 0.813 / 0.818 / 0.889 / 0.751
HE / 0.725 / 0.878 / 0.840 / 0.836 / 0.818 / 0.824 / 0.878
HWE / 0.003 / ns / ns / ns / ns / ns / 0.001
HD / 0.015 / ns / ns / ns / ns / ns / < 0.001
FIS / 0.261 / 0.012 / 0.014 / 0.060 / 0.023 / -0.050 / 0.147*
Mean
/ A / 6.8 / 9.2 / 9.7 / 6.7 / 6.5 / 8.2 / 19.17R / 4.78 / 5.83 / 6.33 / 5.14 / 4.58 / 5.55 / -
Overall / HO / 0.531 / 0.692 / 0.742 / 0.635 / 0.659 / 0.620 / 0.625
HE / 0.622 / 0.681 / 0.748 / 0.663 / 0.637 / 0.720 / 0.731
HWE / <0.001 / ns / ns / 0.014 / ns / ns / < 0.001
HD / 0.008 / ns / ns / ns / ns / 0.009 / < 0.001
FIS / 0.174* / 0.003 / 0.034 / 0.074 / -0.012 / 0.166* / 0.148*
Table S2. Results ofthe MicroChecker analysis for P. arion sampled from 13 sites in Poland. Frequency of null alleles was estimated using the Oosterhout method (van Oosterhout et al., 2004). * - P 0.05
Locality / Locus with‘null alleles’ / Frequency of
‘null alleles’
SOS / – / –
GUG / – / –
PIA / Macu16 / 0.269
SOW / Macu16 / 0.128*
LUZ / Macu16 / 0.254*
Macu17 / 0.150*
TRU / – / –
HOR / – / –
ORC / Macu16 / 0.305
SUK / – / –
HUT / – / –
BAB / Macu16 / 0.288
KLU / Macu16 / 0.167
SRO / Macu16 / 0.345*
Table S3. Pairwise comparison of genetic variability in the sampled locations. Significance of differences in mean number of alleles (below the diagonal) and mean allelic richness (above the diagonal) were tested using the Wilcoxon test. (+)*/(-)* indicated that a locality in a row has significantly higher/lower mean value than a locality in a column (p < 0.05); ns - not significant.
Locality / Sos / GuG / Pia / Sow / Luz / Tru / Hor / Orc / Suk / Hut / Bab / Klu / SROSOS / (+)* / ns / ns / ns / (+)* / (+)* / ns / ns / (-)* / ns / ns / ns
GUG / ns / ns / ns / (-)* / ns / ns / ns / (-)* / (-)* / ns / (-)* / (-)*
PIA / ns / (+)* / ns / ns / (+)* / ns / ns / ns / ns / ns / ns / ns
SOW / ns / ns / ns / ns / ns / ns / ns / (-)* / (-)* / ns / ns / ns
LUZ / ns / (+)* / ns / ns / (+)* / ns / ns / ns / ns / ns / ns / ns
TRU / (-)* / ns / (-)* / ns / (-)* / ns / (-)* / (-)* / (-)* / (-)* / (-)* / (-)*
HOR / ns / ns / ns / ns / ns / ns / ns / ns / (-)* / ns / (-)* / (-)*
ORC / ns / ns / ns / ns / ns / (+)* / ns / (-)* / (-)* / ns / ns / ns
SUK / (+)* / (+)* / ns / (+)* / ns / (+)* / (+)* / (+)* / ns / (+)* / ns / ns
HUT / (+)* / (+)* / ns / (+)* / ns / (+)* / (+)* / (+)* / ns / (+)* / (+)* / ns
BAB / ns / (+)* / ns / ns / ns / (+)* / (+)* / ns / (-)* / (-)* / ns / ns
KLU / ns / (+)* / ns / ns / ns / (+)* / ns / ns / (-)* / (-)* / ns / ns
SRO / (+)* / (+)* / ns / ns / ns / (+)* / (+)* / ns / ns / (-)* / (+)* / ns
Table S4. Genetic differentiation (pairwise FST, below the diagonal) and Nei standard genetic distance (pairwise DS, above the diagonal) among 13 populations of P. arion. All FST values are significant except one shown in bold (B-H correction).
