Living with the genetic signature of Miocene induced change: evidence from the phylogeographic structure of the endemic angulate tortoise Chersina angulata

The phylogeographic structure of the monotypic endemic southern African angulate tortoise Chersina angulata was investigated throughout its distribution with the use of partial sequences from three mtDNA loci (COI, cyt b and ND4). Phylogeographic and phylogenetic structuring obtained for the three mtDNA markers were highly congruent and suggested the presence of two genetically distinct, reciprocally monophyletic evolutionary lineages. Group one contained two subclades with haplotypes from the north-western Cape and south-western Cape, respectively, while haplotypes from the southern Cape comprised group two. The two major clades were separated by nine and eight mutational steps for COI and ND4, respectively. Of the three mtDNA gene regions examined, the ND4 partial sequence contained the most phylogenetic signal. Haplotype diversity was generally low and we recovered 34 haplotypes for the 125 animals sequenced for the ND4 subunit. Nested clade analyses performed on the variable ND4 partial sequences suggested the presence of two major refugial areas for this species. The demographic history of the taxon was characterised by range expansion and prolonged historical fragmentation. Divergence time estimates suggest that the temporal and spatial distribution of the taxon was sculpted by changes in temperature and rainfall patterns since the late Miocene. Corroborative evidence from other reptiles is also suggestive of a late Miocene divergence, indicating that this was a major epoch for cladogenesis in southern Africa. Apart from the genetic differences between the two major clades, we also note morphometric and behavioural differences, alluding to the presence of two putative taxa nested within C. angulata.     

A molecular phylogeny of aquatic gastropods provides a new perspective on biogeographic history of the Snake River Region

Mitochondrial DNA sequences of aquatic gastropods of the subgenus Pyrgulopsis (Natricola) were analyzed to test a commonly accepted hypothesis concerning the early history of the Snake River in the northwestern US. Distributions of Natricola and other regional biota were previously used to infer that the Snake River flowed to the Pacific through southeastern Oregon and northern California during the Neogene prior to its capture by the Columbia River in the late Pliocene (2 Ma). A molecular phylogeny based on partial sequences of COI and NDI (1149 bp) indicates that the Natricola clade is restricted to the modern Snake-Columbia River Basin and the Oregon Lakes region whereas northern California populations previously assigned to this subgenus belong to other lineages. The Natricola clade is not deeply subdivided into Oregon Lakes and Snake River Basin units consistent with late Pliocene fragmentation of the hypothesized paleodrainage, but instead is shallowly structured and contains multiple transitions among these two geographic areas. The strongly supported sister relationship between Natricola and a species from northwest Nevada (P. imperialis) is consistent with a recent proposal that the ancestral Snake River did not flow through southeast Oregon but instead flowed south to the Humboldt River. Within the context of this hypothesis, the multiple transitions between the Snake River Basin and the Oregon Lakes region that occurred within Natricola may be attributed to a late Pleistocene connection between these areas that was unrelated to the early course of the Snake River.     

Biogeography and speciation patterns of the golden orb spider genus Nephila (Araneae: Nephilidae) in Asia

The molecular phylogeny of the globally distributed golden orb spider genus Nephila (Nephilidae) was reconstructed to infer its speciation history, with a focus on SE Asian/W Pacific species. Five Asian, two Australian, four African, and one American species were included in the phylogenetic analyses. Other species in Nephilidae, Araneidae, and Tetragnathidae were included to assess their relationships with the genus Nephila, and one species from Uloboridae was used as the outgroup. Phylogenetic trees were reconstructed from one nuclear (18S) and two mitochondrial (COI and 16S) markers. Our molecular phylogeny shows that the widely distributed Asian/Australian species, N. pilipes, and an African species, N. constricta, form a clade that is sister to all other Nephila species. Nested in this Nephila clade are one clade with tropical and subtropical/temperate Asian/Australian species, and the other containing African and American species. The estimated divergence times suggest that diversification events within Nephila occurred during mid-Miocene to Pliocene (16 Mya-2 Mya), and these time periods were characterized by cyclic global warming/cooling events. According to Dispersal and Vicariance Analysis (DIVA), the ancestral range of the Asian/Australian clade was tropical Asia, and the ancestral range of the genus Nephila was either tropical Asia or Africa. We conclude that the speciation of the Asian/Australian Nephila species was driven by Neogene global cyclic climate changes. However, further population level studies comparing diversification patterns of sister species are needed to determine the mode of speciation of these species.     

