Ioana C Chintauan-Marquier

Ed Eric Legendre

Sylvain Hugel

Tony Robillard

Philippe Grandcolas

Andr E Nel

Dario Zuccon

Laure Desutter-Grandcolas

Laying the foundations of evolutionary and systematic studies in crickets (Insecta, Orthoptera): a multilocus phylogenetic analysis

Orthoptera have been used for decades for numerous evolutionary questions but several of its constituent groups, notably crickets, still suffer from a lack of a robust phylogenetic hypothesis. We propose the first phylogenetic hypothesis for the evolution of crickets sensu lato, based on analysis of 205 species, representing 88% of the subfamilies and 71% tribes currently listed in the database Orthoptera Species File (OSF). We reconstructed parsimony, maximum likelihood and Bayesian phylogenies using fragments of 18S, 28SA, 28SD, H3, 12S, 16S, and cytb (~3600 bp). Our results support the monophyly of the cricket clade, and its subdivision into two clades: mole crickets and ant-loving crickets on the one hand, and all the other crickets on the other (i.e. crickets sensu stricto). Crickets sensu stricto form seven monophyletic clades, which support part of the OSF families, "subfamily groups", or subfamilies: the mole crickets (OSF Gryllotalpidae), the scaly crickets (OSF Mogoplistidae), and the true crickets (OSF Gryllidae) are recovered as monophyletic. Among the 22 sampled subfamilies, only six are monophyletic: Gryllotalpinae, Trigonidiinae, Pteroplistinae, Euscyrtinae, Oecanthinae, and Phaloriinae. Most of the 37 tribes sampled are paraor polyphyletic. We propose the best-supported clades as backbones for future definitions of familial groups, validating some taxonomic hypotheses proposed in the past. These clades fit variously with the morphological characters used today to identify crickets. Our study emphasizes the utility of a classificatory system that accommodates diagnostic characters and monophyletic units of evolution. Moreover, the phylogenetic hypotheses proposed by the present study open new perspectives for further evolutionary research, especially on acoustic communication and biogeography.

Introduction

For more than 30 years now, phylogenetics has become the reference system in a tree-thinking perspective for evolutionary biology [START_REF] Eldredge | Phylogenetic Patterns and the Evolutionary Process, Method and Theory in Comparative Biology[END_REF][START_REF] Carpenter | Testing scenarios: wasp social behavior[END_REF]Brooks andMcLennan, 1991, 2002;[START_REF] O'hara | Telling the tree: narrative representation and the study of evolutionary history[END_REF][START_REF] Larsen | Phylogenetic systematics of adaptation[END_REF]. Phylogenetic trees are currently used to propose and test evolutionary hypotheses in all domains where evolution may interplay with systems studied. As a consequence, phylogenies have also become the backbone of modern classifications. Both fields of application are important and require robust phylogenetic hypotheses based on relevant and broad taxonomic sampling.

Studies on orthopteran insects have investigated many important aspects of animal behaviour, from the neural basis of behaviour to the ecology of acoustic communication [START_REF] Gwynne | Orthopteran Mating Systems[END_REF][START_REF] Huber | Cricket Behavior and Neurobiology[END_REF][START_REF] Bailey | The Tettigoniidae, Biology, Systematics and Evolution[END_REF][START_REF] Field | The Biology of Wetas, King Crickets and their Allies[END_REF][START_REF] Gwynne | Katydids and Bush-crickets, Reproductive Behavior and Evolution of Tettigoniidae[END_REF]. However, the lack of phylogenetic foundation has made it difficult to interpret the results of these studies in an evolutionary or comparative framework. In the past, taxonomy has been used as a proxy for phylogeny, with hypotheses of "lower" (i.e. "primitive") and "higher" taxa, and varied assumptions about "ancestral" character states and evolutionary series [START_REF] Baccetti | Spermatozoa and phylogeny in orthopteroid insects[END_REF][START_REF] Bailey | Acoustic Behaviour of Insects. An Evolutionary Perspective[END_REF][START_REF] Otte | Evolution of cricket songs[END_REF][START_REF] Field | Structure and evolution of stridulatory mechanisms in New Zealand wetas (Orthoptera: Stenopelmatidae)[END_REF]. This applies also to the numerous, and frequently well preserve, orthopteran fossils, which are currently not classified in a satisfactory, phylogenetic manner, resulting in a majority of fossil families and subfamilies not correctly defined on the basis of clear synapomorphies. Only one preliminary cladistic analysis was made on the basis of the forewing venation [START_REF] Nel | Venation pattern and revision of Orthoptera sensu nov. and sister groups. Phylogeny of Palaeosoic and Mesozoic Orthoptera sensu nov[END_REF], but it would need significant improvement by the addition of the numerous fossil taxa more recently described and a better comparison with the recent taxa.

Earlier attempts to reconstruct the molecular phylogeny of Orthoptera and their suborders took into account too few taxa and markers to achieve any stability and robustness (Flook andRowell, 1997a, 1998;[START_REF] Rowell | Phylogeny of the Caelifera and the Orthoptera as derived from ribosomal gene sequences[END_REF][START_REF] Flook | Combined molecular phylogenetic analysis of the Orthoptera (Arthropoda, Insecta) and implications for their higher systematics[END_REF][START_REF] Jost | Phylogeny of Ensifera (Hexapoda: Orthoptera) using three ribosomal loci, with implications for the evolution of acoustic communication[END_REF], as shown in [START_REF] Legendre | One hundred years of instability in ensiferan relationships[END_REF] for Ensifera, and most published phylogenies have been performed at a limited taxonomic scale [START_REF] Chapco | A molecular biogeographic analysis of the relationship between North American melanoploid grasshoppers and their Eurasian and South American relatives[END_REF][START_REF] Litzenberger | Molecular phylogeny of selected Eurasian Podismine grasshoppers (Orthoptera: Acrididae)[END_REF][START_REF] Bugrov | Molecular phylogeny of palaearctic genera of Gomphocerinae grasshoppers (Orthoptera, Acrididae)[END_REF]references in Heller, 2006;[START_REF] Allegrucci | Testing phylogenetic hypotheses for reconstructing the evolutionary history of Dolichopoda cave crickets in the eastern Mediterranean[END_REF][START_REF] Ullrich | Secondary structure and phylogenetic analysis of the internal transcribed spacers 1 and 2 of bush crickets (Orthoptera: Tettigoniidae: Barbistini)[END_REF][START_REF] Chintauan-Marquier | Evolutionary history and taxonomy of a short-horned grasshopper subfamily, the Melanoplinae (Orthoptera, Acrididae)[END_REF][START_REF] Gorochov | Classification of the Phalangopsinae subfamily group, and new taxa from the subfamilies Phalangopsinae and Phaloriinae (Orthoptera: Gryllidae)[END_REF]; but see Nattier et al., 2011a;[START_REF] Song | 300 million years of diversification: elucidating the patterns of orthopteran evolution based on comprehensive taxon and gene sampling[END_REF]. Only recently, phylogenies relying on large taxonomic and/or character samples have been performed, which suggest that a comprehensive phylogeny for the Orthoptera could be achieved in the near future: [START_REF] Leavitt | Searching for the optimal data partitioning strategy in mitochondrial phylogenomics: a phylogeny of Acridoidea (Insecta: Orthoptera: Caelifera) as a case study[END_REF] explored large dataset partitions while studying the phylogeny of caeliferan Acridoidea, and [START_REF] Mugleston | A century of paraphyly: a molecular phylogeny of katydids (Orthoptera: Tettigoniidae) supports multiple origin of leaf-like wings[END_REF] studied the phylogeny of Tettigonioidea. However, crickets, a large group of Orthoptera (~5000 species), are missing in this global achievement of orthopteran phylogeny and this is the goal of our study.

As crickets have been used as a model system for studies on speciation and acoustic communication [START_REF] Huber | Cricket Behavior and Neurobiology[END_REF][START_REF] Gerhardt | Acoustic Communication in Insects and Anurans[END_REF], the first phylogenies of crickets focused on restricted groups based on model species. For example, hypotheses about speciation have been tested in the Hawaiian genus Laupala [START_REF] Shaw | Conflict between nuclear and mitochondrial DNA phylogenies of a recent species radiation: what mtDNA reveals and conceals about modes of speciation in Hawaiian crickets[END_REF], the Caribbean genus Amphiacusta [START_REF] Oneal | Testing for biogeographic mechanisms promoting divergence in Caribbean crickets (genus Amphiacusta)[END_REF] and the New Caledonian genus Agnotecous (Nattier et al., 2011b[START_REF] Nattier | Secondary sympatry caused by range expansion informs on the dynamics of microendemism in a biodiversity hotspot[END_REF]. Acoustic signal evolution has been analysed in the North American species of Gryllus [START_REF] Huang | Phylogenetic relationships of North American field crickets inferred from mitochondrial DNA data[END_REF]Desutter-Grand-colas and Robillard, 2003), and the diversification of stridulatory structures and calling songs in the cricket subfamily Eneopterinae has been studied first in a phylogenetic perspective [START_REF] Robillard | Phylogeny and the modalities of acoustic diversification in extant extant Eneopterinae (Insecta, Orthoptera, Grylloidea, Eneopteridae)[END_REF], 2006, 2011a), and later taking into account functional data [START_REF] Robillard | A shift toward harmonics for high-frequency calling shown with phylogenetic study of frequency spectra in Eneopterinae crickets (Orthoptera, Grylloidea, Eneopteridae)[END_REF][START_REF] Robillard | Mechanisms of high-frequency song generation in brachypterous crickets and the role of ghost frequencies[END_REF].

