Supporting Information for online publication
Supplementary Material and Methods. Taxonomic notes on the identification of the Nasutitermes
species. The following abbreviations are used to indicate termite collections: AMNH = American
Museum of Natural History, New York; MCZ = Museum of Comparative Zoology, Harvard
University, Cambridge; UnB = University of Brasília.
Nasutitermes acajutlae (Holmgren, 1910)
Soldiers of this species are similar to those of N. nigriceps (Haldeman, 1853) and N. macrocephalus
(Silvestri, 1903). Specimens from French Guiana were identified based on the diagnosis presented by
Thorne et al. (1994) and comparison with non-type specimens (UnB).
Nasutitermes acangussu Bandeira & Fontes, 1979
Compared to the original description and to non-type specimens (UnB). Soldiers of this species are
very distinctive and can be recognized easily by their size, shape of head capsule and pilosity.
Nasutitermes callimorphus Mathews, 1977
Compared to the original description and specimens from several localities (UnB), including samples
from the type-locality (topotypes).
Nasutitermes corniger (Motschulsky, 1855)
This is a very common neotropical species and was described many times under different names.
Identification was based primarily on the comparison to specimens from several localities of Central
and South America (UnB), including samples from the type-locality (topotypes). Soldiers of N.
corniger can be distinguished from those of N. ephratae based on their pilosity, especially the short
hairs on the surface of the tergites. It is necessary to examine the specimens under high magnification
and good illumination. Imagoes of these two species are very different.
Nasutitermes coxipoensis (Holmgren, 1910)
Compared to the original description, to Mathews's (1977) description, to images of syntypes
(AMNH) and to specimens from many different localities (UnB). This is a very common species in
South American savannas and can be recognized by the elongate nasus and the distribution of hairs on
thoracic nota and tergites.
Nasutitermes ephratae (Holmgren, 1910)
Compared to the descriptions presented by Holmgren (1910), Emerson (1925), and Mathews (1977)
and to specimens from several localities (UnB). Soldiers of this species are somewhat similar to those
of N. corniger and N. coxipoensis, and can be distinguished based on their pilosity, but need to be
examined under high magnification and good illumination.
Nasutitermes guayanae (Holmgren, 1910)
See comments under N. similis. Compared to specimens from many different localities (UnB).
Nasutitermes intermedius Banks, 1919
Compared to the Banks's (1919) description, to images of soldiers from the type-colony (MCZ and
AMNH) and to specimens form other localities (UnB). The combination of gut morphology and
distribution of hairs on the head capsule are very distinct from any other species of neotropical
Nasutitermes.
Nasutitermes obscurus Snyder & Emerson, 1949
Compared to Holmgren's (1910) description of Eutermes (Eutermes) obscurus, to images of syntype
soldiers (AMNH) and to specimens from several localities (UnB). Soldiers of this species can be
distinguished by their dark color, thick nasus and hairy tergites.
Nasutitermes octopilis Banks, 1918
Compared to the descriptions presented by Banks (1918) and Emerson (1925), to images of syntypes
(AMNH) and to specimens from several localities (UnB). Soldiers of this species have a distinct head
shape.
Nasutitermes similis Emerson, 1935
Compared to Emerson's (1935) description and to specimens from several different localities (UnB).
According to Emerson (1935), soldiers of this species are very similar to those of N. guayanae and
cannot be reliably identified based on morphology. Soldiers of N. guayanae are usually darker
colored, but due to natural variation and to color change due to preservation in alcohol, this character
is unreliable.
Nasutitermes surinamensis (Holmgren, 1910)
Compared to the descriptions presented by Holmgren (1910) and Emerson (1925), to images of
syntypes (AMNH) and to specimens from several different localities (UnB). Soldiers of this species
have a distinct head shape and hairy tergites. Gut morphology is also distinct.
Nasutitermes unduliceps Mathews, 1977
Compared to the original description, specimens from several localities (UnB), and images of a
paratype soldier from the type-colony.
Nasutitermes wheeleri Emerson, 1925
Compared to Emerson's (1925) original description, to images of a paratype soldier (type colony,
AMNH) and to specimens from different localities (UnB). Soldiers of this species have a distinct
pilosity on the head capsule and are easily separated from other neotropical Nasutitermes.