Systematics, biogeography, and evolution of the Neotropical peacock basses Cichla (Perciformes: Cichlidae)

To investigate forces influencing diversification in Neotropical fishes, the phylogenetic relationships among species and populations of the cichlid genus Cichla were examined. Mitochondrial DNA was sequenced for 454 individuals of the 5 nominal Cichla species and several putative undescribed species. Phylogenetic analyses support the distinction of two major clades of Cichla. Clade A includes C. temensis and two undescribed species from the lower Amazonas and Xingu Rivers. Clade B includes C. orinocensis, C. monoculus, C. ocellaris. C, intermedia, and an undescribed species from the upper Madeira River. Species boundaries were relatively well-circumscribed for clade B, while incomplete lineage sorting was inferred for clade A. Three probable instances of introgression were observed, including a regional population of C. orinocensis from the Negro River that shows a history of introgression. Biogeographic patterns from Cichla are partially congruent with those seen in several other Neotropical fish clades, and the diversification of Cichla species is inferred to result from both vicariance and sympatric divergence.     

Extensive diversification of pebblesnails (Lithoglyphidae: Fluminicola) in the upper Sacramento River basin, northwestern USA

Mitochondrial DNA sequences from the cytochrome c oxidase subunit I (COI) and cytochrome b (cytb) genes were obtained from the nine extant, previously described species of the northwestern North American freshwater gastropod genus Fluminicola (commonly known as pebblesnails) and from a large number of taxonomically undescribed populations of these animals from the upper Sacramento River basin, California and Oregon, which is composed of the Sacramento River headwaters, and the McCloud and Pit Rivers. Phylogenetic analyses of separate and combined molecular datasets yielded well‐supported and largely congruent trees delineating 13 genetically divergent and morphologically distinctive upper Sacramento basin lineages, which we describe as new species. These include two groups of closely related and geographically proximal species that are further united by unique radular or shell features. Most of these novelties have narrow geographical distributions and are restricted to headspring areas, whereas several are more wide ranging and typically occupy larger, well‐integrated habitats. The highly endemic fauna of upper Sacramento River pebblesnails is not a single species flock, but instead a polyphyletic assemblage spread among four separate clades. Our phylogeny, together with the application of a COI molecular clock for Fluminicola, suggests that upper Sacramento River clades originated as a result of late Neogene separation of this basin from neighbouring regions (northwestern Great Basin, Klamath River basin), which is consistent with previous biogeographical hypotheses based on the distributions of fishes. The upper Sacramento River pebblesnails evolved in association with the complex late Cenozoic history of regional landscape and drainage and diversification was also facilitated by the invasion of and adaptation to insular spring habitats. Our findings are consistent with the generally limited dispersal ability and geologically ancient (mid‐Tertiary) age of this genus and imply that other portions of northwestern North America may also harbour a large number of undescribed pebblesnail species. Journal compilation © 2007 The Linnean Society of London, Zoological Journal of the Linnean Society, 2007, 149 , 371–422. No claim to original US government works.     

