Older than New Caledonia emergence? A molecular phylogenetic study of the eneopterine crickets...

ORIGINALARTICLE

Older than New Caledonia emergence?A molecular phylogenetic study of theeneopterine crickets (Orthoptera:Grylloidea)

Romain Nattier1*, Tony Robillard1, Laure Desutter-Grandcolas1,

Arnaud Couloux2 and Philippe Grandcolas1

INTRODUCTION

New Caledonia is a hotspot of biodiversity, the origin of which

has traditionally been traced back to the fragmentation of

Gondwana, specifically the separation of Australia around

80 Ma (Raven & Axelrod, 1972; Morat et al., 1986; Jaffre, 1992;

Chazeau, 1993; de Laubenfels, 1996; Lowry, 1998). This

assumption of an old origin of the local biota was based on

both the antiquity of New Caledonia’s geological basement and

the presence of many so-called relict groups (Raven & Axelrod,

1Museum national d’Histoire naturelle,

Departement Systematique et Evolution, UMR

7205 CNRS OSEB, Case postale 50

(Entomologie), 57 rue Cuvier, 75231 Paris

Cedex 05, France, 2Genoscope, Centre national

de Sequencage, 2 rue Gaston Cremieux, Case

postale 5706, 91057 Evry Cedex, France

*Correspondence: Romain Nattier, Museum

national d’Histoire naturelle, Departement

Systematique et Evolution, UMR 7205 CNRS

OSEB, Case postale 50 (Entomologie), 57 rue

Cuvier, 75231 Paris Cedex 05, France.

E-mail: [email protected]

ABSTRACT

Aim A New Caledonian insect group was studied in a world-wide phylogenetic

context to test: (1) whether local or regional island clades are older than 37 Ma,

the postulated re-emergence time of New Caledonia; (2) whether these clades

show evidence for local radiations or multiple colonizations; and (3) whether

there is evidence for relict taxa with long branches in phylogenetic trees that relate

New Caledonian species to geographically distant taxa.

Location New Caledonia, south-west Pacific.

Methods We sampled 43 cricket species representing all tribes of the subfamily

Eneopterinae and 15 of the 17 described genera, focusing on taxa distributed in

the South Pacific and around New Caledonia. One nuclear and three

mitochondrial genes were analysed using Bayesian and parsimony methods.

Phylogenetic divergence times were estimated using a relaxed clock method and

several calibration criteria.

Results The analyses indicate that, under the most conservative dating scenario,

New Caledonian eneopterines are 5–16 million years old. The largest group in the

Pacific region dates to 18–29 Ma. New Caledonia has been colonized in two

phases: the first around 10.6 Ma, with the subsequent diversification of the

endemic genus Agnotecous, and the second with more recent events around

1–4 Ma. The distribution of the sister group of Agnotecous and the lack of

phylogenetic long branches in the genus refute an assumption of major extinction

events in this clade and the hypothesis of local relicts.

Main conclusions Our phylogenetic studies invalidate a simple scenario of local

persistence of this group in New Caledonia since 80 Ma, either by survival on the

New Caledonian island since its rift from Australia, or, if one accepts the

submergence of New Caledonia, by local island-hopping among other subaerial

islands, now drowned, in the region during periods of New Caledonian

submergence.

Keywords

Biogeographical tests, calibration points, dating, divergence times, emergence,

endemic clade, Eneopterinae, New Caledonia, relict.

Journal of Biogeography (J. Biogeogr.) (2011) 38, 2195–2209

ª 2011 Blackwell Publishing Ltd http://wileyonlinelibrary.com/journal/jbi 2195doi:10.1111/j.1365-2699.2011.02563.x

1972; Raven, 1979; Morat et al., 1986). This widely accepted

Gondwanan paradigm and continental characterization of

New Caledonia was recently questioned in the light of

geological and phylogenetic evidence (Murienne et al., 2005,

2008; Grandcolas et al., 2008).

First, geological studies do not support a very old origin for

the subaerial land but instead indicate several recent and long

episodes of total submergence related to major regional

tectonic events in the Palaeocene and then in the Eocene,

lasting until 37 Ma (Paris, 1981; Aitchison et al., 1995; Cluzel

et al., 2001; Crawford et al., 2003; Pelletier, 2006). Indeed, the

last of these complete Tertiary submergences, although rarely if

ever commented on in the biogeographical literature, had

substantial geological consequences that have long been

recognized by sedimentologists. In fact, the famous New

Caledonian nickel mines are located on lateritic soils derived

from ultramafic rocks that were obducted onto most of the

New Caledonian mainland, known as Grande Terre (Chevil-

lotte et al., 2006), when it was situated deeper than the

lithospheric ocean floor around 40 Ma (Paris, 1981; Aitchison

et al., 1995; Cluzel et al., 2001; Crawford et al., 2003; Pelletier,

2006). Several biologists have, however, expressed doubt that

the local biota could be more recent than these generalized

submergences and invoked island refuges and that colonization

occurred by stepping-stone dispersal, in an effort to rescue the

assumption of an old origin for the New Caledonian biota

(Morat et al., 1986; Heads, 2005, 2008, 2009a, 2010; Ladiges &

Cantrill, 2007; Ladiges, 2008). These scenarios assume that

some groups evolved by island-hopping before the emergence

of the present-day islands, involving islands that are no longer

emergent as potential refuges, and thereby reviving an old

dispersalist model (Gressitt, 1956; Darlington, 1957; Carlquist,

1965, 1974; Axelrod, 1972).

Second, the old or relict groups on New Caledonia were

often loosely defined, and an assumption that they occupy

‘basal’ phylogenetic positions (Heads, 2009b) was used in a

circular fashion to infer the biota’s antiquity (see the critique

in Waters & Craw, 2006). Recent work has shown instead that

the few groups adequately diagnosed as relict actually provide

evidence of important regional extinctions rather than of local

persistence of the biota (Grandcolas et al., 2008; Sharma &

Giribet, 2009).

Most New Caledonian groups have not yet been studied in

any detail from a phylogenetic perspective. Their local

antiquity should now be assessed by disentangling the possible

old age of the clade to which they belong from the age of the

local New Caledonian taxa. The age of Pacific groups in which

New Caledonian taxa may be nested should also be considered

in order to test the regional island-hopping dispersal scenario.

Several studies have already brought to light the problem of the

origin of New Caledonia and the possibility of multiple

colonizations (Swenson et al., 2001; Bartish et al., 2005;

Murienne et al., 2005, 2008; Page et al., 2005; Desutter-

Grandcolas & Robillard, 2006; Balke et al., 2007a,b; Espeland

& Johanson, 2010). However, more analyses are needed to

enable accurate biogeographical tests (Grandcolas et al., 2008),

and the following key questions need to be resolved indepen-

dently of knowledge of geology. First, are New Caledonian or

regional island clades older than 37 Ma, that is, older than the

most recent emergence of New Caledonia, the oldest island of

the region? Second, do these clades show evidence for local

radiations or for multiple colonizations? Third, do phyloge-

netic trees relate New Caledonian species to geographically

distant taxa with long branches, providing evidence that they

may be local relicts (Grandcolas et al., 2008; Sharma & Giribet,

2009)?

To answer these questions, we analyse a group, the cricket

subfamily Eneopterinae, that includes both endemic New

Caledonian species and close relatives distributed at a regional

or larger scale (Robillard & Desutter-Grandcolas, 2004a;

Desutter-Grandcolas & Robillard, 2006). Eneopterinae have a

world-wide distribution but are particularly diversified in the

south-western Pacific islands. They are represented in New

Caledonia by one genus endemic to Grande Terre (Agnotecous)

and by two species belonging to two genera more widely

distributed in the region (Desutter-Grandcolas & Robillard,

2006; Robillard & Ichikawa, 2009). Here we investigate the age

and origin of these taxa in New Caledonia using a calibrated

molecular phylogeny.

Molecular dating is often seen as problematic because of the

use of poor or mistrusted calibrations, which cast doubt on the

biogeographical interpretation of the results (e.g. Heads, 2005;

Ho et al., 2008). We therefore carry out our tests using the

most conservative approach, calibrating the dating with the

oldest palaeogeographical evidence available for territories

largely independent from New Caledonia and the neighbour-

ing islands, and also by placing emphasis on the geological age

of the land rather than on the strict date of land emergence. To

complete this conservative approach, we calibrated the dating

with the most ancient fossil attributed to Eneopterinae

independent of any assumptions of evolution by vicariance

as implied by the palaeogeographical calibrations. We also

performed a ‘reverse’ dating approach to control for which

palaeogeographical scenarios should be endorsed for the whole

group if we were to assume that taxa from New Caledonia and

the Loyalty Islands were older than the island on which they

occur. In all these cases, dating is done by allowing rates to

vary across tree branches (Drummond et al., 2006) because

strict clock-like assumptions are not tenable (Pybus, 2006;

Rutschmann, 2006; Pulquerio & Nichols, 2007).

