Table S1. Summary of the nine locations used to measure clone diversity and spatial structure in diploid and tetraploid
Chamerion angustifolium. Lab code and lat/long coordinates correspond to the closest population previously identified and
studied by the Husband lab. Also listed are the grid dimensions (Grid dim.), final sample sizes after ploidy and AFLP analysis
(N Final), and number of diploids (2x) and tetraploids (4x) each location. The populations used for statistical analyses are
bolded. Ten triploids were also identified from location C (108 = 31 + 67 + 10).
Location Lab code
Elev. (m)
Latitude
Longitude
Grid dim. (m)
N Sampled
N Final 2x
4x
C
K-100
1732
N 50°18'38.88"
W 114°36'45.66"
10 x 10
119
108
31
67
H
K-020
2197
N 50°35'38.71"
W 114°58''56.51"
10 x 10
117
99
88
11
M
B-067
1442
N 51°15'23.16"
W 115°52'3.36"
12 x 8
114
80
1
79
P
K-015
1760
N 50°30'35.59"
W 114°49'29.23"
12 x 10
117
87
2
85
S1
K-004
1503
N 51°02'29.46"
W 114°59'8.76"
10 x 10
103
73
1
72
S2
K-005
1531
N 51°03'9.12"
W 114°56'59.58"
8 x 10
121
90
0
90
SM
B-082
2178
N 51°08'34.68"
W 115°34'24.48"
18 x 7
87
71
70
1
WC1
J-047
2045
N 52°13'3.3"
W 117°11'10.02"
10 x 14
109
10
1
9
WC2
J-047
2042
N 52°12'59.64"
W 117°10'53.4"
9x9
117
79
74
5
1004
697
268
419
Overall
Table S2. The number of plant samples, number of AFLP fragments, frequency of fragments, and repeatability of fragments.
Values are given for each fluorescently labeled primer pair used: MseI-CAG with EcoRI-ACG (blue), MseI-CAG with EcoRI-ACA
(green), and MseI-CGA with EcoRI-AAG (yellow).
Primer pair
Blue
Green
Yellow
Overall
Number of plant samples
766
880
948
697
Number of fragments
144
136
153
433
Frequency of fragments
0.0721
0.0452
0.0849
0.0696
Repeatability of fragments
0.9802
0.9763
0.9608
0.9721
Table S3. Summary of locations, sample sizes, and ploidy frequencies for seed sources used to test the effect of ploidy on root bud
production in Chamerion angustifolium in a greenhouse study. Initial sample sizes are the number of plants and maternal seed families
initially germinated and transplanted. The final sample size includes only families and plants that survived to the end of the study and
that were also successfully screened for ploidy.
Initial sample size
Final sample size
Ploidy freq.
Lab code
Elevation (m)
Latitude
Longitude
Families
Plants
Families
Plants
2x
4x
J-077
1768
N 52°48'6.00''
W 118°04'57.36''
32
51
16
26
25
1
K-038
2010
N 50°49'32.04''
W 115°12'3.12''
25
51
21
38
31
7
B-060
2090
N 51°27'15.54''
W 116°08'36.30''
12
20
4
11
0
11
WY-093
2110
N 44°23'57.07''
W 104°57'14.86''
10
25
8
20
0
20
K-003
1365
N 51°03'21.00''
W 115°00'50.16''
16
46
15
39
4
35
97
193
64
134
60
74
Total
Table S4. Clone diversity and extent of clonality in four diploid and five tetraploid populations of Chamerion angustifolium.
Genotype number is the number of distinct AFLP clones (G). The “percentage of genotypes that are clonal” is the proportion of
genotypes that are assigned to more than one ramet. The “percentage of ramets in the largest genotype” is the proportion of ramets in
the genotype with the most ramets.
