Systematic Entomology (2001) 26, 241±254

Phylogeny of the water strider genus Gerris Fabricius

(Heteroptera: Gerridae) based on COI mtDNA, EF-1a
nuclear DNA and morphology
J A K O B D A M G A A R D and F E L I X A . H . S P E R L I N G
Zoological Museum and Zoological Institute, University of Copenhagen, Copenhagen, Denmark and Department of
Environmental Science, Policy and Management, University of California, Berkeley, California, U.S.A.

Abstract. Phylogenetic relationships between water striders (Heteroptera: Gerridae)

of genus Gerris Fabricius were examined using molecular and morphological
characters. The molecular dataset was 820 bp DNA from the 3¢ half of the
mitochondrial gene encoding cytochrome oxidase subunit I and 515 bp DNA from
the nuclear gene encoding elongation factor 1 alpha. The morphological dataset was
a slightly modi®ed version of a previously published dataset. Representatives from
all eight recognized species groups of Gerris, as well as six species from three
related genera, including Gigantometra gigas, Limnoporus esakii, L. rufoscutellatus,
Aquarius najas, A. conformis and A. paludum, were included. Unweighted parsimony
analyses of the COI sequences gave a topology with strong support for only those
nodes that were already recognized as closely related based on morphological
characters. Similar analyses of EF-1a gave a cladogram with a topology quite
different from that based on morphology and COI. Unweighted parsimony analyses
of the `total evidence' dataset largely supports the traditional view of Gerris
phylogeny. Finally, the implications of the reconstructed phylogeny in relation to
biogeography and ecological phylogenetics of Gerris is discussed.

Introduction company with Limnoporus species. Unlike Aquarius, Gerris is

Water striders are familiar inhabitants of the surface ®lm of
different types of aquatic habitats all over the world. These
bugs are conspicuously adapted for life on the water surface,
especially with respect to locomotion, feeding, reproductive
behaviour and life history (Andersen, 1982; Spence, 1989;
Spence & Andersen, 1994). The pond skater genus Gerris
Fabricius includes forty-two species with a predominantly
Holarctic distribution. A few species extend into the
Afrotropical Region, and a single species, G. ®rmus Drake &
Hottes, is recorded from Mexico (Andersen, 1993a). Fossils in
Baltic amber (Eocene/Oligocene) have been assigned to
Gerris, giving a minimum age of 40±35 my for the genus
(Andersen, 1998). Gerris can be distinguished from the related
genera Limnoporus StaÊl and Aquarius Schellenberg by the
relatively short ®rst antennal segment, nonspinous connexival
corners and details in the male phallus (Andersen, 1993a).
Species of Gerris are found in lakes and ponds, often in
Correspondence: Jakob Damgaard, Zoological Museum and
Zoological Institute, University of Copenhagen, Universitetsparken
15, DK-2100 Copenhagen, Denmark. E-mail: jdamgaard@zi.ku.dk
generally found to inhabit more temporary habitats, and no
Gerris species are exclusively con®ned to lotic waters.
Gerris is one of the best studied genera of water striders in
terms of wing polymorphism, habitat selection, mating
systems, etc. (Spence, 1989; Andersen, 1993b, 1997; Spence
& Andersen, 1994). The striking pattern of permanent and
seasonal wing polymorphism found in Gerris has intrigued
natural historians for decades, and in its integration with
systematics we are starting to understand the evolution of such
traits, even though the environmental and genetic mechanisms
behind them are still poorly understood.
Andersen (1993a) reviewed the character complexes in
genitalia and metasternal scent apparatus of particular
phylogenetic and taxonomic importance, and presented the
®rst attempt to classify Gerris and its subgenera into
monophyletic species-groups. He demonstrated that subgenus
Gerriselloides (Hungerford & Matsuda), as de®ned by
Andersen (1975) and Kanyukova (1982), was a paraphyletic
assemblage and restricted the subgenus to the Palearctic G.
lateralis group. He elevated the eastern Palearctic G.
gracilicornis group to a new subgenus, Macrogerris, and
grouped the remaining species in presumed monophyletic

# 2001 Blackwell Science Ltd 241

242 J. Damgaard and F. A. H. Sperling

species groups in subgenus Gerris s. str. Finally, Andersen
(1993a) discussed the biogeographical and ecological implica-
tions of the reconstructed phylogeny of Gerris.
Water strider phylogenies based on morphological datasets
have repeatedly faced challenges from phylogenetic recon-
structions based on DNA sequences. Examples are the
Holarctic genus Limnoporus (Sperling et al., 1997), the
Pantropical marine Halobates Eschscholtz (Damgaard et al.,
2000a) and the cosmopolitan genus Aquarius (Damgaard et al.,
2000b). Although these studies have not greatly changed the
traditional view of water strider phylogenies, the inclusion of
new molecular data has provided a way to test the established
morphologically based phylogenies and have led to a better
understanding of phylogenetic relationships among water
striders. These new analyses have questioned the taxonomic
validity of several closely related species pairs, as well as
deeper relationships between more distantly related species,
genera and higher level taxa. Furthermore, the use of
molecular markers to evaluate well established morphological
phylogenies adds new insight into the limitations of different
kinds of data when analysed separately or in combination. The
focus of this study was to test the morphologically based
phylogeny of Gerris with new DNA sequence data, and to
review and re-evaluate the biogeographical implications and
evolution of ecological and behavioural traits in the context of
a well corroborated phylogeny.
Material and methods
DNA sequences and protocols
DNA was sequenced from twenty-two species of Gerris
selected to represent all subgenera and species-groups
recognized by Andersen (1993a). In addition, DNA was
sequenced from species belonging to closely related genera,
including Aquarius conformis Drake & Hottes, A. najas (De
Geer), A. paludum (Fabricius), Limnoporus esakii (Miyamoto),
L. rufoscutellatus (Latreille) and Gigantometra gigas (China).
The last species has been proposed as the closest relative of the
three above-mentioned genera (Andersen, 1993a, 1995). The
target sequences were 830 bp from the 3¢ half of the
mitochondrial gene encoding cytochrome oxidase subunit I
(COI), corresponding to position 2184±3013 in Drosophila
yakuba (Genbank accession no. NP006902) and 523 bp of the
nuclear gene encoding elongation factor-1 alpha (EF-1a),
corresponding to position 2413±2935 in D. melanogaster
(Genbank accession no. X06870). Some water strider DNA
sequences were available from studies by Sperling et al. (1997)
(Genbank accession nos U83337±U83345) and Damgaard
et al. (2000b) (Genbank accession nos AF200255±AF200737).
See Table 1 for collection data and accession numbers.
For most species, DNA was extracted from alcohol-
preserved adults, but for a few species live specimens were