Locality / Sos / Gug / Pia / Sow / Luz / Tru / Hor / Orc / Suk / Hut / Bab / Klu / SROSOS / 0.347 / 0.257 / 0.256 / 0.250 / 0.371 / 0.187 / 0.284 / 0.352 / 0.259 / 0.448 / 0.462 / 0.415
GUG / 0.117 / 0.352 / 0.301 / 0.277 / 0.343 / 0.350 / 0.251 / 0.357 / 0.405 / 0.368 / 0.743 / 0.459
PIA / 0.054 / 0.124 / 0.215 / 0.101 / 0.205 / 0.287 / 0.126 / 0.247 / 0.241 / 0.283 / 0.349 / 0.381
SOW / 0.067 / 0.118 / 0.061 / 0.100 / 0.293 / 0.251 / 0.146 / 0.130 / 0.239 / 0.184 / 0.242 / 0.257
LUZ / 0.054 / 0.101 / 0.011 / 0.019 / 0.231 / 0.196 / 0.137 / 0.224 / 0.263 / 0.222 / 0.293 / 0.359
TRU / 0.114 / 0.151 / 0.063 / 0.106 / 0.074 / 0.372 / 0.125 / 0.243 / 0.399 / 0.238 / 0.446 / 0.453
HOR / 0.048 / 0.142 / 0.086 / 0.084 / 0.055 / 0.145 / 0.314 / 0.379 / 0.310 / 0.387 / 0.564 / 0.435
ORC / 0.074 / 0.100 / 0.028 / 0.043 / 0.031 / 0.031 / 0.106 / 0.165 / 0.258 / 0.151 / 0.299 / 0.368
SUK / 0.070 / 0.120 / 0.055 / 0.032 / 0.051 / 0.077 / 0.106 / 0.043 / 0.194 / 0.207 / 0.240 / 0.220
HUT / 0.038 / 0.122 / 0.045 / 0.059 / 0.052 / 0.110 / 0.080 / 0.063 / 0.028 / 0.344 / 0.363 / 0.268
BAB / 0.099 / 0.131 / 0.070 / 0.050 / 0.053 / 0.079 / 0.117 / 0.036 / 0.044 / 0.066 / 0.344 / 0.370
KLU / 0.093 / 0.201 / 0.081 / 0.066 / 0.070 / 0.136 / 0.147 / 0.082 / 0.049 / 0.061 / 0.080 / 0.357
SRO / 0.082 / 0.146 / 0.086 / 0.068 / 0.083 / 0.135 / 0.119 / 0.098 / 0.042 / 0.042 / 0.084 / 0.074
Table S5. Analysis of molecular variance for different grouping patterns (see Tab. 1). Variation among groups (Va), variation among populations within group (Vb), variation within populations (Vc).
Level of analysis (number of groups/populations) / Component of variance / % of variance / P-valueGeographical distribution 1: micro-scale (7) / Va / 0.08 / 0.4232
Vb / 7.36 / ≤ 0.0001
Vc / 92.56 / ≤ 0.0001
Geographical distribution 2: macro-scale (2) / Va / 1.93 / 0.0009
Vb / 6.32 / ≤ 0.0001
Vc / 91.75 / ≤ 0.0001
Host plants (2) / Va / 1.15 / 0.0635
Vb / 6.92 / ≤ 0.0001
Vc / 91.94 / ≤ 0.0001
Host ants (3) / Va / 2.86 / 0.0450
Vb / 5.89 / ≤ 0.0001
Vc / 91.24 / ≤ 0.0001
Table S6. Proportion of membership of each pre-defined population of P. arion in each of the four clusters. The TRU population was excluded because of too small sample size. The highest proportion in each location is shown in bold.
Locality / N / Inferred ClustersI / II / III / IV
SOS / 16 / 0.079 / 0.140 / 0.602 / 0.179
GUG / 14 / 0.027 / 0.900 / 0.037 / 0.036
PIA / 20 / 0.131 / 0.251 / 0.427 / 0.191
SOW / 26 / 0.313 / 0.137 / 0.347 / 0.203
LUZ / 17 / 0.160 / 0.301 / 0.296 / 0.243
HOR / 13 / 0.043 / 0.073 / 0.782 / 0.101
ORC / 18 / 0.182 / 0.362 / 0.186 / 0.270
SUK / 26 / 0.227 / 0.203 / 0.191 / 0.379
HUT / 20 / 0.104 / 0.131 / 0.223 / 0.542
BAB / 16 / 0.327 / 0.148 / 0.140 / 0.384
KLU / 22 / 0.777 / 0.049 / 0.081 / 0.094
SRO / 18 / 0.398 / 0.102 / 0.188 / 0.312
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