Explosive evolution of an ancient group of Cyphophthalmi (Arachnida: Opiliones) in the Balkan Peninsula

Aim  To investigate the phylogeny of the genus Cyphophthalmus in the Balkan Peninsula and to test the current recognition of 'phyletic lines' and phylogenetic groups proposed in previous studies in order to elucidate the biogeographical history of the region.
Location  Europe, Balkan Peninsula, Adria microplate.
Methods  Two mitochondrial (cytochrome c oxidase subunit I and 16S rRNA) and two nuclear (28S rRNA and 18S rRNA) markers were used to infer the phylogenetic history of the group. Molecular dating with relaxed molecular clocks was used to elucidate the relative time of diversification within the genus Cyphophthalmus and its constituent lineages.
Results  Our analyses confirm the monophyly of the genus Cyphophthalmus , and that of the Aegean and gjorgjevici lineages, whereas the 'Dinaric lineage' appears paraphyletic.
Main conclusions  We show that the genus Cyphophthalmus is an old endemic from the Balkan biogeographical region, which gave origin to at least three main lineages. Those lineages have diversified within overlapping ranges. According to our molecular dating, they have also diversified within the same timeframe. The Dinaric Alps, although presenting a large number of species, cannot be inferred as the centre of origin of the group. Instead, the biogeographical evolution of the genus could be related to the palaeogeographic history of the Adria microplate.     

Molecular phylogeny of the Greek legless skink Ophiomorus punctatissimus (Squamata: Scincidae): the impact of the mid-Aegean trench in its phylogeography

Sequence data derived from three mitochondrial markers (cytochrome b, 16S rRNA and 12S rRNA genes) were used to infer the evolutionary history of several insular and mainland populations of the Greek legless skink (Ophiomorus punctatissimus), covering most of its distributional range. All phylogenetic analyses produced topologically identical trees that revealed a well-resolved phylogeny. These trees support two O. punctatissimus clades, which are geographically separated (west and east of the mid-Aegean trench). The assumption of a clock-like evolution could not be rejected, and thus a local clock was calibrated for the O. punctatissimus lineages. The non-overlapping geographic distributions of the major clades suggest a spatial and temporal sequence of diversification that coincides with paleogeographic separations during the geological history of the Aegean region. It seems that O. punctatissimus is an old eastern Mediterranean species that has been differentiating in this region at least from middle Miocene. It is possible that the ancestral form of O. punctatissimus invaded the Aegean region from Anatolia before the complete formation of the mid-Aegean trench, when the Aegean was still a uniform landmass, while other vicariant events have led to its present distribution.     

Molecular phylogeography of Troglophilus cave crickets (Orthoptera,Rhaphidophoridae): A combination of vicariance and dispersal drove diversification in the East Mediterranean region

In this study, we investigated the molecular phylogenetic divergence and historical biogeography of cave crickets belonging to the genus Troglophilus (Orthoptera, Rhaphidophoridae) from caves in eastern Mediterranean and Anatolia regions. Three mitochondrial DNA genes (COI, 12S rDNA, and 16S rDNA) and two nuclear ones (18S rDNA and 28S rDNA) were amplified and partially sequenced to reconstruct phylogenetic relationships among most of the known Troglophilus species. Results showed a well‐resolved phylogeny with three main clades representing the Balkan, the Anatolian, and the Cycladian–Cretan lineages. Based on Bayesian analyses, we applied a relaxed molecular clock model to estimate the divergence times between these three lineages. Dating estimates indicate that radiation of the ingroup might have been triggered by the opening of the Mid‐Aegean trench, while the uplift of the Anatolian Plateau in Turkey and the changes of relief, emergence, and disappearance of orographic and hydrographical barriers in the Balkan Peninsula are potential paleogeographic events responsible for the initial diversification of the genus Troglophilus. A possible biogeographic scenario, reconstructed using S‐DIVA with RASP software, suggested that the current distribution of Troglophilus species can be explained by a combination of both dispersal and vicariance events that occurred in particular in the ancestral populations. The radiation of Troglophilus species likely started from the Aegean and proceeded eastward to Anatolia and westward to the Balkan region. Results are additionally compared to those available for Dolichopoda, the only other representative genus of Rhaphidophoridae present in the Mediterranean area.     