These studies did not aim to analyse evolutionary questions at the scale of the entire cricket clade, neither in terms of evolutionary origin nor in terms of diversification. [START_REF] Gwynne | Phylogeny of the Ensifera (Orthoptera): a hypothesis supporting multiple origins of acoustical signalling, complex spermatophores and maternal care in crickets, katydids and wetas[END_REF][START_REF] Gwynne | The evolution of edible 'sperm sacs' and other forms of courtship feeding in crickets, katydids and their kin (Orthoptera: Ensifera)[END_REF] gathered all available information from local phylogenetic trees and classificatory hypotheses to derive hypotheses about cricket mating behaviour, acoustic communication, and habitat. The lack of a wide-scale phylogeny of crickets weakened, however, any evolutionary interpretation, not only because of the lack of a historical framework to follow character transformations, but also because of a deficient sampling of cricket diversity [START_REF] Gerhardt | Acoustic Communication in Insects and Anurans[END_REF][START_REF] Desutter-Grandcolas | Acoustic evolution in crickets: need for phylogenetic study and a reappraisal of signal effectiveness[END_REF]. For example, studies on cricket acoustic communication have been based mostly on just a few species of Gryllus, Teleogryllus and Acheta, which has given rise to a straightforward, narrow view of cricket acoustics [START_REF] Nocke | Physiological aspects of sound communication in crickets (Gryllus campestris L.)[END_REF][START_REF] Elliott | The clockwork cricket[END_REF][START_REF] Otte | Evolution of cricket songs[END_REF][START_REF] Bennet-Clark | Wing resonances in the Australian field cricket Teleogryllus oceanicus[END_REF]. However, original sound emission and apparatus, including non-resonant or high-frequency calls, and asymmetrical or complex sound apparatus were discovered in Phalangopsidae and Eneopterinae crickets [START_REF] Desutter-Grandcolas | Les Phalangopsidae de Guyane franc ßaise (Orthopt eres, Grylloidea): syst ematique, el ements de phylog enie et de biologie[END_REF], 1997[START_REF] Desutter-Grandcolas | Broad-frequency modulation in cricket (Orthoptera, Grylloidea) calling songs: two convergent cases and a functional hypothesis[END_REF][START_REF] Robillard | Phylogeny and the modalities of acoustic diversification in extant extant Eneopterinae (Insecta, Orthoptera, Grylloidea, Eneopteridae)[END_REF]. Their actual diversity and the complexity of the stridulum functioning mechanism [START_REF] Robillard | Mechanisms of high-frequency song generation in brachypterous crickets and the role of ghost frequencies[END_REF] escaped the classic "cricket model", and a possible explanation has emerged only when a sound phylogenetic reference became available [START_REF] Robillard | A shift toward harmonics for high-frequency calling shown with phylogenetic study of frequency spectra in Eneopterinae crickets (Orthoptera, Grylloidea, Eneopteridae)[END_REF][START_REF] Robillard | Mechanisms of high-frequency song generation in brachypterous crickets and the role of ghost frequencies[END_REF]Robillard and Desutter-Grandcolas, 2011a). Therefore, cricket phylogenetic relationships need to be investigated with state-of-theart phylogenetic tools.

The lack of a phylogeny for crickets goes along with the lack of a stable and well-supported classification for this group. Defined on various morpho-anatomical datasets, different classifications have been proposed [START_REF] Brunner Von Wattenwy | Syst eme des Gryllides[END_REF][START_REF] De Saussure | Family Gryllidae[END_REF][START_REF] De Saussure | M elanges orthopt erologiques, V eme fascicule. Gryllides (1 ere partie)[END_REF][START_REF] Scudder | Guide to the Genera and Classification of the North American Orthoptera[END_REF][START_REF] Bruner | South American crickets, Gryllotalpoidea and Achetoidea[END_REF][START_REF] Blatchley | Orthoptera of Northeastern America with Especial Reference to the Faunas of Indiana and Florida[END_REF][START_REF] Chopard | Ordre des Orthopt eres[END_REF][START_REF] Chopard | The Fauna of India and Adjacent Countries. Orthoptera[END_REF][START_REF] Vickery | Taxon ranking in Grylloidea and Gryllotalpoidea[END_REF][START_REF] Gorochov | System and morphological evolution of crickets from the family Gryllidae (Orthoptera) with description of new taxa[END_REF][START_REF] Gorochov | System and evolution of the suborder Ensifera (Orthoptera). Parts 1, 2[END_REF]emended in Storozhenko, 1997;[START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 1 ere partie[END_REF][START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 2 eme partie[END_REF]. These classifications had various internal structures and were a mix of stable groups, well characterized by their morphology and used by a majority of authors, and of artificial, vaguely defined groups. This situation resulted in much confusion about cricket taxonomy, increased by the assignment of newly described taxa to higher taxonomic categories according to ambiguous diagnoses. Today, the most widely used classification system for crickets is the internet reference Orthoptera Species File (OSF; http://orthoptera.speciesfile.org), but this classification system has not been subject to recent and thorough phylogenetic hypotheses. It is now an important challenge to build a robust phylogeny of crickets, not only to understand their evolution and that of Orthoptera in general, but also to establish a stable classification for this diverse clade.

Here we present the first attempt to reconstruct a phylogeny of crickets using a wide taxonomic and character sample (i.e. 205 species representative of cricket diversity; four nuclear and three mitochondrial markers), analysed with parsimony, maximum likelihood, and Bayesian inference. Our aims are: (i) to test the monophyly of a putative cricket clade comprising true crickets, mole crickets, ant-loving crickets, and scaly crickets; (ii) to determine the phylogenetic relationships between these groups, (iii) to determine their main phylogenetic subdivisions and compare them with the OSF; and (iv) to draw from this phylogenetic hypothesis the backbone of a phylogenetic classification of crickets and the framework for future evolutionary studies. We discuss some general outcomes about acoustic communication and biogeography in crickets, which may help guide future evolutionary studies.

Material and methods

Taxon sampling

Taxon sampling was designed to lay the foundation of a phylogenetic classification of crickets sensu lato, that is, true crickets, mole crickets, ant-loving crickets, and scaly crickets. As no clear hypothesis of cricket classification exists today, we referred to the Internet database OSF. We included species belonging to as many suprageneric groups as possible among those listed in the OSF on 1 September 2014. To improve the phylogenetic diversity of the ingroup, and to avoid sampling biases, we also sampled genera that are not currently classified in a suprageneric category in the OSF, and we took into account the genera that were previously classified in tribal or subfamilial groups no longer in use in the OSF.

According to the OSF (Table 1), crickets sensu lato are classified into one superfamily, the Grylloidea, comprising four families, the Mogoplistidae (Mogoplistinaeor scaly cricketsand the Malgasiinae with the Malagasy genus Malgasia Uvarov, 1940), the Gryllotalpidae (Gryllotalpinae and Scapteriscinaemole crickets), the Myrmecophilidae (Myrmecophilinaeant-loving crickets) and the Gryllidae (true crickets). Gryllidae include three "subfamily groups" (the Gryllinae, Phalangopsinae, and Podoscirtinae "subfamily groups", comprising 16 subfamilies) and four unclassified subfamilies (Nemobiinae, Trigonidiinae, Oecanthinae, Eneopterinae).

Our ingroup includes a total of 205 species (Tables 1 and2), belonging to 152 genera (~23% of all known genera). These samples represent 22 of the 25 subfamilies (~88%) and 37 of the 52 tribes (~71%) listed in the OSF. All OSF families and "subfamily groups" are represented.

In addition, two species of Schizodactylidae were included: Schizodactylus monstrosus Drury, 1770 and Comicus campestris Irish, 1986. Sand crickets have been considered potentially close to crickets, on a morphological [START_REF] Gwynne | Phylogeny of the Ensifera (Orthoptera): a hypothesis supporting multiple origins of acoustical signalling, complex spermatophores and maternal care in crickets, katydids and wetas[END_REF] or molecular [START_REF] Jost | Phylogeny of Ensifera (Hexapoda: Orthoptera) using three ribosomal loci, with implications for the evolution of acoustic communication[END_REF][START_REF] Legendre | One hundred years of instability in ensiferan relationships[END_REF] basis (see also [START_REF] Blanchard | Histoire des insectes: traitant de leurs moeurs et de leurs m etamorphoses en g en eral et comprenant une nouvelle classification fond ee sur leurs rapports naturels[END_REF] for S. monstrosus). They are classified as a distinct superfamily in the OSF, that is the Schizodactyloidea, but constitute the sister group of the leafroller crickets (Stenopelmatoidea, Gryllacridinae) in [START_REF] Gorochov | System and evolution of the suborder Ensifera (Orthoptera). Parts 1, 2[END_REF][START_REF] Gorochov | The higher classification, phylogeny and evolution of the superfamily Stenopelmatoidea[END_REF]. Outgroup taxa were taken within the two suborders of Orthoptera, that is, Caelifera and Ensifera (Table 2). Caelifera were represented by two well-documented grasshopper species, Schistocerca gregaria Forsk al, 1775 and Locusta migratoria Linnaeus, 1758. A wider panel of species was chosen for Ensifera, as the monophyly and phylogenetic structure of this clade is unclear [START_REF] Legendre | One hundred years of instability in ensiferan relationships[END_REF]. It included representatives of the families Anostostomatidae (one species), Rhaphidophoridae (two species) and Gryllacrididae (two species).

We assigned a voucher for number all the specimens we used. Most of them are recorded in the specimen database of the Mus eum national d'Histoire naturelle (MNHN), where they can be found, together with all available geographical information at http://science.mnhn.fr/institution/mnhn/collection/eo/search; the others are deposited in the institutions and collections listed in Table 2.

