1

The macroevolution of size and complexity in insect male

genitalia

Andrey Rudoy and Ignacio Ribera

Institute of Evolutionary Biology (CSIC-Universitat Pompeu Fabra), Passeig Maritim

de la Barceloneta 37-49, 08003 Barcelona, Spain

Electronic Supplemental Information

Table S1. Specimens used, with locality, collector, voucher, species group according to

Rudoy et al. (in press) and genbank accession numbers. Nf and Nm, number of females

and males measured respectively; Ng, number of male genitalias measured; lf and lm,

length of females and males respectively; stv.lf and stv.lm, standard deviation of lf and

lm (for less than seven specimens the total range is given); lg, length of the male

genitalia; stv.lg, standard deviation of lg.

Table S2. Reconstructed values of the measured variables in all branches of the

phylogeny (see Figure S2 for the location of the branch numbers). The raw values of the

initial and final nodes of each branch are given, together with the amount of phenotypic

change, the absolute phenotypic change, and the change measured in darwins (x10^6).

In pink, composite branch after deleting terminals with missing males. In yellow,

reconstructed values.

2

Figure S1. Aedeagus of L. gerhardti, with A) maximum length (lg); B) perimeter,

outlined by hand and measured in ImageJ; and C) log-log plot of the regression between

the scale of the box and the number of boxes necessary to cover the image, obtained

with the Fractal Dimension Estimator (see Methods). The absolute value of the slope of

the regression line is the fractal dimension of the image (in this case, the outline of the

aedeagus).

A" B" C"

lg#

3

Figure S2. Phylogeny of Limnebius, with the branch numbers used in Table S2.

4.0

L_lusitanus_AI899

L_aguilerai_AI910

L_myrmidon_AI703

L_pilicauda_AR60

L_wewalkai_RA108

Laeliana_sahlb_HI19

L_arenicola_AI466

L_monfortei_AF199

L_zaerensis_AC14

L_Philippines_AR12

L_nakanei_AC30

L_atomus_AI1217

L_sinuatus_AR9

L_millani_AI920

L_hieronymi_AR22

L_furcatus_AI987

L_piceus_RA1120

L_India_AF193

L_oblongus_AI942

L_murentinus_AN53

L_hispanicus_AI1309

L_mucronatus_AI704

L_gracilipes_AI1094

L_levantinus_RA731

L_nitiduloides_AR16

L_Madagascar_AI549

L_corybus_AI1280

L_minoricensis_AR23

L_ibericus_RA88

L_pararabicus_RA761

L_evanescens_AI915

L_mesatlanticus_AI913

L_acupunctus_AI582

L_simplex_AC39

L_schoemanni_RA1000

L_bacchus_PA274

L_China_RA627

L_reuvenortali_AN45

L_extraneus_RA89

L_maurus_AI1317

L_stagnalis_AI824

L_rubropiceus_AN32

L_gerhardti_AI891

L_corfidius_AN33

L_endroedyi_AN78

L_India_AF190

L_cordobanus_AI1024

L_ordunyai_AI854

L_parvulus_AI706

L_irmelae_AI949

L_fretalis_AI1307

L_nitidus_AI377

L_dioscoridus_RA728

L_doderoi_AI1174

L_montanus_AI936

L_alibei_RA90

L_cordobanus_PA275

L_crassipes_RA68

L_crinifer_AR55

L_feuerborni_AR11

L_truncatellus_AI451

L_pollex_RA1019

L_kocheri_RA91

L_spinosus_AN54

L_hilaris_AI919

L_aluta_AI953

L_papposus_RA84

L_ignarus_AI993

L_fallaciosus_RA1121

L_Bhutan_AI1271

L_thery_AI950

L_perparvulus_AI428

1

2

3

4

5

6

7

8

9

10

11

1213

1415

16

17

18

19

20

21

2223

2425

26

27

28

29

3031

3233

34

35

36

37

38

39

4041

42

43

44

45

46

47

48

49

50

51

5253

5455

56

57

58

59

60

61

62

63

64

65

66

67

6869

70

71

72

73

74

75

76

77

78

79

80

81

8283

8485

86

87

88

89

90

91

92

93

94

95

96

97

98

99

100101

102103

104

105

106

107

108

109

110

111

112113

114115

116

117

118

119

120

121

122

123

124

125

126

127

128

129

130

131

132

133

135

136

137

141138

139

140

134

142

4

Figure S3. Kernel density estimates (KDE) of the probability density function of the

PAD measurements and the branch lengths, for all and the terminal branches; and for

the subgenera Bilimneus and Limnebius s.str. KDEs were used for illustrative purposes

only; see Table 4 for the parameters of the comparison between the average and

variance of the different variables.