Table 1. Species, geographic localities and GenBank accession numbers.

Subgenus/species-group Species Locality Collector COI EF-1a

Macrogerris Gerris gracilicornis (Horvath) Japan, Kochi T. Harada AF251100 AF251080

Gerriselloides G. lateralis Schummel Denmark, Grib Skov J. Damgaard AF251102 AF251082
G. asper (Fieber) Switzerland, Berne J.R. Spence AF251101 AF251081
G. gillettei-group G. sphagnetorum Gaunitz Sweden, BjaÈdjesjoÈ J. Damgaard AF251103 AF251083
G. incognitus Drake & Hottes USA, New Mexico G. Arnqvist AF251104 AF251084
G. pingreensis Drake & Hottes Canada, Alberta J.R. Spence U83345 AF200275
G. gillettei Lethierry & Severin USA, New Mexico G. Arnqvist AF251105 AF251085
G. argenticollis-group G. argenticollis Parshley USA, Virginia J.T. Polhemus AF251111 AF251092
G. nepalensis-group G. nepalensis Distant Japan, Kochi T. Harada AF251106 AF251086
G. odontogaster-group G. argentatus Schummel Denmark, Gl. Holte J.R. Spence AF251107 AF251087
G. buenoi Kirkaldy Canada, Ottawa F.A.H. Sperling U83343 AF251089
G. babai Miyamoto China, Tianjin J.R. Spence AF251108 AF251090
G. odontogaster (Zetterstedt) Denmark, Gl. Holte J.R. Spence AF251109 AF251088
`G. thoracicus-group' G. thoracicus Schummel Denmark, TaÊstrup J. Damgaard AF251114 AF251093
G. costae poissoni Wagner & Zimmermann Spain, Pyrenees V. Michelsen AF251113 AF251094
G. swakopensis-group G. swakopensis StaÊl Kenya, Masai Mara J.R. Spence AF251112 AF251097
G. lacustris-group G. lacustris L. Denmark, Gl. Holte J.R. Spence AF200735 AF200278
G. maculatus Tamanini Tunesia, Tamara ZMUC expedition AF251116 AF251099
G. gibbifer Schummel Denmark, Bornholm J. Damgaard AF251117 AF251098
`G. marginatus-group' G. latiabdominis Miyamoto Japan, Sakurai T. Harada AF251110 AF251091
G. comatus Drake & Hottes Canada, Ottawa F.A.H. Sperling U83344 AF251096
G. alacris Hussey U.S.A., Pennsylvania J.R. Spence AF251115 AF251095
Outgroup taxa Gigantometra gigas (China) Viet-Nam Lab culture D. Currie AF200245 AF200280
Limnoporus esakii (Miyamoto) Japan, Honshu J.R. Spence U83341 AF200263
L. rufoscutellatus (Latreille) Finland, Hanko J.R. Spence U83337 AF200268
Aquarius najas (De Geer) Denmark, Lellinge A Ê N.M. Andersen AF200736 AF200262
A. conformis (Uhler) U.S.A., Kentucky J. Krupa AF200255 AF200272
A. paludum (Fabricius) Denmark, Agersù F.A.H. Sperling AF200737 AF200266