Diploid populations
Tetraploid populations
Location code
C
H
SM
WC2
C
M
P
S1
S2
Ramet number (N)
31
88
70
74
67
79
85
72
90
Genotype number (G)
10
5
6
3
14
11
8
13
8
Effective genotype number (Ge)
4.23
1.62
3.61
1.25
5.30
1.66
1.50
3.23
3.35
Genotypic richness (G-1) / (N-1)
0.30
0.05
0.07
0.03
0.20
0.13
0.08
0.17
0.08
Ramets per genotype (N/G)
3.10
17.60
11.67
24.67
4.79
7.18
10.63
5.54
11.25
Ramets per effective genotype (N/Ge)
7.32
54.42
19.40
59.29
12.64
47.73
56.70
22.30
26.89
Percentage of genotypes that are clonal
40.0
60.0
100.0
100.0
57.1
36.4
50.0
46.2
75.0
Percentage of ramets in the largest genotype
38.7
77.3
38.6
89.2
32.8
77.2
81.2
52.8
44.4
Clonal diversity
Clone size
Table S5. Moran’s I values of spatial autocorrelation for AFLP genotypes at each of eight distance classes and correlogram slopes for
nine populations of diploid (2x) and tetraploid (4x) Chamerion angustifolium. Moran’s I values that are significantly greater than zero
are shaded in dark grey and values significantly less than zero are shaded light grey. Asterisks indicate significance level (P <0.05*,
<0.01**, <0.001***).
Distance interval (maximum)
Location
Ploidy
1.4
2.8
4.2
5.7
7.1
8.5
9.9
13.0
Slope
C
2x
0.127
0.082
0.016
-0.099
-0.109
-0.157*
-0.047
0.127
-0.038**
H
2x
0.186 *
0.110
0.116*
0.083
-0.047
-0.107
-0.154
-0.350
-0.048*
SM
2x
0.519***
0.409***
0.178***
-0.007
-0.147***
-0.169***
-0.157**
-0.211***
-0.049***
WC2
2x
0.627***
0.123
-0.144
-0.309* 0.047
0.210*
0.173
-0.288
0.004
C
4x
0.083
-0.000
0.032 *
-0.015
-0.043
-0.034
-0.061
-0.109
-0.014*
M
4x
0.034
-0.036
0.015
-0.054
-0.062
-0.055
0.112
0.719**
-0.012
P
4x
0.054
0.020
-0.088
-0.053
0.001
0.087
-0.083
0.034
0.006
S1
4x
-0.030
-0.000
0.006
-0.019
0.029
-0.045
-0.011
-0.090
0.006
S2
4x
0.537***
0.467***
0.209***
-0.050
-0.248***
-0.312***
-0.391***
-0.385***
-0.118***
80
70
Sample count
60
50
40
30
20
10
0
50
60
70
80
90
100
110
120
130
140
FL2 – Area (relative fluorescence)
150
160
170
Fig S1. Distribution of relative fluorescence values for all samples analyzed via flow cytometry. The distributions of diploid (first
peak) and tetraploid (last peak) Chamerion angustifolium are distinct from each other. Diploid and tetraploid values are also distinct
from the distribution of the standard, Solanum lycopersicum, and the nine triploids identified (middle peak). Samples within 10% of
means for 2x, 3x and 4x were automatically classified as such (shaded regions). Samples that deviate by more than 10% were assigned
to the nearest ploidy (illustrated by arrows).
220
Number of pairwise comparisons
165
C
110
H
SM
WC2
55
0
0
5
10
15
20
25
30
35
40
45
50
Number of differences between AFLP genotypes
55
60
65
70
220
Number of pairwise comparisons
165
C
PJ
110
S1
S2
M
55
0
0
5
10
15
20
25
30
35
40
45
50
Number of differences between AFLP genotypes
55
60
65
70
Fig S2. Pairwise distance distribution of AFLP genotypes for the A) four diploid
populations and B) five tetraploid populations. The first major valley (highlighted by red
circles) in each the pairwise distance distribution for each population (excluding the
tetraploid populations M and S1, which did not have a well defined valley) was averaged
across all populations to determine the overall threshold for clone identification (18
differences, indicated by the red lines).
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Fig S3. Spatial distribution of ALFP genotypes and ploidy for individual Chamerion
angustifolium plants sampled along grids with 1 meter intervals at eight locations: A) C Coleman Clear-Cut, B) H - Highwood Pass, C) M - Moose Meadows, D) P - Pickel Jar,
E) S1 - Sibbald Road 1, F) S2 - Sibbald Road 2, G) SM - Sulphur Mountain, and H) WC2
- Wilcox Creek 2. At each location, numbers indicate samples assigned to the same
genotype. Diploids are shown with black squares, triploids with purple squares and
tetraploids with pink squares.
15