Molecular methods

We generated 1219 sequences (95.6% of the total dataset) for this study (Table 2). When necessary, additional sequences were obtained from the vouchers used in previous phylogenetic studies [START_REF] Robillard | Phylogeny of the cricket subfamily Eneopterinae (Orthoptera, Grylloidea, Eneopteridae) based on four molecular loci and morphology[END_REF]Nattier et al., 2011b[START_REF] Nattier | Secondary sympatry caused by range expansion informs on the dynamics of microendemism in a biodiversity hotspot[END_REF][START_REF] Jaiswara | Testing concordance in species boundaries using acoustic, morphological, and molecular data in the field cricket genus Itaropsis (Orthoptera, Grylloidea, Gryllidae: Gryllinae)[END_REF]. No sample could be obtained for Schizodactylus monstrosus and Comicus campestris. We therefore relied on sequences already published in GenBank (Benson et al., 2005) for these taxa.

We extracted total genomic DNA from middle or hind femora of dried, alcohol-preserved, or newly collected specimens. We used QIAamp DNA tissue microkit (QIAGEN) according to the manufacturer's instructions. Molecular work was performed at the Service de Syst ematique Mol eculaire of the MNHN.

We sequenced seven markers, three mitochondrial and four nuclear, used in previous phylogenetic studies on insects (Table 3). These are the small (12SrRNA, ~400 bp) and large (16srRNA, ~500 bp) mitochondrial ribosomal subunits, the mitochondrial gene coding for cytochrome b protein (cytb, ~400 bp), a fragment of the small nuclear ribosomal subunit (18SrRNA, ~650 bp), two fragments of the large nuclear ribosomal unit (28SA, ~400 bp and 28SD, ~900 bp rRNA) and the gene coding for H3 protein (H3, ~330 bp). Primers and annealing temperatures are given in Table 3. Sequencing reactions were carried out on both DNA strands. Ambiguous results were checked by multiple sequencing either on a different DNA extraction from the same individual, or of an extraction from another conspecific individual. The quality of museum-preserved specimens varied considerably and DNA degradation did not allow the amplification of all target sequences for each species: however, 85% of the whole matrix was completed (Table 2).

Sequence analyses

Newly generated sequences were edited in Sequencher V. 4.9 (Gene Codes Corporation, Ann Arbor, MI, USA), blasted with NCBI blast tools, and submitted to GenBank (Table 2).

Multiple alignments were generated for each locus using Muscle 3.8 [START_REF] Edgar | MUSCLE: multiple sequence alignment with high accuracy and high throughput[END_REF] with default parameters, and refined manually in terminal regions. The positional homology of nucleotides in protein-coding genes was constrained manually in BioEdit 7.0.5.3 [START_REF] Hall | BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT[END_REF] using the codon reading frame information. For rDNA, we checked that our alignments were congruent with published models of secondary structure in insects (Flook and Rowell, 1997b;[START_REF] Buckley | Secondary structure and conserved motifs of the frequently sequenced domains IV and V of the insect mitochondrial large subunit rRNA gene[END_REF][START_REF] Page | Comparative analysis of secondary structure of insect mitochondrial small subunit ribosomal RNA using maximum weighted matching[END_REF][START_REF] Kjer | Aligned 18S and insect phylogeny[END_REF], which was the case for very conserved stems. Nevertheless, following the method proposed by [START_REF] Lutzoni | Integrating ambiguously aligned regions of DNA sequences in phylogenetic analyses without violating positional homology[END_REF], three hypervariable and ambiguously aligned regions (AARs), totalling 421 bp (positions 129-456, 548-556, and 628-711 of the 28SD initial alignment) were identified in the 28SD fragment. We coded the AARs as single multistate characters with the INAASE package [START_REF] Lutzoni | Integrating ambiguously aligned regions of DNA sequences in phylogenetic analyses without violating positional homology[END_REF] for the parsimony analysis and removed it from the likelihood and Bayesian analyses.

We concatenated the different partitions with Se-quenceMatrix 1.7.7 [START_REF] Vaidya | SequenceMatrix: concatenation software for the fast assembly of multi-gene datasets with character set and codon information[END_REF], which resulted in an alignment of ~3600 bp (online Supporting Information).

Base composition, substitution rates, and rate heterogeneity across sites (Table 4) were estimated for each dataset under the most likely model of evolution suggested by the Akaike Information Criterion (AIC; [START_REF] Akaike | Information theory as an extension of the maximum likelihood principle[END_REF][START_REF] Posada | Model selection and model averaging in phylogenetics: advantages of Akaike information criterion and Bayesian approaches over likelihood ratio tests[END_REF]) in jmoldeltest 2.1.3 [START_REF] Guindon | A simple, fast and accurate method to estimate large phylogenies by maximum-likelihood[END_REF][START_REF] Darriba | jModelTest 2: more models, new heuristics and parallel computing[END_REF]. The constancy of base frequencies across taxa was evaluated with a chi-square (v 2 ) "goodness of fit" test using PAUP 4.0b10 [START_REF] Swofford | PAUP*. Phylogenetic Analysis Using Parsimony (*and other methods)[END_REF]. Tests were performed with all sites and variable sites only (Table 4) in order to assess the potential confounding effects of invariant sites (e.g. [START_REF] Buckley | Exploring amongsite rate variation models in a maximum likelihood framework using empirical data, effects of model assumptions on estimatesof topology, branch lengths, and bootstrap support[END_REF].

Phylogenetic analyses

We conducted maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI) analyses on separate and combined datasets to address questions regarding the levels of homoplasy, tree incongruence, and information content within and among loci. We also conducted separate analyses to check for obvious artefacts or contaminations. In addition, we performed combined analyses (MP, BI) of the whole dataset without the poorly documented taxa Schizodactylus monstrosus and Comicus campestris, to test their influence on the resultant topology. Trees were visualized with FigTree 1.4.0. (http://tree.bio.ed.ac.uk/software/figtree/).

In a probabilistic framework, we used jModelTest 2.1.3 under the AIC to select the best model for each marker among the models tested (Table 4). For all markers, the best model was GTR + I + Γ. We did not use though models mixing a proportion of invariant sites (I) with a gamma distribution shape parameter (Γ) because these two parameters are strongly correlated [START_REF] Sullivan | The effect of taxon sampling on estimating rate heterogeneity parameters of maximum-likelihood models[END_REF][START_REF] Yang | Computational Molecular Evolution[END_REF]. Therefore each partition (i.e. marker) was analysed under a GTR + Γ model. We performed 500 ML replicates under the rapid hill-climbing algorithm as implemented in RAxML 7.4.2. [START_REF] Stamatakis | RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models[END_REF][START_REF] Silvestro | raxmlGUI: a graphical front-end for RAxML[END_REF], branch lengths being estimated together for all markers, and we selected the optimal solution (with and without Schizodactylus monstrosus and Comicus campestris). We ran two similar analyses for the nuclear and mitochondrial partitions, respectively. For the combined analyses, we also computed bootstrap support values with 500 pseudoreplicates. All these analyses were performed on a HP Z800 Workstation with 17.9 Gbits RAM and an Intel Xeon CPU E5520, using seven threads.

The BI analyses (with and without Schizodactylus monstrosus and Comicus campestris) were conducted using the same model strategy with MrBayes 3.1.2 [START_REF] Ronquist | MrBayes 3: Bayesian phylogenetic inference under mixed models[END_REF] on the MNHN cluster. We ran the analyses for 50 millions of generations with the command stoprule = yes, sampling tree every 10 000 generations. We used Tracer 1.5 [START_REF] Rambaut | Tracer, Version 1.5[END_REF] to check that our effective sample size was large enough for a meaningful estimation of parameters and to assess the burn-in. Finally, we checked for convergence of our results ensuring that the potential scale-reduction factor approached 1.0 for all parameters.

Heuristic searches, on nuclear and mitochondrial data, separated or combined (with and without Schizodactylus monstrosus and Comicus campestris), with or without the coded AAR, were implemented in PAUP* under the MP criteria with unordered (Fitch) characters, 100 random sequence addition replicates, the TBR branch-swapping, and gaps treated as missing data. Strict consensus trees were then reconstructed. Support for internal branches was assessed in PAUP* by nonparametric bootstrapping [START_REF] Felsenstein | Confidence limits on phylogenies, an approach using the bootstrap[END_REF][START_REF] Efron | Bootstrap confidence levels for phylogenetic trees[END_REF] with 100 pseudoreplicates, using full heuristic searches with 10 random addition sequence replicates, TBR branch swapping, and one tree held at each step during stepwise addition (e.g. [START_REF] Debry | A simulation study of reduced tree-search effort in bootstrap resampling analysis[END_REF].

Taxonomic hypotheses

Although we sampled many cricket species, we consider that we did not sample enough genera to accurately represent cricket diversity below the subfamilial level. The subfamilial rank is the classification category that has been most commonly retained by the successive authors of classificatory systems for crickets, although they have been used in various systems comprising from one to thirteen families according to the authors. For the sake of stability, and as a first contribution to cricket classification, we consequently chose to test the monophyly of the subfamilies currently used in the OSF and to propose a new, phylogeny-based, classification of crickets from and above the subfamilial level.

Additional studies will be required to build comprehensive phylogenies below the subfamilial level. A detailed analysis of the present results in terms of cricket classification will be published elsewhere (L. Desutter-Grandcolas, unpublished data).

Results

Sequence analyses

The sequences used have a high content of G/C (60-80%) and A/T (70%) for the nuclear and mitochondrial DNA respectively (Table 4), in agreement with values previously reported for other orthopterans (e.g. [START_REF] Chintauan-Marquier | Evolutionary history and taxonomy of a short-horned grasshopper subfamily, the Melanoplinae (Orthoptera, Acrididae)[END_REF][START_REF] Gorochov | Classification of the Phalangopsinae subfamily group, and new taxa from the subfamilies Phalangopsinae and Phaloriinae (Orthoptera: Gryllidae)[END_REF] and insects in general [START_REF] Simon | Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved Polymerase Chain Reaction primers[END_REF][START_REF] Simon | Incorporating molecular evolution into phylogenetic analysis, and a new compilation of conserved polymerase chain reaction primers for animal mitochondrial DNA[END_REF]. Base frequencies were constant for all nuclear partitions taking all sites, and for the ribosomal nuclear partitions taking variable sites only. Across all fragments, 2262 sites (62.8%) were variable and 1849 sites (51.4%) were parsimony-informative. The nuclear and mitochondrial partitions represent 47.1% and 52.9% of the informative sites, respectively. Models of evolution and associated parameters for the probabilistic analyses are given in Table 4.