# 2001 Blackwell Science Ltd, Systematic Entomology, 26, 241±254


killed in a freezer at ± 70 °C and DNA was extracted later.
Heads, abdomens and legs were stored in 70% alcohol as
voucher specimens, and are deposited in the Zoological
Museum, University of Copenhagen.
A phenol/chloroform extraction method was used for Gerris
alacris, G. argentatus, G. asper, G. babai, G. gibbifer, G.
odontogaster and G. swakopensis. Later, the CTAB extraction
protocol (Doyle & Doyle, 1987) was used for Gerris lateralis
and G. sphagnetorum. Because this method only gave 30 mL
DNA in solution, template DNA for G. sphagnetorum was
exhausted and DNA had to be extracted from another
specimen, although from the same collection. For all
remaining species, the QiaAmp tissue kit protocol (QIAGEN
Inc., Santa Clara, California) was used, which included at least
2 h digestion of tissue with Proteinase K and gave a volume of
300 mL DNA in solution.
PCR ampli®cations for both gene regions were carried out in
a thermal cycler in 51 mL of a cocktail containing 2 mL template,
5 mL of each primer (5 mM), 14 mL ddH2O, 20 mL dNTP (GATC
0.5 mM each) and 5 mL 103 Promega PCR-reaction buffer
(15 mM MgCl2). After a `hot start' with 2 min of denaturation at
94 °C, the reaction was paused at 72 °C and 0.2 mL Taq-
polymerase (5 U/mL) was added. Ampli®cation parameters for
each of the following 35 cycles were as follows: 94 °C for 1 min
(denaturation), 45 °C for 1 min (annealing) and 72 °C for
1.5 min (extension). For EF-1a the annealing temperature was
raised to 50±55 °C in cases where there were multiple bands.
The target segment of COI was delimited by the primers C1-
J-2183 to TL2-N-3014 (reproduced from Simon et al., 1994).
Because the segments were too long to be ampli®ed and sequen-
ced in one step, two internal primers were used for ampli®cation
with the end-primers, namely C1-N-2609 (from Damgaard
et al., 2000a) to work with C1-J-2183, and a new primer, C1-J-
2456 (5¢TTAGCAAATTCTTC AATTGA 3¢), to work with
Tl2-N-3014. These two sequences had an overlap of 153 bp.
To amplify and sequence EF-1a, the primers M2412 and
rcM52.6 from Damgaard et al. (2000b) were used. The PCR
product was electrophoresed on a 2% NuSieve gel, stained with
ethidium bromide and sized against a fX174/HaeIII
(Boehringer Mannheim, Mannheim, Germany) DNA ladder
under UV light. PCR products were cleaned with a QIAquick
PCR Puri®cation Kit (QIAGEN Inc.). Cycle sequencing was
carried out using a Perkin Elmer/ABI Dye Terminator Cycle
Sequencing Kit and run on a thermocycler using the pro®les
recommended by the manufacturers. Cycle sequencing products
were cleaned using Centrisep columns or ethanol precipitation
and sequenced using a Perkin Elmer ABI377 Automated
Sequencer (Applied Biosystems Inc., Foster City, California).
DNA sequence for each species was con®rmed with both sense
and anti-sense strands. Because both gene segments are protein
coding and relatively conserved in amino acid sequences,
alignment was unproblematically performed in the program
Sequencher (Gene Codes Corporation, Ann Arbor, Michigan).
Morphological characters
Characters used in the morphological analyses were those
used by Andersen (1993a) for twenty-two species of Gerris,
# 2001 Blackwell Science Ltd, Systematic Entomology, 26, 241±254
Phylogeny of Gerris 243
three species of Aquarius, Limnoporus rufoscutellatus and
Gigantometra gigas. The data matrix is shown in
Table 2. Andersen (1993a) should be consulted for de®nitions
of the characters and their states. Andersen used Limnoporus
canaliculatus (Say) among the outgroup taxa because this
species could be scored for characters only found in wingless
morphs. Because L. esakii is found to be the sister species to all
other species of Limnoporus, this species was used as outgroup
instead of L. canaliculatus. The two species are scored
identically, except for character 29, which is scored as
character 19 in Andersen & Spence (1992), and characters
related to the wingless morph of L. canaliculatus. Some
characters (20, 25, 30, 32, 42, 48 and 63) are conditionally
de®ned and can only be scored if certain structures are present.
These characters are scored as inapplicable (denoted by a
question mark). All multistate characters were treated as non-
additive (states unordered).
Phylogenetic analyses
Phylogenetic reconstructions were obtained by both max-
imum parsimony and maximum likelihood. Unweighted
parsimony analyses of various datasets were performed using
PAUP* 4.0 b2 (Swofford, 1998) in combination with MacClade
3.05 (Maddison & Maddison, 1992). As the number of taxa
and the size of the data matrix often precluded more thorough
searches, heuristic searches were carried out with twenty
random-taxon-addition replicates. In case of multiple equally
parsimonious cladograms, a successive weighting procedure
(Farris, 1969) was applied to reduce the number of cladograms.
The resulting cladograms were only considered if they were a
subset of the most parsimonious cladograms.
Maximum likelihood analyses were conducted in PAUP*.
The 50% parsimony bootstrap search was used as the starting
point for NNI branch swapping under likelihood. The
Hasegawa±Kishino±Yano (Hasegawa et al., 1985) model of
sequence evolution was implemented using observed nucleo-
tide frequencies, two substitution types: TI/TV (transition/
transversion) ratio initially estimated by MacClade from the
50% bootstrap cladogram, and estimation of the rate hetero-
geneity, a, according to a G distribution. When a was
estimated, this value was used for estimation of a new TI/TV
ratio. At the completion of this search, the estimated a and TI/
TV values were used for a heuristic search using NNI branch
swapping to ®nd the cladogram with the highest ln-likelihood
(Swofford et al., 1996).
To determine whether signi®cant incongruence exists
between the nucleotide datasets and the morphological dataset,
Incongruence Length Difference (ILD) tests (Farris et al.,
1995) were conducted by excluding all invariant sites and
using the partition homogeneity test in PAUP* with 100
iterations.
Clade stability was estimated using two different para-
meters: bootstrap and branch support (a.k.a. Bremer support or
decay index; Bremer, 1994). Bootstrap values were generated
in PAUP* from 500 replicates, each with ten random-addition
heuristic searches. Branch support values were obtained in
244 J. Damgaard and F. A. H. Sperling

Table 2. Data matrix of sixty-three morphological characters scored for twenty-two species of Gerris, three species of Aquarius, two species of
Limnoporus and Gigantometra gigas. De®nitions of characters and character states taken from Andersen (1993a). A questionmark (?) denotes
inapplicable or missing character scores.

