Insights into the molecular phylogeny of Rhaphidophoridae, an ancient, worldwide lineage of Orthoptera.

We investigated the molecular phylogenetic divergence and historical biogeography of cave crickets belonging to the family Rhaphidophoridae (Orthoptera, Ensifera). We used taxa representative of most of the regions embraced by the family, considering samples of Macropathinae from Gondwana land (i.e., Tasmania, Australia, New Zealand, South Africa, and South America); Aemodogryllinae and Rhaphidophorinae from Southern-eastern Asia (i.e., India, Bhutan, China, Philippines and the Sulawesi islands); Dolichopodainae and Troglophilinae from the Mediterranean region and Ceuthophilinae from North America. Based on previous papers, we carried out an analysis of both mitochondrial and nuclear DNA sequences considering the ribosomal RNA units 12S, 16S, 18S, and 28S. To reconstruct phylogeny, we use cladistics, Maximum Likelihood (ML), and Bayesian analyses. All phylogenetic analyses showed the same highly supported topology generally congruent with the classical systematic arrangement at the level of each sub-family but strongly disagree with previous affinity hypotheses between sub-families based on morphological characters. Our results reveal a close affinity between Asiatic and Gondwanian taxa from one hand and between North American and Mediterranean ones from the other hand. Dating estimates indicated that Rhaphidophoridae originated in the Cretaceous period during the Mesozoic era with the ancestral area located both in the northern and southern hemisphere. A possible biogeographic scenario, reconstructed using S-DEC with RASP software, suggested that the current distribution of Rhaphidophoridae might be explained by a combination of both dispersal and vicariance events occurred especially in the ancestral populations. The radiation of Rhaphidophoridae started within the Pangaea, where the ancestor of Rhaphidophoridae occurred throughout an ancestral area including Australia, North America, and the Mediterranean region. The opening of the Atlantic Ocean promoted the divergence of North American and Mediterranean lineages while the differentiation of the southern lineages, spread from Australia, appears to be related to the fragmentation of Gondwana land.

Testing Classical Species Properties with Contemporary Data: How "Bad Species" in the Brassy Ringlets (Erebia tyndarus complex, Lepidoptera) Turned Good.

All species concepts are rooted in reproductive, and ultimately genealogical, relations. Genetic data are thus the most important source of information for species delimitation. Current ease of access to genomic data and recent computational advances are blooming a plethora of coalescent-based species delimitation methods. Despite their utility as objective approaches to identify species boundaries, coalescent-based methods (1) rely on simplified demographic models that may fail to capture some attributes of biological species, (2) do not make explicit use of the geographic information contained in the data, and (3) are often computationally intensive. In this article, we present a case of species delimitation in the Erebia tyndarus species complex, a taxon regarded as a classic example of problematic taxonomic resolution. Our approach to species delimitation used genomic data to test predictions rooted in the biological species concept and in the criterion of coexistence in sympatry. We (1) obtained restriction-site associated DNA (RAD) sequencing data from a carefully designed sample, (2) applied two genotype clustering algorithms to identify genetic clusters, and (3) performed within-clusters and between-clusters analyses of isolation by distance as a test for intrinsic reproductive barriers. Comparison of our results with those from a Bayes factor delimitation coalescent-based analysis, showed that coalescent-based approaches may lead to overconfident splitting of allopatric populations, and indicated that incorrect species delimitation is likely to be inferred when an incomplete geographic sample is analyzed. While we acknowledge the theoretical justification and practical usefulness of coalescent-based species delimitation methods, our results stress that, even in the phylogenomic era, the toolkit for species delimitation should not dismiss more traditional, biologically grounded, approaches coupling genomic data with geographic information.

The evolutionary jigsaw puzzle of the surviving trout (Salmo trutta L. complex) diversity in the Italian region. A multilocus Bayesian approach.