Den

sity

longitud

Lim.txt - longitud KDEterminals.log - longitud KDE

-10 0 10 20 30 40 500

2.5E-2

5E-2

7.5E-2

0.1

0.125

0.15

0.175

Den

sity

per

Lim.txt - per KDEterminals.log - per KDE

-2 -1 0 1 2 30

0.25

0.5

0.75

1

1.25

1.5

Den

sity

f d

Lim.txt - fd KDEterminals.log - fd KDE

-7.5E-2 -5E-2 -2.5E-2 0 2.5E-2 5E-2 7.5E-2 0.10

5

10

15

20

25

30

35

40

Den

sity

Lf

Lim.txt - Lf KDEterminals.log - Lf KDE

-0.3 -0.2 -0.1 0 1E-1 0.2 0.3 0.4 0.50

1

2

3

4

5

6

7

8

9

10D

ensi

ty

Lm

Lim.txt - Lm KDEterminals.log - Lm KDE

-0.4 -0.3 -0.2 -1E-1 0 1E-1 0.2 0.3 0.4 0.5 0.6 0.70

1

2

3

4

5

6

Den

sity

Lg

Lim.txt - Lg KDEterminals.log - Lg KDE

-0.2 -0.1 0 0.1 0.2 0.3 0.40

2.5

5

7.5

10

12.5

branchlength per fd

lf lm lg

A)Branchlengthsandphenotypicchange,terminalsvsallbranches

Den

sity

longitud

sstr.log - longitud KDEbilim.log - longitud KDE

-5 0 5 10 15 20 25 300

2.5E-2

5E-2

7.5E-2

0.1

0.125

0.15

0.175

0.2

Den

sity

per

sstr.log - per KDEbilim.log - per KDE

-2 -1 0 1 2 30

0.5

1

1.5

2

2.5

3

3.5

Den

sity

f d

sstr.log - fd KDEbilim.log - fd KDE

-7.5E-2 -5E-2 -2.5E-2 0 2.5E-2 5E-2 7.5E-2 0.10

10

20

30

40

50

60

70

Den

sity

Lf

sstr.log - Lf KDEbilim.log - Lf KDE

-0.3 -0.2 -0.1 0 1E-1 0.2 0.3 0.4 0.50

2.5

5

7.5

10

12.5

15

17.5

Den

sity

Lm

sstr.log - Lm KDEbilim.log - Lm KDE

-0.4 -0.3 -0.2 -1E-1 0 1E-1 0.2 0.3 0.4 0.5 0.6 0.70

1

2

3

4

5

6

7

8

9

Den

sity

Lg

sstr.log - Lg KDEbilim.log - Lg KDE

-0.2 -0.1 0 0.1 0.2 0.30

2.5

5

7.5

10

12.5

branchlength per fd

lf lm lg

B)Branchlengthsandphenotypicchange,BilimneusvsLimnebiuss.str.

Den

sity

Dper

terminals_darwin.log - Dper KDELimnebius_darwin.log - Dper KDE

-0.1 0 0.1 0.2 0.3 0.40

2.5

5

7.5

10

12.5

15

17.5

Den

sity

Dfd

terminals_darwin.log - Dfd KDELimnebius_darwin.log - Dfd KDE

-1E-2 0 1E-2 2E-2 3E-2 4E-2 5E-20

25

50

75

100

125

150

Den

sity

DLf

terminals_darwin.log - DLf KDELimnebius_darwin.log - DLf KDE

-5E-2 0 5E-2 0.1 0.15 0.20

5

10

15

20

25

30

35

40

Den

sity

DLm

terminals_darwin.log - DLm KDELimnebius_darwin.log - DLm KDE

-5E-2 0 5E-2 0.1 0.15 0.2 0.25 0.30

5

10

15

20

25

30

35

Den

sity

DLg

terminals_darwin.log - DLg KDELimnebius_darwin.log - DLg KDE

-0.1 0 0.1 0.2 0.3 0.4 0.5 0.60

2.5

5

7.5

10

12.5

15

per fd

lf lm lg

C)darwins,terminalsvsallbranches

Den

sity

Dper

sstr_darwin.log - Dper KDEBil_darwin.log - Dper KDE

-0.1 0 0.1 0.2 0.3 0.40

10

20

30

40

50

60

Den

sity

Dfd

sstr_darwin.log - Dfd KDEBil_darwin.log - Dfd KDE

-1E-2 0 1E-2 2E-2 3E-2 4E-2 5E-20

100

200

300

400

500

Den

sity

DLf

sstr_darwin.log - DLf KDEBil_darwin.log - DLf KDE

-5E-2 0 5E-2 0.1 0.15 0.20

10

20

30

40

50

60

70

80

90

Den

sity

DLm

sstr_darwin.log - DLm KDEBil_darwin.log - DLm KDE

-5E-2 0 5E-2 0.1 0.15 0.2 0.250

10

20

30

40

50

60

Den

sity

DLg

sstr_darwin.log - DLg KDEBil_darwin.log - DLg KDE

-0.1 0 0.1 0.2 0.3 0.4 0.5 0.60

5

10

15

20

25

30

35

per fd

lf lm lg

D)darwins,BilimneusvsLimnebiuss.s.tr.