Phylogenetic analyses

For all phylogenetic methods, combining the nuclear and mitochondrial data improved tree resolution and nodal support (Figs 123456) by comparison with separate analyses (e.g. Figs S1-S7). The nuclear and mitochondrial partitions yielded very similar trees (Figs S11-S14), except for a few taxa (namely sand crickets and Pteroplistinae), the positions of which remain unsupported in the combined analysis. The addition of the coded AAR (in the MP analysis) had no significant effect on tree topologies. The MP, ML, and BI methods yielded similar topologies for most of the main clades (see below and Figs S8 andS10). Removing Schizodactylus monstrosus and Comicus campestris from the data did not modify the results of the ML and BI analyses significantly (see Figs S15 and S16). In MP, it resulted in a less resolved strict consen-sus that still shows most of the main clades (i.e. clades A, B, C, D, E and F+G); a trifurcation between ensiferan outgroups, clade A and the rest of the crickets forbids finding crickets sensu lato monophyletic.

Support values (posterior probabilities and nonparametric bootstrap values) are given in Fig. 1 for the deeper nodes, and in Figs 2-6 for the other nodes.

Our results with the complete data set (Fig. 1) suggest that, within the monophyletic Ensifera, the crickets sensu lato form a clade strongly supported in BI (PP = 1.00) and ML analyses (ML = 93), provided they include Schizodactylus. They are the sister group to the other sampled Ensifera (i.e. Rhaphidophoridae, Gryllacrididae, Anostostomatidae). The sand crickets are not recovered as a monophyletic group: the genus Comicus gathered with the Rhaphidophoridae (ML, BI -Fig. 2; MP, Fig. S10), while Schizodactylus comes within the ant-loving cricket clade. The position of these two taxa may be, however, an artefact due to incomplete data. Among the outgroups, the Gryllacrididae are always monophyletic and sistergroup to the Anostostomatidae. All these relationships are very well supported (PP ≥ 0.99). The analyses per- formed without the partition 28SD gave the same relationships with a weaker support (0.5 < PP < 0.7) (Fig. S9).

Within the cricket clade (Figs 2-6), three main clades are identified. They correspond to (i) ant-loving crickets and mole crickets (Clade A), (ii) scaly crickets (plus the Malagasy genus Malgasia) (clade B), and (iii) true crickets, separated into five clades named C to G, with the clade C being the sister group to all the others. These clades and this structure are recovered in all analyses: all the clades have a very high PP, but clades E, F and G are not so well supported in MP and ML, as are their internal relationships. The main differences concern the clades F and G in MP (see infra) and in the position of clade D: in ML, the clade D is the sister group of all true crickets (Fig. S8), while it is the sister group of the clade (E + F + G) in MP and BI (PP = 0.69, Fig. 1; Fig. S10); in ML analysis performed without the marker 28SD, the clade D is the sister group of the clade C (PP = 0.58, Fig. S9).

Within clade A (Fig. 2), the mole crickets and the ant-loving crickets (+Schizodactylus) form two very well-supported clades (A1 and A2, PP = 1.00 for both). Clade B, gathering the scaly crickets and Malgasia (Fig. 1, PP = 1.00), is subdivided into two sister groups, with Malgasia nested within one of them, which invalidates the separation of Malgasia as a separate subfamily. Clade C (Fig. 3, PP = 1.00) is made of two very well-supported sister groups, which comprise all the sampled species of the subfamilies Trigonidiinae (clade C1, PP = 1.00) and Nemobiinae (clade C2, PP = 1.00), at the exception of the genus Hemigryllus Saussure, 1877, which is nested in clade G. The current tribes of Trigonidiinae are not found to be monophyletic, as the Phylloscyrtini, represented by Cranistus sp., are nested within the Trigonidiini. Nemobiinae separates into two groups (PP = 1.00, both): the first one includes mostly Pteronemobiini, whereas the second includes mostly Nemobiini. This result supports the monophyly of these two tribes, providing that Absonemobius is transferred from the Pteronemobiini to the Nemobiini. Also, the nested position of the genus Thetella in the Nemobius group invalidates the Thetellini tribe. The genus Cophonemobius, not classified in the OSF, is nested within the Nemobius group. In MP topology, the pteronemobiine genus Allonemobius is the sister group of the Nemobius clade, while Absonemobius is nested within the pteronemobiine clade (Fig. S10).

Clade D (Fig. 4) comprises the crickets belonging to the tribe Pteroplistini of the subfamily Pteroplistinae. This small clade is very well supported, unlike its exact position within crickets (PP < 0.7 -Fig. 1; Figs S8-S10). Clade E (Fig. 4, PP = 0.93) includes mostly taxa presently classified in the "Phalangopsinae subfamily group". It comprises five well-supported clades (E1 to E5), recovered in all analyses (except E1 in MP). The species Parendacustes lifouensis is the sister group of Trigonidiinae + Nemobiinae on ML topology, and nested with taxa of clade E2 on MP topology: this position maybe due to a lack of data, this taxon being here documented for 12S and cytb only. The clades E1 to E5 do not fit with the subfamilies of the "Phalangopsinae subfamily group", which are all but one (viz. Phaloriinae) recovered as polyphyletic. Moreover, the genera Gryllomorpha and Petaloptila, which are currently classified in the tribes Gryllomorphini and Petaloptelini of the Gryllomorphinae subfamily inside the "Gryllinae subfamily group", are clearly nested within clade E.

Clade E1 (PP = 0.99) comprises genera classified in the Phaloriinae (i.e. Afrophaloria, Phaloria, Pseudotrigonidium and Tremellia), together with the genera Megacris, not assigned to a subfamily in the OSF but originally described as a Phaloriinae.

Clade E2 (PP = 1.00) comprises two sister clades. The first one corresponds to the wide Neotropical genus Uvaroviella (Phalangopsinae, Phalangopsini), and the second includes taxa classified in the polyphyletic Endacustini tribe (Paragryllodes, Protathra) and all the sampled taxa presently classified in the Paragryllinae. This second clade (PP = 1.00) could be congruent with the definition of several taxonomic groups described as tribes or subtribes, that is, Paragryllini (Paragryllus), Benoistellini (Silvastella), Rumeini (Rumea), and a mix of Neoaclini + Strogulomorphini (Aclella, Superacla, Kevanacla, Ectecous, Unithema, Strogulomorphini sp, Yoyuteris).

Clade E3 (PP = 0.97) includes all sampled Gryllomorphini except Eugryllodes (nested in clade G4), in addition to Phaeogryllus (classified in Phalangopsinae, Heterogryllini), Meloimorpha and Homoeogryllus (Cachoplistinae, Homoeogryllini), and Socotracris (Phalangopsinae).

Clade E4 (PP = 0.93) gathers Stalacris and Opiliosina (respectively classified within the "Phalangopsinae subfamily group" and the Phalangopsinae subfamily), with Parendacustes (Phalangopsinae, Phalangopsini), Kempiola (Cachoplistinae, Homoeogryllini), Leptopedetes, Amphiacusta and Noctivox (Luzarinae, Amphiacustina), and the wide African genus Phaeophilacris ("Phalangopsinae subfamily group", Endacustini). Internal relationships of clade E4 are strongly supported, except for the sister relationship between the neotropical Amphiacustina and Phaeophilacris (PP < 0.5).

Clade E5 includes all the sampled taxa presently classified in the subfamily Luzarinae, except Amphiacusta, Leptopedetes, and Noctivox (in clade E4). Apart from Lernecella and Miogryllodes, the positions of which within the clade are weakly supported (PP < 0.8), the internal structure of E5 is strongly supported: a clade made of Phalangopsis and Philippopsis (PP = 1.00; Phalangopsinae, Phalangopsini) is the sister group of a clade comprising on one hand Aracamby, Lerneca, Microlerneca, and Prostama (PP = 0.97), and on the other hand Luzara, Luzarida, Acantoluzarida, Luzaridella, Niquirana, and Amusodes (PP = 1.00) (all Luzarinae).

In the MP topology (Fig. S10), clades E4 and E5 vary for the position of Lernecella and Miogryllodes.

Clade F (Fig. 5, PP = 1.00) broadly corresponds to the "Podoscirtinae subfamily group"; it is composed of five main clades, which correspond roughly to five subfamilies used in the OSF, provided their content and definition are slightly modified. Clade F1 (PP = 1.00) is the sister-group of the other clades and corresponds to the Euscyrtinae ("Podoscirtinae subfamily group"). Clades (F2 + F3) and (F4 + F5) are sister-groups (PP = 0.95), with a weak (PP < 0.5) and strong (PP = 1.00) support, respectively. Clade F2 (PP = 0.97) includes the Oecanthinae (not classified), whereas clade F3 (PP = 1.00) includes the genera Neometrypus, Paroecanthus, Tafalisca, and Diatrypa, which are today scattered in three different tribes of the Podoscirtinae subfamily (Podoscirtini Neometrypina, Paroecanthini Paroecanthina and Tafaliscina, and Aphonoidini Diatrypina, respectively).

Clade F4 (PP = 1.00) includes the Podoscirtinae (minus the taxa listed above), and clade F5 (PP = 1.00) corresponds to the Hapithinae plus some of the taxa presently classified in the Eneopterinae.