Salenthydrobia gen. nov. (Rissooidea: Hydrobiidae): a potential relict of the Messinian salinity crisis

The Messinian salinity crisis (MSC) occurred synchronously throughout the Mediterranean basin about 5.96 ± 0.02 Mya and represents one of the most dramatic oceanic changes since the early Miocene. It is thought that the concomitant environmental changes brought about isolation of faunas and the development of endemism. As part of the search for possible speciation events triggered by the MSC, the author studied 38 populations of hydrobiine snails from the Mediterranean and Black Sea, including three populations from the Salentina Peninsula, Italy. Partial sequences (COI, 16S) and anatomical data were used to test the taxonomic and phylogenetic status of the peninsular populations. A maximum likelihood analysis of 11 hydrobiine taxa revealed five clades and lineages, four of which corresponded to previously recognized genera: Adriohydrobia , Hydrobia , Peringia , Ventrosia . The fifth clade was formed by haplotypes of the peninsular populations, which are characterized by distinct male and female reproductive systems. Based on molecular and anatomical data, these populations are considered to represent a new species, Salenthydrobia ferrerii , belonging to a new genus, Salenthydrobia . Ecological and biogeographical data for S. ferrerii strongly support a correlation between its origin and the MSC. Based on an island age of 5.33 Myr and a population divergence of 0.0973 ± 0.0114, the COI molecular clock rate for the Salenthydrobia and Peringia clades would be 1.83 ± 0.21% population divergence per Myr. The genetic diversity of S. ferrerii , its phylogenetic relationships, and the validity of the proposed local molecular clock rate are discussed.     

Testing phylogenetic hypotheses for reconstructing the evolutionary history of Dolichopoda cave crickets in the eastern Mediterranean

Aim To investigate the molecular phylogenetic divergence and historical biogeography of cave crickets belonging to the genus Dolichopoda (Orthoptera, Rhaphidophoridae). Location Caves in continental and insular Greece. Methods We sequenced 1967 bp of mitochondrial DNA, corresponding to three fragments of the small and large subunit of the ribosomal RNA (16S and 12S rRNA, respectively) and to the subunit I of cytochrome oxidase (COI), to reconstruct phylogenetic relationships among all 30 known Greek species of Dolichopoda. Alternative hypotheses about the colonization of the Hellenic Peninsula by Dolichopoda species were tested by comparing the degree of discordance between species trees and gene trees under four plausible biogeographical scenarios. Results The present study revealed a rather well resolved phylogeny at species level, identifying a number of clades that represent long‐separated lineages and diverse evolutionary histories within the genus Dolichopoda. Two main clades were revealed within Hellenic–Aegean species, identifying a north‐western and a south‐eastern species group. Based on Bayesian analysis, we applied a relaxed molecular clock to estimate the divergence times between the lineages. The results revealed that the origins of eastern Mediterranean lineages are much older than those of previously studied western Mediterranean Dolichopoda. Tests of alternative biogeographical hypotheses showed that a double colonization of the Hellenic Peninsula, following separate continental and trans‐Aegean routes during the Messinian stage, best accounts for the present distribution of Greek Dolichopoda species. Main conclusions Reconstruction and biogeographical hypothesis testing indicated that the colonization of Greece by Dolichopoda species comprised two episodes and two different routes. The southern lineage probably arose from a trans‐Aegean colonization during the Messinian salinity crisis (5.96–5.33 Ma). The northern lineage could be the result of dispersal from the north through the Balkan Peninsula. The opening of the Mid‐Aegean Trench could have promoted an initial diversification within the uprising Anatolian Plateau, while the Messinian marine regression offered the conditions for a rapid dispersal through the whole Aegean–Hellenic region. In addition, climatic events during the Plio‐Pleistocene may have been responsible for the speciation within each of the two different phylogeographical units, principally attributable to vicariance events.     