1 1111111112 2222222223 3333333334 4444444445 5555555556 666

1234567890 1234567890 1234567890 1234567890 1234567890 1234567890 123

G. gigas 1111111111 112121112? 1111131221 1111111111 1?11111?11 1111111112 111

L. esakii 1112221112 1122122112 111112222? 1111111211 1?11111?11 1112121112 21?
L. rufoscutellatus 1112221112 1122122112 112112222? 1111111211 1?12111?12 1112111112 21?
A. najas 2121212312 2122211111 1222?21211 1111111111 1?11111?12 1111121112 111
A. paludum 2121212312 1122211111 1222?22211 1111111211 1?12122221 1121111112 113
A. conformis 2121212312 1122211211 1222?3122? 1?11111211 1?11112231 1121111112 11?
G. lateralis 2211222112 122211112? 1221111221 1121111111 2121122111 1211111122 112
G. asper 2211222112 122211112? 1221111221 1121111111 2121122111 1211111122 112
G. gracilicornis 2211222112 112221112? 2221121222 1121212111 2222122111 1111212222 113
G. sphagnetorum 2211212212 1122111111 2221111221 1121121111 2121122111 1111212222 122
G. incognitus 2211212222 1122111111 2221111221 1122121111 2121122111 1211212222 122
G. pingreensis 2211212222 1122111111 2221111221 1122121111 2121122111 1211212222 122
G. gillettei 2211212222 1122111111 2221111221 1122121111 2121122111 1211212222 122
G. argenticollis 2211212212 112221312? 2221221222 2221221111 2121222312 2211312222 113
G. nepalensis 2211212312 112211112? 2222?21221 1222121111 2121122112 2111312222 122
G. latioabdominis 2211212222 112221312? 2221121222 2221221111 2121222312 1111312222 123
G. argentatus 2211212212 112221312? 2221221222 2221321111 2221222312 3211312222 123
G. babai 2211212212 112221312? 2221221222 2221321111 2221222312 3211312222 123
G. buenoi 2211212212 112221312? 2221221222 2221321111 2221222312 3211312222 123
G. thoracicus 2211222212 112221212? 2221111222 2221221111 2121222312 1111312222 123
G. odontogaster 2211212212 112221312? 2221221222 2221321111 2221222312 3211312222 123
G. alacris 2211212312 112231312? 2221221222 2221221112 2121222312 1111312222 123
G. comatus 2211212322 112221312? 2221121222 2221221112 2121222312 1111312222 123
G. swakopensis 2211212212 112221312? 2221121212 2222322123 2121222312 3211312222 123
G. costae poissoni 2211222212 112231212? 222111122? 2221221111 2121222312 1111312222 12?
G. lacustris 2211212213 112221312? 2221121222 2221121111 2121222312 1111312222 113
G. gibbifer 2211212313 112231312? 2221121222 2221121111 2121222312 1111312222 113
G. maculatus 2211212313 112231312? 2221121222 2221121111 2121222312 1111312222 113

PAUP* by using the `converse constraints' approach to obtain
branch support for the most stable clades (Bremer, 1994). In
order to assess the degree of support provided by each dataset
when analysed together, the partitioned branch support (a.k.a.
partitioned Bremer support; Baker & DeSalle, 1997; Baker
et al., 1998) was calculated for all datasets with reference to
the combined cladogram.
Results
Comparison of the two genes
A 820 bp segment of COI for twenty-two species of Gerris
and Limnoporus and a 780 bp segment for Gigantometra gigas
and species of Aquarius were sequenced. A total of 263 bp
(32.07%) varied between all taxa and 210 bp (25.61%) were
phylogenetically informative, with twenty-®ve (11.9%) in ®rst,
four (1.90%) in second and 181 (86.19%) in third codon
positions. The ratio of transitions to transversions was 1.72 for
all sites (9.13 in ®rst + second codon positions and 1.48 in third
codon positions). The predominance of transitions has been
documented widely for insect mtDNA (Simon et al., 1994),
including gerrine water striders of genera Limnoporus (Sperling
et al., 1997) and Aquarius (Damgaard et al., 2000b). The
average base composition was 35.0% A, 14.6% C, 13.2% G
and 37.1% T. For the different positions, the average base
composition for ®rst codon position was 35.9% A, 12.7% C,
22.9% G and 28.4% T; second codon position was 20.5% A,
23.7% C, 14.9% G and 40.8% T; and third codon composition
was 48.7% A, 7.4% C, 1.9% G and 42.1% T. The sequence is
therefore A/T rich with an overall average of 64.3% A + T. The
third codon position was extremely A/T rich with 90.8% A + T.
A 515 bp segment of EF-1a was sequenced for twenty-eight
species of gerrine water striders. EF-1a sites that consistently
displayed double peaks on electropherograms after multiple
sequencing runs were assumed to be heterozygous (for Papilio
see Reed & Sperling, 1999; for Aquarius see Damgaard et al.,
2000b). In the EF-1a matrix, twenty-eight sites were scored as
dimorphic, and thirteen individuals were scored as hetero-
zygous. Most of the dimorphisms were synonymous third
position variations. Of these, fourteen were C/T, ®ve were A/
G, two were A/T and one was G/T. Two dimorphisms were
non-synonymous in second position with one A/G and one G/T
variation. Finally, one A/G and one A/T non-synonymous
variation occurred in ®rst positions.
Of the 515 bp sequenced, 128 (24.85%) varied between all
taxa and seventy-nine (15.34%) were phylogenetically in-