MATERIALS AND METHODS

Sampling

Taxon sampling

The subfamily Eneopterinae comprises a diverse assemblage of

tropical crickets that live in a wide array of habitats (e.g. leaf

litter and tree canopy in forests, low bushes in open areas, trees

at forest edges, coastal areas, savannas), have diverse potential

vagility (one genus is apterous, the others have either short or

R. Nattier et al.

2196 Journal of Biogeography 38, 2195–2209ª 2011 Blackwell Publishing Ltd

long wings), and that are known for their diverse calling songs

(Robillard & Desutter-Grandcolas, 2004b, 2011a,b). They have

a world-wide distribution, with most of the diversity located in

Pacific islands. The subfamily has been the subject of several

phylogenetic studies focused on the evolution of acoustic

communication and behaviour (Robillard & Desutter-Grand-

colas, 2004a,b, 2006, 2011b). Previous studies were based on

both morphological (Robillard & Desutter-Grandcolas, 2004a)

and limited molecular data sets (Robillard & Desutter-

Grandcolas, 2006), and did not discuss distributional evidence.

The ingroup consists of 43 eneopterine species (see Appen-

dix S1 in the Supporting Information), representing all tribes

of the subfamily and 15 of the 17 genera (sensu Robillard &

Desutter-Grandcolas, 2008; Robillard, 2011). Given the region

of interest for the study, the sampling is concentrated on taxa

from the South Pacific and in particular around New

Caledonia (Fig. 1), which corresponds to the tribe Lebinthini

and encompasses 28 of the 43 species. Acheta domesticus,

Anurogryllus muticus, Gryllus bimaculatus (Gryllinae) and

Gryllitara sp. (Itarinae) were chosen as outgroups, based on

previous studies (Robillard & Desutter-Grandcolas, 2006).

Gene sampling

Molecular markers were selected following previous studies of

the Eneopterinae (Robillard & Desutter-Grandcolas, 2006) and

available sequences in GenBank: three mitochondrial gene

fragments: cytochrome b (cyt b, 750 bp) and the large (16S

rRNA, c. 530 bp) and small (12S rRNA, c. 430 bp) ribosomal

sub-units, as well as a fragment of the nuclear large ribosomal

sub-unit gene (18S rRNA, c. 650 bp). All sequences have been

deposited in GenBank (Appendix S1).

DNA extraction, amplification and sequencing

The right hind femur of specimens was collected and DNA

extractions were performed using a QIAamp DNA MicroKit

(QIAGEN, Courtaboeuf, France) following the manufacturer’s

instructions. Molecular work was carried out at the Museum

national d’Histoire naturelle (MNHN), Service de Systema-

tique Moleculaire. The oligonucleotide primers used for

polymerase chain reaction (PCR) and sequencing are listed

in Appendix S2. Amplifications were performed in a 25-lL

reaction volume with 0.4 lL of each 10-pM primer, 19.2 lL of

H20, 2.5 lL of buffer, 1.25 lL of dimethyl sulfoxide (DMSO),

1 lL of MIX, 0.15 lL of Taq polymerase and 1 lL of DNA.

The PCR consisted of an initial denaturing step at 94 �C for

4 min, 40 amplification cycles [denaturation at 94 �C for 30 s,

annealing at between 48 and 55 �C (Appendix S2) for 40 s,

and extension at 72 �C for 40 s], and a final step at 72 �C for

7 min. PCR products were checked on agarose gels and

sequenced in both directions with the same primers at

Genoscope (Evry, France). Sequences were cleaned, and coding

sequences were translated using the invertebrate mitochondrial

genetic code to check for the absence of stop codons using

Sequencher v. 4.8 (GeneCodes Corporation, Ann Arbor, MI,

USA). All genes were screened for potential contamination

using the BlastX algorithm on GenBank.

Phylogenetic analyses

DNA sequences were aligned under Muscle 3.8.31 (Edgar,

2004) using default parameters, and the complete molecular

dataset was concatenated in order to use all available evidence

(Kluge, 1989; Nixon & Carpenter, 1996). Bayesian and

parsimony analyses were conducted.

The substitution model of evolution was estimated using

jModelTest 0.1.1 (Posada, 2008), and the Akaike information

criterion (AIC; Akaike, 1973, 1974) was used to select the

GTR+I+G model. Bayesian analyses were conducted with

MrBayes 3.1.2 (Ronquist & Huelsenbeck, 2003). Four Markov

chains were run simultaneously for 15 million generations,

sampling every 100 generations to ensure independence of

samples. The first 1.5 million trees generated were discarded as

burn-in and determined empirically from the log-likelihood

values using Tracer 1.4 (Rambaut & Drummond, 2007). The

remaining trees were used to construct 50% majority-rule

consensus trees. Two independent runs were performed to

check whether convergence on the same posterior distribution

was reached and whether the final trees converged on the same

topology. The statistical confidence of each node was evaluated

by posterior probability.

Parsimony analyses were performed with tnt 1.1 (Goloboff

et al., 2008). The search strategy consisted first of 1000

replications of random addition sequence and tree bisection–

reconnection (TBR). Then, to avoid local optima (Maddison,

1991), we added 100 iterations of tree fusing (Goloboff, 1999),

each iteration being swapped with TBR and subtree pruning

and regrafting (SPR); and 20 iterations of ratchet (Nixon,

1999), in weighting characters with a factor of four. tnt was

also used to calculate bootstrap support (BS; Felsenstein, 1985)

values with 1000 replicates.

Molecular dating analyses

Detecting among-lineage rate heterogeneity

Prior to estimating divergence times using relaxed molecular

clock methods, we used a likelihood-ratio test (Huelsenbeck &

Crandall, 1997) to assess rate homogeneity among taxa. This

test compares a molecular clock-constrained tree and an

unconstrained tree in paup* 4.0b10 (Swofford, 2002) with a

null hypothesis of a homogeneous rate of evolution among all

branches in the phylogeny. In the present study, the test

rejected the null hypothesis, which indicates that rates of

substitution varied significantly among branches and that a

molecular clock model would be inappropriate.

Calibration points

Molecular dating requires calibration data such as a known

substitution rate, dated fossil records or palaeogeographical

Older than New Caledonia emergence?

Journal of Biogeography 38, 2195–2209 2197ª 2011 Blackwell Publishing Ltd

Figure 1 Localities of outgroups and eneopterine species as studied in the phylogeny. Acheta domesticus is a domestic species with a

world-wide distribution. The number of terminals in each area is indicated in parentheses.

R. Nattier et al.


events, or date estimates from previous studies (Magallon,

2004; Sanderson et al., 2004; Drummond et al., 2006; Ho,

2007; Ho et al., 2008), which have different strengths and

drawbacks (Heads, 2005; Kodandaramaiah, 2011). In the

present study, the use of substitution rates would be inappro-

priate because our data do not follow a molecular clock model

and there are no recent calibration points from previous

studies.

We used two kinds of palaeogeographical events to estimate

divergence times: (1) formation of islands, and (2) continental

break-up or ocean seaway closure. Each calibration point was

chosen from regions outside the region of interest (New

Caledonia and neighbouring islands). To prevent the inaccu-

rate estimation of node ages, we used multiple calibration

points (Lee, 1999; Wang et al., 1999) on basal and apical

nodes. We also checked whether all calibration points were

compatible with the reference topology, and adopted a

conservative approach by using the oldest age estimate for

each calibration (Table 1). In a second round of estimation,

and in order to be fully independent from regional geological

knowledge, dating was obtained by incorporating calibration

information only from outside the Pacific region and the

results compared with the first round that incorporated all

calibration.

(1) Island formation: the geological age of an island can

provide a maximum age estimate for the species divergence

time on that island (Fleischer et al., 1998), but islands resulting

from geological hotspots should be considered with respect to

the geological age of the hotspot itself (Price & Clague, 2002;

Heads, 2005; Parent et al., 2008) because islands form in

chronological sequence such that lineages may have diversified

on long-submerged islands, that is, long before the formation

of the present islands. The Fiji islands were formed by

subduction, and the oldest rocks are from a volcanic island-arc

of late Eocene age, that is, 34–37 Ma (Neall & Trewick, 2008).

Volcanism in the Samoa Islands is consistent with a plume-

driven hotspot model, with at least some of the volcanic timing

controlled by the Tonga Trench (Natland, 1980; Hart et al.,

2004). The oldest Samoa island in the Samoan archipelago,

Savai’i, is dated to 5 Ma, but basalts on the Alexa oceanic

plateau (West of Savai’i) were formed 24 Ma (Hart et al.,

2004). This latter age was retained as calibration point D.

(2) Continental break-up or ocean seaway closure: separa-

tion between Australia and eastern Antarctica, namely the

opening of the South Tasman Sea, is dated at around 35.5 Ma

(Veevers et al., 1991; McLoughlin, 2001; Sanmartın & Ron-

quist, 2004), but Woodburne & Chase (1996) pointed out

earlier biotic separation, with climatic deterioration at 46 Ma

(calibration point A).

The opening of the Drake Passage, which separates South

America and western Antarctica, is dated to 28–35.5 Ma

(Veevers et al., 1991; Sanmartın & Ronquist, 2004; Livermore

et al., 2005) and corresponds to calibration point B.