Molecular biogeography of the Mediterranean lizards Podarcis Wagler, 1830 and Teira Gray, 1838 (Reptilia, Lacertidae)

Aim   We discuss biogeographical hypotheses for the Mediterranean lizard species Podarcis and Teira within a phylogenetic framework based on partial mitochondrial DNA sequences.
Methods   We derived the most likely phylogenetic hypothesis from our data set (597 aligned positions from the 12S rDNA and phenyl tRNA) under parsimony, distance and maximum likelihood assumptions.
Results   The species usually included in Teira do not form a strongly monophyletic clade. In contrast, the monophyly of the genus Podarcis is rather well supported. Seven lineages are identified in the genus; in order of appearance within the tree, these are: the Balearic pityusensis and lilfordi pair, the sicula complex, a Tyrrhenian tiliguerta and raffonei pair, muralis , the Siculo-Maltese filfolensis and wagleriana pair, the Balkan group ( erhardi , peloponnesiaca , milensis , melisellensis and taurica ), and the Ibero-Maghrebian group ( bocagei , atrata , hispanica and vaucheri ).
Conclusions   The origin of the three European genera of lacertid assayed ( Lacerta , Teira and Podarcis ) is hypothesized to have occurred in the Oligocene. For Podarcis , a possible scenario of a Miocene diversification is derived from the sequence data, and the zoogeography of the lineages are discussed in relation to the palaeogeography of the Mediterranean. It is hypothesized that in the early history of the genus the main lineages separated by rapid, numerous and close events that produced a starting point very similar to a polytomy, hard to resolve by parsimony analysis of the data set.     

Molecular phylogeny and phylogeography of the Greek populations of the genus Orthometopon (Isopoda, Oniscidea) based on mitochondrial DNA sequences

We infer phylogenetic relationships among isopod species of the genus Orthometopon distributed in the Greek area, comparing partial mitochondrial DNA sequences for cytochrome oxidase I (COI). All phylogenetic analyses produced topologically identical trees that revealed a well-resolved phylogeny. These trees support the monophyly of the genus Orthometopon , and suggest two clades that correspond to separate geographical regions (west and east of the mid-Aegean trench). However, the phylogenetic relationships among Greek populations of Orthometopon spp. are different from the presumed pattern on the basis of morphological evidence. The distinct geographical distribution of the major clades of the phylogenetic tree and its topology suggest a spatial and temporal sequence of phylogenetic separations, which coincide with some major palaeogeographical separations during the geological history of the Aegean Sea. The results stress the need for a reconsideration of the evolutionary history of Orthometopon species, which will help overcome difficulties encountered in classical taxonomy at the species level. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society , 2008, 152 , 707–715.     

Fluvial basin history in the northeastern Mediterranean region underlies dispersal and speciation patterns in the genus Dugesia (Platyhelminthes,Tricladida, Dugesiidae)