Finally, Clade G (Fig. 6) is strongly supported (PP = 0.96), but its internal relationships are not all well supported. Four clades proposed by our results partially cover the "Gryllinae subfamily group" used in the OSF, but do not support the monophyly of the subfamilies. Only two clades (G3, G4) are well supported (PP ≥ 0.96), the other two (G1, G2) having a support < 0.7. Clade G1 includes taxa currently classified either in the Pentacentrinae ("Podoscirtinae subfamily group") or in the Landrevinae ("Gryllinae subfamily group"). Clade G2 includes taxa once classified in the tribe Odontogryllini (since then synonymized with the Landrevini), together with the genus Hemigryllus, currently classified in the Nemobiinae (see clade C2). Clade G3 includes the Eneopterinae sensu stricto. Four tribes among the five acknowledged today are recovered as monophyletic, the Eneopterini being polyphyletic, and the genus Swezwilderia, classified as incertae sedis in the OSF, is nested within the Lebinthini. Clade G4 includes two monophyletic groups: the first includes some Landrevinae (but see above, clade G1) and the taxa presently classified in the Gryllinae, Sclerogryllinae, and Gryllomorphinae pro parte (p.p.) (genus Eugryllodes); the second shows a sistership relation between Eugryllodes and all the other taxa (minus the grylline genus Taciturna). Finally, none of the sampled tribes of the Gryllinae subfamily (Cephalogryllini, Cophogryllini, Gryllini, Modicogryllini, Sciobiini, Turanogryllini) are recovered as monophyletic, the genera Cophogryllus, Sciobia, and Turanogryllus being nested among the other genera.

In the MP topology (Fig. S10), the clades F and G are not sister-groups, but they form a paraphyletic assemblage, where the clades F1 (Euscyrtinae), F3 (Tafaliscinae), F4 (Podoscirtinae) and F5 (Hapithinae) are recovered, while the clades F2 (Oecanthinae), G1 (Pentacentrinae + Landrevinae p.p.) and G4 (Gryllinae + Sclerogryllus + Eugryllodes) are almost recovered.

Figure 7 and Table 5 sum up how our results support the currently used classification of crickets. We can distinguish four different situations, according to the congruence between the OSF classification and our phylogenetic hypothesis (Table 5): (i) groups recovered as monophyletic in their current meaning; (ii) groups that could be recovered as monophyletic providing minor changes in cricket classification (most often, this means to move a few genera from one clade to another); (iii) groups recovered as polyphyletic; (iv) groups (most often isolated genera) nested within larger clades. Six subfamilies are thus monophyletic (Euscyrtinae, Oecanthinae, Phaloriinae, Pteroplistinae, Trigonidiinae, Gryllotalpinae), while five others are almost recovered as monophyletic (Gryllinae, Paragryllinae, Nemobiinae, Mogoplistinae, Myrmecophilinae).

Discussion

Monophyly of the cricket clade, and its main subdivisions

Our results are congruent with the current grouping of crickets and their allies: in all analyses, crickets, scaly crickets, ant-loving crickets, and mole crickets are grouped together, attesting this long-lasting hypothesis. Schizodactylus and Comicus, currently gathered in the ensiferan family Schizodactylidae (sand crickets), are not monophyletic here, a result that could be an artefact, ensuing from an incomplete data set for the sand crickets (Table 2). Overall, our data confirm the phylogenetic affinity of sand crickets with crickets sensu lato, but are insufficient to propose a clear phylogenetic pattern of basal relationships between the cricket clade and its potential sister group (but see [START_REF] Song | 300 million years of diversification: elucidating the patterns of orthopteran evolution based on comprehensive taxon and gene sampling[END_REF].

The cricket clade is composed of seven main monophyletic groups (Fig. 1, clades A to G). These clades, together with their main subdivisions, support part of the currently hypothesized families, "subfamily groups", or subfamilies (Fig. 7 and Table 5).

The higher rank categories, that is, families or "subfamily groups", correspond to the broad morphological categories used since the beginning of modern cricket classification. Three of the four families used in the OSF are recovered as monophyletic (Table 5, Fig. 7), but their relationships are not consistent with a similar taxonomic level (see below). These families are: the mole crickets (OSF Gryllotalpidae), the scaly crickets (OSF Mogoplistidae), and the true crickets (OSF Gryllidae). The monophyly of the fourth family, the Myrmecophilidae, sampled here for the Myrmecophilini tribe only, is also well supported in the analyses without Schizodactylus (see above and Figs S15-S17). Out of the three "subfamily groups" used in the OSF, only one corresponds to a well-supported clade ("Phalangopsinae subfamily group", clade E); the "Podoscirtinae subfamily group" and the "Gryllinae subfamily group" are included in our clades F and G respectively.

Among the 22 sampled subfamilies, only six are monophyletic, that is, the Gryllotalpinae, Trigonidiinae, Pteroplistinae, Euscyrtinae, Oecanthinae (sampled here for the Oecanthini tribe only), and Phaloriinae. They correspond to the smallest groups currently recognized in terms of cricket classification, except the Trigonidiinae, and the most homogeneous morphologically. All the other subfamilies are para-or polyphyletic. Some "nearly monophyletic" subfamilies correspond to morphologically well-characterized groups, the definitions for which have been made confused by the wrong placement of a few genera: their monophyly may be obtained with slight taxonomic changes, that is, provided some particular genera are moved from their current definition in the OSF. This is the case of the Nemobiinae, which form a well-supported clade, from which the genus Hemigryllus is excluded. [START_REF] Gorochov | System and morphological evolution of crickets from the family Gryllidae (Orthoptera) with description of new taxa[END_REF] and [START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 1 ere partie[END_REF][START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 2 eme partie[END_REF] previously suggested that Hemigryllus is not a Nemobiinae: [START_REF] Gorochov | System and morphological evolution of crickets from the family Gryllidae (Orthoptera) with description of new taxa[END_REF] had erected the Hemigryllinae subfamily within an "Eneopteridae group" comprising the Eneopterinae, Landrevinae, and Phalangopsinae, and [START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 1 ere partie[END_REF][START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 2 eme partie[END_REF] had tentatively placed Hemigryllus within the newly erected Tafaliscinae subfamily comprising the Neotropical Tafaliscini, Paroecanthini, Neometrypini, and Diatrypini. In the same way, the genus Malgasia is nested within the Mogoplistinae from which it cannot be separated as a distinct subfamily. The Pentacentrinae are monophyletic provided they include some genera currently classified within the Landrevinae, a reorganization suggested by several morphological characters (S. Hugel, unpublished data). However, because these two subfamilies belong to different "subfamily groups", this modification implies their complete reassessment. Finally, the Eneopterinae are monophyletic provided that they are narrowed according to [START_REF] Robillard | Clarification of the taxonomy of extant crickets of the subfamily Eneopterinae (Orthoptera, Grylloidea, Eneopteridae)[END_REF]; the genera added in this group by [START_REF] Otte | Caribbean Crickets[END_REF] must be transferred to the Hapithinae.

The other subfamilies should be more deeply reassessed: the Gryllinae should include the Sclerogryllinae and some of the Gryllomorphinae (at least Eugryllodes among the sampled genera). The Luzarinae, Phalangopsinae, Cachoplistinae, and Paragryllinae of the "Phalangopsinae subfamily group" should be fully redefined and the genera reclassified. The subfamilies proposed by [START_REF] Gorochov | Classification of the Phalangopsinae subfamily group, and new taxa from the subfamilies Phalangopsinae and Phaloriinae (Orthoptera: Gryllidae)[END_REF] for the "Phalangopsinae subfamily group" on a taxonomic basis are not vali- dated here, his "Phalangopsinae" gathering taxa belonging to our clades E2, E4, and E5. The position of the taxa yet to be classified within the Podoscirtinae and the Hapithinae of the "Podoscirtinae subfamily group" should be revised.

At a lower taxonomic level, most of the 37 sampled tribes are para-or polyphyletic. These non-monophyletic tribes belong to the "Phalangopsinae subfamily group", the "Podoscirtinae subfamily group", and the "Gryllinae subfamily group", which are scattered across our clades E, F and G, but also to the Trigonidiinae and the Nemobiinae (clade C).

The vast majority of the phylogenetic relationships discussed above are well supported (PP > 0.80), except for the definition of the clades F and G (see above). These results will imply deep modifications in the current cricket classification, even if the general framework looks preserved. The bulk of cricket diversity will have to be reorganized to design a phylogenybased classification of crickets, which also calls for a reassessment of the morphological characters used to define the different taxa.

Laying the foundation of a phylogenetic classification

Our results (Fig. 7) support the monophyly of the cricket clade (AE Schizodactylus), and its subdivision into two main clades: mole crickets and ant-loving crickets on the one hand, and all the other crickets on the other. In the framework of the OSF classification of Ensifera, the separation of (Gryllotalpidae + Myrmecophilidae) from the Grylloidea justifies the erection of a superfamily Gryllotalpoidea Leach, 1815, while the Grylloidea Laicharting, 1781 is restricted to scaly crickets and true crickets. The whole cricket clade should consequently be elevated at the infraorder level, that is, the Gryllidea, as proposed by several authors in the past [START_REF] Vickery | Taxon ranking in Grylloidea and Gryllotalpoidea[END_REF][START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 1 ere partie[END_REF][START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 2 eme partie[END_REF][START_REF] Gorochov | System and evolution of the suborder Ensifera (Orthoptera). Parts 1, 2[END_REF]; see also [START_REF] Song | 300 million years of diversification: elucidating the patterns of orthopteran evolution based on comprehensive taxon and gene sampling[END_REF]. The monophyly of Gryllidea, which is ascertained here for extant taxa, will have to be checked relative to ensiferan fossils, with a reanalysis of morphological characters in fossils and extant taxa (see B ethoux and [START_REF] Nel | Venation pattern and revision of Orthoptera sensu nov. and sister groups. Phylogeny of Palaeosoic and Mesozoic Orthoptera sensu nov[END_REF] for fossil taxa only).

