Mario Vargas-Ramírez

Deep genealogical lineages in the widely distributed African helmeted terrapin: Evidence from mitochondrial and nuclear DNA (Testudines: Pelomedusidae: Pelomedusa subrufa)

We investigated the phylogeographic differentiation of the widely distributed African helmeted terrapin Pelomedusa subrufa based on 1503 base pairs of mitochondrial DNA (partial cyt b and ND4 genes with adjacent tRNAs) and 1937 bp of nuclear DNA (partial Rag1, Rag2, R35 genes). Congruent among different analyses, nine strongly divergent mitochondrial clades were found, representing three major geographical groupings: (1) A northern group which includes clades I from Cameroon, II from Ghana and Ivory Coast, III from Benin, Burkina Faso and Niger, IV from the Central African Republic, and V from Kenya, (2) a northeastern group consisting of clades VI from Somalia, and VII from Saudi Arabia and Yemen, and (3) a southern group comprising clade VIII from Botswana, the Democratic Republic of Congo, Madagascar and Malawi, and clade IX from South Africa. Malagasy and continental African populations were not clearly differentiated, indicating very recent arrival or introduction of Pelomedusa in Madagascar. The southern group was in some phylogenetic analyses sister to Pelusios, rendering Pelomedusa paraphyletic with respect to that genus. However, using partitioned Bayesian analyses and sequence data of the three nuclear genes, Pelomedusa was monophyletic, suggesting that its mitochondrial paraphyly is due to either ancient introgressive hybridization or phylogenetic noise. Otherwise, nuclear sequence data recovered a lower level of divergence, but corroborated the general differentiation pattern of Pelomedusa as revealed by mtDNA. This, and the depth of the divergences between clades, indicates ancient differentiation. The divergences observed fall within, and in part exceed considerably, the differentiation typically occurring among chelonian species. To test whether Pelomedusa is best considered a single species composed of deep genealogical lineages, or a complex of up to nine distinct species, we suggest a future taxonomic revision that should (1) extend the geographical sampling of molecular data, specifically focusing on contact zones and the possible sympatric occurrence of lineages without admixture, and (2) evaluate the morphology of the various genealogical lineages using the type specimens or topotypical material of the numerous junior synonyms of P. subrufa.

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., Široký, 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.

Extremely low genetic diversity and weak population differentiation in the endangered Colombian river turtle Podocnemis lewyana (Testudines: Podocnemididae)

Podocnemis lewyana is an endangered endemic river turtle of Colombia. Using ten unlinked polymorphic microsatellite loci and a 691-bp-long DNA fragment corresponding to the more variable portion of the mitochondrial control region, we investigated genetic diversity and population structure throughout its range. Both neutral markers showed extremely low diversity and weak population differentiation. Our data indicate that the genetic history of P. lewyana has been impacted by multiple bottlenecks and population expansion since the Pleistocene. The observed differentiation pattern is most likely the result of historically low genetic variation resulting from restricted geographic range and aggravated by recent anthropogenically induced bottlenecks. Based on slight differences in allele frequencies among populations, we suggest that three regions should be treated as demographically independent Management Units in order to preserve maximal genetic diversity: (1) the Upper Magdalena River Basin, (2) the Lower Magdalenaxa0+xa0Lower Caucaxa0+xa0San Jorge River Basins, and (3) the Sinú River Basin. Among the Management Units, only low to moderate levels of gene flow were detected; these are largely unidirectional from Management Units 1 and 3 into Management Unit 2.ResumenPodocnemis lewyana es una tortuga endémica y amenazada de Colombia. Usando diez loci polimórficos de microsatélites y un fragmento de 691 pares de bases correspondiente a la porción más variable de la región control mitocondrial, investigamos la diversidad genética y estructura poblacional a través de su rango de distribución. Ambos marcadores neutrales revelaron una diversidad extremadamente baja y débil diferenciación poblacional. Nuestros datos indican que la historia genética de P. lewyana ha sido impactada por múltiples cuellos de botella y expansión poblacional desde el Pleistoceno. El patrón de diferenciación observado es más probable el resultado de la variación genética historica baja derivada del rango de distribución restringido, agravada por cuellos de botella recientes resultado de intervención antrópica. Basados en diferencias tenues de frecuencias alélicas entre poblaciones, sugerimos que tres regiones deben ser tratadas como Unidades de Manejo (UM) demográficamente independientes con el fin de preservar al máximo la diversidad genética. (1) Cuenca del alto río Magdalena, (2) Cuencas del bajo río Magdalenaxa0+xa0Bajo río Caucaxa0+xa0río San Jorge, y (3) Cuenca del río Sinú. Entre las UM se detectaron niveles bajos a moderados de flujo genético que es principalmente unidireccional de las UM 1 y 3 a la UM 2.