In this study we analyzed the phylogenetic relationships of eastern Mediterranean freshwater planarians of the genus Dugesia, estimated divergence times for the various clades, and correlated their phylogeographic patterns with geological and paleoclimatic events, in order to discover which evolutionary processes have shaped the present-day distribution of these animals. Specimens were collected from freshwater courses and lakes in continental and insular Greece. Genetic divergences and phylogenetic relationships were inferred by using the mitochondrial gene subunit I of cytochrome oxidase (COI) and the nuclear ribosomal internal transcribed spacer-1 (ITS-1) from 74 newly collected individuals from Greece. Divergence time estimates were obtained under a Bayesian framework, using the COI sequences. Two alternative geological dates for the isolation of Crete from the mainland were tested as calibration points. A clear phylogeographic pattern was present for Dugesia lineages in the Eastern Mediterranean. Morphological data, combined with information on genetic divergences, revealed that eight out of the nine known species were represented in the samples, while additional new, and still undescribed species were detected. Divergence time analyses suggested that Dugesia species became isolated in Crete after the first geological isolation of the island, and that their present distribution in the Eastern Mediterranean has been shaped mainly by vicariant events but also by dispersal. During the Messinian salinity crisis these freshwater planarians apparently were not able to cross the sea barrier between Crete and the mainland, while they probably did disperse between islands in the Aegean Sea. Their dependence on freshwater to survive suggests the presence of contiguous freshwater bodies in those regions. Our results also suggest a major extinction of freshwater planarians on the Peloponnese at the end of the Pliocene, while about 2 Mya ago, when the current Mediterranean climate was established, these Peloponnese populations probably began to disperse again. At the end of the Pliocene or during the Pleistocene, mainland populations of Dugesia colonized the western coast, including the Ionian Islands, which were then part of the continent.     

Sister species within the Triops cancriformis lineage (Crustacea, Notostraca)

We investigated the phylogenetic relationships among the three presently recognized subspecies of the tadpole shrimp, Triops cancriformis , using mitochondrial 16S and 12S rDNA sequences. Our results indicate that the taxon is divided into two distinct lineages. One lineage is formed of T. c. cancriformis populations and samples from northern Spain that had been classified as T. c. simplex in the most recent literature. The second lineage comprises all populations of T. c. mauritanicus and northern African populations of T. c. simplex . These two main lineages separated 2.3 to 8.9 million years ago, based on the range of inferred molecular clocks recognized for crustacean mtDNA sequence divergence. Percentages of divergence are in the range reported for recognized species in other notostracan lineages and we therefore propose to recognize them as two species, Triops cancriformis and Triops mauritanicus . The latter would comprise two subspecies in northern Africa, one consisting of the Moroccan populations of the former T. c. mauritanicus , the other comprising the African populations of the former T. c. simplex . It also includes three as-yet unnamed lineages. A comparison of morphological characters with the molecular data revealed that the former T. c. simplex cannot be reliably separated from T. c. cancriformis , using morphological characters that have hitherto been used to distinguish among subspecies of T. cancriformis . Our investigation is the first to demonstrate the presence of T. c. cancriformis in Africa (Tunisia). The genetic haplotypes of these populations are identical with haplotypes also occurring in Central and Western Europe, as well as in Sicily. Therefore, we hypothesize that the African populations of T. c. cancriformis represent a result of repeated long-distance dispersal across the Mediterranean Sea.     

Molecular systematics and biogeography of the western Mediterranean stonefly genus Tyrrhenoleuctra (Insecta, Plecoptera)

The stonefly genus Tyrrhenoleuctra includes species living in western Mediterranean temporary freshwater streams, sometimes also at sea level, a very unusual habitat for most Plecoptera. Traditional morphological approaches proved unsuccessful in drawing both taxonomic and phylogenetic patterns, thus hampering discussion of biogeographical patterns for this interesting group. We aimed at: (a) assessing the taxonomic status of populations of Tyrrhenoleuctra covering the geographic range of the genus; (b) studying the phylogenetic relationships among the recognized species; and (c) describing biogeographic patterns. We used phylogenetic analyses to infer the phylogenetic history of this group of stoneflies based on a combined data set of 1666 bp including fragments of the 12S ribosomal (12S) and cytochrome oxidase I (CO‐I) mtDNA genes, with maximum likelihood and Bayesian methods. Two main clades have been identified: a Sardo‐Corsican one, including Tyrrhenoleuctra zavattarii, and an Ibero‐Maghrebian one including four lineages of unkown taxonomic rank from the Balearic Islands (Maiorca), from northern Africa (Ceuta) and southernmost Spain (Algeciras), and a complex preliminarily referred herein to T. minuta (Klapálek, 1901), which includes two lineages, one from Cordoba, and one from Sierra de Grazalema (El Cerro) and Portugal (Tellhares) respectively. Dating the nodes by fixing the split of the Ibero‐Maghrebian clade from the Sardo‐Corsican one at 29 million years ago (Mya), yielded dates referring to the major geological events in the Mediterranean basin. Estimated molecular evolutionary rates ranged from 0.02–0.09% per million years (my) in the T. zavattarii lineages, to 0.2–0.7% per MY in the Ibero‐Maghrebian clade. The phylogenetic pattern emerged from the present study is congruent with the known paleo‐history of the western Mediterranean basin, with the divergence of the two main Tyrrhenoleuctra lineages corresponding to the split of the Sardo‐Corsican microplate from the Iberian block. Vicariance events have characterized the history of this stonefly group along its entire biogeographical history. Surprisingly low evolutionary rates, previously supposed by Fochetti (1991, 1994) and Fochetti et al. (2004) based on nuclear markers (allozymes), have been herein found also in mitochondrial markers.     