Asia and Africa were isolated by the Tethys Sea, which acted

as a dispersal barrier and a climatic filter (Gheerbrant & Rage,

2006) until the Neogene. A connection between them

Table 1 beast prior probability distributions (Ma) for calibration nodes (A–F) of the Eneopterinae used in this study. For each node, the

geological event is indicated with various age estimates, the oldest in bold, and the divergence event based on the tree topology (Fig. 2).

Node Geological event Age (Ma) Reference Divergence event on the topology

A South Tasman

Sea opens

35 Sanmartın & Ronquist (2004) Between Australian species and other

Eneopterinae35.5 McLoughlin (2001)

35.5 Veevers et al. (1991)

46 Woodburne & Chase (1996)

B Drake Passage opens 30–28 Sanmartın & Ronquist (2004) Between Eneoptera (South America)

and the other eneopterine species34–30 Livermore et al. (2005)

35.5 Veevers et al. (1991)

C Africa and Eurasia

connected

23 Kosuch et al. (2001) Between Xenogryllus transversus

(India) and Xenogryllus

eneopteroides (East Africa)

Cox & Moore (1993)

Bernor et al. (1987)

D Fiji emerges 25–20 Lucky & Sarnat (2010) Between Swezwilderia sp. (Fiji) and

Swezwilderia bryani (Samoa)37–34 Neall & Trewick (2008)

Samoa emerges 5 Hart et al. (2004)

23.9–22.9 Hart et al. (2004)

E Isthmus of Panama forms 2.95 Bartoli et al. (2005) Between Ponca (Central America)

and Ligypterus (South America)3.5–3.1 Burnham & Graham (1999)

4.6–2.7 Haug & Tiedemann (1998)

F Passage through the

North Atlantic

land bridge opens

37–33 Tiffney & Manchester (2001) Between Swezwilderia (Fiji, Samoa)

and Ponca–Ligypterus (Central

and South America)



(calibration point C) was established in the early Miocene

(23 Ma) through the Arabic peninsula, which permitted the

interchange of tropical biotas for the first time (Bernor et al.,

1987; Cox & Moore, 1993; Kosuch et al., 2001).

The age of the emergence of the Isthmus of Panama, namely

the formation of the land connection between North and

South America, is dated at a maximum of 4.6 Ma (Haug &

Tiedemann, 1998; Burnham & Graham, 1999; Bartoli et al.,

2005) and is used here as calibration point E.

As an additional calibration point (F), we used a break in the

connection of land in the Northern Hemisphere through the

North Atlantic land bridge, that is, the break in the direct land

connection between South Greenland and Europe dated to 33–

37 Ma (Tiffney & Manchester, 2001), even though further

exchange has been shown for some temperate taxa (Tiffney &

Manchester, 2001).

The problem with the palaeogeographical criterion for

dating, although strongly preferred by some authors (e.g.

Heads, 2005), is that it makes assumptions of past ancestral

distributions and vicariance of taxa in terminal areas docu-

mented for the present-day taxa of the outgroup (Ho et al.,

2008; Crisp et al., 2011). In this situation, the earliest

dichotomy between areas at the deepest node in the tree

particularly constrains the dating analysis and determines the

dates of the subsequent nodes.

To make the biogeographical test even more conservative by

pushing back the dating as far as possible and to escape this

constraining assumption, we therefore employ the only fossil

presumptively attributed to the cricket subfamily Eneopteri-

nae, namely Proecanthus anatolicus Sharov, 1968, which

Gorochov (1985) dated at 96 Ma, putting this date at the base

of the tree. Although this assignment can be strongly doubted

because of the poor state of conservation of diagnostic body

parts (T. Robillard, pers. obs.), it is used here within the

context of our conservative framework because the age of this

ambiguous fossil (96 Ma) is older than any palaeogeographical

event taken into consideration.

We also made another attempt at escaping the assumption

of vicariance by using a ‘reverse’ approach. We constrained the

clade of New Caledonian and Loyalty taxa to be older than

37 Ma (the postulated re-emergence time of New Caledonia)

and examined how the dating obtained for the deepest nodes

of the tree is compatible with palaeogeography, given the tree

topology and the distribution of taxa.

Dating methods

Bayesian methods are among the least sensitive of the

commonly used molecular dating methods to undersampling

(Linder et al., 2005). To estimate the relative age of divergence

of the lineages studied, we used the Bayesian relaxed phylo-

genetic approach implemented in beast 1.4.8 (Drummond &

Rambaut, 2007), which allows for variation in substitution

rates among branches (Drummond et al., 2006) and does not

assume that these rates are autocorrelated across the tree. We

used the best-fitting model, as estimated by jModelTest 0.1.1

(Posada, 2008), using the entire data set. Substitution rates

were estimated using an uncorrelated lognormal relaxed

molecular clock model and the Yule process of speciation

was assumed, as recommended for species-level phylogenies

(Drummond & Rambaut, 2007).

We used a normal distribution for the tree prior to

calibrating nodes, which is particularly suitable for modelling

biogeographical events (Ho, 2007), with a standard deviation

of 1%. Calibration nodes were constrained on the topology to

match with the most strongly supported nodes obtained in

MrBayes (Fig. 2). All other relationships were left free to vary

so that topological uncertainty was incorporated into posterior

estimates of divergence dates. We confirmed the results by

using two independent analyses over 25 million generations,

and sampled every 2500th generation to obtain a maximum of

10,000 samples, as recommended by Drummond & Rambaut

(2007). The two analyses converged on similar posterior

estimates. Then we used Tracer 1.4.1 (Drummond &

Rambaut, 2007) to assess convergence, measure the effective

sample size of each parameter, and calculate the mean and 95%

highest posterior density (HPD) interval for divergence times.

We assessed whether a sample size greater than 200 was

achieved for all parameters after the analyses. Results of the

two runs were combined with LogCombiner 1.4.7 (Drum-

mond & Rambaut, 2007), and the consensus tree was compiled

with TreeAnnotator 1.4.7 (Drummond & Rambaut, 2007).

RESULTS

Alignment and phylogenetic relationships

Our data matrix consisted of 174 sequences (12S: 45, 16S: 46,

18S: 37, cyt b: 46) from 47 taxa (Appendix S1). All coding

sequences could be translated into amino acids with no

evidence of pseudogenes (Song et al., 2008).

The Bayesian and parsimony analyses resulted in very similar

relationships amongst taxa. Differences in topology did not

affect the position of calibration points, and thus the results of

the dating events, with the exception of calibration point F.

In both analyses, all the clades of interest are well supported

(Fig. 2 and Fig. S1 in Appendix S3) and belong to the mono-

phyletic tribe Lebinthini. There are slight differences in the

relationships of three Lebinthus species, but the phylogenetic

positions of New Caledonian taxa do not vary. Cardiodactylus

novaeguineae from Lifou and Vanuatu are sister groups in both

analyses. The position of C. novaeguineae from Grande Terre is

unresolved in the Bayesian tree, whereas a sister relationship

between C. novaeguineae from Grande Terre and from Papua

New Guinea is obtained in the parsimony analysis. Lebinthus

lifouensis is nested within a clade of Lebinthus species from

Vanuatu. The genus Agnotecous is monophyletic and is sister to

the Lebinthus species from Vanuatu and Lifou.

Our phylogenetic results support at least four independent

colonizations of Lebinthini in New Caledonia: C. novaeguineae

on Grande Terre and Lifou independently, L. lifouensis on

Lifou, and Agnotecous on Grande Terre (Fig. 2).

R. Nattier et al.


0.1 substitution/site

Acheta domesticusGryllus bimaculatus

Anurogryllus muticus Gryllitaria sp.

Myara sordidaMyara unicolorArilpa gidya

Eurepella mjobergiEurepella moojerraEurepella torowatta

Salmanites taltantrisSalmanites wittilliko

Eurepa wirkuttaEurepa marginipennis

Eurepa nurndinaEneoptera guyanensisEneoptera gracilis

Eneoptera surinamensisNisitrus vittatus

Paranisitra longipesXenogryllus sp.