A revision of African helmeted terrapins (Testudines: Pelomedusidae: Pelomedusa), with descriptions of six new species.

Using nearly range-wide sampling, we analyze up to 1848 bp of mitochondrial DNA of 183 xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 helmeted terrapins and identify a minimum of 12 deeply divergent species-level clades. Uncorrected p distances of these clades equal or clearly exceed those between the currently recognized species of Pelusios, the genus most closely related to Pelomedusa. We correlate genetic discontinuities of Pelomedusa with data on morphology and endoparasites and describe six new Pelomedusa species. Moreover, we restrict the name Pelomedusa subrufa (Bonnaterre, 1789) to one genetic lineage and resurrect three further species from its synonymy, namely P. galeata (Schoepff, 1792), P. gehafie (Rüppell, 1835), and P. olivacea (Schweigger, 1812). In addition to these ten Pelomedusa species, we identify two further clades from Cameroon and Sudan with similar levels of genetic divergence that remain unnamed candidate species. We also note that some problematical terrapins from South Africa and Somalia may represent two additional candidate species. Some of the Pelomedusa species are morphologically distinctive, whilst others can only be identified by molecular markers and are therefore morphologically cryptic taxa.

Red- and yellow-footed tortoises, Chelonoidis carbonaria and C. denticulata (Reptilia: Testudines: Testudinidae), in South American savannahs and forests: do their phylogeographies reflect distinct habitats?

Using sequence data of the mitochondrial cytochrome b gene, we investigated phylogeographic differentiation of the Amazonian tortoise species Chelonoidis carbonaria and C. denticulata. While C. carbonaria is generally restricted to savannah habitats and adjacent forests, C. denticulata is associated with wet tropical and subtropical forests. Our study suggests a correlation between distinct habitat preferences and phylogeography of the two species. In Maximum Parsimony, Maximum Likelihood and Bayesian analyses, haplotypes of C. carbonaria cluster in several distinct clades reflecting the species’ patchy distribution in savannah habitats. By contrast, haplotypes of C. denticulata are only weakly differentiated; a finding also confirmed by parsimony network analysis. This suggests that the contiguous Amazonian rainforest allows gene flow between populations of the forest-dwelling C. denticulata throughout the range, but significantly impedes gene flow in C. carbonaria. The phylogeographic structure and extant distribution pattern of C. carbonaria is supportive of former Amazonian rainforest fragmentation, enabling the dispersal of savannah species. Based on fossil calibration, we dated divergence times for the C. carbonaria clades using a relaxed molecular clock, resulting in average estimates ranging from 4.0–2.2xa0mya. This implies that the onset of rainforest fragmentation could predate the Pleistocene considerably. Furthermore, our findings call for further research on geographic and taxonomic variation in C. carbonaria and for a reassessment of the conservation status of the distinct genetic units.

Northern genetic richness and southern purity, but just one species in the Chelonoidis chilensis complex

Fritz, U., Alcalde, L., Vargas‐Ramírez, M., Goode, E.V., Fabius‐Turoblin, D.U. & Praschag, P. (2012). Northern genetic richness and southern purity, but just one species in the Chelonoidis chilensis complex. —Zoologica Scripta, 41, 220–232.