Phylogeography and morphological variability in land snails: the Sicilian Marmorana (Pulmonata, Helicidae)

Land snails have long been recognized as suitable organisms for studying phenotypic differentiation and phylogeny in relation to geographical distribution. Morphological data (shell and anatomy biometry on different geographical scales) and partial sequences from mitochondrial genes (cytochrome oxidase subunit I , 16S rDNA) were used to test whether morphological patterns match phylogeny in a diversified group of Sicilian rock-dwelling land snails belonging to the genus Marmorana . The taxonomic implications of the three character sets (shell and anatomical biometry and molecular data) were also considered. The inferred phylogenetic relationships do not match morphological (shell and genitalia) patterns. This result may significantly modify the current taxonomy. Mitochondrial based reconstructions define several supported clades well correlated with geographic distribution and populations were found to be distributed parapatrically. The progressive decline in mitochondrial DNA sequence similarity over a distance of 250 km is consistent with a model of isolation by distance, a pattern previously recognized for other groups of land snails. For one clade of Marmorana , colonization along Mediterranean trade routes appears to be a possibility.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 809–823.     

Phylogenetic relationships in the orchid genus Serapias L. based on noncoding regions of the chloroplast genome

A molecular phylogenetic analysis was performed on 14 species of the Mediterranean unrewarding orchid genus Serapias using sequences of four noncoding regions of chloroplast DNA. This study has led to a new interpretation of the evolutionary relationships in this genus. The well-defined phylogenetic tree supports a division of taxa into two main clades, each including two minor groups. The molecular relationships found in this study differ from those defined by traditional systematic morphological assessments. By comparing the variation in sequence to variations in floral traits, we propose that the split in the two main lineages reflects an early differentiation of flower size, perhaps due to the shift from allo- to self-pollination. Conversely, the relationships within each minor group do not reflect floral size variation; therefore, we presume that this diversification resulted from genetic drift, local selection forces, and multiple, independent transitions towards self-pollination and polyploidy.     

Phylogeographical and speciation patterns in subterranean worm lizards of the genus Blanus (Amphisbaenia: Blanidae)

The peculiar lifestyle of subterranean reptiles must determine their modes of speciation and diversification. To further understand the evolutionary biology of subterranean reptiles, we studied the phylogeny of worm lizards of the genus Blanus and the phylogeography of its Iberian representatives. We used mitochondrial (ND4 and 16S rRNA) and nuclear (anonymous) partial gene sequences to resolve phylogenetic relationships within Blanus. The Eastern Mediterranean Blanus strauchi was recovered as sister group of Western Mediterranean species. Iberian and North African Blanus were recovered as reciprocally monophyletic groups. The same genes were used to determine phylogeography of 47 populations of Blanus cinereus. Mitochondrial and nuclear sequence data recovered two highly supported Iberian clades. Parapatry and high sequence divergences between them suggest that these clades may represent independent taxonomic units. A molecular clock was calibrated considering that the split between Iberian and North African Blanus was due to the re-opening of the Betic Strait in the Upper Tortonian (8-9 million years ago). Differentiation between the two Iberian clades was estimated to date back to 5.2 million years ago. The Central Iberian clade included five mitochondrial haplotype lineages (A-E). Geographical ranges of two of them broadly overlap in the central Iberian plateau. After testing alternative hypotheses, the most likely explanation for this striking phylogeographical pattern involves recent dispersal of one of the lineages (C) over the geographical range of the other (B). The inferred recent dispersal of this fossorial reptile is explained in terms of demographic advantages associated to underground lifestyle.     