# 2001 Blackwell Science Ltd, Systematic Entomology, 26, 241±254


formative with three (3.8%) in ®rst and seventy-six (96.2%) in
third positions. The ratio of transitions to transversions was
2.96 for all sites (0.78 for ®rst + second positions and 3.49 for
third positions). The TI/TV ratio is lower than the 4.25 found
in heliothine moths (Cho et al., 1995) and the 3.81 found in
Aquarius (Damgaard et al., 2000b), but higher than the 2.8
found in Ips bark beetles (Cognato, 1998). The average base
composition was 26.0% A, 23.3% C, 25.7% G and 25.0% T.
For the different codon positions, the average base composi-
tion for ®rst codon position was 31.4% A, 17.9% C, 38.4% G
and 12.4% T; second codon position was 30.1% A, 22.7% C,
18.7% G and 28.5% T; and third codon composition was
16.3% A, 29.5% C, 19.9% G and 34.4% T. The average
composition of each base is therefore roughly equal overall as
well as between the different positions.
Phylogenetic analyses
COI. An unweighted parsimony analysis of the COI dataset
gave a single MPC (most parsimonious cladogram) of 950
steps (Table 3 summarizes cladogram statistics). This clado-
gram is shown with branch lengths, bootstrap and branch
support values in Fig. 1. The cladogram includes clade
numbers 1, 4, 6, 10, 12, 14 and 15 (Table 4). Surprisingly,
G. argenticollis was placed as the sister species to all other
Gerris. To rule out the possibility of contamination, COI from
another specimen of G. argenticollis from Ontario, Canada,
was sequenced, giving an identical DNA sequence. To test for
saturation of substitutions in the gene, the number of inferred
steps from the MPT was plotted against pairwise comparisons
of the uncorrected genetic differences (Philippe et al., 1994).
The result shown in Fig. 2 was an almost straight line, only
indicating some degree of saturation of COI among the most
distantly related taxa. An unweighted parsimony analysis of
only transversions gave ®ve MPCs, each of 381 steps
(CI = 0.3255, RI = 0.5105). A successive weighting procedure
did not reduce the number of MPCs. A strict consensus
contains clade numbers 1, 2, 4, 7, 10 and 12 (Table 4). Finally,
the COI sequence data was used in generating a cladogram
based on maximum likelihood. A TI/TV ratio of 2.4 and an
among site variation of 0.13 gave the ln-likelihood ±
5302.1399. The cladogram is sixteen steps longer than the
Table 3. Summary of parsimony analyses of molecular, morphological and variously combined datasets.
PIC = phylogenetically informative characters.
No. of No. of Cladogram
Dataset characters PICs length
COI 820 210 950
EF-1a 515 79 287
Morphology 63 54 119
COI + EF-1a 1335 289 1252
COI + morphology 883 264 1088
EF-1a + morphology 578 133 418
`Total evidence' 1398 343 1388
# 2001 Blackwell Science Ltd, Systematic Entomology, 26, 241±254
Phylogeny of Gerris 245
MPC, and contains clade numbers 2, 4, 7, 9±12, 14 and 15
(Table 4).
EF-1a. Sixty-four MPCs (each of 287 steps) were obtained
from an unweighted parsimony analysis of the EF-1a
nucleotide dataset, which by successive weighting were
reduced to eight cladograms (see Table 3 for cladogram
statistics). Figure 3 shows the strict consensus cladogram with
bootstrap and branch support values, and includes clade
numbers 1, 4, 5, 10, 12 and 13 (Table 4). The EF-1a dataset
was tested for saturation, and a relatively straight line was
obtained that indicated virtually no saturation (Fig. 4). An
unweighted parsimony analysis of only transversions gave 515
MPCs, each of sixty-®ve steps (CI = 0.7385; RI = 0.7733),
which by successive weighting were reduced to twenty-eight
cladograms. A strict consensus of these includes clade numbers
1, 4, 9±12, 14 and 15 (Table 4). Finally, the EF-1a sequence
data were used in generating a cladogram based on maximum
likelihood. A TI/TV ratio of 3.65 and an among site variation
of 0.134 gave the highest likelihood (2177.8308). This is seven
steps longer than the MPC and includes clade numbers 1, 4, 5,
9, 10, 12 and 15 (Table 4).
Morphology
An unweighted parsimony analysis of the morphological
dataset gave twelve MPCs, each of 119 steps, which by
successive weighting were reduced to four cladograms (see
Table 3 for cladogram statistics). Figure 5 shows the strict
consensus of these cladograms with bootstrap and branch
support, and includes clade numbers 1±4, 7±12, 14 and 15
(Table 4). The cladogram is congruent with the established
morphologically based phylogeny (Andersen, 1993a: Fig. 23).
Combined analyses
Unweighted parsimony analyses were performed on the
morphology and nucleotide sequence datasets in different
combinations, as well as in `total evidence' analyses. An
unweighted parsimony analysis of the `total nucleotide' dataset
gave eight MPCs, each of 1252 steps, which by successive
weighting were reduced to a single cladogram that includes
No. of
cladograms CI RI
1 0.3811 0.4463
64 0.5296 0.5846
12 0.6050 0.8374
8 0.4105 0.4679
2 0.3989 0.5159
5 0.5359 0.6840
3 0.4222 0.5215
246 J. Damgaard and F. A. H. Sperling

Fig. 1. Single most parsimonious cladogram resulting from an unweighted parsimony analysis of 820 bp of the 3¢ half of COI conducted in
PAUP* using a heuristic search with twenty random addition replicates. Numbers above branches indicate branch length. The ®rst number below
each branch indicates bootstrap support from 500 replicates with ten random-addition heuristic searches per replicate. The second number below
each branch is branch support.

clade numbers 1, 2, 4, 6, 7 and 9±15 (Table 4). An unweighted
parsimony analysis for only transversions gave ®ve MPCs,
each of 456 steps (CI = 0.3772; RI = 0.5267), which by
successive weighting were reduced to a single cladogram that
includes clade numbers 1±4, 7, 10 and 12 (Table 4). The ILD
test gave P = 0.27, which indicates that the two molecular
partitions are not signi®cantly incongruent. When the pairwise
uncorrected genetic distances from the two datasets were
plotted against each other (Fig. 6), the uncorrected genetic
distances from EF-1a were found to be generally smaller than
for COI, and the majority of variable characters for closely
related taxa arose from COI, whereas for the more distantly
related species the dominance of COI levelled off. When the
two genes are corrected for differences in length, COI contains
about 25% more variable sites and 40% more phylogenetically
informative characters. An unweighted parsimony analysis of
the COI and morphology datasets gave two MPTs, each of
1088 steps, which by successive weighting were reduced to a
single cladogram containing clade numbers 1, 2, 4, 7±12, 14
and 15 (Table 4). The ILD test gave P = 0.85, which indicates
no signi®cant incongruence between the two partitions. An
unweighted parsimony analysis of EF-1a + morphology gave
®ve MPCs, each of 418 steps, which by successive weighting
were reduced to two cladograms that include clade numbers 1±
4, 6±12 and 14. The ILD test gave P = 0.03, which indicates
signi®cant incongruence between EF-1a and morphology.
Finally, an unweighted `total evidence' analysis of the three
datasets gave three MPCs, each of 1388 steps. A successive