Mediterranean trout populations display a diversity of phenotypes, representing a valuable model for the study of adaptation and a puzzling dilemma for taxonomists and biogeographers, which is further entangled by the widespread introgression of allochthonous genes. In this paper we analysed DNA polymorphism at multiple loci (sequence variation of the mitochondrial control region and eight nuclear fragments and length variation at eleven nuclear microsatellite loci) in representative samples of the autochthonous taxonomic diversity described in Italian trout populations (Salmo marmoratus, S. carpio, S. cenerinus, S. cettii and S. fibreni) and in samples from hatchery-originated strains of Atlantic S. trutta. We employed model-based clustering and Approximate Bayesian Computation in order to: (i) describe the phylogeographic structure of Italian autochthonous trout populations; (ii) evaluate a set of evolutionary/biogeographic models. The inclusion of hatchery-originated strains allowed to account for man-mediated allochthonous introgression in Italian populations. Our results (i) showed that the analysed sample consists of two main autochthonous evolutionary lineages, including the marble trout populations on one side ('marble' lineage) and the three peninsular populations of S. cettii, S. cenerinus and S. fibreni on the other side ('peninsular' lineage); (ii) indicated that S. carpio originated from a 'peninsular' population, with a possible, limited contribution from the 'marble' lineage; (iii) pointed out that the 'marble' lineage started diverging before the separation of the 'peninsular' lineage from Atlantic S. trutta; (iv) suggested that a model of divergence involving gene flow from the 'peninsular' population into the ancestral gene pool of 'marble' trout is most consistent with the genetic data; (v) provided evidence that the autochthonous trout gene pools in the Tyrrhenian and Adriatic basins of the Italian peninsula started diverging very recently (most likely after the last glacial maximum).

Tempo and mode of species diversification in Dolichopoda cave crickets (Orthoptera, Rhaphidophoridae).

This study focuses on the phylogenetic relationships among ninety percent of known Dolichopoda species (44 out of 49); primarily a Mediterranean genus, distributed from eastern Pyrenees to Caucasus. A total of 2490 base pairs were sequenced corresponding to partial sequences of one nuclear (28SrRNA) and three mitochondrial genes (12S, 16S and COI). A relaxed molecular clock, inferred from Bayesian analysis was applied to estimate the divergence times between the lineages using well dated palaeoevents of the study areas. Molecular substitution rates per lineage per million years were also obtained for each analyzed gene. Based on the nearly complete species phylogeny, temporal patterns of diversification were analyzed using Lineage-Through-Time plots and diversification statistics. Alternative hypotheses about the colonization of present range by Dolichopoda species were tested by means of Approximate Bayesian Computation analysis. Results from this analysis carried out on the 90% of known Dolichopoda species confirmed the previous ones based on subgroups of species, suggesting the ABC analysis as a remarkable tool in biogeographic studies. Based on these results, the distribution of Dolichopoda species appears to have been shaped by the palaeogeographic and climatic events that occurred from Late Miocene up to the Plio-Pleistocene. Both vicariance and dispersal events appear to have influenced Dolichopoda species distributions, with many processes occurring in ancestral epigean populations before the invasion of the subterranean environment.

New data on Weddell seal (Leptonychotes weddellii) colonies: A genetic analysis of a top predator from the Ross Sea, Antarctica.

In this paper, we studied the genetic variability in Weddell seal from colonies in Terra Nova Bay and Wood Bay, both sites located in the Ross Sea area, Antarctica. Two mitochondrial genes and one nuclear gene, with different mutation rates, were sequenced to investigate the haplotype diversity of the colonies and to test for a possible recent expansion. Fifteen microsatellites were used to analyze their genetic structure. Sequenced genes and microsatellites were also used to estimate the effective population size of the studied colonies and the Ross Sea seal population. The Ross Sea has a high density population of Weddel seals, with an estimated effective number of 50,000 females, and 1,341 individuals for the sampling area, possibly due to its high primary production. The colonies showed high diversity (Hd > 0.90) and many exclusive haplotypes (> 75%), likely a consequence of the surprisingly high site fidelity of Weddell seals, despite the proximity of the colonies. Nevertheless, there was low microsatellite differentiation between colonies, suggesting that they are part of a single larger population. Their expansion seemed to have started during the last glacial cycle (around 58,000 years ago), indicating that the Ross Sea seal populations have been present in the area for long time, probably due to the lack of hunting by humans and terrestrial predation. As a top predator, the role of Weddell seals in the Ross Sea ecology is crucial, and its demographic dynamics should be monitored to follow the future changes of such an important ecosystem.