Xenogryllus transversusXenogryllus eneopteroides

Swezwilderia sp.Swezwilderia bryani

Ponca venosa Ligypterus fuscus

Ligypterus linharensisCardiodactylus enkraussi

Cardiodactylus tankaraCardiodactylus guttulusCardiodactylus sp.Cardiodactylus novaeguineae

Cardiodactylus novaeguineaeCardiodactylus novaeguineaeCardiodactylus novaeguineae

Lebinthus sp.Lebinthus villemantae

Lebinthus bitaeniatusLebinthus nattawa

Lebinthus lifouensisLebinthus santoensis

Agnotecous albifronsAgnotecous robustus

Agnotecous tapinopusAgnotecous sarramea

Agnotecous yahoue

*

0.99

1

1

1

0.97

1

1

0.52

1

0.95

**

*

**

1

1

1

10.95

1

0.98 *

*

*

*

0.95

1

1

0.57

1

*

*

0.86

0.990.95

1

11

*1

1

1

0.961*

1

0.69

0.52

1

1

1

0.8

0.81

1*

*

* Australia

S. America

MalaysiaPhilippines

Indi

a

E. AfricaFijiSamoa

C. America

S. America

Grande Terre

VanuatuLoyalty Is.VanuatuSingaporeSulawesiPNGVanuatuLoyalty Is.Grande TerrePNGSulawesiJapanVanuatuVanuatu

Pac

ific

A

B

C

FE

D

Eneopterinae

Lebinthini

Figure 2 The 50% majority-rule consensus tree of the Eneopterinae obtained from the Bayesian analysis of the combined data set (18S, 12S,

16S and cytochrome b). Bootstrap support from 1000 pseudo-replicates is indicated above branches by a star (only if ‡ 70%), and the

Bayesian posterior probability is indicated below branches. The geographical distribution is given to the right of the taxa names, and

calibration points are marked on the topology. Species present in New Caledonia are highlighted in bold.



Setting calibration points on the phylogenetic tree

We used the divergence between taxa to set calibration nodes

on the resulting Bayesian phylogeny (Fig. 3 and Table 1). An

exception was node F, which we set at 33 Ma to avoid

incompatibility with node B.

Divergence time estimates

The two combined beast runs yielded high effective sample

sizes (> 200) for all relevant parameters, indicating adequate

sampling of the posterior distribution. Taking the most

conservative approach to dating (i.e. including calibration

6.0 substitutions/site

0 Ma10 Ma20 Ma30 Ma40 Ma50 Ma60 Ma

Gryllus bimaculatus

Eurepa wirkutta

Nisitrus vittatus

Eurepella torowatta

Xenogryllus eneopteroides

Xenogryllus sp.

Eneoptera surinamensis

Cardiodactylus enkraussi

Agnotecous robustus

Ligypterus fuscus

Lebinthus bitaeniatus

Myara unicolor

Arilpa gidya

Ponca venosa

Lebinthus santoensis

Myara sordida

Anurogryllus muticus

Eurepella moojerra

Lebinthus nattawa

Agnotecous yahoue

Ligypterus linharensis

Cardiodactylus tankara

Cardiodactylus sp.

Cardiodactylus novaeguineae (Loyalty Is.)

Eneoptera gracilis

Cardiodactylus novaeguineae (GT)

Cardiodactylus novaeguineae (PNG)

Salmanites wittilliko

Eurepa nurndina

Lebinthus sp.

Xenogryllus transversus

Swezwilderia sp.

Agnotecous sarramea

Paranisitra longipes

Acheta domesticus

Eurepa marginipennis

Eurepella mjobergi

Swezwilderia bryani

Cardiodactylus novaeguineae (Vanuatu)

Lebinthus villemantae

Cardiodactylus guttulus

Gryllitaria sp.

Lebinthus lifouensis

Agnotecous albifrons

Salmanites taltantris

Eneoptera guyanensis

Agnotecous tapinopus

1.9

4.9

2.4

35.6

4.3

20

22.9

18.1

31.7

45.9

19.2

24.5

18

10.6

25.2

2.7

23

7

4.6

7.4

3.3

2.1

13.3

13.7

10.5

0.2

12.6

14.6

23.8

29.5

50.9

17.6

27.8

12.1

6.2

17.6

7.7

15.3

33.6

10.6

8

9.1

6.9

23.9

37 Ma

Lebinthini

36.3

**

******

Eneopterinae

Figure 3 Chronogram representing the divergence times of the principal lineages of Eneopterinae. Divergence times estimated using beast

are shown on the Bayesian consensus topology obtained from the analysis of the concatenated data set. Node positions indicate mean

estimated divergence times, and node bars indicate the 95% confidence intervals. Species present in New Caledonia are highlighted in bold

and with an asterisk. Geographical localization for Cardiodactylus novaeguineae is specified (PNG, Papua New Guinea; GT, Grande Terre).

The bold dotted line indicates the emergence time according to geological evidence.

R. Nattier et al.


node F at 33 Ma and the highest confidence interval), a

maximum age for Lebinthini is estimated as 32.9 Ma (mean:

26.9 Ma; 95% confidence interval: 20.24–32.9 Ma; Table 2).

Without calibration point F, this estimate is 29.7 Ma as a

maximum age (mean: 24.5 Ma; 95% confidence interval: 18.5–

29.7 Ma). Two main clades within this tribe originated

between 19 and 20 Ma: Cardiodactylus at 19.2 Ma (11.8–

26.2 Ma) and Lebinthus–Agnotecous at 20 Ma (13.6–26.3 Ma).

Dating results from the second round of analyses, obtained

with calibration information from outside the Pacific area

(calibration points A, B, C and E), are very close to those from

the first round, which incorporated all calibration points

outside New Caledonia and neighbouring islands (Table 2).

By using the oldest fossil related to Eneopterinae as a

calibration point, the New Caledonian eneopterine clade is

estimated as dating to 24.7 Ma (95% confidence interval: 18.3–

37.8 Ma), the clade containing species from New Caledonia

and the Loyalty Islands to 37.6 Ma (95% confidence interval:

29.9–45.6 Ma), and the whole Pacific clade to 45.6 Ma (95%

confidence interval: 37.7–53.6 Ma) (Fig. S2 in Appendix S3).

By constraining the oldest clade for New Caledonia and the

Loyalty Islands to be 37 Myr old (the postulated re-emergence

time of New Caledonia), we obtain that the Eneopterinae are

92.9 Myr old (95% confidence interval: 76.5–111.7 Ma)

(Fig. S3 in Appendix S3). This means that accepting that the

New Caledonian taxa survived the submersion of New

Caledonia by hopping between the Loyalty Islands and New

Caledonia implies that the eneopterine crickets are close to

100 Myr old.

Our results highlight two phases in the colonization of New

Caledonia. The first is dated at 16.3 Ma as a maximum age

(mean: 10.6 Ma; 95% confidence interval: 5.3–16.3 Ma) with

subsequent diversification of the endemic genus Agnotecous.

The second involves more recent events and is dated around

3–4 Ma as a maximum age: on Grande Terre after 3.3 Ma

(C. novaeguineae), and on Lifou at 4.8 Ma (mean: 2.1 Ma; 95%

confidence interval: 0.5–4.8 Ma) (L. lifouensis) and at 0.6 Ma

(mean: 0.2 Ma; 95% confidence interval: 0.01–0.6 Ma)

(C. novaeguineae). The largest lineage occurring in the Pacific

region, including the Loyalty Islands, Vanuatu, Fiji, Samoa and

New Caledonia, is dated back to 29.5 Ma (27–32 Ma),

notwithstanding the fact that it includes some taxa outside

the Pacific region (Japan, Singapore, South America).

DISCUSSION

Could New Caledonian taxa be dated to more than

37 Ma, prior to the island’s emergence time?

The phylogenetic analysis of the subfamily Eneopterinae shows

that the species of New Caledonia and neighbouring geo-

graphic localities belong to the monophyletic tribe Lebinthini

(Fig. 2). This tribe is sister to a clade containing species from

the Pacific (Fiji, Samoa) and from Central and South America,

but none from Australia. Within Lebinthini, all the species are

from the Pacific region, and New Caledonian lineages are

related to lineages of species from Vanuatu or Papua New

Guinea.

Taking the most conservative dating approach, including

using the upper (oldest) 95% confidence interval, our results

show that the oldest New Caledonian eneopterine clade is no

more than 16 Myr old and is represented by the genus

Agnotecous. This result is consistent with studies on other

insect groups, as well as on other arthropods and plants, which

have dated New Caledonian groups to younger than 32 Ma

and therefore after emergence (Bartish et al., 2005; Page et al.,

2005; Balke et al., 2007b; Harbaugh & Baldwin, 2007; Smith

et al., 2007; Buckley et al., 2010; Espeland & Johanson, 2010).

Even by using the oldest fossil possibly related to Eneopterinae

(96 Ma, and therefore much older than the palaeogeography

estimates) as a calibration point in a conservative approach, we

still obtain the result that the oldest New Caledonian

eneopterine clade is not older than 24.7 Ma, and that the

previous clade linking New Caledonia and the Loyalty Islands

is only 37.6 Myr old.

Some authors have suggested that New Caledonian clades

have been able to maintain a regional presence, despite the

submergence of New Caledonia until 37 Ma, by invoking

hopping among nearby, temporary islands, some of which

have disappeared as a result of erosion, subsidence and/or

subduction (Morat et al., 1986; Heads, 2005, 2008, 2009a,

2010; Ladiges & Cantrill, 2007; Ladiges, 2008). According to

Table 2 Bayesian posterior distributions of the Eneopterinae: mean and 95% highest posterior density for different combinations of

calibration points, as obtained from beast. Results of the first round, incorporating all calibration points outside New Caledonia and

neighbouring islands, are indicated in the first and second columns. Results of the second round, with datings obtained by incorporating

calibration information only from outside the whole Pacific area, are indicated in the third column.