# 2001 Blackwell Science Ltd, Systematic Entomology, 26, 241±254

 Phylogeny of Gerris 247

Table 4. Summary of phylogenetic analyses of the three datasets (morphology, EF-1a and COI). Tot. nuc. = COI + EF-1a data, Tot.
ev. = COI + EF-1a + morphology, MP = maximum parsimony, TV = transversions only, ML = maximum likelihood, Mor. = morphology.

Tot. nuc.
COI COI COI EF1a EF1a EF1a COI EF1a Tot.
Clade no. MP TV ML MP TV ML Mor. MP TV + Mor. + Mor. Ev.

(1) G. gibbifer + G. maculatus X X ± X X X X X X X X X

(2) G.lacustris-group ± X X ± ? ± X X X X X X
(3) G. costae + G. thoracicus ± ? ± ? ± ± X ± X ± X ±
(4) G. alacris + G. comatus X X X X X X X X X X X X
(5) 2 + 3 + 4 ± ± ± X ± X ± ± ± ± ± ±
(6) 5 + G. swakopensis X ± ± ± ± ± ± X ± X X X
(7) G. odontogaster group ± X X ± ± ± X X X X X X
(8) `derived Gerris' ± ± ± ± ± ± X ± ± X X X
(9) 8 + G. nepalensis ± ± X ± X X X X ± X X X
(10) G. gillettei-group X X X X X X X X X X X X
(11) Gerris s. str. ± ± X ± X ± X X ± X X X
(12) Gerriselloides X X X X X X X X X X X X
(13) 11 + 12 ± ± ± X ? ± ± X ± ± ± X
(14) Gerris s.l. X ± X ± X ± X X ± X X X
(15) Gerris + Aquarius X ± X ± X X X X ± X ± X

Fig. 2. Number of inferred steps based on Fig. 1 against uncorrected (p) distances for pairwise comparisons between 820 bp nucleotide sequences
of COI in PAUP*.

weighting procedure was not successful in reducing the test for the data partitions gave P = 0.27, which indicates that
number of cladograms. Figure 7 shows the strict consensus the three datasets are not signi®cantly incongruent. Table 5
cladogram with bootstrap and branch support values, and shows the partitioned branch support for the consensus
includes clade numbers 1, 2, 4 and 6±15 (Table 4). The ILD cladogram (Fig. 7).

# 2001 Blackwell Science Ltd, Systematic Entomology, 26, 241±254

248 J. Damgaard and F. A. H. Sperling

Fig. 3. Strict consensus of eight equally parsimonious cladograms obtained by successive weighting of sixty-four equally parsimonious
cladograms resulting from parsimony analysis of 515 bp of EF-1a conducted in PAUP* using a heuristic search with twenty random-addition
replicates. Format for numbers as in Fig. 1.

Discussion from these partitions that failed to resolve numerous relation-

Combination of datasets
The data partitions yielded three different cladograms of
widely varying topology, and produced a consensus cladogram
ships. Based on the ILD tests, it is evident that the
phylogenetic signals from COI and morphology show no
signi®cant incongruence. With the inclusion of EF-1a, the
incongruence increases, and the combination of EF-1a and
morphology is signi®cantly incongruent. Damgaard et al.

# 2001 Blackwell Science Ltd, Systematic Entomology, 26, 241±254


Fig. 4. Number of inferred steps based on Fig. 2 against uncorrected (p) distances for pairwise comparisons between 515 bp nucleotide sequences
of EF-1a in PAUP*.
(2000b) Aquarius found no signi®cant incongruence between
these two datasets in their study.
The different signal in EF-1a is poorly supported and
largely dissolves when combined with the other partitions.
Figure 4 shows that the poor phylogenetic signal of EF-1a
cannot be explained as a result of saturation, and is more likely
to be caused by the restricted number of phylogenetically
informative characters related to the slow mutation rate in this
gene. Therefore, its signal is probably swamped by the greater
amount of information yielded by COI with its faster mutation
rate. Phylogenetic studies of > 1000 bp of EF-1a have shown
the marker to be excellent for solving intrageneric relation-
ships in Lepidoptera (Cho et al., 1995; Reed & Sperling, 1999)
and a longer strand would probably have increased its utility in
the present study. The `total nucleotide analysis' resolved
many of the clades found in the `total evidence analysis', that
were probably caused by the combination of COI with a
relatively high mutation rate and EF-1a with a slower rate. As
shown in Fig. 5, the phylogenetic signal for closely related taxa
mostly arises from COI, whereas EF-1a becomes increasingly
supportive, probably because the information yieded by EF-1a
becomes more signi®cant with increasing saturation of
information yielded by COI at higher taxonomic levels.
In the `total evidence' cladogram (Fig. 7), COI supplies
37.5%, EF-1a 35.4% and morphology 27.2% of the total
branch support (Table 5). Therefore, we conclude that even
though analyses of EF-1a gave a topology different than the
other partitions, this topology is poorly supported in other
# 2001 Blackwell Science Ltd, Systematic Entomology, 26, 241±254
Phylogeny of Gerris 249
datasets, and all three partitions contribute equally to the
clades found in the `total evidence' cladogram.
The species groups of Gerris
All of the datasets strongly support the sister-species
relationship between G. asper and G. lateralis, and thereby
subgenus Gerriselloides. Andersen (1993a) omitted the G.
gracilicornis-group from Gerriselloides and elevated it to
subgenus Macrogerris, a belief supported by all present
analyses. Strong support was also found for the Nearctic sister
species G. gillettei and G. pingreensis, and their inclusion in a
monophyletic group together with the Nearctic G. incognitus
and the Palearctic G. sphagnetorum. Andersen (1993a)
recognized convincing, morphological similarities between
the species and named the assemblage the G. gillettei-group.
The data strongly support the sister-group relationship
between the Palearctic G. babai and G. odontogaster and the
Nearctic G. buenoi. Andersen (1993a) grouped these three
species in the G. odontogaster-group along with the Palearctic
G. argentatus, and this topology is recognized in all of the
combined analyses, even though the support is relatively low.
All of the analyses show a relationship between G. alacris and
G. comatus. Andersen (1993a) included these and three
additional species in the Nearctic G. marginatus-group, and
assigned the members of the group to two different clades
based on differences in the male genitalia. Even though G.
250 J. Damgaard and F. A. H. Sperling