Is radon emission in caves causing deletions in satellite DNA sequences of cave-dwelling crickets?

The most stable isotope of radon, 222Rn, represents the major source of natural radioactivity in confined environments such as mines, caves and houses. In this study, we explored the possible radon-related effects on the genome of Dolichopoda cave crickets (Orthoptera, Rhaphidophoridae) sampled in caves with different concentrations of radon. We analyzed specimens from ten populations belonging to two genetically closely related species, D. geniculata and D. laetitiae, and explored the possible association between the radioactivity dose and the level of genetic polymorphism in a specific family of satellite DNA (pDo500 satDNA). Radon concentration in the analyzed caves ranged from 221 to 26,000 Bq/m3. Specimens coming from caves with the highest radon concentration showed also the highest variability estimates in both species, and the increased sequence heterogeneity at pDo500 satDNA level can be explained as an effect of the mutation pressure induced by radon in cave. We discovered a specific category of nuclear DNA, the highly repetitive satellite DNA, where the effects of the exposure at high levels of radon-related ionizing radiation are detectable, suggesting that the satDNA sequences might be a valuable tool to disclose harmful effects also in other organisms exposed to high levels of radon concentration.

Phylogeography and systematics of the westernmost Italian Dolichopoda species (Orthoptera, Rhaphidophoridae).

The genus Dolichopoda (Orthoptera; Rhaphidopohoridae) is present in Italy with 9 species distributed from northwestern Italy (Piedmont and Liguria) to the southernmost Apennines (Calabria), occurring also in the Tyrrhenian coastal areas and in Sardinia. Three morphologically very close taxa have been described in Piedmont and Liguria, i.e., D. ligustica ligustica, D. ligustica septentrionalis and D. azami azami. To investigate the delimitation of the northwestern species of Dolichopoda, we performed both morphological and molecular analyses. Morphological analysis was carried out by considering diagnostic characters generally used to distinguish different taxa, as the shape of epiphallus in males and the subgenital fig in females. Molecular analysis was performed by sequencing three mitochondrial genes, 12S rRNA, 16S rRNA, partially sequenced and the entire gene of COI. Results from both morphological and molecular analyses highlighted a very homogeneous group of populations, although genetically structured. Three haplogroups geographically distributed could be distinguished and based on these results we suggest a new taxonomic arrangement. All populations, due to the priority of description, should be assigned to D. azami azami Saulcy, 1893 and to preserve the names ligustica and septentrionalis, corresponding to different genetic haplogroups, we assign them to D. azami ligustica stat. n. Baccetti & Capra, 1959 and to D. azami septentrionalis stat. n. Baccetti & Capra, 1959.

Molecular phylogeography of Dolichopoda cave crickets (Orthoptera, Rhaphidophoridae): a scenario suggested by mitochondrial DNA.

This study focuses on the phylogenetic relationships among a number of West-Mediterranean cave crickets species belonging to Dolichopoda; primarily a Mediterranean genus, distributed from eastern Pyrenees to Caucasus. In this paper, 11 Dolichopoda species from the French Pyrenees (D. linderi), the island of Corsica (D. bormansi and D. cyrnensis), and northern, central, and southern Italy (D. ligustica, D. schiavazzii, D. aegilion, D. baccettii, D. laetitiae, D. geniculata, D. capreensis, and D. palpata) were studied. Two more species, one from the Caucasus, D. euxina, and one from Greece, D. remyi, were also included in the analyses, together with more distant species within the same family to be used as outgroups. Fifteen hundred base pairs of mitochondrial DNA, corresponding to the small subunit of the ribosomal RNA (16S rRNA) and to the subunit I of the cytochrome oxidase I (COI), were sequenced in order to clarify the phylogenetic relationships and biogeography of this group of Mediterranean cave crickets. The molecular data are congruent with a phylogeographic pattern; with the geographically close species also the most related ones. Based on mtDNA divergence, the present-day distribution of genetic diversity seems to have been impacted by climatic events due to glacial and interglacial cycles that have characterized the Pleistocene era.