Molecular phylogeny of African hinged and helmeted terrapins (Testudines: Pelomedusidae: Pelusios and Pelomedusa)

Fritz, U., Branch, W. R., Hofmeyr, M. D., Maran, J., Prokop, H., Schleicher, A., ?iroký, P., Stuckas, H., Vargas‐Ramírez, M., Vences, M. & Hundsdörfer, A. K. (2010). Molecular phylogeny of African hinged and helmeted terrapins (Testudines: Pelomedusidae: Pelusios and Pelomedusa). —Zoologica Scripta, 40, 115–125. With 18 currently recognised species, Pelusios is one of the most speciose chelonian genera worldwide, even though the taxonomy of some species is contentious. Recent investigations suggested that the closely related, but morphologically distinct genus Pelomedusa is paraphyletic with respect to Pelusios, and that Pelomedusa consists of nine deeply divergent lineages. Using three mitochondrial and three nuclear DNA fragments (2054 bp mtDNA, 2025 bp nDNA), we examined for the first time the phylogeny of Pelusios by molecular means. Our analyses included all Pelusios species, except the probably extinct P. seychellensis, as well as the nine Pelomedusa lineages. The results showed that Pelusios and Pelomedusa are reciprocally monophyletic. Limited sampling of Pelusios species and homoplasy introduced by remote outgroups most likely explain the paraphyly of Pelomedusa in previous studies. The distinctiveness of most Pelusios species was confirmed, but none of the currently recognised species groups within Pelusios was monophyletic. In Pelusios rhodesianus and P. sinuatus distinct genetic lineages were discovered, suggestive of cryptic taxa. In contrast, the recognition of the weakly differentiated P. castaneus and P. chapini as full species is doubtful, as is the validity of the Malagasy and Seychellois subspecies of P. castanoides. GenBank sequences of P. williamsi were nested within P. castaneus, but the morphological distinctiveness of the two species makes it likely that the GenBank sequences (derived from a turtle from the pet trade) are misidentified. Divergence among the distinct genetic lineages of Pelomedusa equals or exceeds the differences among Pelusios species, supporting the view that Pelomedusa is a species complex.     

Comparative phylogeographies of six species of hinged terrapins (Pelusios spp.) reveal discordant patterns and unexpected differentiation in the P. castaneus/P. chapini complex and P. rhodesianus

Using up to 2117 bp of mitochondrial DNA and up to 2012 bp of nuclear DNA, we analysed phylogeographic differentiation of six widely distributed species of African hinged terrapins (Pelusios spp.) representing different habitat types. Two taxa each live in savannahs or in forests and mesic savannahs, respectively, and the remaining two species occur in intermediate habitats. The species living in forests and mesic savannahs do not enter dry savannahs, whereas the savannah species may occur in dry and wet savannahs and even in semi‐arid steppe regions. We found no obvious correlation between habitat type and phylogeographic pattern: one savannah species (P. rhodesianus) shows phylogeographic structure, i.e. pronounced genetic differences among geographically distinct populations, and the other (P. nanus) not. One species inhabiting forests and mesic savannahs (P. carinatus) has phylogeographic structure, the other (P. gabonensis) not. The same pattern is true for the two ecologically intermediate species, with phylogeographic structure present in P. castaneus and absent in P. chapini. Nuclear evidence suggests that the latter two taxa with abutting and partially overlapping ranges are distinct, while mtDNA is only weakly differentiated. Pelusios castaneus shows pronounced phylogeographic structure, which could reflect Pleistocene range interruptions correlated with the fluctuating forest cover in West and Central Africa. Our results do not support the recognition of an extinct subspecies of P. castaneus for the Seychelles. Pelusios carinatus contains two well supported clades, which are separated by the Congo River. This species is closely related to P. rhodesianus, a taxon consisting of two deeply divergent mitochondrial clades. One of these clades is paraphyletic with respect to P. carinatus, but the two clades of P. rhodesianus are not differentiated in the studied nuclear markers and, again, paraphyletic with respect to P. carinatus. Using mtDNA sequences from the type material of P. rhodesianus, we were able to allocate this name to one of the two clades. However, owing to the confusing relationships of P. rhodesianus and P. carinatus, we refrain from taxonomic decisions.     