A preliminary revision of the genus Plecotus (Chiroptera, Vespertilionidae) based on genetic and morphological results

The phylogenetic relationships within the genus Plecotus were assessed using molecular as well as morphological methods. With only three species missing, our study is based on an almost comprehensive taxonomic sampling. The genetic analysis comprised 151 individuals from throughout the range. Sequences of two mitochondrial sections, parts of the 16S rRNA gene (16S) and of the control region (CR) were analysed. The morphological analysis of cranial and external characters comprised 697 individuals, including 10 holotypes and one lectotype. Data from 15 craniometric characters of 442 specimens were used in the multivariate analyses. The molecular data identified nine primary clades representing 11 species, 10 of which could be assigned to described taxa, whereas one was described as a new species, Plecotus strelkovi Spitzenberger sp. nov. The tree based on 16S revealed two major lineages, one consisting of only one primary clade restricted to the Mediterranean, the other consisting of eight primary clades representing Eurasian taxa. The morphological analysis revealed five additional species, two of them not described. Together with the recently described P. taivanus, P. sardus and P. balensis, which were not included in our analysis, the genus Plecotus comprises at least 19 more or less cryptic species. Phylogenetic and phenetic analyses resulted in similar but not completely concordant arrangements of the species. The proposed classification relies mainly on the tree based on 16S sequences. The current distribution indicates that 16 species can be linked to arboreal refugia, three to eremial refugia. We assume that speciation within the gleaning, rather slow flying long‐eared bats is due to a multitude of disruption and isolation processes within a formerly continuous range of the broad‐leaved Arcto‐Tertiary forest in which Plecotus probably originated. An exact calibrated molecular dating of the splits is not possible. The Early Oligocene age of the presumed ancestor of the Plecotini and a correlation of the molecular diversifications with palaeogeographic reconstructions suggest that the divergence of the two major lineages may have occurred already during the Middle Miocene, 14.5 Mya.     

Waves of dispersal in island-hopping Chondrina species (Gastropoda, Pulmonata, Chondrinidae)

The aim of this study was to unravel the historical biogeography of the speciose land snail genus Chondrina. To this end phylogenetic hypotheses were tested using mitochondrial DNA sequence data.Mitochondrial DNA sequences of the Cytochrome Oxidase subunit I region were obtained for 89 individuals, representing just over 70% of the extant Chondrina species. The extent of molecular genetic diversity and phylogeographical patterns were investigated by using neighbour joining, parsimony and bayesian methods for phylogeny reconstructions. The resulting data were used to infer historical biogeographical patterns for the genus Chondrina.The three phylogenetic methods yielded congruent topologies for the phylogeny reconstruction. Six clades were identified, each of which with at least one taxon that is known from the Iberian peninsula. The most parsimonious scenario indicates at least three waves of dispersal out of the Iberian peninsula into the North and East of Europe and Northern Africa.The phylogenetic relationships combined with the distributional patterns of the various species, indicate that only vicariance events cannot explain the actual situation. Apparently, separate waves of dispersal and subsequent speciation occurred, each time starting from the southwestern part of the present generic range. Until recently, this was obscured by repetitive cases of parallel or convergent evolution in shell characters, as became evident with the use of molecular methods.     