Clades/Taxa Calibration points A–E Calibration points A–F Calibration points A, B, C and E

Lebinthini 24.45 (18.51–29.73) 26.9 (20.24–32.87) 19.86 (13.8–26)

Cardiolactylus 19.24 (11.84–26.24) 21.17 (13.09–29.62) 16.07 (9.9–22.5)

Lebinthus–Agnotecous 20.04 (13.6–26.3) 21.67 (14.65–29.34) 16.6 (11.4–22.4)

Cardiolactylus novaeguineae 3.28 (1.06–6.32) 3.48 (1.09–6.81) 2.87 (1.5–2.9)

Cardiolactylus novaeguineae Lifou-Santo 0.23 (0.01–0.63) 0.25 (0.01–0.7) 0.2 (0.01–0.53)

Lebinthus lifouensis–Lebinthus santoensis 2.1 (0.48–4.79) 2.24 (0.53–5) 1.76 (0.5–3.8)

Agnotecous 10.56 (5.26–16.3) 11.34 (6.08–17.89) 9 (4.9–13.7)



this hypothesis, these clades would be older than the oldest

existing emerged island in the region, namely New Caledonia

itself. Using the most conservative approach, we have tested

this assumption by dating the eneopterine taxa distributed

within the Pacific region: these taxa are related to New

Caledonian Eneopterinae and could have hopped among

Pacific islands during the time New Caledonia was submerged.

Even using this approach, the oldest Eneopterinae in the

Pacific dates back to 32 Ma as a maximum age (mean:

29.5 Ma; 95% confidence interval: 27–32 Ma); that is, several

million years after the emergence of the oldest regional island,

New Caledonia (37 Ma). If we artificially support the island-

hopping hypothesis by constraining the eneopterine clade for

New Caledonia and the Loyalty Islands to be older than 37 Ma,

we obtain the result that the subfamily of Eneopterinae should

be older than 93 Ma. This would mean that the first speciation

and vicariance-like event in eneopterine crickets between

species from Australia and species from other lands should be

47 Myr older than the closest corresponding palaeogeograph-

ical disjunction (the connection between Australia and Ant-

arctica weakened by climate change at 46 Ma). At 93 Ma, the

global configuration of land areas was different from that at

46 Ma (Sanmartın & Ronquist, 2004), with positions of land

masses inconsistent with our phylogenetic results. For exam-

ple, the final separation between Africa and Antarctica

occurred 90–85 Ma, South America was still connected to

North America during the Middle Cretaceous (100 Ma), and

the ‘Tasmania’ block began to drift away and became isolated

c. 80 Ma. All these events could be consistent with our

phylogeny only by the addition of numerous ad hoc hypoth-

eses and by rejecting most vicariance-like speciation events in

the eneopterine clade.

Thus regional island-hopping is neither necessary nor

helpful to explain the presence of eneopterine species in New

Caledonia. In this respect, when using the paleogeographical

calibration deep in the trees, all subsequent dated nodes of the

tree are also consistent with the ages of the various islands.

Therefore, there is no indication for any lineage within the

eneopterine clade that these taxa could be older than any island

in the region. This result is similar to age estimates of taxa

examined in other studies focusing on Pacific island groups.

For example, the freshwater shrimps of Norfolk Island and

New Caledonia were dated as no older than 8.5 Ma (Page

et al., 2005), and the scincid lizards of New Caledonia and New

Zealand (the so-called Tasmantis of the authors) as no older

than 12.7 Ma (Smith et al., 2007). However, our result must

also be compared with two recent regional studies in which a

group was found that was older than the island using

molecular datings with fossil calibrations. In one study, Giribet

et al. (2010) recently found an older date of 77 Ma for a New

Caledonian opilionid clade previously dated at 28–49 Ma with

a smaller taxon sampling (Boyer et al., 2007). In a second

study, scincid lizards were found to be older (16–22.6 Ma)

than Norfolk and Lord Howe Islands (7 and 3 Ma, respec-

tively), as sister groups to New Zealand species, according to a

molecular phylogeny calibrated with a Miocene New Zealand

fossil (Smith et al., 2007; Chapple et al., 2009). These are the

only two cases known from the region, in contrast to the high

number of groups dated older than the islands of geological

hotspots (e.g. Price & Clague, 2002; Parent et al., 2008).

However, it must be noted that such datings for opilionids and

lizards do not necessarily corroborate an island-hopping

scenario that implies several short-distance dispersal events

among neighbouring islands. An age older than the island

could also be explained by a long-distance dispersal event from

a continental source, now extinct, near to the island. In fact,

there is no way to determine a scenario of island-hopping and

short-distance dispersal in preference to a scenario of a long-

distance dispersal event (P. Grandcolas et al., in prep.). In

contrast, when the group is dated as younger than the island,

as in the present case of eneopterine crickets, the island-

hopping scenario is clearly refuted for that group.

Do New Caledonian taxa show evidence for local

radiations or for multiple colonizations?

Our results indicate an initial eneopterine colonization of New

Caledonia involving Agnotecous at 16.3 Ma as a maximum

age (mean: 10.6 Ma; 95% confidence interval: 5.3–16.3 Ma).

The local radiation of Agnotecous took place approximately at

the midpoint between the older diversifications, as found in the

Sapotaceae (< 32.4 Ma, Bartish et al., 2005) and Trichoptera

(28.2 Ma, Espeland & Johanson, 2010), and the more recent

radiations, as found in Angustonicus cockroaches (2 Ma,

Murienne et al., 2005), the sandalwood genus Santalum

(1–1.5 Ma, Harbaugh & Baldwin, 2007) and the freshwater

shrimp Paratya (3.5–8.5 Ma, Page et al., 2005).

This initial colonization event involving Eneopterinae was

followed by at least three colonizations within the tribe

Lebinthini during the last 4 Myr. Two events concern the

widespread species C. novaeguineae (later than 3.3 Ma on

Grande Terre and around 0.2 Ma on Lifou). The wide

distribution of this species could be correlated with key

biological attributes such as vagility (long wings) or use of

secondary habitats (mostly coastal). Similarly, the ancestor of

L. lifouensis colonized Lifou no earlier than 2.1 Ma, apparently

from Vanuatu (Fig. 2).

Multiple colonizations of New Caledonia within a single

regional group have previously been shown in arthropods

(Balke et al., 2007b; Edgecombe & Giribet, 2009) and plants

(Bartish et al., 2005; Duangjai et al., 2009) but have always

concerned ancient events. In the present case, a single clade of

eneopterine crickets exhibits both an old diversification and

multiple, very recent dispersal events to New Caledonia and

the neighbouring islands.

From a regional biogeographical point of view, our results

also favour the scenario that the eneopterine crickets of New

Caledonia and the islands of the South Pacific have a

proximate origin in Indo-Malaysia rather than in Australia,

confirming the interpretation, based on morphological anal-

ysis, of Desutter-Grandcolas & Robillard (2006). From a New

Caledonian point of view, this does not contradict earlier views

R. Nattier et al.


because many groups were often supposed to have largely

Australian or Indo-Malaysian origins (e.g. Morat et al., 1986).

Do phylogenetic trees indicate relationships between

New Caledonian and geographically distant taxa,

suggesting that they are local relicts?

The ancient origin of the New Caledonian biota was most

often assumed, based on the presence of supposed relict taxa

that had survived there and gone extinct everywhere else

(Raven & Axelrod, 1972; Morat et al., 1986; Lowry, 1998).

However, only a few of these taxa have recently been validated

as true relicts, being phylogenetically related by long branches

to geographically distant or globally distributed relatives

(Grandcolas et al., 2008), for example Amborella, the sister

group of all other flowering plants (Parkinson et al., 1999), the

kagu (Rhynochetos jubatus, Cracraft, 2001), and some troglos-

ironid harvestmen (Sharma & Giribet, 2009). Could Agnote-

cous, an endemic and diversified genus of Grande Terre, be

considered a local relict? The sister group of Agnotecous,

composed of three Lebinthus species, has a restricted distribu-

tion in the geographic region of New Caledonia (Vanuatu for

L. nattawa and L. santoensis, and the Loyalty Islands for

L. lifouensis). Moreover, the genus is not connected to the rest

of the phylogeny by a long branch (Fig. 2). None of these

relationships supports a hypothesis of major extinction events

in this clade.

CONCLUSIONS

The number of phylogenetic studies focusing on New Cale-

donian groups is increasing rapidly, and this should allow

testing of existing biogeographical models in the next few

years. The first step towards this objective was to revisit the

biogeography of New Caledonia, emphasizing which tests

would be especially relevant (Grandcolas et al., 2008). It was

necessary to determine whether some regionally endemic

groups have dates of origin and relationships to taxa of other

areas consistent with a Gondwanan origin. Unsurprisingly,

most if not all phylogenetic studies invalidate a simplistic

scenario of local survival since 80 Ma, the date of the rift of the

geological basement from Australia (Grandcolas et al., 2008).