References
Banks N (1918) The termites of Panama and British Guiana. Bulletin of the American Museum of
Natural History 38, 659–667.
Banks N (1919) Antillean Isoptera. Bulletin of the Museum of Comparative Zoology 26, 475-489.
Emerson AE (1925) The termites of Kartabo, Bartica District, British Guiana. Zoologica 6, 291-459.
Emerson AE (1935) Termitophile distribution and quantitative characters as indicators of
physiological speciation in British Guiana termites (Isoptera). Annals of the Entomological
Society of America 28, 369-395.
Holmgren N (1910) Versuch einer Monographie der amerikanische Eutermes - Arten. Jahrbuch der
Hamburgischen Wissenschaftlichen Anstalten 27, 171-325.
Mathews AGA (1977) Studies on termites from the Mato Grosso State, Brazil Academia Brasileira de
Ciências, Rio de Janeiro, Brazil.
Thorne BL, Haverty MI, Collins MS (1994) Taxonomy and biogeography of Nasutitermes acajutlae
and N. nigriceps (Isoptera: Termitidae) in the Caribbean and Central America. Annals of the
Entomological Society of America 87, 762-770.
Supplementary Table 1. Sequences of primers and PCR profiles used
Genes
Primer names
Sequences (5′–3′)
References
COII
Modified A-tLeu
CAGATAAGTGCATTGGATTT
(Miura et al. 2000)
B-tLys
GTTTAAGAGACCAGTACTTG
(Liu & Beckenbach 1992; Simon et al. 1994)
LCO1490F
GGTCAACAAATCATAAAGATATTGG
(Folmer et al. 1994)
HCO2198R
TAAACTTCAGGGTGACCAAAAAATCA
(Folmer et al. 1994)
16S-F1
TTACGCTGTTATCCCTAA
(Kambhampati & Smith 1995)
16S-R4
TCTGGTTTTTCAAGAAATGA
(Miura et al. 2000)
ITS2F
TGTGAACTGCAGGACACAT
(Jenkins et al. 2001)
ITS2Rnas
GCTTAAATTCAGMGGGTAGTC
modified from Jenkins et al. 2001 for this study
Genes
Initial denaturation
Denaturation
Annealing
Extension
Final extension
Cycles
COII
94°C (3min)
94°C (30s)
45°C (1min)
72°C (3min)
72°C (10min)
35
COI
94°C (2min)
94°C (1min)
49.5-57.5°C (1min)
72°C (1min15s)
72°C (7min)
40
16S
94°C (3min)
94°C (30s)
45°C (1min)
72°C (3min)
72°C (10min)
35
ITS2
96°C (5min)
96°C (30s)
50°C (1min)
72°C (1min30s)
72°C (5min)
45
COI
16S
ITS2
References
Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R (1994) DNA primers for amplification of
mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular
Marine Biology and Biotechnology 3, 294-299.
Jenkins TM, Dean RE, Verkerk R, Forschler BT (2001) Phylogenetic analyses of two mitochondrial
genes and one nuclear intron region illuminate European subterranean termite (Isoptera:
Rhinotermitidae) gene flow, taxonomy, and introduction dynamics. Molecular Phylogenetics
and Evolution 20, 286-293.
Kambhampati S, Smith PT (1995) PCR primers for the amplification of four insect mitochondrial gene
fragments. Insect Molecular Biology 4, 233-236.
Liu H, Beckenbach AT (1992) Evolution of the mitochondrial cytochrome oxidase II gene among 10
orders of insects. Molecular Phylogenetics and Evolution 1, 41-52.
Miura T, Roisin Y, Matsumoto T (2000) Molecular phylogeny and biogeography of the nasute termite
genus Nasutitermes (Isoptera: Termitidae) in the Pacific tropics. Molecular Phylogenetics and
Evolution 17, 1-10.
Simon C, Frati F, Beckenbach A, et al. (1994) Evolution, weighting, and phylogenetic utility of
mitochondrial gene sequences and a compilation of conserved polymerase chain reaction
primers. Annals of the Entomological Society of America 87, 651-701.