Fig. 5. Strict consensus of twelve equally parsimonious cladograms resulting from an unweighted parsimony analysis of sixty-three
morphological characters conducted in PAUP* using a heuristic search with twenty random-addition replicates. Format for numbers as in Fig. 1.

alacris and G. comatus were assigned to different clades in the
G. marginatus-group, strong support was found here for their
monophyly, and hence the monophyly of the G. marginatus-
group. Andersen (1993a) further included the Palearctic G.
latiabdominis in the G. marginatus-group based on the
presence of lateral stripes of silvery pubescence on the
pronotum, but none of the analyses here supports this, and
this similarity to members of the group must be due to
convergence.
Strong support was found for the relationship between G.
gibbifer and G. maculatus. Andersen (1993a) included these
along with G. lacustris in the G. lacustris-group, but the group
is poorly supported in the analyses of morphological data, and
was not found in analysis of molecular data. On the other hand,
it was found in analyses of all combinations of data, and was
well supported in the `total evidence' analysis; therefore, we
accept the G. lacustris-group. Finally, Andersen (1993a)
suggested a relationship between G. thoracicus and G. costae
on the basis of two supposed synapomorphies (rufous or
yellowish brown pronotal lobe and relatively short, stout legs),
but interpreted them both as reversals. Based on the analysis of
molecular data, when analysed alone or in combination with

# 2001 Blackwell Science Ltd, Systematic Entomology, 26, 241±254


Fig. 6. Uncorrected (p) distances for pairwise comparison between 820 bp nucelotide sequences for COI against 515 bp nucleotide sequences of
EF-1 a.
morphological data, there is no support for a close relationship
between these species, and we therefore consider the `G.
thoracicus-group' to be a polyphyletic assemblage.
Phylogeny of Gerris
Analysis of both molecular datasets indicates a relationship
between the G. lacustris-group, the G. marginatus-group, G.
thoracicus and G. costae. The analyses of COI further link G.
swakopensis to this group as the sister species to G. costae,
although this condition is contradicted by analyses of both EF-
1a and the morphological data. Andersen (1993a) placed these
species along with G. argenticollis, G. latiabdominis and the
G. odontogaster-group, in a `derived Gerris' clade. This clade
is not supported by the molecular data, but is present, though
weakly supported, in the combined analyses of morphological
and molecular data. Hypotheses about the relationships could
well be tested against sequencing data in order to validate or
contradict them. Andersen (1993a) suggested that G. nepa-
lensis is the sister taxon to the `derived Gerris'. This
relationship is not supported in the molecular analyses, but it
is recognized and relatively well supported in the `total
evidence' analysis. The analyses of COI and the combined data
indicate a sister-group relationship between the G. gillettei-
group and the `derived Gerris + G. nepalensis' clade, except
for the exclusion of G. argenticollis. Andersen (1993a)
recognized this basal position of the G. gillettei-group and
elevated the entire assemblage to subgeneric rank, Gerris s. str.
Whether subgenera Gerriselloides or Macrogerris is the sister
# 2001 Blackwell Science Ltd, Systematic Entomology, 26, 241±254
Phylogeny of Gerris 251
group to Gerris s. str. is ambiguous in all analyses. Although
the analyses of the morphological data gave relatively strongly
support for Andersen's (1993a) belief that Gerriselloides is the
most basal clade in Gerris, this is contradicted in analyses of
both COI and the combined data. We therefore suggest that
Macrogerris is the basal subgenus in Gerris.
Biogeographical implications
The distributions of Gerriselloides and the G. lacustris-
group cover most of the Palearctic Region, and several species
have broadly overlapping ranges. Closely related species,
however, are usually allopatric, showing both north±south and
east±west vicariance, e.g. the sister species G. lateralis + G.
asper and G. gibbifer + G. maculatus. Andersen (1993a)
included other pairs of sister species, but these ideas need to be
veri®ed on the basis of molecular data. Subgenus Macrogerris
and the monotypic G. nepalensis-group are endemic to eastern
Asia, and the most widespread species of Macrogerris, G.
gracilicornis, is sympatric with G. nepalensis (Andersen,
1993a). Andersen (1993a) included the Mexican G. ®rmus and
the eastern Palearctic G. latiabdominis in the otherwise
Nearctic G. marginatus-group. The analysis here shows that
the af®nity with G. latiabdominis probably is due to
convergence. The inclusion of G. ®rmus still needs to be
tested with DNA sequence data, but until then, the G.
marginatus-group and the monotypic G. argenticollis-group
are the only strictly Nearctic species-groups in Gerris. The G.
swakopensis-group occurs in all sub-Saharan Africa, including
252 J. Damgaard and F. A. H. Sperling
Fig. 7. Strict consensus of three equally parsimonious cladograms resulting from unweighted parsimony analyses of the combined molecular and
morphological data conducted in PAUP* using a heuristic search with twenty random-addition replicates. Format for numbers as in Fig. 1.
Madagascar (Andersen, 1993a), and is therefore endemic to the
Afrotropical Region. Finally, we can con®rm Andersen's
(1993a) hypothesis of `trans Beringian' relationships in the
G. gillettei-group and the G. odontogaster-group.
Ecological phylogenetics
Andersen (1993a,b) used his reconstructed phylogeny of
Gerris based on morphological characters to interpret the
evolution of morphological, developmental and ecological
patterns. Andersen (1993b) showed that the ancestral state for
Gerris is wing dimorphism, even for non-diapausing adults.
The replacement of Gerriselloides with Macrogerris as the
most basal subgenus in Gerris does not require one to postulate
any additional changes in character states. Andersen (1993a)
stated that species of Macrogerris, including Chinese G.
gracilicornis are wing-dimorphic, whereas Andersen (1993b:
Fig. 2) stated that Japanese G. gracilicornis are monomorphic
long-winged. Therefore, additional work should be performed
on this subgenus to resolve whether the monomorphic or
dimorphic long-winged condition is ancestral. Changes toward
the monomorphic long-winged condition in the diapausing
generation were found to have arisen twice, once in G.
gracilicornis and once in all `derived Gerris', except for
reversals to wing dimorphism in the G. lacustris-group.
Changes toward the monomorphic long-winged condition in
the non-diapausing generation were found to have arisen twice,
once in the `G. thoracicus-group', and once in G. latiabdo-
minis (Andersen, 1993b). Because G. costae and G. thoracicus
no longer are considered to be sister taxa, the changes toward
the monomorphic long-winged condition in the non-diapausing
generation must have arisen three times. The evolution of wing
dimorphism in Gerris is correlated with the habitat preferences
of the species, in which the basal, permanently wing dimorphic
species Gerriselloides and in the G. gillettei- and G.
nepalensis-groups are restricted to more stable habitats,
whereas the `derived Gerris' are either permanently or
# 2001 Blackwell Science Ltd, Systematic Entomology, 26, 241±254
 Phylogeny of Gerris 253