Biogeography in the Death Valley region: evidence from springsnails (Hydrobiidae: Tryonia)

Allozyme and mitochondrial DNA variation were analysed to examine evolution of the nine species of springsnails (genus Tryonia) living in the Death Valley system (Owens and Amargosa basins) of southeastern California and southwestern Nevada. Both allozyme and mtDNA evidence indicate that this highly endemic fauna is non-monophyletic. Species from the upper Amargosa basin comprise a clade most closely related to snails living in the Colorado basin. Snails from the lower Amargosa basin (Death Valley trough) reflect a complex evolutionary history and two of these species are more closely related to an estuarine species from western California than to other snails of the region. These results indicate a commonality of pattern with the well-studied Death Valley pupfishes (Cyprinodon), which also are non-monophyletic and include species that are most closely related to Colorado basin congeners. These biogeographic patterns are interpreted within the context of a recently proposed model for the early history of the lower Colorado River.     

Phylogeny of the Neotropical Alibertia group (Rubiaceae), with emphasis on the genus Alibertia, inferred from ITS and 5S ribosomal DNA sequences

Phylogenetic relationships of 38 species of the Alibertia group (Rubiaceae) and two outgroup species were investigated using the nuclear ribosomal 5S nontranscribed spacer (5S-NTS) and the internal transcribed spacers (ITS). Analysis of the data sets separately and in combination resulted in several well-supported and congruent groupings. However, the three analyses yielded different results as to the branching order of the basal clades. With the exception of Alibertia hispida, the species in the genus Alibertia appear in one weakly to moderately supported clade. This clade is in turn composed of two strongly supported subclades. One comprises several Alibertia species, including the type (A. edulis), three Borojoa species, and Randia tessmannii. The other subclade consists of Alibertia species only. This division is also generally supported morphologically by fruit size, corolla size, number of corolla lobes, and pollen aperture (porate vs. colporate). The sister group to the Alibertia clade comprises Duroia with Amaioua species internested. The close relationship of Ibetralia and Kutchubaea is corroborated. In addition, Alibertia hispida is a member of this strongly supported clade. Likewise, the two "Genipa" species are supported as a monophyletic group in 100% of the bootstrap replicates. It is concluded that the 5S spacer is superior to the commonly used ITS region in terms of resolution and robustness among closely related taxa.     

RAD sequencing resolved phylogenetic relationships in European shrub willows (Salix L. subg. Chamaetia and subg. Vetrix) and revealed multiple evolution of dwarf shrubs

The large and diverse genus Salix L. is of particular interest for decades of biological research. However, despite the morphological plasticity, the reconstruction of phylogenetic relationships was so far hampered by the lack of informative molecular markers. Infrageneric classification based on morphology separates dwarf shrubs (subg. Chamaetia) and taller shrubs (subg. Vetrix), while previous phylogenetic studies placed species of these two subgenera just in one largely unresolved clade. Here we want to test the utility of genomic RAD sequencing markers for resolving relationships at different levels of divergence in Salix. Based on a sampling of 15 European species representing 13 sections of the two subgenera, we used five different RAD sequencing datasets generated by Ipyrad to conduct phylogenetic analyses. Additionally we reconstructed the evolution of growth form and analyzed the genetic composition of the whole clade. The results showed fully resolved trees in both ML and BI analysis with high statistical support. The two subgenera Chamaetia and Vetrix were recognized as nonmonophyletic, which suggests that they should be merged. Within the Vetrix/Chamaetia clade, a division into three major subclades could be observed. All species were confirmed to be monophyletic. Based on our data, arctic‐alpine dwarf shrubs evolved four times independently. The structure analysis showed five mainly uniform genetic clusters which are congruent in sister relationships observed in the phylogenies. Our study confirmed RAD sequencing as a useful genomic tool for the reconstruction of relationships on different taxonomic levels in the genus Salix.     