Molecular evidence for species level divergence in African Nile Crocodiles Crocodylus niloticus (Laurenti, 1786)

Relationships of the newly discovered dwarf crocodiles from Mauritania were inferred from mitochondrial 12S sequences. Specimens from 13 different Crocodylus niloticus populations (from East Africa, West Africa and Madagascar) were compared. Additional representatives of the genus Crocodylus (one from Africa and one from Australia), the African genus Osteolaemus and the South American alligatorid Paleosuchus palpebrosus (as outgroup) were included in the analysis. Maximum-likelihood and Bayesian analyses yielded relationships that were strikingly different from currently prevailing phylogenetic hypotheses. Both analyses consistently revealed two groups, one consisting of the monophyletic West- and Central African populations and the other of a paraphyletic group containing the East African and Madagascan populations. High genetic divergence between those groups indicates separation on the species level. Furthermore ‘C’ cataphractus is clearly shown not to be a member of the genus Crocodylus. The resulting nomenclatural changes are discussed. To cite this article: A. Schmitz et al., C. R. Palevol 2 (2003).     

Phylogenetic position of African and Malagasy Pimpinella species and related genera (Apiaceae, Pimpinelleae)

The phylogenetic position of the African and Malagasy species of Pimpinella is assessed using nrDNA ITS sequence data and a representative sampling of the genus, including 16 species from Africa and Madagascar and 26 species from Eurasia. The results of maximum parsimony and Bayesian analyses of these data show that the African and Malagasy species ally with their Eurasian counterparts in Pimpinelleae. The genus Pimpinella is rendered paraphyletic by the inclusion of African Cryptotaenia and the small African and Malagasy endemic genera Frommia and Phellolophium. Within a paraphyletic Pimpinella, three major clades are recovered, with the African species occupying two of these clades. The current sectional classification of the genus, based predominantly on fruit vestiture, is largely artificial. Chromosome base number, however, was found to be consistent with the groupings recovered in the molecular analyses. Those African and Malagasy Pimpinella species with a chromosome base number of x = 11 and largely glabrous petals and fruits, form the earliest diverging clade together with Frommia, which also has a base count of n = 11 and glabrous petals and fruits. The remaining African species ally with several Eurasian species of Pimpinella and share a chromosome base number of x = 9 and usually hairy petals and fruits.     

Phylogenetic relationships among New Caledonian Sapotaceae (Ericales): molecular evidence for generic polyphyly and repeated dispersal

The phylogeny of a representative group of genera and species from the Sapotaceae tribe Chrysophylleae, mainly from Australia and New Caledonia, was studied by jackknife analyses of sequences of nuclear ribosomal DNA. The phylogeny conflicts with current opinions on generic delimitation in Sapotaceae. Pouteria and Niemeyera, as presently circumscribed, are both shown to be nonmonophyletic. In contrast, all species currently assigned to these and other segregate genera confined to Australia, New Caledonia, or neighboring islands, form a supported clade. Earlier classifications in which more genera are recognized may better reflect relationships among New Caledonian taxa. Hence, there is need for a revision of generic boundaries in Chrysophylleae, and particularly within the Pouteria complex, including Leptostylis, Niemeyera, Pichonia, Pouteria pro parte (the main part of section Oligotheca), and Pycnandra. Section Oligotheca have been recognized as the separate genus Planchonella, a monophyletic group that needs to be resurrected. Three clades with strong support in our jackknife analysis have one Australian species that is sister to a relatively large group of New Caledonian endemics, suggesting multiple dispersal events between this small and isolated tropical island and Australia. The phylogeny also suggests an interesting case of a relatively recent and rapid radiation of several lineages of Sapotaceae within New Caledonia.     

Genetic evidence for female-biased dispersal in the bullfrog, Rana catesbeiana (Ranidae)

Sex-biased dispersal is an important but unexplored area of amphibian ecology. We predicted female-biased dispersal in the bullfrog (Rana catesbeiana) based on aspects of their mating system and tested this prediction using data from seven polymorphic DNA microsatellite loci. Allelic (F-statistics) and genotypic [assignment index, (AIc)] frequencies from nine Ontario populations support our prediction, although significant sex differences in inbreeding and variance of AIc were not detected. The diversity of mating systems found in amphibians represents an important avenue for investigating the relationship between reproductive systems, dispersal and phylogeny.     