The second step requires more sophisticated tests to

investigate whether some old regional groups have survived

by island-hopping in spite of the submergence of various land

masses (Morat et al., 1986; Heads, 2005, 2008, 2009a, 2010;

Ladiges & Cantrill, 2007; Ladiges, 2008). The present study

focuses only on this aim, within a conservative testing

framework, and refutes the island-hopping model in the case

of an old radiation of eneopterine crickets. It would be

interesting to perform similar analyses on other taxonomic

groups, especially those including relict taxa, to detect possible

presence in the region pre-dating the emergence of New

Caledonia at 37 Ma. A first analysis pointing in this direction is

found in the general study of opilionids by Giribet et al.

(2010).

More work is clearly needed in this respect, especially to

distinguish between island-hopping sensu stricto and long-

distance dispersal events. This is also why it is important to

explore further the possibility that many groups may be relict

in New Caledonia, an old intuitive assumption that remains

poorly supported, with only three documented cases (Parkin-

son et al., 1999; Cracraft, 2001; Sharma & Giribet, 2009). In

conclusion, it is clear that New Caledonia is a remarkable

natural laboratory for conducting a range of tests of evolution,

being a very old oceanic island harbouring relicts, palaeoen-

demics and neoendemics in a regionally well-known tectonic

context.

ACKNOWLEDGEMENTS

We are grateful to the following institutions or programmes

that made our work possible: the French Agence Nationale de

la Recherche (ANR) for funding via the Biodiversity research

grant BIONEOCAL, to P.G.; the Museum national d’Histoire

naturelle (MNHN); the Ministry of Research for grants to

Philippe Janvier; the Gouvernement de Nouvelle-Caledonie for

funding R.N.’s PhD work; and also the Prevost Fund of the

Ecole Doctorale of the Museum, and the Directions de

l’Environnement of New Caledonia’s Province Nord and

Province Sud for permits to work in natural parks and forest

reserves. This work was supported by the Consortium National

de Recherche en Genomique’, and the MNHN (CNRS UMS

2700). It was carried out under agreement no. 2005/67 between

Genoscope and the MNHN for the project ‘Macrophylogeny of

life’ directed by Guillaume Lecointre. Herve Jourdan, Christian

Mille, Judith Najt and Fabrice Colin provided valuable help

and are sincerely thanked. We are also grateful to Robert

Cowie, Porter P. Lowry, Marianne Elias and Herve Jourdan for

their critical reading of the paper.

REFERENCES

Aitchison, J.C., Clarke, G.L., Meffre, S. & Cluzel, D. (1995)

Eocene arc–continent collision in New Caledonia and

implications for regional southwest Pacific tectonic evolu-

tion. Geology, 23, 161–164.

Akaike, H. (1973) Information theory and an extension of the

maximum likelihood principle. Second International Sym-

posium on Information Theory (ed. by B. Petran and

F. Csaki), pp. 267–281. Akademiai Kiado, Budapest.

Akaike, H. (1974) A new look at the statistical model identi-

fication. IEEE Transactions on Automatic Control, 19, 716–

723.

Axelrod, D.I. (1972) Ocean-floor spreading in relation to

ecosystematic problems. Symposium on Ecosystematics (1971:

University of Arkansas) (ed. by R.T. Allen and F.C. James),

pp. 15–68. Occasional Paper No. 4, University of Arkansas

Museum, Fayetteville, AR.

Balke, M., Pons, J., Ribera, I., Sagata, K. & Vogler, A.P. (2007a)

Infrequent and unidirectional colonization of hyperdiverse



Papuadytes diving beetles in New Caledonia and New Gui-

nea. Molecular Phylogenetics and Evolution, 42, 505–516.

Balke, M., Wewalka, G., Alarie, Y. & Ribera, I. (2007b)

Molecular phylogeny of Pacific island Colymbetinae: radi-

ation of New Caledonian and Fijian species (Coleoptera,

Dytiscidae). Zoologica Scripta, 36, 173–200.

Bartish, I.V., Swenson, U., Munzinger, J. & Anderberg, A.A.

(2005) Phylogenetic relationships among New Caledonian

Sapotaceae (Ericales): molecular evidence for generic

polyphyly and repeated dispersal. American Journal of

Botany, 92, 667–673.

Bartoli, G., Sarnthein, M., Weinelt, M., Erlenkeuser, H., Garbe-

Schonberg, D. & Lea, D.W. (2005) Final closure of Panama

and the onset of Northern Hemisphere glaciation. Earth and

Planetary Science Letters, 237, 33–44.

Bernor, R.L., Brunet, M., Ginsburg, L., Mein, P., Pickford, M.,

Rogl, F., Sen, S., Steininger, F. & Thomas, H. (1987) A

consideration of some major topics concerning Old World

Miocene mammalian chronology, migrations and paleoge-

ography. Geobios, 20, 431–439.

Boyer, S. L., Clouse, R. M., Benavides, L. R., Sharma, P.,

Schwendinger, P. J., Karunarathna, I. & Giribet, G. (2007)

Biogeography of the world: a case study from cyphoph-

thalmid opiliones, a globally distributed group of arachnids.

Journal of Biogeography, 34, 2070–2085.

Buckley, T.R., Attanayake, D., Nylander, J.A.A. & Bradler, S.

(2010) The phylogenetic placement and biogeographical

origins of the New Zealand stick insects (Phasmatodea).

Systematic Entomology, 35, 207–225.

Burnham, R.J. & Graham, A. (1999) The history of neotropical

vegetation: new developments and status. Annals of the

Missouri Botanical Garden, 86, 546–589.

Carlquist, S. (1965) Island life. The Natural History Press, New

York.

Carlquist, S. (1974) Island biology. Columbia University Press,

New York.

Chapple, D.G., Ritchie, P.A. & Daugherty, C.H. (2009) Origin,

diversification, and systematics of the New Zealand skink

fauna (Reptilia: Scincidae). Molecular Phylogenetics and

Evolution, 52, 470–487.

Chazeau, J. (1993) Research on New Caledonian terrestrial

fauna: achievements and prospects. Biodiversity Letters, 1,

123–129.

Chevillotte, V., Chardon, D., Beauvais, A., Maurizot, P. & Colin,

F. (2006) Long-term tropical morphogenesis of New Cale-

donia (Southwest Pacific): importance of positive epeirogeny

and climate change. Geomorphology, 81, 361–375.

Cluzel, D., Aitchison, J.C. & Picard, C. (2001) Tectonic

accretion and underplating of mafic terranes in the Late

Eocene intraoceanic fore-arc of New Caledonia (Southwest

Pacific): geodynamic implications. Tectonophysics, 340, 23–

59.

Cox, B. & Moore, P.D. (1993) Biogeography: an ecological and

evolutionary approach, 5th edn. Blackwell Science, Oxford.

Cracraft, J. (2001) Avian evolution, Gondwana biogeography

and the Cretaceous–Tertiary mass extinction event.

Proceedings of the Royal Society B: Biological Sciences, 268,

459–469.

Crawford, A.J., Meffre, S. & Symonds, P.A. (2003) Chapter 25:

120 to 0 Ma tectonic evolution of the Southwest Pacific and

analogous geological evolution of the 600 to 220 Ma Tas-

man fold belt system. Geological Society of Australia Special

Publication, 22, 377–397.

Crisp, M., Trewick, S.A. & Cook, L. (2011) Hypothesis testing

in biogeography. Trends in Ecology and Evolution, 26, 66–72.

Darlington, P.J. (1957) Zoogeography: the geographical distri-

bution of animals. Wiley, New York.

Desutter-Grandcolas, L. & Robillard, T. (2006) Phylogenetic

systematics and evolution of Agnotecous in New Caledonia

(Orthoptera: Grylloidea, Eneopteridae). Systematic Ento-

mology, 31, 65–92.

Drummond, A.J. & Rambaut, A. (2007) Beast: Bayesian evo-

lutionary analysis by sampling trees. BMC Evolutionary

Biology, 7, 214.

Drummond, A.J., Ho, S.Y.W., Phillips, M.J. & Rambaut, A.

(2006) Relaxed phylogenetics and dating with confidence.

PLoS Biology, 4, 699–710.

Duangjai, S., Samuel, R., Munzinger, J., Forest, F., Wallnofer, B.,

Barfuss, M.H.J., Fischer, G. & Chase, M.W. (2009) A multi-

locus plastid phylogenetic analysis of the pantropical genus

Diospyros (Ebenaceae), with an emphasis on the radiation and

biogeographic origins of the New Caledonian endemic spe-

cies. Molecular Phylogenetics and Evolution, 52, 602–620.

Edgar, R.C. (2004) MUSCLE: multiple sequence alignment

with high accuracy and high throughput. Nucleic Acids

Research, 32, 1792–1797.

Edgecombe, G.D. & Giribet, G. (2009) Phylogenetics of Scut-

igeromorph centipedes (Myriapoda: Chilopoda) with

implications for species delimitation and historical biogeo-

graphy of the Australian and New Caledonian faunas.

Cladistics, 25, 406–427.