Table 5. Partitioned branch support for the most parsimonious cladogram resulting from the
`total evidence' analysis.

Clade no. COI EF-1a Morphology Total

(1) G. gibbifer + G. maculatus 2.56 6 2.44 11

(2) G. lacustris-group 0.5 0.5 3 4
(3) G. costae + G. swakopensis 3 1 ±3 1
(4) 2 + 3 0.5 0.55 ± 0.05 1
(5) G. alacris + G. comatus 12.5 8.17 1.33 22
(6) 5 + G. thoracicus 2.43 1.05 ± 2.48 1
(7) 4 + 6 ±1 4 0 3
(8) G. buenoi/G. odontogaster/G. babai 6.8 4.53 5.67 17
(9) G. odontogaster-group 3 0.72 ± 0.72 3
(10) `derived Gerris' ± 6.5 ± 0.17 7.67 1
(11) 10 + G. nepalensis 0 1.33 2.67 4
(12) G. pingreensis + G. gillettei 9 4 9 13
(13) 12 + G. incognitus ± 0.27 1.79 3.48 5
(14) G. gillettei-group 4.25 2.08 1.67 8
(15) Gerris s.str. ± 2.8 2.93 3.87 7
(16) G. asper + G. lateralis 24.5 10.33 1.67 36
(17) 15 + 16 0.5 2.83 ± 2.33 1
(18) Gerris s.l 0.5 3.83 4.67 10
(19) A. conformis + A. paludum ± 0.67 5.73 1.07 6
(20) Aquarius 2.5 ± 2.17 1.67 2
(21) Aquarius + Gerris 2.5 ± 2.17 1.67 2
(22) Limnoporus 0.5 3.83 3.67 8
Sum 64.3 60.69 46.64 166*

* The difference between the sum of the partitions (171.63) and the sum of partitioned branch
support is due to the restricted number of decimals in each partition.

seasonally wing-dimorphic or monomorphic long-winged and was supported by grants from the Danish Natural Science
occupy more unstable habitats. Research Council (grant no. 9502155 to J.D.) and the
California Agriculture Experiment Station (to F.A.H.S).

Acknowledgements
This study was carried out in the Department of Entomology,
Zoological Museum (ZMUC) and the Department of
Evolutionary Biology, Zoological Institute (ZIUC), both in
the University of Copenhagen, and the Department of
Environmental Sciences, Policy and Management, University
of California, Berkeley. We are indebted to Anthony Cognato,
Mike Caterino and May Kuo, University of California,
Berkeley, and Pia Friis, Peter Gravlund and Sheila Tang (all
at ZIUC) for technical advice and support; and Nils Mùller
Andersen (ZMUC), Henrik Glenner and Bo Vest Pedersen
(both at ZIUC) for valuable comments on the manuscript. We
are also thankful to the following persons for collecting
specimens for this study: GoÈran Arnquist (Department of
Animal Ecology, University of UmeaÊ, Sweden), Doug Currie
(Toronto, Canada), Tetsuo Harada (Department of Biology,
Osaka City University, Japan), James J. Krupa (University of
Kentucky, Lexington, U.S.A.), Verner Michelsen
(Entomological Department, Zoological Museum, University
of Copenhagen, Denmark), John T. Polhemus (Colorado
Entomological Museum, Englewood, Colorado, U.S.A.), B.S.
Smith (Ithaca College, Ithaca, New York, U.S.A.) and John R.
Spence (University of Alberta, Edmonton, Canada). This work
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