New evidence for parallel evolution of colour patterns in Malagasy poison frogs (Mantella)

Malagasy poison frogs of the genus Mantella are diurnal and toxic amphibians of highly variable and largely aposematic coloration. Previous studies provided evidence for several instances of homoplastic colour evolution in this genus but were unable to sufficiently resolve relationships among major species groups or to clarify the phylogenetic position of several crucial taxa. Here, we provide cytochrome b data for 143 individuals of three species in the Mantella madagascariensis group, including four newly discovered populations. Three of these new populations are characterized by highly variable coloration and patterns but showed no conspicuous increase of haplotype diversity which would be expected under a scenario of secondary hybridization or admixture of chromatically uniform populations. Several populations of these variable forms and of M. crocea were geographically interspersed between the distribution areas of Mantella aurantiaca and Mantella milotympanum. This provides further support for the hypothesis that the largely similar uniformly orange colour of the last two species evolved in parallel. Phylogenies based on over 2000 bp of two nuclear genes (Rag-1 and Rag-2) identified reliably a clade of the Mantella betsileo and Mantella laevigata groups as sister lineage to the M. madagascariensis group, but did not support species within the latter group as monophyletic. The evolutionary history of these frogs might have been characterized by fast and recurrent evolution of colour patterns, possibly triggered by strong selection pressures and mimicry effects, being too complex to be represented by simple bifurcating models of phylogenetic reconstruction.     

New nuclear evidence for the oldest divergence among neognath birds: the phylogenetic utility of ZENK (i)

To date, there is little consensus concerning the phylogenetic relationships among neognath orders, which include all extant birds except ratites and tinamous. Different data sets, both molecular and morphologic, have yielded radically different and often unresolved ordinal topologies, especially within the neoaves clade. This lack of resolution and ongoing conflict indicates a need for additional phylogenetic characters to be applied to the question of higher-level avian phylogeny. In this study, sequences of a single-copy nuclear gene, ZENK, were used to reconstruct an ordinal-level phylogeny of neognath birds. Strong support was indicated for the oldest divergence within Neognathae; the chicken- and duck-like birds formed a clade that was sister to all other modern birds. In addition, many families of traditional taxonomic orders clustered together in the ZENK tree, indicating the gene's general phylogenetic reliability. However, within the neoaves clade, there was little support for relationships among orders, which is a result similar to all other recent molecular studies of higher-level avian phylogeny. This similarity among studies suggests the possibility of a rapid radiation of the major neoaves lineages. Despite the ongoing lack of neoaves resolution, ZENK's sequence divergence and base composition patterns indicate its general utility as a new phylogenetic marker for higher-level avian systematics.     

Recent oceanic long-distance dispersal and divergence in the amphi-Atlantic rain forest genus Renealmia L.f. (Zingiberaceae)

Renealmia L.f. (Zingiberaceae) is one of the few tropical plant genera with numerous species in both Africa and South America but not in Asia. Based on phylogenetic analysis of nuclear ribosomal internal transcribed spacer (ITS) and chloroplast trnL-F DNA, Renealmia is shown to be monophyletic with high branch support. Low sequence divergence found in the two genome regions (ITS: 0-2.4%; trnL-F: 0-1.9%) suggests recent diversification within the genus. Molecular divergence age estimates give further support to the recent origin of the genus and show that Renealmia has attained its amphi-Atlantic distribution by an oceanic long-distance dispersal event from Africa to South America during the Miocene or Pliocene (15.8-2.7 My ago). Some support is found for the hypothesis that speciation in neotropical Renealmia was influenced by the Andean orogeny. Speciation has been approximately simultaneous on both sides of the Atlantic, but increased taxon sampling is required to compare the speciation rates between the New World and Old World tropics.     