Espeland, M. & Johanson, K.A. (2010) The effect of environ-

mental diversification on species diversification in New

Caledonian caddisflies (Insecta: Trichoptera: Hydropsychi-

dae). Journal of Biogeography, 37, 879–890.

Felsenstein, J. (1985) Confidence-limits on phylogenies: an

approach using the bootstrap. Evolution, 39, 783–791.

Fleischer, R.C., Mcintosh, C.E. & Tarr, C.L. (1998) Evolution

on a volcanic conveyor belt: using phylogeographic recon-

structions and K–Ar-based ages of the Hawaiian islands to

estimate molecular evolutionary rates. Molecular Ecology, 7,

533–545.

Gheerbrant, E. & Rage, J.C. (2006) Paleobiogeography of

Africa: how distinct from Gondwana and Laurasia? Palaeo-

geography, Palaeoclimatology, Palaeoecology, 241, 224–246.

Giribet, G., Vogt, L., Gonzalez, A.P., Sharma, P. & Kury, A.B.

(2010) A multilocus approach to harvestman (Arachnida:

Opiliones) phylogeny with emphasis on biogeography and

the systematics of Laniatores. Cladistics, 26, 408–437.

Goloboff, P.A. (1999) Analyzing large data sets in reasonable

times: solutions for composite optima. Cladistics, 15, 415–

428.

R. Nattier et al.


Goloboff, P.A., Farris, J.S. & Nixon, K.C. (2008) TNT, a free

program for phylogenetic analysis. Cladistics, 24, 774–786.

Gorochov, A.V. (1985) Mesozoic crickets (Orthoptera, Gryl-

loidea) of Asia. Paleontologicheskii Zhurnal, 2, 59–68.

Grandcolas, P., Murienne, J., Robillard, T., Desutter-Grandc-

olas, L., Jourdan, H., Guilbert, E. & Deharveng, L. (2008)

New Caledonia: a very old Darwinian island? Philosophical

Transactions of the Royal Society B: Biological Sciences, 363,

3309–3317.

Gressitt, J.L. (1956) Some distribution patterns of Pacific

island faunae. Systematic Zoology, 5, 11–32.

Harbaugh, D.T. & Baldwin, B.G. (2007) Phylogeny and bio-

geography of the sandalwoods (Santalum, Santalaceace):

repeated dispersals throughout the Pacific. American Journal

of Botany, 94, 1028–1040.

Hart, S.R., Coetzee, M., Workman, R.K., Blusztajn, J., Johnson,

K.T.M., Sinton, J.M., Steinberger, B. & Hawkins, J.W.

(2004) Genesis of the Western Samoa seamount province:

age, geochemical fingerprint and tectonics. Earth and

Planetary Science Letters, 227, 37–56.

Haug, G.H. & Tiedemann, R. (1998) Effect of the formation of

the isthmus of Panama on Atlantic Ocean thermohaline

circulation. Nature, 393, 673–676.

Heads, M. (2005) Dating nodes on molecular phylogenies: a

critique of molecular biogeography. Cladistics, 21, 62–78.

Heads, M. (2008) Panbiogeography of New Caledonia, south-

west Pacific: basal angiosperms on basement terranes,

ultramafic endemics inherited from volcanic island arcs and

old taxa endemic to young islands. Journal of Biogeography,

35, 2153–2175.

Heads, M. (2009a) Globally basal centres of endemism: the

Tasman-Coral Sea region (south-west Pacific), Latin

America and Madagascar/South Africa. Biological Journal of

the Linnean Society, 96, 222–245.

Heads, M. (2009b) Inferring biogeographic history from

molecular phylogenies. Biological Journal of the Linnean

Society, 98, 757–774.

Heads, M. (2010) The endemic plant families and the palms of

New Caledonia: a biogeographical analysis. Journal of Bio-

geography, 37, 1239–1250.

Ho, S.Y.W. (2007) Calibrating molecular estimates of substi-

tution rates and divergence times in birds. Journal of Avian

Biology, 38, 409–414.

Ho, S.Y.W., Saarma, U., Barnett, R., Haile, J. & Shapiro, B.

(2008) The effect of inappropriate calibration: three case

studies in molecular ecology. PLoS ONE, 3, e1615.

Huelsenbeck, J.P. & Crandall, K.A. (1997) Phylogeny estima-

tion and hypothesis testing using maximum likelihood.

Annual Review of Ecology and Systematics, 28, 437–466.

Jaffre, T. (1992) Floristic and ecological diversity of the vege-

tation on ultramafic rocks in New Caledonia. The vegetation

of ultramafic (serpentine) soils (ed. by A.J.M. Baker, J. Proc-

tor and R.D. Reeves), pp. 101–107. Intercept Ltd., Andover.

Kluge, A.G. (1989) A concern for evidence and a phylogenetic

hypothesis of relationships among Epicrates (Boidae, Ser-

pentes). Systematic Zoology, 38, 7–25.

Kodandaramaiah, U. (2011) Tectonic calibrations in molecular

dating. Current Zoology, 57, 116–124.

Kosuch, J., Vences, M., Dubois, A., Ohler, A. & Bohme, W.

(2001) Out of Asia: mitochondrial DNA evidence for an

oriental origin of tiger frogs, genus Hoplobatrachus. Molec-

ular Phylogenetics and Evolution, 21, 398–407.

Ladiges, P.Y. (2008) Synthesizing biotic patterns and geology

for New Caledonia. Journal of Biogeography, 35, 2151–2152.

Ladiges, P.Y. & Cantrill, D. (2007) New Caledonia–Australian

connections: biogeographic patterns and geology. Australian

Systematic Botany, 20, 383–389.

de Laubenfels, D.J. (1996) Gondwanan conifers on the Pacific

rim. The origin and evolution of Pacific islands biotas, New

Guinea to Eastern Polynesia: patterns and processes (ed. by A.

Keast and S.E. Miller), pp. 261–265. SPB Academic Pub-

lishing, Amsterdam.

Lee, M.S.Y. (1999) Molecular clock calibrations and metazoan

divergence dates. Journal of Molecular Evolution, 49, 385–

391.

Linder, H.P., Hardy, C.R. & Rutschmann, F. (2005) Taxon

sampling effects in molecular clock dating: an example from

the African Restionaceae. Molecular Phylogenetics and Evo-

lution, 35, 569–582.

Livermore, R., Nankivell, A., Eagles, G. & Morris, P. (2005)

Paleogene opening of Drake passage. Earth and Planetary

Science Letters, 236, 459–470.

Lowry, P.P. (1998) Diversity, endemism, and extinction in the

flora of New Caledonia: a review. Proceedings of the Inter-

national Symposium: Rare, Threatened, and Endangered

Floras of Asia and the Pacific (ed. by C.I. Peng and P.P.

Lowry), pp. 181–206. Institute of Botany, Academica Sinica,

Taipei, Taiwan.

Lucky, A. & Sarnat, E. M. (2010) Biogeography and diversifi-

cation of the Pacific ant genus Lordomyrma Emery. Journal

of Biogeography, 37, 624–634.

Maddison, D.R. (1991) The discovery and importance of

multiple islands of most-parsimonious trees. Systematic

Zoology, 40, 315–328.

Magallon, S.A. (2004) Dating lineages: molecular and pale-

ontological approaches to the temporal framework of clades.

International Journal of Plant Sciences, 165, S7–S21.

McLoughlin, S. (2001) The breakup history of Gondwana and

its impact on pre-Cenozoic floristic provincialism. Austra-

lian Journal of Botany, 49, 271–300.

Morat, P., Jaffre, T., Veillon, J.M. & MacKee, H.S. (1986)

Affinites floristiques et considerations sur l’origine des

maquis miniers de la Nouvelle-Caledonie. Adansonia, 2, 133–

182.

Murienne, J., Grandcolas, P., Piulachs, M., Belles, X., D’Haese,

C., Legendre, F., Pellens, R. & Guilbert, E. (2005) Evolution

on a shaky piece of Gondwana: is local endemism recent in

New Caledonia? Cladistics, 21, 2–7.

Murienne, J., Pellens, R., Budinoff, R.B., Wheeler, W.C. &

Grandcolas, P. (2008) Phylogenetic analysis of the endemic

New Caledonian cockroach Lauraesilpha. Testing competing

hypotheses of diversification. Cladistics, 24, 802–812.



Natland, J.H. (1980) The progression of volcanism in the

Samoan linear volcanic chain. American Journal of Science,

280, 709–735.

Neall, V.E. & Trewick, S.A. (2008) The age and origin of the

Pacific islands: a geological overview. Philosophical Trans-

actions of the Royal Society B: Biological Sciences, 363, 3293–

3308.

Nixon, K.C. (1999) The parsimony ratchet, a new method for

rapid parsimony analysis. Cladistics, 15, 407–414.

Nixon, K.C. & Carpenter, J.M. (1996) On simultaneous anal-

ysis. Cladistics, 12, 221–241.

Page, T.J., Baker, A.M., Cook, B.D. & Hughes, J.M. (2005)

Historical transoceanic dispersal of a freshwater shrimp: the

colonization of the South Pacific by the genus Paratya

(Atyidae). Journal of Biogeography, 32, 581–593.