Molecular phylogeny of the spiny lobster genus Palinurus (Decapoda: Palinuridae) with hypotheses on speciation in the NE Atlantic/Mediterranean and SW Indian Ocean

Sequence data derived from the mitochondrial DNA 16S rRNA and COI genes were used to determine the phylogenetic relationships among six Palinurus spiny lobster species. Three species (P. charlestoni, P. elephas, and P. mauritanicus) occur in the northeastern Atlantic/Mediterranean, and the others (P. barbarae, P. delagoae and P. gilchristi) inhabit the southwestern Indian Ocean. Parsimony and model based phylogenetics strongly supported the monophyly of the genus. A combined parsimony analysis based on 1001bp and 274 parsimony informative characters recovered the most resolved phylogeny with >70% bootstrap support for associations among species. The Atlantic P. charlestoni consistently clusters nested within the Indian Ocean clade, and the mtDNA sequence divergence between the two most distant species is 8.24%. If the northward collision of Africa with Eurasia in the Miocene caused the final physical separation between the Atlantic and Indian Ocean taxa, then the Palinurus mtDNA (COI and 16S combined) evolved no faster than 0.18% (lower bound) to 0.36% (upper bound) per lineage per million years. The six extant species occur in the pathways of the North Atlantic and South Indian Ocean gyres, and hypotheses on their radiation are developed relative to the strengthening of boundary currents in the Miocene and life history traits congruent with survival in strong ocean currents.     

Evolution of 4-coumarate:coenzyme A ligase (4CL) gene and divergence of Larix (Pinaceae)

The evolutionary dynamics of the 4CL gene encoding 4-coumarate:coenzyme A ligase was investigated in the genus Larix (Pinaceae) by comparing copy number, GC content and codon usage, sequence divergence, and phylogenetic analysis. All 4CL clones of Larix formed a strongly supported monophyletic group, in which two robust clades (4clA and 4clB) derived from an ancient gene duplication event in the common ancestor of Larix were identified. Further gene duplication in the 4clA clade gave rise to two subclades 4clA(1) and 4clA(2). Frequent duplication/deletion appears to be a common evolutionary phenomenon in the 4CL gene family and paralogous genes differ greatly in their evolution rate. The existence of L. speciosa in subclades 4clA(1) and 4clA(2) suggests that this species may represent a primitive form of Larix or the closest relative of the common ancestor of the Eurasian Sect. Multiserialis. In addition, cpDNA and nrDNA ITS analyses support the hypothesis of an early separation of Larix into a North American and a Eurasian clade, which is congruent with the results of previous allozyme and very recent AFLP analyses. The unexpected close relationship between North American larches and the short-bracted species L. gmelinii in East Asia, based on the 4CL gene tree, may stem from lineage sorting.     

Molecular phylogeny of the subterranean genus Niphargus (Crustacea: Amphipoda) in the Middle East: a comparison with European Niphargids

The subterranean genus Niphargus is one of the most species‐rich genera among freshwater amphipods in the world, distributed in the Western Palearctic. Thus far, taxonomic and phylogenetic research has focused mainly on the European half of the genus range. In this study, 25 populations of Niphargus from Iran, Lebanon and the Crimean Peninsula were investigated. Bayesian inference based on 28S, H3 and COI gene sequences suggests that populations from the area belong to four different clades. Three species from Crimea and one species from Iran are nested at basal nodes, indicating their rather ancient origin. The rest of the species are younger and belong to two separate clades. One Crimean species is a sister‐species to east Romanian species. The second clade includes one species from Lebanon and all but one population from Iran. The origin of this clade corresponds to marine transgression between the Black Sea and Mediterranean approximately 12 Mya. This clade was further investigated taxonomically. Revision of qualitative morphological traits and unilocus species delimitation methods using COI suggest that this clade comprises 12–16 species, of which only three have been described so far. Multilocus coalescence delimitation methods (using fragments of COI, 28S, H3 and ITS) strongly supported 11 of these species. The remaining populations comprise at least two species complexes that require further and more detailed taxonomic research. © 2015 The Linnean Society of London     

Glomus, the largest genus of the arbuscular mycorrhizal fungi (Glomales), is nonmonophyletic.