Genetic relationships among American species of the genus Prosopis (Mimosoideae, Leguminosae) inferred from ITS sequences: evidence for long-distance dispersal

Aim  The genus Prosopis includes 44 species and has a pseudoamphitropical, disjunct distribution. We aimed to determine whether American Prosopis sections arose in North or South America, and to explain the current distribution of their species on the basis of their genetic relationships.
Location  South-western USA, Mexico, Caribbean Antilles, Peru–Ecuador, central and northern Argentina, south-western Argentina (Patagonia) and Cuyo, south-western Asia and northern Africa.
Methods  Internal transcribed spacer fragments from 21 species of Prosopis were sequenced and the data were used to analyse the phylogenetic relationships using Microlobius and Mimosa as outgroups. Genetic distances were calculated to estimate the degree of divergence. Dispersal–vicariance (DIVA) analysis was conducted to help understand the biogeographical history of the genus.
Main conclusions  The sections Strombocarpa and Algarobia are not monophyletic. Prosopis argentina (section Monilicarpa ) and the species of Algarobia are included in single clade. The phylogeny, DIVA analysis, and the pattern of genetic distances indicate that the ancestral area for the American species was wide, from south-western USA to Central and northern Argentina. Successive vicariance events split this area, and long-distance dispersal episodes (perhaps mediated by birds) led to recolonizations from North to South America, and vice versa .     

Allozyme divergence among four species ofPodaechmea s. l. and the status ofUrsulaea (Bromeliaceae, Bromelioideae)

The genetic relationships ofAechmea mexicana, A. lueddemanniana, A. macvaughii andA. tuitensis were investigated using starch gel electrophoresis. Eight enzyme systems encoded by ten putative gene loci were resolved in seventeen populations.Nei's (1978) genetic distances were obtained from allelic frequencies and used with UPGMA algorithm. Results indicate that some populations belonging to different species display genetic similarities closer to each other than to some conspecific populations. Our results do not support the proposed genusUrsulaea (Read & Baensch 1994), sinceA. tuitensis was closer toA. lueddemanniana andA. mexicana than toA. macvaughii.     

Molecular and cytogenetic evidence for cryptic speciation within a rare endemic Malagasy lemur, the Northern Sportive Lemur (Lepilemur septentrionalis)

Evolutionary relationships of different populations of the threatened malagasy lemur Lepilemur septentrionalis were assessed by sequence analysis of mitochondrial DNA (D-loop region and partial Cyt b gene). One hundred and fifty nine samples were collected from five main different localities in the northern part of Madagascar. We applied the phylogenetic species concept based on fixed diagnostic differences to determine the status of different geographical populations. No nucleotide site diagnoses Ankarana from Andrafiamena or Analamera. However, numerous fixed differences separate Sahafary from all other populations. These results were corroborated by phylogenetic trees. As previous cytogenetic studies, our molecular data suggest that two cryptic species of Lepilemur occur in the extreme north of Madagascar. This speciation is probably caused by chromosomal rearrangements in at least one of the evolutionary lineages. Our study comprises another striking example of how molecular genetic assay can detect phylogenetic discontinuities that are not reflected in traditional morphologically based taxonomies. Our study indicates that the Sahafary population is a hitherto undescribed endangered endemic species which urgently needs conservation efforts.     

Genetic evidence for sex-biased dispersal in resident bottlenose dolphins (Tursiops aduncus)

In most mammals males usually disperse before breeding, while females remain in their natal group or area. However, in odontocete cetaceans behavioural and/or genetic evidence from populations of four species indicate that both males and females remain in their natal group or site. For coastal resident bottlenose dolphins field data suggest that both sexes are philopatric to their natal site. Assignment tests and analyses of relatedness based on microsatellite markers were used to investigate this hypothesis in resident bottlenose dolphins, Tursiops aduncus, from two small coastal populations of southeastern Australia. Mean corrected assignment and mean relatedness were higher for resident females than for resident males. Only 8% of resident females had a lower probability than average of being born locally compared to 33% of resident males. Our genetic data contradict the hypothesis of bisexual philopatry to natal site and suggest that these bottlenose dolphins are not unusual amongst mammals, with females being the more philopatric and males the more dispersing sex.     