Parent, C.E., Caccone, A. & Petren, K. (2008) Colonization

and diversification of Galapagos terrestrial fauna: a phy-

logenetic and biogeographical synthesis. Philosophical

Transactions of the Royal Society B: Biological Sciences, 363,

3347–3361.

Paris, J.P. (1981) Geologie de la Nouvelle-Caledonie. Un essai

de synthese (Memoire pour servir notice explicative a la

carte geologique de la Nouvelle-Caledonie a l’echelle du

1/200000). Memoires du Bureau de Recherches Geologiques

et Minieres, 113, 1–278.

Parkinson, C.L., Adams, K.L. & Palmer, J.D. (1999) Multigene

analyses identify the three earliest lineages of extant flow-

ering plants. Current Biology, 9, 1485–1488.

Pelletier, B. (2006) Geology of the New Caledonia region and

its implications for the study of the New Caledonian bio-

diversity. Compendium of marines species from New Cale-

donia, Forum Biodiversite des Ecosystemes Coralliens, 30

octobre–4 novembre 2006, Noumea, Nouvelle-Caledonie (ed.

by C.E. Payri and B. Richer de Forges), pp. 17–30. Institut de

Recherche pour le Developpement, Noumea, France.

Posada, D. (2008) JModelTest: phylogenetic model averaging.

Molecular Biology and Evolution, 25, 1253–1256.

Price, J.P. & Clague, D.A. (2002) How old is the Hawaiian

biota? Geology and phylogeny suggest recent divergence.

Proceedings of the Royal Society B: Biological Sciences, 269,

2429–2435.

Pulquerio, M.J.F. & Nichols, R.A. (2007) Dates from the

molecular clock: how wrong can we be? Trends in Ecology

and Evolution, 22, 180–184.

Pybus, O.G. (2006) Model selection and the molecular clock.

PLoS Biology, 4, 686–688.

Rambaut, A. & Drummond, A.J. (2007) Tracer v1.4. Available

at: http://beast.bio.ed.ac.uk/tracer.

Raven, P.H. (1979) Plate tectonics and Southern Hemisphere

biogeography. Tropical botany (ed. by K. Larsen and L.B.

Holm-Nielsen), pp. 3–24. Academic Press, London.

Raven, P.H. & Axelrod, D.I. (1972) Plate tectonics and Aus-

tralasian paleobiogeography. Science, 176, 1379–1386.

Robillard, T. (2011) Centuriarus, a new genus of Eneopterinae

crickets from Papua (Insecta, Orthoptera, Grylloidea).

Zoosystema, 33, 49–60.

Robillard, T. & Desutter-Grandcolas, L. (2004a) Phylogeny

and the modalities of acoustic diversification in extant

Eneopterinae (Insecta, Orthoptera, Grylloidea, Eneopteri-

dae). Cladistics, 20, 271–293.

Robillard, T. & Desutter-Grandcolas, L. (2004b) High-

frequency calling in Eneopterinae crickets (Orthoptera,

Grylloidea, Eneopteridae): an adaptive radiation revealed by

phylogenetic analysis. Biological Journal of the Linnean

Society, 83, 577–584.

Robillard, T. & Desutter-Grandcolas, L. (2006) Phylogeny of

the cricket subfamily Eneopterinae (Orthoptera, Grylloidea,

Eneopteridae) based on four molecular loci and morphol-

ogy. Molecular Phylogenetics and Evolution, 40, 643–661.

Robillard, T. & Desutter-Grandcolas, L. (2008) Clarification of

the taxonomy of extant crickets of the subfamily Eneop-

terinae (Orthoptera: Grylloidea; Gryllidae). Zootaxa, 1789,

66–68.

Robillard, T. & Desutter-Grandcolas, L. (2011a) The complex

stridulatory behavior of the cricket Eneoptera guyanensis

Chopard (Orthoptera: Grylloidea: Eneopterinae). Journal of

Insect Physiology, 57, 694–703.

Robillard, T. & Desutter-Grandcolas, L. (2011b) Evolution of

calling songs as multicomponent signals in crickets (Orthop-

tera: Grylloidea: Eneopterinae). Behaviour, 148, 627–672.

Robillard, T. & Ichikawa, A. (2009) Redescription of two

Cardiodactylus species (Orthoptera, Grylloidea, Eneopteri-

nae): the supposedly well-known C. novaeguineae (Haan,

1842), and the semi-forgotten C. guttulus (Matsumura,

1913) from Japan. Zoological Science, 26, 878–891.

Ronquist, F. & Huelsenbeck, J.P. (2003) MrBayes 3: Bayesian

phylogenetic inference under mixed models. Bioinformatics,

19, 1572–1574.

Rutschmann, F. (2006) Molecular dating of phylogenetic trees:

a brief review of current methods that estimate divergence

times. Diversity and Distributions, 12, 35–48.

Sanderson, M.J., Thorne, J.L., Wikstrom, N. & Bremer, K.

(2004) Molecular evidence on plant divergence times.

American Journal of Botany, 91, 1656–1665.

Sanmartın, I. & Ronquist, F. (2004) Southern Hemisphere

biogeography inferred by event-based models: plant versus

animal patterns. Systematic Biology, 53, 216–243.

Sharma, P. & Giribet, G. (2009) A relict in New Caledonia:

phylogenetic relationships of the family Troglosironidae

(Opiliones: Cyphophthalmi). Cladistics, 25, 279–294.

Sharov, A.G. (1968) The phylogeny of the orthopteroid insects.

Trudy Paleontologicheskogo Instituta, 118, 184–189.

Smith, S.A., Sadlier, R.A., Bauer, A.M., Austin, C.C. & Jack-

man, T. (2007) Molecular phylogeny of the scincid lizards of

New Caledonia and adjacent areas: evidence for a single

origin of the endemic skinks of Tasmantis. Molecular Phy-

logenetics and Evolution, 43, 1151–1166.

Song, H., Buhay, J.E., Whiting, M.F. & Crandall, K.A. (2008)

Many species in one: DNA barcoding overestimates the

number of species when nuclear mitochondrial pseudogenes

are coamplified. Proceedings of the National Academy of

Sciences USA, 105, 13486–13491.

R. Nattier et al.


Swenson, U., Backlund, A., McLoughlin, S. & Hill, R.S. (2001)

Nothofagus biogeography revisited with special emphasis on

the enigmatic distribution of subgenus Brassospora in New

Caledonia. Cladistics, 17, 28–47.

Swofford, D.L. (2002) PAUP*: phylogenetic analysis using par-

simony (*and other methods), 4.0 edn. Sinauer, Sunderland,

MA.

Tiffney, B.H. & Manchester, S.R. (2001) The use of geological

and paleontological evidence in evaluating plant phylogeo-

graphic hypotheses in the Northern Hemisphere Tertiary.

International Journal of Plant Sciences, 162, S3–S17.

Veevers, J.J., Powell, C.M. & Roots, S.R. (1991) Review of sea-

floor spreading around Australia. 1. Synthesis of the patterns

of spreading. Australian Journal of Earth Sciences, 38, 373–

389.

Wang, D.Y.C., Kumar, S. & Hedges, S.B. (1999) Divergence

time estimates for the early history of animal phyla and the

origin of plants, animals and fungi. Proceedings of the Royal

Society B: Biological Sciences, 266, 163–171.

Waters, J.M. & Craw, D. (2006) Goodbye Gondwana? New

Zealand biogeography, geology, and the problem of circu-

larity. Systematic Biology, 55, 351–356.

Woodburne, M.O. & Chase, J.A. (1996) Dispersal, vicariance,

and the Late Cretaceous to early Tertiary land mammal

biogeography from South America to Australia. Journal of

Mammalian Evolution, 3, 121–161.

SUPPORTING INFORMATION

Additional Supporting Information may be found in the

online version of this article:

Appendix S1 List of taxa for the Eneopterinae and

outgroups.

Appendix S2 Polymerase chain reaction primers.

Appendix S3 Supplementary figures (Figs S1–S3).

As a service to our authors and readers, this journal provides

supporting information supplied by the authors. Such mate-

rials are peer-reviewed and may be re-organized for online

delivery, but are not copy-edited or typeset. Technical support

issues arising from supporting information (other than

missing files) should be addressed to the authors.

BIOSKETCH

Romain Nattier is a PhD student at the Museum national

d’Histoire naturelle (Paris, France). His interests include the

evolution and the diversification of the New Caledonian biota,

with a special focus on orthopteroid insects.

Author contributions: R.N. analysed and interpreted the data;

T.R. contributed to the phylogenetic sampling and framework;

A.C. helped to obtain DNA sequences; L.D.-G. contributed to

the biogeographical and phylogenetic framework; and P.G.

conceived the biogeographical context of the study.

Editor: Robert Whittaker



Date post:	18-Nov-2023
Category:	Documents
Upload:	mnhn
View:	0 times
Download:	0 times

Older than New Caledonia emergence? A molecular phylogenetic study of the eneopterine crickets...

Documents