Arbuscular mycorrhizal (AM) fungi form a widespread and ecologically important symbiosis with plants in the land ecosystem. The phylogeny of the largest presently accepted genus, Glomus, of the monogeneric family Glomaceae (Glomales; AM fungi) was analyzed. Phylogenetic trees were computed from nearly full-length SSU rRNA gene sequences of 30 isolates, and show that "Glomus" is not monophyletic. Even after the very recent separation of Archaeospora and Paraglomus from "Glomus," the genus further separates into two suprageneric clades. One of them diverges further into two subclades, differing by phylogenetic distances equivalent to family level. The other, comprising Glomus versiforme, G. spurcum, and a species morphologically similar to G. etunicatum, is not closely related to the Glomaceae, but clusters together with the Acaulosporaceae and Gigasporaceae in a monophyletic clade. Based on the molecular evidence, a new family, separate from the Glomaceae, is required to accommodate this group of organisms, initially named Diversisporaceae fam. ined. The current taxonomic concept of the recently erected family Archaeosporaceae also requires future emendation, because Geosiphon pyriformis (Geosiphonaceae) renders Archaeospora, the sole genus formally included in this family, paraphyletic. The suborders Gigasporineae and Glominaeae are not congruent with the natural phylogeny of the AM fungi. Our data necessitate a general reexamination of the generic concepts within the Glomales. In addition to the new family structure hypothesized herein, establishment of at least three new genera will be necessary in the future.     

Molecular studies of photobionts of selected lichens from the coastal vegetation of Brazil

A light microscopic and molecular analysis of photobionts in Ramalina and Cladonia from coastal habitats of Brazil is presented. A Bayesian phylogenetic analysis of ITS rDNA sequences suggests a Trebouxia lineage which is preferentially tropical in geographic distribution. This highly diverse clade also includes the morphological similar species Trebouxia higginsiae and galapagensis. Within the predominantly tropical clade of Trebouxia we distinguish several subclades, three of which are represented in our samples of Ramalina species. Since sexuality has not been recognized in coccal lichenised photobionts until recently, we cannot apply a biological species concept, but when compared with the sequence diversity between known species we conclude that several new species need to be described in this clade. The mutually exclusive presence of other Trebouxia lineages in temperate samples of Ramalina suggests an evolution towards higher selectivity in this genus. A strictly tropical lineage is not conspicuous in the photobionts of the genus Asterochloris sampled from Cladonia so far.     

Phylogeny of Paramysis (Crustacea: Mysida) and the origin of Ponto-Caspian endemic diversity: resolving power from nuclear protein-coding genes

The Ponto-Caspian (Black and Caspian seas) brackish-water fauna represents a special case of the endemic diversification in world's ancient lakes; it also involves a hotspot of continental diversity in the predominantly marine mysid crustaceans. We explored the origins and history of the mysid diversification in a phylogenetic analysis of some 20 endemic Ponto-Caspian species mainly of the genus Paramysis and their marine congeners, using sequences of two nuclear protein-coding genes, two nuclear rRNA genes, the mitochondrial COI gene and morphological data. A nearly completely resolved phylogeny was recovered, with no indication of rapid diversification bursts. Deep divergences were found among the main endemic clades, attesting to a long independent faunal history in the continental Paratethys waters. The current marine Paramysis species make a monophyletic cluster secondarily derived from the continental Paratethyan (Ponto-Caspian) Paramysis ancestors. The good phylogenetic resolution was mainly due to the two nuclear protein-coding genes, opsin and EPRS, here for the first time applied to peracarid systematics. In contrast, 'conventional' mtDNA and nuclear rRNA genes provided poor topological resolution and weak congruence of divergence rates. The two nuclear protein-coding genes had more congruent rates of evolution, and were about 10-15 times slower than the mitochondrial COI gene.     