A very similar classification was proposed by [START_REF] Vickery | Taxon ranking in Grylloidea and Gryllotalpoidea[END_REF], who kept, however, the Myrmecophilidae within the Grylloidea. [START_REF] Gorochov | System and evolution of the suborder Ensifera (Orthoptera). Parts 1, 2[END_REF] separated the Gryllotalpidae and possibly the Myrmecophilidae from the Mogoplistidae and all other crickets within the Gryllidea, but put Malgasia in a subfamily of the Myrmecophilidae. Also his Gryllidea included both extant and fossil taxa, but some of his clades were paraphyletic (see [START_REF] Gorochov | System and evolution of the suborder Ensifera (Orthoptera). Parts 1, 2[END_REF]Gorochov, , his fig. 1149)).

Within the Grylloidea sensu stricto, we propose to consider four families: the Mogoplistidae Brunner von Wattenwy, 1873 (clade B), the Trigonidiidae Saussure, 1874 (clade C), the Phalangopsidae Blanchard, 1845 (clade E), and the Gryllidae Laicharting, 1781 (clade F + G). The first three are robust and highly supported, while the latter has variable support and its inner structure needs to be studied more deeply. Clade D (Pteroplistinae Chopard, 1936), which is supported by all analyses but its phylogenetic position remains unstable, should be kept incertae sedis within Grylloidea. These families fit variously with morphological characters.

The monophyly of the Trigonidiidae, and of their two traditional subfamilies, the Trigonidiinae and the Nemobiinae (without Hemigryllus), is corroborated by many morphological characters [START_REF] Gorochov | System and morphological evolution of crickets from the family Gryllidae (Orthoptera) with description of new taxa[END_REF][START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 1 ere partie[END_REF][START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 2 eme partie[END_REF]. The morphological definition of the Phalangopsidae, which has been recognized as a main cricket group since [START_REF] Blanchard | Histoire des insectes: traitant de leurs moeurs et de leurs m etamorphoses en g en eral et comprenant une nouvelle classification fond ee sur leurs rapports naturels[END_REF], will have to be checked to take into account at least Gryllomorpha and Petaloptila, and possibly other genera of the Petaloptilini tribe. Finally, the Gryllidae, as defined here, is not compatible with the morphological characters currently used: it will have to be reanalysed to clarify the situation. Except for the Trigonidiinae and Nemobiinae [START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 1 ere partie[END_REF][START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 2 eme partie[END_REF], the Eneopterinae [START_REF] Robillard | Clarification of the taxonomy of extant crickets of the subfamily Eneopterinae (Orthoptera, Grylloidea, Eneopteridae)[END_REF], and the Pentacentrinae (S. Hugel, unpublished data), the putative subfamilies will have to be reconsidered from the point of view of their morphological diagnosis, a valid concern for nearly all the groups of crickets below the familial rank (L. Desutter-Grandcolas, unpublished data).

The classification system proposed here fits the structure of the phylogeny better than the OSF, and uses the best-supported clades as a backbone for a future classification of crickets. It is intermediary between the very split system proposed by several authors, with up to 12 or 13 hypothesized families [START_REF] Bruner | South American crickets, Gryllotalpoidea and Achetoidea[END_REF][START_REF] Chopard | Ordre des Orthopt eres[END_REF][START_REF] Vickery | Taxon ranking in Grylloidea and Gryllotalpoidea[END_REF][START_REF] Mce Kevan | Orthoptera[END_REF][START_REF] Desutter | Structure et evolution du complexe phallique des Gryllidea (Orthoptera) et classification des genres n eotropicaux de Grylloidea. 2 eme partie[END_REF], and the lumped system with all crickets gathered in one family subdivided in as many subfamilies [START_REF] Scudder | Guide to the Genera and Classification of the North American Orthoptera[END_REF][START_REF] Otte | The Australian crickets[END_REF]). This system is not incompatible with the OSF, but beyond an apparent consistency, the classification of crickets needs to be deeply revised to achieve a classification system that accommodates monophyletic units of evolution.

Laying the foundation of future evolutionary studies

The proposed phylogenetic hypothesis brings new perspectives for understanding cricket evolution, especially on communication, for which these insects are important models [START_REF] Gerhardt | Acoustic Communication in Insects and Anurans[END_REF].

Acoustic evolution in Ensifera should be reconsidered, as previous analyses of acoustic evolution in Ensifera focused on the structural homology between crickets sensu lato on the one hand, and katydids (Tet-tigonioidea) on the other. The question was then to decide whether acoustic communication has evolved once (ancestral communication) or twice (convergent evolution) within Ensifera as a whole: "crickets" being considered an acoustically homogeneous group (see e.g. [START_REF] Bailey | Acoustic Behaviour of Insects. An Evolutionary Perspective[END_REF]. Structural analyses of forewing venation revealed an unsuspected diversity in crickets sensu lato, supporting a hypothesis of convergence between the mole crickets and the crickets sensu stricto; in other words, the primary homology hypothesis was not supported [START_REF] Desutter-Grandcolas | Phylogeny and the evolution of acoustic communication in extant Ensifera (Insecta, Orthoptera)[END_REF][START_REF] Desutter-Grandcolas | Convergence and parallelism: is a new life ahead of old concepts?[END_REF]but see B ethoux, 2012). By definition, such a hypothesis cannot be tested by a phylogeny, but it may imply more or less complex patterns of character change according to the phylogenetic topology. Our present phylogeny is parsimoniously consistent with the convergence hypothesis between crickets and mole crickets, and is also consistent with the fact that crickets sensu lato may have structurally diverse acoustic structure (see below); their early acoustic evolution may then not have been straightforward, especially as acoustic communication may have developed in Ensifera within a general framework of communication by vibration and/or sound (Desutter-Grandcolas, 2003 and references therein;Strauss and Lakes-Harlan, 2008a,b;[START_REF] Stritih | Mating behaviour and vibratory signalling in non-hearing cave crickets reflect primitive communication of Ensifera[END_REF].

Crickets sensu stricto moreover present a wide acoustic diversity, both in terms of structures and in terms of functional properties. The few species used to generate the basic model of sound production in crickets, that is, Acheta, Gryllus, or Teleogryllus grylline species [START_REF] Nocke | Physiological aspects of sound communication in crickets (Gryllus campestris L.)[END_REF][START_REF] Elliott | The clockwork cricket[END_REF][START_REF] Jonsson | Sound radiation and wing mechanics in stridulating field crickets (Orthoptera: Gryllidae)[END_REF]but see Bennet-Clark and Bailey, 2002), all belong to a small apical group nested within our clade G. Recent bioacoustic analyses of more distantly related Eneopterinae species questioned this basic cricket model: Eneopterinae emit high-frequency calls using the second or third harmonic as a dominant frequency [START_REF] Robillard | Mechanisms of high-frequency song generation in brachypterous crickets and the role of ghost frequencies[END_REF], or produce several kinds of syllables using complex stridulatory behaviours and structures (Robillard and Desutter-Grandcolas, 2011b). Yet, Eneopterinae are quite close to Gryllinae, both belonging to clade G, and the two groups taken together do not reflect the high diversity of sound-production structures in crickets. For example, the sound generating system of an Oecanthus species analysed with laser Doppler vibrometry proved recently really different from the usual cricket model when considering the vibratory properties of the wings [START_REF] Mhatre | Changing resonator geometry to boost sound power decouples size and song frequency in a small insect[END_REF]. To further study cricket bioacoustics, good targets to begin with could be the Trigonidiidae, the sister group of all other crickets, and non-mute Phalangopsidae, the sister group of our Gryllidae. These two groups have very peculiar stridulums that do not fit well with the usual acoustic structures of crickets according to the current basic model of sound production. Trigonidiidae are characterized by a wide harp area, crossed by one vein and bordered by a high diagonal vein that may extend the resonant area of the wing to almost the whole forewing surface [START_REF] Desutter-Grandcolas | Chant et appareil stridulatoire de deux Trigonidiinae originaires d'Equateur (Orthoptera: Grylloidea: Trigonidiidae)[END_REF]. Yet, Trigonidiidae emit either very loud calls (Trigonidiinae), or soft, courting-like calls (Nemobiinae): this situation reflects the presence versus absence of the mirror on the forewings, whereas this structure is supposed to be inactive in sound radiation (e.g. [START_REF] Nocke | Physiological aspects of sound communication in crickets (Gryllus campestris L.)[END_REF][START_REF] Bennet-Clark | Wing resonances in the Australian field cricket Teleogryllus oceanicus[END_REF][START_REF] Jonsson | Sound radiation and wing mechanics in stridulating field crickets (Orthoptera: Gryllidae)[END_REF]. In addition, many Phalangopsidae have a corneous, stiff right forewing, and a soft and fragile left forewing, and they are able to emit calling songs, which are either loud and high-pitched, or soft and non-resonant. Such a highly asymmetrical stridulum may not vibrate the same way as a typical grylline stridulum (e.g. [START_REF] Simmons | Symmetry in the songs of crickets[END_REF][START_REF] Bennet-Clark | Wing resonances in the Australian field cricket Teleogryllus oceanicus[END_REF][START_REF] Jonsson | Sound radiation and wing mechanics in stridulating field crickets (Orthoptera: Gryllidae)[END_REF], and this situation is not as yet fully understood. To study cricket call evolution deeper, or even predict cricket calls in extant and fossil taxa from the study of their stridulatory apparatus, as undertaken for katydids [START_REF]Scale effects and constraints for sound production in katydids (Orthoptera: Tettigoniidae): generator size constrains signal parameters[END_REF][START_REF] Gu | Wing stridulation in a Jurassic katydid (Insecta, Orthoptera) produced low-pitched musical calls to attract females[END_REF], it is now essential to study the large diversity of cricket bioacoustics within a phylogenetic perspective, considering the explicit diversity documented by behavioural observations, and the implicit diversity suggested by the phylogeny.