Fluvial range expansion, allopatry, and parallel evolution in a Danubian snail lineage (Neritidae: Theodoxus)

The snails Theodoxus danubialis and Theodoxus prevostianus form a single clade native to freshwaters of south-eastern Europe whose inter- and intraspecific relationships remain unresolved. The present study utilized a phylogeographical approach to clarify the relationship of these species as well as to reconstruct the evolutionary and demographic history of populations in the western portion of their range. Phylogenetic, population genetic, and nested clade analyses reveal a clade that has distributed itself upriver from a lower Danube River source population and become genetically distinct primarily through range expansion and localized allopatric divergence. Notably, this geographical pattern is replicated phylogenetically in the form of two cytochrome c oxidase subunit I (CO I) lineages that are present simultaneously in individual snails. Haplotypes from polymorphic individuals form two distinct clades, both of which show phylogenetic and nucleotide substitution patterns consistent with a mitochondrial origin, and whose common ancestor must have occurred in a lower Danube source population. Separated allopatrically from their Danubian relatives, populations of T. danubialis in northern Italy have also undergone substantial range expansion, much more recently than Danube watershed lineages. In addition to repeated patterns of range expansion, parallelism is found in T. prevostianus , which is shown to be a nonmonophyletic taxon of remarkable morphological and ecological similarity.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 603–617.     

Phylogenetic relationships among the Ibero-African cobitids (Cobitis, cobitidae) based on cytochrome b sequence data

We estimated the phylogenetic relationships of all Ibero-African spined loaches of the genus Cobitis using the complete mitochondrial cytochrome b gene (1140bp). We analysed 93 individuals of seven cobitid species found in all the principal drainages of the Iberian Peninsula and North Africa. A molecular phylogeny was used to revise current systematics of the Ibero-African Cobitis species and to infer a biogeographical model for Cobitis in the Western Mediterranean area during the Cenozoic period. Phylogenetic analysis provided support for the monophyly of two mtDNA clades: Clade A or Italian Clade with the Italian species (C. bilineata, C. zanandreai), and Clade B or the Ibero-African Clade. The Ibero-African Clade included all species endemic for the Iberian Peninsula (C. vettonica, C. calderoni, and C. paludica) and North Africa (C. maroccana). The species C. paludica does not constitute a natural group, and could be divided into at least four monophyletic mtDNA lineages with moderate to high bootstrap values and posterior probability support. Phylogenetic relationships of the Ibero-African species were not resolved satisfactorily, but in all analyses C. calderoni from Northern Iberian Peninsula was basal. We have re-calibrated a molecular clock for the genus Cobitis (0.68% per million year by pairwise) using populations inhabiting both sides of the Gibraltar Strait. Application of this Cobitis mtDNA clock provides evidence that the Messinian salinity crisis played a primary role in the diversification of some Ibero-African cobitid species. The basal polytomies of the Ibero-African Clade might suggest that all mtDNA lineages diversified rapidly.     

Relationships of the Macaronesian and Mediterranean floras: molecular evidence for multiple colonizations into Macaronesia and back-colonization of the continent in Convolvulus (Convolvulaceae)

A molecular phylogenetic analysis of the Macaronesian endemic species of Convolvulus was undertaken using data from the nuclear ribosomal internal transcribed spacer (ITS) regions. The results of the analysis support two introductions into Macaronesia from distantly related clades within Convolvulus and a subsequent back-colonization to the continent from within one of the clades. Hypothesized relationships between Macaronesian species and New World taxa and between the Canarian endemic C. caput-medusae and the Moroccan C. trabutianus are refuted. Both Macaronesian clades are shown to have Mediterranean sister groups although one is predominantly western Mediterranean and the other predominantly eastern Mediterranean in distribution. The patterns of colonization into Macaronesia demonstrated by Convolvulus and also by other multiple colonizing genera conform to either a pattern of phylogenetic distinctiveness or a checkerboard distribution of island lineages. Both are consistent with the hypothesis that niche preemption is responsible for the limited number of colonizations into the region. A review of sister group relationships demonstrates that, in common with Convolvulus, most Macaronesian groups have sister groups distributed in the near-continent (i.e., western Mediterranean). Disjunct sister group relationships (including Eastern Mediterranean disjunctions) occur in only 18% of groups.     