Evolutionary relationships within the Neotropical, eusporangiate fern genus Danaea (Marattiaceae)

Genera within the eusporangiate fern family Marattiaceae have long been neglected in taxonomic and systematic studies. Here we present the first phylogenetic hypothesis of relationships within the exclusively Neotropical genus Danaea based on a sampling of 60 specimens representing 31 species from various Neotropical sites. We used DNA sequence data from three plastid regions (atpB, rbcL, and trnL-F), morphological characters from both herbarium specimens and live plants observed in the field, and geographical and ecological information to examine evolutionary patterns. Eleven representatives of five other marattioid genera (Angiopteris, Archangiopteris, Christensenia, Macroglossum, and Marattia) were used to root the topology. We identified three well-supported clades within Danaea that are consistent with morphological characters: the "leprieurii" clade (containing species traditionally associated with the name D. elliptica), the "nodosa" clade (containing all species traditionally associated with the name D. nodosa), and the "alata" clade (containing all other species). All three clades are geographically and ecologically widely distributed, but subclades within them show various distribution patterns. Our phylogenetic hypothesis provides a robust framework within which broad questions related to the morphology, taxonomy, biogeography, evolution, and ecology of these ferns can be addressed.     

Molecular systematics of a speciose, cosmopolitan songbird genus: defining the limits of, and relationships among, the Turdus thrushes

The avian genus Turdus is one of the most speciose and widespread of passerine genera. We investigated phylogenetic relationships within this genus using mitochondrial DNA sequence data from the ND3, ND2 and cytochrome b genes. Our sampling of Turdus included 60 of the 65 extant species currently recognized, as well as all four species from three genera previously shown to fall inside Turdus (Platycichla, Nesocichla, and Cichlherminia). Phylogenetic trees based on maximum likelihood and maximum parsimony algorithms were congruent. Most of the Turdus taxa sampled fall into one of four clades: an African clade, a Central American-Caribbean clade, a largely South American clade, and a Eurasian clade. Still other taxa are placed either at the base of Turdus, or as links between clades. In no instance is any continent reciprocally monophyletic for the species distributed on it. A general lack of nodal support near the base of the phylogeny seems related to a rapid intercontinental establishment of the major clades within Turdus very early in the history of the genus. The monotypic genus Psophocichla is distantly related to, but clearly the sister of, Turdus rather than a constituent member of it.     

A molecular phylogeny of <Emphasis Type="Italic">Haptoglossa</Emphasis> species,terrestrial peronosporomycetes (oomycetes) endoparasitic on nematodes

Phylogenetic relationships of seven Haptoglossa isolates were analyzed by using mitochondrial COII amino acid sequences with a data set of 34 peronosporomycetes. Haptoglossa isolates formed a single clade and appeared to be basal to the clade consisting of all other peronosporomycetes. The Haptoglossa clade was divided into two subclades: one clade consisted of five aplanosporic isolates and the other included one aplanosporic and one zoosporic isolate. These results indicate that the genus Haptoglossa is monophyletic, and patterns of infection cell formation reflect more the phylogenetic relationship between the species than patterns of sporogenesis.     

Phylogenetic relationships of Ansonia from Southeast Asia inferred from mitochondrial DNA sequences: Systematic and biogeographic implications (Anura: Bufonidae)

We investigated the phylogenetic relationships and estimated the history of species diversification and biogeography in the bufonid genus Ansonia from Southeast Asia, a unique organism with tadpoles adapted to life in strong currents chiefly in montane regions and also in lowland rainforests. We estimated phylogenetic relationships among 32 named and unnamed taxa using 2461 bp sequences of the mitochondrial 12S rRNA, tRNA^val, and 16S rRNA genes with equally-weighted parsimony, maximum likelihood, and Bayesian methods of inference. Monophyletic clades of Southeast Asian members of the genus Ansonia are well-supported, allowing for the interpretation of general biogeographic conclusions. The genus is divided into two major clades. One of these contains two reciprocally monophyletic subclades, one from the Malay Peninsula and Thailand and the other from Borneo. The other major clade primarily consists of Bornean taxa but also includes a monophyletic group of two Philippine species and a single peninsular Malaysian species. We estimated absolute divergence times using Bayesian methods with external calibration points to reconstruct the relative timing of faunal exchange between the major landmasses of Southeast Asia.     