In addition to acoustics, the Gryllotalpoidea includes taxa inhabiting specialized acoustic burrows (mole crickets), taxa living in symbiosis with ants (Myrmecophilinae, Myrmecophilini), and taxa found in deserts (Myrmecophilinae, Bothriophylacini). These habitats and life habits require specific morphological, physiological, and behavioural adaptations, for which the main lines of evolution could be better deciphered in a tree-thinking perspective.

Finally, cricket biogeography and diversification have never been analysed in a wide-scale context due to the lack of properly defined monophyletic groups, but our study brings valuable insights in this respect. Whereas familial and most subfamilial cricket clades are distributed worldwide, some subfamilial taxa, as modified according to our phylogenetic hypothesis, show interesting new biogeographical patterns. This is the case, for example, of the sister-groups Podoscirtinae and Hapitinae in clade F, which now both show a biogeographical coherence: the former is distributed in the Paleotropics and in Oceania, while the latter is present in the Neotropics only. In the same way, the nested positions of Gryllomorpha and Petaloptila within the Phalangopsidae attest for the first time the presence, in the Mediterranean region, of cricket taxa that diversified from a wide intertropical group. These examples suggest new biogeographical scenarios, which should be tested further as typically undertaken for all organisms, with well-supported phylogenetic hypotheses [START_REF] Sanmartin | Patterns of animal dispersal, vicariance and diversification in the Holarctic[END_REF][START_REF] Sanmartin | Dispersal vs. vicariance in the Mediterranean: historical biogeography of the Palearctic Pachydeminae (Coleoptera, Scarabaeoidea)[END_REF][START_REF] Crisp | Hypothesis testing in biogeography[END_REF].

  Fig. 1. Phylogeny of crickets based on 205 terminals and seven DNA markers, obtained by Bayesian inference. Support values (BI/ML/MP) are indicated for the deeper nodes and the clades A to G. Taxa codes (i.e. voucher codes) as in Table 2, and made explicit in Figs 2-6.

Fig. 2 .

 2 Fig. 2. Phylogeny of crickets based on 205 terminals and seven DNA markers in Bayesian inference: detailed topology for clades A and B (see Fig. 1). Support values (BI/ML/MP) are indicated for all nodes. Ensiferan outgroups are displayed to show the position of the two schizodactylid species, Comicus campestris Irish, 1986 and Schizodactylus monstrosus Drury, 1770 (squared). The analyses without these two taxa are provided as Supplementary Figs 15-17.

Fig. 3 .

 3 Fig. 3. Phylogeny of crickets based on 205 terminals and seven DNA markers in Bayesian inference: detailed topology for clade C (see Fig. 1). Support values as in Fig. 2.

Fig. 4 .

 4 Fig. 4. Phylogeny of crickets based on 205 terminals and seven DNA markers in Bayesian inference: detailed topology for clades D and E (see Fig. 1). Support values as in Fig. 2.

Fig. 5 .Fig. 6 .

 56 Fig. 5. Phylogeny of crickets based on 205 terminals and seven DNA markers in BI: detailed topology for clade F (see Fig. 1). Support values as in Fig. 2.

Fig. 7 .

 7 Fig. 7. Phylogeny of crickets based on 205 terminals and seven DNA markers in Bayesian inference, and derived classification of crickets, with two superfamilies (Gryllotalpoidea, Grylloidea), including, respectively, two (Gryllotalpidae, Myrmecophilidae) and four families (Mogoplistidae, Trigonidiidae, Phalangopsidae, Gryllidae). The current position of each terminal species in the subfamilies used by the OSF is indicated on the right of the tree.

Table 1

 1 Taxonomic sampling used in this study, with taxonomic assignment according to the internet database Orthoptera Species File

			Sampled	Sampled
	Families/Subfamilies	Tribes	genera	species
	GRYLLIDAE
	"Gryllinae subfamily group"
	Gryllinae	6 of 7		31
	Gryllomiminae	No tribe	-	-
	Gryllomorphinae	2 of 2		4
	Itarinae	No tribe	-	-
	Landrevinae	1 of 2		6
	Sclerogryllinae	0 of 1		1
	"Phalangopsinae subfamily group"
	Cachoplistinae	1 of 2		4
	Luzarinae	No tribe		17
	Paragryllinae	1 of 1		15
	Phalangopsinae	2 of 3		11
	Phaloriinae	1 of 2		9
	Pteroplistinae	1 of 1		2
	No subfamily	1 of 2		7
	No subfamily	No tribe		2
	"Podoscirtinae subfamily group"
	Euscyrtinae	No tribe		3
	Hapithinae	2 of 2		5
	Podoscirtinae	3 of 3		17
	Pentacentrinae	1 of 5		2
	No hypothesized "subfamily group"
	Eneopterinae	5 of 5		19
	Nemobiinae	4 of 6		18
	Oecanthinae	1 of 2		3
	Trigonidiinae	2 of 2		13
	GRYLLOTALPIDAE
	Gryllotalpinae	No tribe		4
	Scapteriscinae	No tribe	-	-
	MOGOPLISTIDAE
	Malgasiinae	No tribe		1
	Mogoplistinae	2 of 2		9
	MYRMECOPHILIDAE
	Myrmecophilinae	1 of 2		2
	4 of 4 families	37 of 52		205
	22 of 25 subfamilies
	-, not sampled.

Table 2

 2 Specimens use for molecular analysis, with voucher data and GenBank accession numbers (sequences generated for this study are in bold)

  Sci, Sciobiini; Scl, Sclerogryllini; The, Thetellini; Tri, Trigonidiini; Tur, Turanogryllini; Xen, Xenogryllini. Repository abbreviations: ANIC, Australian National Insect Collection, Australia; ES, Collection E. Sardet, France; IAC, Institut Agronomique de Nouvelle-Cal edonie, La Foa, New Caledonia; INBio, Instituto Nacional de Biodiversidad, Santo Domingo de Heredia, Costa Rica; ENSIF, Ensifera collection of the MNHN, Paris; MEUV, Museo de Entomologia del Universidad de la Valle, Cali, Colombia; RF, Collection Rob Felix, The Netherlands; SH, Collection Sylvain Hugel, Strasbourg, France; ZSI, Zoological Survey of India, Calcutta, India.

				12S 16S cyt b 18S 28SA 28SD H3	KR903918 KR903745 KR903406 KR904105 KR903579 KR903068 ND		KR903976 ND KR903452 KR904160 KR903631 KR903109 KR903283				Number Final	Elongation of cycles elongation Reference	°C 40 s, 30 s at 72 °C 10, 40 7 min at 72 °C Kamphambati (1995)	40 s at 72 °C 45 5 min at 72 °C Robillard and	Desutter-Grandcolas (2006)	50 s at 72 °C 50 7 min at 72 °C Giribet et al. (1999)		40 s at 72 °C 45 7 min at 72 °C Svenson and Whiting (2004)		60 s at 72°C 40 7 min at 72 °C Whiting (2002)	Jarvis et al. (2004)	50 s at 72°C 40 7 min at 72 °C This study		40 s at 72°C 50 7 min at 72 °C Robillard and Desutter-Grandcolas	(2006)
	Table 2	(Continued)	Voucher/ Molecular	Locality repository codes Genus Species Subfam/tribe	France ENSIF2923 LDG 115 Tachycines asynamora (Adelung, OG-RH	1902)	West Indies, ENSIF2160 LDG 195 Xanthogryllacris punctipennis (Walker, OG-GC	Guadeloupe 1869)	Voucher codes as indicated in Fig. 1 ND	Table 3	PCR profiles and primers used, with their sources	Gene Primer Denaturation Annealing	°C 30 s, 30 s 12SF TACTATGTTACGACTTAT 30 s at 94	at 47 °C, 48 12SR AAACTAGGATTAGATACCC	°C °C 40 s at 55 16S-AG CGCCTGTTTATCAAAAACATGT 30 s at 94	16S-BG AGATCACGTAAGAATTTAATGGTC	°C °C 50 s at 52 18S-A2 ATGGTTGCAAAGCTGAAAC 50 s at 94	18S-9R GATCCTTCCGCAGGTTCACCTAC	°C °C 40 s at 58 HisF ATGGCTCGTACCAAGCAGACGGC 50 s at 94	HisR ATATCCTTGGGCATGATGGTGAC	°C °C 40 s at 50 28S--R1.2a CCC SSG TAA TTT AAG CAT ATT A 40 s at 94	28S-Rd3b CCY TGA ACG GTT TCA CGT ACT	°C °C 35 s at 57 28SF4 CGA CAC GCC CCG ATC CTC AGA GCC A 35 s at 94	28SR4 GATTCTGACGTGCAAATCGATC	°C °C 40 s at 48 30 s at 94 cytb427F YTWGTWCAATGARTMTGAGG	cytb800R CCYARTTTATTAGGAATTGATCG

= No data. Subfamily abbreviations: CACH, Cachoplistinae; ENEO, Eneopterinae; EUSC, Euscyrtinae; GRMP, Gryllomorphinae; GRTL, Gryllotalpinae; GRYL, Gryllinae; HAPI, Hapithinae; LAND, Landrevinae; LUZA, Luzarinae; MALG, Malgasiinae; MOGO, Mogoplistinae; MYRM, Myrmecophilinae; NEMO, Nemobiinae; OECA, Oecanthinae; PARA, Paragryllinae; PENT, Pentacentrinae; PHAL, Phalangopsinae; PHOR, Phaloriinae; PODO, Podoscirtinae; PTER, Pteroplistinae; SCLE, Sclerogryllinae; TRIG, Trigonidiinae. Tribe abbreviations: Aph, Aphonoidini; Ara, Arachnocephalini; Cae, Cearacesaini; Cep, Cephalogryllini; Cop, Cophogryllini; End, Endacustini; Ene, Eneopterini; Eur, Euripini; Grm, Gryllomorphini; Gry, Gryllini; Hap, Hapithini; Hem, Hemigryllini; Het, Heterogryllini; Hom, Homoeogryllini; Lan, Landrevini; Leb, Lebinthini; Mod, Modicogryllini; Mog, Mogoplistini; Myr, Myrmecophilini; Nem, Nemobiini; Nis, Nisitrini; Oec, Oecanthini; Pac, Paroecanthini; Par, Paragryllini; Pet, Petaloptilini; Pha, Phalangopsini; Pho, Phaloriini; Phy, Phylloscirtini; Pod, Podoscirtini; Pte, Pteronemobiini; Ptr, Pteroplistini;