Phylogenetic relationships and divergence times among New World monkeys (Platyrrhini, Primates)

Orthologous sequences of six nuclear genes were obtained for all recognized genera of New World monkeys (Primates: Platyrrhini) and outgroups to evaluate the phylogenetic relationships and to estimate divergence times. Phylogenetic relationships were reconstructed by maximum parsimony, maximum likelihood, and Bayesian approaches. All methods resolved with 100% branch support genus-level relationships, except for the grouping of Aotus as a sister taxa of Cebus and Saimiri, which was supported by low bootstrap percentages and posterior probability. All approaches depict three monophyletic New World monkey families: Atelidae, Cebidae, and Pitheciidae; also within each family, all approaches depict the same branching topology. However, the approaches differ in depicting the relationships of the three families to one another. Maximum parsimony depicts the Atelidae and Cebidae as sister families next joined by the Pitheciidae. Conversely, likelihood and Bayesian phylogenetic trees group families Atelidae and Pitheciidae together to the exclusion of Cebidae. Divergence time estimations using both local molecular clock and Bayesian approaches suggest the families diverged from one another over a short period of geological time in the late Oligocene-early Miocene.     

Genetic divergence among species of the section Strombocarpa,genusProsopis (Leguminosae)

We studied degree of variability and relationships among populations belonging to five species of the section Strombocarpa of the family Leguminosae, namelyProsopis ferox, P. torquata, P. pubescens, P. strombulifera andP. reptans, by isozyme electrophoresis. The genetic similarity betweenP. reptans andP. strombulifera is high enough for the two to be considered subspecies or geographic races ofP. strombulifera instead of good taxonomic species. As expected these species cluster withP. pubescens, all belonging to series Strombocarpae. This group is different isoenzymatically from the remaining species,P. torquata andP. ferox. Comparison of our results with those obtained earlier for species of the section Algarobia showed that genetic variability in the section Strombocarpa is lower than in the section Algarobia, while the species divergence is much higher. These results suggest different adaptive strategies between these sections.     

Relaxed molecular clock provides evidence for long-distance dispersal of Nothofagus (southern beech)

Nothofagus (southern beech), with an 80-million-year-old fossil record, has become iconic as a plant genus whose ancient Gondwanan relationships reach back into the Cretaceous era. Closely associated with Wegener's theory of “Kontinentaldrift”, Nothofagus has been regarded as the “key genus in plant biogeography”. This paradigm has the New Zealand species as passengers on a Moa's Ark that rafted away from other landmasses following the breakup of Gondwana. An alternative explanation for the current transoceanic distribution of species seems almost inconceivable given that Nothofagus seeds are generally thought to be poorly suited for dispersal across large distances or oceans. Here we test the Moa's Ark hypothesis using relaxed molecular clock methods in the analysis of a 7.2-kb fragment of the chloroplast genome. Our analyses provide the first unequivocal molecular clock evidence that, whilst some Nothofagus transoceanic distributions are consistent with vicariance, trans-Tasman Sea distributions can only be explained by long-distance dispersal. Thus, our analyses support the interpretation of an absence of Lophozonia and Fuscospora pollen types in the New Zealand Cretaceous fossil record as evidence for Tertiary dispersals of Nothofagus to New Zealand. Our findings contradict those from recent cladistic analyses of biogeographic data that have concluded transoceanic Nothofagus distributions can only be explained by vicariance events and subsequent extinction. They indicate that the biogeographic history of Nothofagus is more complex than envisaged under opposing polarised views expressed in the ongoing controversy over the relevance of dispersal and vicariance for explaining plant biodiversity. They provide motivation and justification for developing more complex hypotheses that seek to explain the origins of Southern Hemisphere biota.     