Phylogenetic relationships of assimineid gastropods of the Death Valley–lower Colorado River region: relicts of a late Neogene marine incursion?

Aim A small fauna of amphibious snails (genus Assiminea Fleming, 1828) living in association with highly mineralized springs in the Death Valley–lower Colorado River region (DVLCR) is thought to be a relict of the Bouse Embayment, a putative late Miocene–early Pliocene transgression of the ancestral Gulf of California along the lower Colorado River valley. We analysed the phylogenetic relationships of this fauna using mtDNA sequence data (1171 bp) to determine whether, as would be consistent with this hypothesis, it forms a substantially divergent unit sister to marine coastal congeners. Location South‐western Great Basin and lower Colorado River region, USA. Methods Two genes [mitochondrial cytochrome c oxidase subunit I (COI) and the mitochondrial 16S ribosomal RNA gene] were sequenced for 10 populations of DVLCR assimineas (Assiminea infima Berry, 1947 ; Assiminea sp.). We also sequenced an undescribed population from a spring in the Colorado River delta; western North American Pacific Coastal Assiminea californica (Tryon, 1865); the three other congeners that live on the continent; and three Old World assimineids (outgroups). Phylogenies based on the combined data set were obtained using Bayesian methods, and divergence times were estimated using a COI molecular clock for related gastropods. Results Composite haplotypes of the DVLCR assimineas, together with that observed in the Colorado River delta population, formed a weakly supported clade that was sister to a clade composed of populations of North American Pacific and Atlantic coastal species. The genetic distance between members of these two clades was 3.46 ± 0.47% for COI and 1.69 ± 0.38% for 16S. The former clade was composed of five subunits that differed from each other by 1.29–2.84% (COI) and 0.52–1.98% (16S) sequence divergence. Main conclusions Application of the COI clock suggests that progenitors of the DVLCR fauna diverged from coastal ancestors 2.13–1.89 Ma (late Pliocene), several million years after the Bouse Embayment would have been terminated by the establishment of the lower (freshwater) Colorado River. This finding, together with shallow genetic structuring of several DVLCR lineages that are widely distributed across the topographically complex regional landscape, suggests that the Assiminea fauna of this inland area was more likely to have been founded by coastal colonists transported on water birds than through a direct connection with the sea.     

Phylogeny and systematics of the Pachychilidae of mainland South‐East Asia – novel insights from morphology and mitochondrial DNA (Mollusca,Caenogastropoda, Cerithioidea)

Pachychilid gastropods are a conspicuous element of the freshwater macro‐invertebrate fauna of South‐East Asia. In this region, three spatially separated groups of pachychilids can be differentiated mostly by means of their brooding strategy. The largest group comprises about 100 described species and displays an extended Sundaland distribution that ranges from north‐east India in the west to southern China in the east and Borneo in the south‐east. The most conspicuous synapomorphy shared by females is a subhaemocoelic brood pouch in which developing embryos are retained. Mitochondrial phylogenies based on partial sequences of the genes cytochrome oxidase C subunit I (COI) and 16S rRNA (16S) support the monophyly of this group of subhaemocoelic brooders. Amongst them, the two genera Brotia and Paracrostoma form a sister pair of clades both together forming the sister taxon of a third clade. This third clade itself is split into three subclades, which occupy separate ranges except for the overlap of two of them in Borneo. The subclades comprise species from (A) Thailand, Cambodia, and northern Borneo; (B) Java and south‐western Borneo; and (C) Vietnam and southern China. In contrast with their spatial structuring, all species share a widely corresponding anatomy. Characteristic key features are a pallial oviduct with a seminal receptacle and an embryonic shell with a smooth, dome‐like inflated apex. Differences in the histology of the pallial oviduct are not congruent with phylogenetic patterns as suggested by the mitochondrial trees. Given the lack of obvious distinguishing morphological characteristics, all species of the subclades A to C are assigned to a single genus, Sulcospira, with Adamietta as a junior synonym.