Table 4

 4 Descriptive statistics for the different sequence datasets with the outgroups included. Statistics were estimated after removal of primer sequences and the best model of evolution

		Average frequencies Constancy of base frequencies	all sites/variable sites (v 2 values/P values/d.f.)		AIC (-) lnL A C G T All sites Variable sites	31301.042 15252.521 0.2/0.2 0.2/0.3 0.3/0.3 0.2/0.3 98.6/1/576 168.9/1/576	29250.050 14219.025 0.3/0.2 0.3/0.3 0.3/0.3 0.2/0.2 320.9/1/546 508.5/1/546	21538.126 10437.063 0.2/0.2 0.3/0.3 0.3/0.3 0.2/0.2 129.6/1/483 234.8/1/483	25791.604 12491.802 0.2/0.1 0.3/0.5 0.3/0.3 0.2/0.1 322.8/1/549 793.6/0/549	66519.833 32831.916 0.4/0.4 0.2/0.2 0.1/0.1 0.3/0.3 626.6/0/615 754.2/0/615	64338.811 31816.405 0.3/0.3 0.1/0.1 0.2/0.2 0.4/0.4 549.6/0.2/519 722.7/0/519	51714.423 25493.211 0.3/0.3 0.2/0.2 0.1/0.1 0.4/0.4 703.6/0/528 1027.3/0/528	312107.864 155617.932 0.3/0.3 0.2/0.2 0.2/0.2 0.3/0.3 2930.8/0/639 3095.7/0/639
				Model of	evolution	TIM2 + I + G	TIM2 + I + G	GTR + I + G	TIM1 + I + G	TIM2 + I + G	TPM2uf + I + G	GTR + I + G	GTR + I + G
	selected with the AIC criterion in jModelTest 2.1.3	No/% parsimony	No/% informative sites	variable (gap as missing No. No.	sites character) Loci taxa characters	368/53.7 304/44.4 18S 192 685	290/60.2 225/46.7 28SA 181 482	304/51.3 213/35.9 28SD 162 593	142/43.3 130/39.6 H3 180 328	469/82.1 382/66.9 12S 206 571	443/74.8 382/64.5 16S 174 592	251/72.5 220/63.6 cytb 177 346	2262/62.8 1849/51.4 Combined 214 3597	d.f., degrees of freedom.

Table 5

 5 Results of the phylogenetic test of the current classification used in the OSF, and its potential match with the phylogenetic topology

	OSF classification	Topology
	OSF Families/Subfamilies	Monophyly status	Resultant clades
	GRYLLIDAE
	"Gryllinae subfamily group"
	Gryllinae	Monophyletic without Gryllomorpha,	Included in clade G4
		Petaloptila, and with Sclerogryllus
	Gryllomiminae	Not tested (1 genus)	-
	Gryllomorphinae	Polyphyletic	Dispatched in clades E3 and G4
	Itarinae	Not tested (2 genera)	-
	Landrevinae	Polyphyletic	Dispatched in clades G1, G2 and G4
	Sclerogryllinae	Nested clade	Included in clade G4
	"Podoscirtinae subfamily group"
	Euscyrtinae	Monophyletic	Equal to clade F1
	Hapithinae	Polyphyletic	Included in clade F5
	Podoscirtinae	Polyphyletic	Dispathed in clades F3, F4 and F5
	Pentacentrinae	Polyphyletic	Included in clade G1
	No hypothesized "subfamily group"
	Eneopterinae	Nested clade	Dispatched in clades F5 and G3
	Oecanthinae	Monophyletic	Equal to clade F2
	"Phalangopsinae subfamily group"
	Cachoplistinae	Polyphyletic	Dispatched in clades E2, E3 and E4
	Luzarinae	Polyphyletic	Dispatched in clades E1, E4 and E5
	Paragryllinae	Monophyletic with Caltathra and Protathra	Included in clade E2
	Phalangopsinae	Polyphyletic	Dispatched in clades E2, E3, E4 and E5
	Phaloriinae	Monophyletic	Included in clade E1
	Pteroplistinae	Monophyletic	Equal to clade D
	Endacustini	Polyphyletic	Dispatched in clades E2 and E4
	No hypothesized "subfamily group"
	Nemobiinae	Monophyletic without Hemigryllus	Dispatched in clades C2 and G2
	Trigonidiinae	Monophyletic	Equal to clade C1
	MOGOPLISTIDAE
	Malgasiinae	Nested clade	Included in clade B
	Mogoplistinae	Monophyletic with Malgasia	Included in clade B
	GRYLLOTALPIDAE
	Gryllotalpinae	Monophyletic	Equal to clade A1
	Scapteriscinae	Not tested (2 genera)	-
	MYRMECOPHILIDAE
	Myrmecophilinae	Paraphyletic due to Schizodactylus, otherwise monophyletic	Included in clade A2

Acknowledgments

We thank Hojun Song (USA) for the opportunity to present this work at the 11th Congress of Orthopterology, Kunming, China,[11][12][13][14][15][16] August 2013. The laboratory work was carried in the Service de Syst ematique mol eculaire (SSM, UMS 2700 CNRS) of the MNHN (Paris, France): we thank M.C. Boisselier, C. Bonillo, D. Gey, J. Lambourdi ere and S. Leprieur for their help in data acquisition. This work was partly supported by agreement no. 2005/67 between the Genoscope (Evry, France) and the MNHN project 'Macrophylogeny of life'; sequencing was also undertaken in the project @SpeedID proposed by F-BoL, the French Barcode of life initiative and the network 'Biblioth eque du vivant' funded by the CNRS, INRA and MNHN. Part of the phylogenetic analyses were carried out using the resources of the MNHN parallel computing cluster (PCIA, UMS 2700). The following institutions and programmes support field work: the Programme Pluriformation DRED/MNHN 'Biodiversit e en Nouvelle-Cal edonie', the ANR 'BioNeoCal' (P. Grandcolas), the IFCPAR (project 3009-1, P.I.s L. Desutter-Grandcolas, MNHN and R. Balakrishnan, IISc), the UMR 7205 CNRS (in respect of L. Deharveng), the European programme ECOFAC (Universit e Rennes I, A. Gautier-Hion); the Mus eum national d'Histoire naturelle, Paris (ATM "Biodiversit e actuelle et fossile", in respect of S. Peign e and P. Janvier; ATM "Formes possibles, forms r ealis ees", V. Bels, P.H. Gouyon), the Agence Nationale de la Recherche's Investissement d'Avenir program referenced ANR-11-INBS-0001AnaEE-Services and Labex CEBA ANR-10-LABX-25-01, the subproject "Connaître pour pr eserver le patrimoine cach e des Comores", part of "Biodiversit e des îles de l'Oc ean indien" (FRB, MNHN, CNRDS), the programme ALAS (OTS, NGS, INBio, J. Longino), The Parc National de la R eunion, the Parc National de la Guadeloupe, the Minist ere de l'Environnement et des forêts (Madagascar), the National Parks and Conservation Service (Mauritius) and the Seychelles Bureau of Standards. We are in debt to the numerous colleagues and institutions who helped us during field work. They can unfortunately not be all named here. The following colleagues provided material for molecular studies: R. Felix (The Netherlands), B. Junger (France), K.-G. Heller (Germany), A. Mohamed Sahnoun (Universit e de Tizi Ouzou, Algeria), E. Sardet (France) and P. Terret (R eserve des Nouragues, Guyane franc ßaise). We thank Hannah ter Hofstede for improving our manu-script. We thank the associate Editors and two anonymous reviewers for their comments.

Supporting Information

Additional Supporting Information may be found in the online version of this article:

Figures S1-S7. Results of the separate phylogenetic analyses in ML for 12S, 16S, 18S, 28SA, 28SD, cytb and H3 markers, respectively. ln L 12S = À32572.585003; ln L 16S = À32702.581814; ln L 18S = À15462.852866; ln L 28SA = À13704.500232; ln L 28SD = À10522.780197; ln L cytb = À25817.127967; ln L H3 = À12141.167417.

Figure S8. Result of the combined phylogenetic analysis in ML (ln L = À150351.480431).

Figure S9. Results of the combined (without marker 28SD) phylogenetic analysis in ML (ln L = À137828.405642).

Figure S10. Result of the combined phylogenetic analysis in parsimony (strict-consensus of four trees of 37341 steps).

Figure S11. Result of the phylogenetic analysis of the nuclear data in MP.

Figure S12. Result of the phylogenetic analysis of the nuclear data in ML.

Figure S13. Result of the phylogenetic analysis of the mitochondrial data in MP.

Figure S14. Result of the phylogenetic analysis of the mitochondrial data in ML.

Figure S15. Result of the combined phylogenetic analysis in ML (ln L = À149666.340159) without Comicus campestris and Schizodactylus monstrosus.

Figure S16. Result of the combined phylogenetic analysis in BI without Comicus campestris and Schizodactylus monstrosus.

Figure S17. Result of the combined phylogenetic analysis in MP (strict-consensus of 333 trees of 37155 steps) without Comicus campestris and Schizodactylus monstrosus.

Data S1. Phylogenetic matrix used in this study.