Global Diversification at the Harsh Sea-Land Interface: Mitochondrial Phylogeny of the Supralittoral Isopod Genus Tylos (Tylidae,Oniscidea)

The supralittoral environment, at the transition between sea and land, is characterized by harsh conditions for life. Nonetheless, evolution of terrestrial isopods (Oniscidea), the only group of Crustacea fully adapted to live on land, appears to have involved a transitional step within the supralittoral. The two most basal oniscidean lineages (Ligiidae and Tylidae) have representatives that successfully colonized the supralittoral. One of them is the genus Tylos, which is found exclusively in supralittoral sandy beaches from tropical and subtropical coasts around the world. Comprehensive phylogenetic hypotheses for this genus are lacking, which are necessary for understanding the evolution and biogeography of a lineage that successfully diversified in the harsh sea-land interface. Herein, we studied the phylogenetic relationships among 17 of the 21 currently recognized species of the genus Tylos, based on sequences from four mitochondrial genes (Cytochrome Oxidase I, Cytochrome b, 16S rDNA, and 12S rDNA). Maximum Likelihood and Bayesian phylogenetic analyses identified several lineages with deep divergences and discrete geographic distributions. Phylogenetic and distributional patterns of Tylos provide important clues on the biogeography and evolution of this group. Large divergences among the most basal clades are consistent with ancient splits. Due to the biological characteristics of Tylos, which likely prevent dispersal of these isopods across vast oceanic scales, we argue that tectonic events rather than trans-oceanic dispersal explain the distribution of Tylos in different continents. Overwater dispersal, however, likely enabled range expansions within some basins, and explains the colonization of volcanic oceanic islands. Present-day distributions were also likely influenced by sea level and climate changes. High levels of allopatric cryptic genetic differentiation are observed in different regions of the world, implying that the dispersal abilities of Tylos isopods are more limited than previously thought. Our results indicate that a taxonomic revision of this group is necessary.     

Molecular inference of a Late Pleistocene diversification shift in Nigella s. lat. (Ranunculaceae) resulting from increased speciation in the Aegean archipelago

Aim  To infer the temporal course and geographical mode of speciation in Mediterranean/Southwest Asian Nigella s. lat.
Location  Mediterranean Basin, Aegean archipelago.
Methods  Phylogenies for Nigella L. and Garidella L. (=  Nigella s. lat.) were obtained from maximum-likelihood analyses of internal transcribed spacer (ITS) sequences. Diversification through time was analysed by log lineages-through-time (LTT) plots and survival analyses. Relative node age estimates were regressed against the degree of sympatry between sister clades to infer the predominant mode of geographical speciation in Nigella s. lat.
Results  The Late Pleistocene radiation of the Nigella arvensis complex in the Aegean region caused a significant departure from a stochastic speciation/extinction process of diversification during the evolution of Nigella s. lat., a lineage of (at least) Late Miocene origin. Speciation within Nigella s. lat. predominantly took place in allopatry.
Main conclusions  No significant effect on diversification rate was found regarding the establishment of a Mediterranean-type climate, or the onset of the Quaternary climatic oscillations. Rather, the accelerated rate of speciation in the N. arvensis complex is plausibly related to increased opportunities for allopatric speciation afforded by the (palaeo)geographical complexity of the Aegean archipelago combined with Late Pleistocene changes in climate and sea level. The evolution of self-pollination and associated changes in habitat preference and flowering time further augmented speciation and niche differentiation within the complex, but these changes did not act as the primary promoters of the radiation process.