Weak genetic divergence between the two South American toad-headed turtles Mesoclemmys dahli and M. zuliae (Testudines: Pleurodira: Chelidae)

Mesoclemmys dahli and M. zuliae are two endangered, little-known toad-headed turtles with small distribution ranges in Colombia and Venezuela, respectively. Using the mitochondrial cytochrome b gene as a marker, we investigate their phylogeographic differentiation. Furthermore, based on 2341 bp of mtDNA and 2109 bp of nDNA of M. dahli, M. zuliae and allied chelid turtles, we infer their divergence time using a fossil-calibrated relaxed molecular clock approach. Mesoclemmys dahli and M. zuliae are closely related species, with an estimated mean divergence time of 10.6 million years. This estimate correlates with the uplift of the Serrania de Perija, an Andean mountain chain separating their distribution ranges, suggesting that this event could have caused the evolution of the two species. Haplotype and nucleotide diversities of M. dahli are markedly higher than in Podocnemis lewyana, another endemic turtle species of Colombia. This pronounced dissimilarity may reflect differences in the phylogeographies and demographic histories of the two species, but also different habitat preferences.

Phylogenetic position of Pelusios williamsi and a critique of current GenBank procedures (Reptilia: Testudines: Pelomedusidae)

We re-examine the phylogenetic position of Pelusios williamsi by merging new sequences with an earlier published data set of all Pelusios species, except the possibly extinct P. seychellensis, and the nine previously identified lineages of the closely allied genus Pelomedusa (2054 bp mtDNA, 2025 bp nDNA). Furthermore, we include new sequences of Pelusios broadleyi, P. castanoides, P. gabonensis and P. marani. Individual and combined analyses of the mitochondrial and nuclear data sets indicate that P. williamsi is sister to P. castanoides, as predicted by morphology. This provides evidence for the misidentification of GenBank sequences allegedly representing P. williamsi. Such mislabelled GenBank sequences contribute to continued confusion, because only the original submitter can revise their identification; an impractical procedure impeding the rectification of obvious mistakes. We recommend implementing another option for revising taxonomic identifications, paralleling the century-old best practice of natural history museums for new determinations of specimens. Within P. broadleyi, P. gabonensis and P. marani, there is only shallow genetic divergence, while some phylogeographic structuring is present in the wide-ranging species P. castaneus and P. castanoides.

Phylogeography of the endangered black-breasted leaf turtle ( Geoemyda spengleri ) and conservation implications for other chelonians

Known-locality samples of Geoemyda spengleri from three Chinese provinces (Guangdong, Guangxi, Hainan) and northern Vietnam provide clear evidence for phylogeographic structure in a southern Chinese chelonian species. Within southern China two clades of mitochondrial haplotypes from east and west of the Xi Jiang together form the sister group to haplotypes from northern Vietnam. Turtles from Hainan Island harbour haplotypes of the same clade as turtles from Guangxi, 400 km distant. These findings suggest that phylogeographic differentiation may have once existed in other co-distributed, highly endangered chelonian species. The possibility of phylogeographic structure should be considered in any in-situ and ex-situ conservation program for Chinese chelonians.

Unexpected hybridization patterns in Near Eastern terrapins (Mauremys caspica, M. rivulata) indicate ancient gene flow across the Fertile Crescent

Recent studies indicate that hybridization in animals occurs more frequently than previously thought and that it may play an important evolutionary role. Chelonians are capable of extensive hybridization, raising the question how chelonian species evolve and maintain genetic integrity despite hybridization. Here, we use two sister species with parapatric distribution, Mauremys caspica and M. rivulata, as our model. These taxa are estimated to have diverged some 5.3–7.0 million years ago. Using rangewide sampling and 13 unlinked polymorphic microsatellite markers, five nuclear loci and one mitochondrial gene, we show that hybridization is rare along the contact zone of the two species in Turkey. However, we discovered an unexpected hybrid swarm in the southern Levant that has been hitherto identified with M. rivulata. This hybrid swarm is separated from the inland species M. caspica by a 700‐km‐wide distribution gap corresponding to the Syrian Desert. Ecological palaeomodelling suggests that during more humid climatic episodes in the Last Glacial Maximum and mid‐Holocene, the current contact zone extended into the southern Levant, facilitating the establishment of the now isolated hybrid swarm. Our results support that there is not necessarily a general hybridization pattern in a given species couple and that the extent of gene flow may differ considerably in different parts of the distribution range. Moreover, our results highlight that studies on hybridization should not focus only on extant contact and hybrid zones, but should use rangewide sampling to detect signals of ancient hybridization that might otherwise be missed.