Unravelling evolutionary lineages among South African velvet worms (Onychophora: Peripatopsis) provides evidence for widespread cryptic speciation

The endemic South African velvet worm genus Peripatopsis currently contains eight recognized species described from variable morphological characters and the current taxonomy is unsatisfactory. In an attempt to investigate evolutionary relationships within Peripatopsis, we collected 137 individuals from 34 sample localities for six of the eight species. Sequence data derived from two partial mitochondrial (mt)DNA gene loci (COI and 12S rRNA), as well as partial sequence data from the ribosomal nuclear 18S rDNA locus in combination with gross morphological characters and scanning electron microscopy (SEM), was used to examine evolutionary relationships. Phylogenetic relationships were investigated using minimum evolution (ME) and Bayesian inferences (BI). Additionally, we also undertook a maximum likelihood (ML) analyses on the combined DNA sequence data set. The combined DNA evidence topologies derived from the ME, BI, and ML was highly congruent and was characterized by the presence of multiple lineages within recognized taxa. Peripatopsis clavigera, Peripatopsis moseleyi, and Peripatopsis sedgwicki each comprised two evolutionary lineages; Peripatopsis capensis comprised three; and Peripatopsis balfouri comprised six operational taxonomic units respectively. Genealogical exclusivity at both mtDNA and nuclear DNA among the geographically coherent groups coupled with pronounced sequence divergence suggested a two‐fold increase in the number of species within Peripatopsis. Previously used gross morphological characters (such as the number of leg pairs and colour) were either highly variable within operational taxonomic units, or were invariant, suggesting that alternative morphological characters are necessary for species discrimination. SEM results revealed potentially useful diagnostic characters that can discriminate between at least discriminate some of the newly‐identified lineages. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society 2009, 97 , 200–216.     

A phylogeny of the oil bee tribe Ctenoplectrini (Hymenoptera: Anthophila) based on mitochondrial and nuclear data: evidence for early Eocene divergence and repeated out-of-Africa dispersal

The bee tribe Ctenoplectrini, with two genera, comprises nine species in tropical Africa and ten in Asia and Australia. Most of them collect floral oil, pollen, and nectar from Cucurbitaceae, but three species are thought to be cleptoparasites. The unusual morphology of Ctenoplectrini has made it difficult to infer their closest relatives, in turn preventing an understanding of these bees' geographic and temporal origin. We used two mitochondrial and two nuclear markers (4741 nucleotides) generated for most of the species to test the monophyly of the tribe, its relationships to other Apidae, and its biogeographic history. Ctenoplectrini are strongly supported as monophyletic and closest to the Long-horned bees, Eucerini. The presumably cleptoparasitic species form a clade (Ctenoplectrina) that is sister to the remaining species (Ctenoplectra), confirming the independent evolution of cleptoparasitism in this tribe. Tree topology and molecular dating together suggest that Ctenoplectrini originated in Africa in the Early Eocene and that Ctenoplectra dispersed twice from Africa to Asia, sometime in the Late Eocene, 30-40 my ago, from where one species reached the Australian continent via Indonesia and New Guinea in the mid-Miocene, c. 13 my ago. Dry and cool mid-Miocene climates also coincide with the divergence between Ctenoplectra bequaerti from West Africa and Ctenoplectra terminalis from East and South Africa, perhaps related to fragmentation of the equatorial African rainforest belt.     

Phylogenetic relationships among Puccinia hemerocallidis, P. funkiae, and P. patriniae (Uredinales) inferred from ITS sequence data

Puccinia hemerocallidis and P. funkiae resemble each other morphologically; however, they are biologically and taxonomically distinct, with telia of the former being restricted to species of Hemerocallis and the latter to Hosta species. However, both fungi share a macrocyclic and heteroecious life cycle with Patrinia villosa as the spermogonial and aecial host. An additional microcyclic rust fungus, P. patriniae, is also known on P. villosa. This microcyclic fungus is similar to the two macrocyclic fungi in its telial structure and teliospore morphology. These similarities in morphology and host relationships suggest the three fungi may also share a close evolutionary relationship. To determine the phylogenetic relationships of the three species, a portion of the nuclear ribosomal DNA repeat encoding the ITS and 5.8S subunit regions was amplified by PCR, sequenced, and analyzed. The resulting phylogenetic trees showed that P. hemerocallidis and P. funkiae share a recent common ancestor and that P. patriniae is closely allied with P. hemerocallidis. The results suggest a possible evolutionary derivation of microcyclic P. patriniae from macrocyclic heteroecious P. hemerocallidis, which fits the evolutionary interpretation of correlated species known as Tranzschel's law.