Supplementary Table 2. Mean corrected p-distances within each PSH (intra-cluster p-distance) for
the COII gene. SE: standard error, N: sample size, n.a.: not available/not calculable
PSH
PSH 1
PSH 2a
PSH 2b
PSH 3
PSH 4
PSH 5
PSH 6
PSH 7
PSH 8
PSH 9
PSH 10
PSH 11
PSH 12
PSH 13
PSH 14
PSH 15
N
8
3
1
11
1
1
10
1
8
3
1
2
6
14
3
6
p
0.0016
0.0010
n.a.
0.0027
n.a.
n.a.
0.0015
n.a.
0.0008
0.0000
n.a.
0.0016
0.0014
0.0004
0.0005
0.0030
SE
0.0007
0.0010
n.a.
0.0010
n.a.
n.a.
0.0006
n.a.
0.0007
0.0000
n.a.
0.0016
0.0008
0.0003
0.0005
0.0010
Supplementary Table 3. Pairwise corrected p-distances between PSHs (inter-cluster p-distance) for
the COII gene (below diagonal) and Standard Errors (above diagonal).
PSH 1
PSH 2a
PSH 2b
PSH 3
PSH 4
PSH 5
PSH 6
PSH 7
PSH 8
PSH 9
PSH 10
PSH 11
PSH 12
PSH 13
PSH 14
PSH 15
PSH 1 PSH 2a
0.008
0.048
0.040 0.013
0.036 0.023
0.066 0.074
0.076 0.075
0.073 0.093
0.074 0.091
0.085 0.083
0.080 0.088
0.068 0.082
0.084 0.090
0.098 0.108
0.096 0.096
0.131 0.135
0.123 0.127
PSH 2b PSH 3
0.007
0.007
0.004
0.006
0.005
0.018
0.070
0.064
0.070
0.066
0.082
0.080
0.082
0.080
0.080
0.079
0.085
0.085
0.080
0.078
0.085
0.082
0.107
0.108
0.102
0.100
0.131
0.131
0.129
0.127
PSH 4
0.010
0.011
0.010
0.010
0.080
0.087
0.087
0.082
0.092
0.086
0.090
0.095
0.091
0.126
0.113
PSH 5
0.012
0.012
0.011
0.011
0.012
0.087
0.089
0.081
0.083
0.083
0.088
0.105
0.081
0.124
0.112
PSH 6
0.010
0.013
0.011
0.011
0.012
0.013
0.024
0.053
0.050
0.047
0.051
0.087
0.084
0.118
0.115
PSH 7
0.011
0.012
0.011
0.011
0.012
0.013
0.006
0.058
0.047
0.050
0.062
0.083
0.089
0.132
0.115
PSH 8
0.012
0.012
0.011
0.011
0.012
0.012
0.009
0.009
0.042
0.048
0.057
0.103
0.083
0.105
0.132
PSH 9
0.012
0.012
0.012
0.012
0.013
0.012
0.008
0.008
0.007
0.032
0.047
0.092
0.084
0.126
0.117
PSH 10
0.010
0.012
0.012
0.011
0.012
0.013
0.008
0.008
0.008
0.007
0.054
0.089
0.079
0.129
0.117
PSH 11
0.012
0.012
0.012
0.011
0.013
0.013
0.009
0.010
0.010
0.009
0.009
0.091
0.092
0.103
0.118
PSH 12
0.013
0.014
0.014
0.014
0.013
0.015
0.012
0.011
0.013
0.012
0.012
0.012
0.089
0.127
0.106
PSH 13
0.012
0.013
0.013
0.013
0.012
0.012
0.012
0.012
0.011
0.012
0.011
0.013
0.013
0.098
0.100
PSH 14
0.016
0.016
0.016
0.016
0.016
0.016
0.016
0.017
0.014
0.016
0.016
0.015
0.016
0.014
0.123
PSH 15
0.015
0.015
0.015
0.015
0.014
0.015
0.014
0.014
0.016
0.015
0.014
0.015
0.014
0.013
0.016
Supplementary Fig. 1. Map of French Guiana, showing the 16 geographic sites of termite sampling.
Figure inset, localization of French Guiana within South America.
Supplementary Fig. 2. Median joining network showing relationships between reconstructed ITS2
haplotypes of Nasutitermes complete dataset (N=75x2). Each PSH is represented with a different
color; pie diagrams showing shared reconstructed alleles between PSHs.