Jörg Plötner

Widespread unidirectional transfer of mitochondrial DNA: a case in western Palaearctic water frogs

Interspecies transfer of mitochondrial (mt) DNA is a common phenomenon in plants, invertebrates and vertebrates, normally linked with hybridization of closely related species in zones of sympatry or parapatry. In central Europe, in an area north of 48°N latitude and between 8° and 22°E longitude, western Palaearctic water frogs show massive unidirectional introgression of mtDNA: 33.7% of 407 Rana ridibunda possessed mtDNA specific for Rana lessonae. By contrast, no R. lessonae with R. ridibunda mtDNA was observed. That R. ridibunda with introgressed mitochondrial genomes were found exclusively within the range of the hybrid Rana esculenta and that most hybrids had lessonae mtDNA (90.4% of 335 individuals investigated) is evidence that R. esculenta serves as a vehicle for transfer of lessonae mtDNA into R. ridibunda. Such introgression has occurred several times independently. The abundance and wide distribution of individuals with introgressed mitochondrial genomes show that R. lessonae mt genomes work successfully in a R. ridibunda chromosomal background despite their high sequence divergence from R. ridibunda mtDNAs (14.2–15.2% in the ND2/ND3 genes). Greater effectiveness of enzymes encoded by R. lessonae mtDNA may be advantageous to individuals of R. ridibunda and probably R. esculenta in the northern parts of their ranges.

A molecular phylogeny of Scrippsiella sensu lato (Calciodinellaceae, Dinophyta) with interpretations on morphology and distribution

The phylogenetic relationships of Scrippsiella sensu lato (including cyst taxa such as Calcigonellum, Calciodinellum, and Pernambugia) were investigated based on sequences from the ribosomal 5.8S rRNA and the Internal Transcribed Spacers ITS1 and ITS2, including interpretations on morphology and distribution. To attach importance to the cyst diversity present in calcareous dinoflagellates, a segregation of Scrippsiella sensu lato into four well-recognizable and monophyletic groups is proposed, corresponding to more- or less-established taxonomic units: (i) Calciodinellum (including Calcigonellum and a few species assigned to Scrippsiella), (ii) Pernambugia (presumptively including Lebessphaera), (iii) S. precaria and S. ramonii, and (iv) Scrippsiella sensu stricto comprising largely the S. trochoidea species complex. The phylogenetic relationships among these four groups are not sufficiently resolved. Molecular data suggest the existence of numerous cryptic taxa showing molecular, but not morphological, variation (especially in Scrippsiella sensu stricto). Closely related strains have a wide range of distribution and occur (at least partly) sympatrically in cold through to tropical seas of the world.

Divergence in mitochondrial DNA of Near Eastern water frogs with special reference to the systematic status of Cypriote and Anatolian populations (Anura, Ranidae)

Water frogs from Anatolia, Syria, Jordan, and central Asia (Turkmenistan, Uzbekistan, Kazakhstan) were compared on the basis of the complete mitochondrial (mt) ND3 gene (340 bp), two flanking mt t-RNA gene fragments (26 bp), and a 374 bp fragment of the mt 12S rRNA gene. A total of 27 haplotypes were found among the investigated individuals. Anatolian water frogs differed from Syrian and Jordanian Rana bedriagae by 2.2-3.4% of the analysed sites. The observed divergence (2.8-4.1%) between the Cypriote water frogs and frogs from the surrounding mainland (southern Turkey, west Syria) was in the same range as between R. bedriagae and European R. ridibunda (3.1-3.9%). These results suggest that neither the Cypriote nor the Anatolian water frogs represent R. bedriagae . Furthermore, maximum parsimony and neighbour-joining trees showed a clear subdivision of Asian water frogs into three Anatolian lineages, two central Asian lineages, a Cypriote lineage and the bedriagae lineage. In all trees the Cypriote lineage branches off first and a clade formed by two Anatolian lineages is placed as the sister group of water frogs from Turkmenistan and Uzbekistan, whereas the phylogenetic positions of R. bedriagae , the Ceyhan lineage, the Kazakhstan lineage and R. ridibunda remain unclear.

Delimitation of the Thoracosphaeraceae (Dinophyceae), Including the Calcareous Dinoflagellates, Based on Large Amounts of Ribosomal RNA Sequence Data

The phylogenetic relationships of the Dinophyceae (Alveolata) are not sufficiently resolved at present. The Thoracosphaeraceae (Peridiniales) are the only group of the Alveolata that include members with calcareous coccoid stages; this trait is considered apomorphic. Although the coccoid stage apparently is not calcareous, Bysmatrum has been assigned to the Thoracosphaeraceae based on thecal morphology. We tested the monophyly of the Thoracosphaeraceae using large sets of ribosomal RNA sequence data of the Alveolata including the Dinophyceae. Phylogenetic analyses were performed using Maximum Likelihood and Bayesian approaches. The Thoracosphaeraceae were monophyletic, but included also a number of non-calcareous dinophytes (such as Pentapharsodinium and Pfiesteria) and even parasites (such as Duboscquodinium and Tintinnophagus). Bysmatrum had an isolated and uncertain phylogenetic position outside the Thoracosphaeraceae. The phylogenetic relationships among calcareous dinophytes appear complex, and the assumption of the single origin of the potential to produce calcareous structures is challenged. The application of concatenated ribosomal RNA sequence data may prove promising for phylogenetic reconstructions of the Dinophyceae in future.

Genetic divergence and evolution of reproductive isolation in eastern Mediterranean water frogs.

Water frogs [genus Pelophylax (Rana)] that occur around the eastern Mediterranean Sea provide an opportunity to study early stages of speciation. The geography of the eastern Mediterranean region has changed dramatically since the Middle Miocene as a result of motions of adjoining lithospheric plates and regional-scale vertical crustal motions (uplift and subsidence). For several hundred thousand years between 6 and 5 million years ago (Mya), the Mediterranean basin was isolated from the Atlantic Ocean, and became desiccated (the Messinian Salinity Crisis; MSC). Geological data suggest that the endemic water frog lineage on Cyprus was isolated by the flooding of the Mediterranean basin by salt water at the end of the MSC, circa 5.5–5.3 Mya. This suggests a rate of uncorrected genetic divergence of approximately 1.1% per million years (My). Divergence time estimates based on this rate are in good agreement with the chronology of events in the history of crustal deformation and landscape development in the eastern Mediterranean region.

Balancing a Cline by Influx of Migrants: A Genetic Transition in Water Frogs of Eastern Greece

Variation patterns of allozymes and of ND3 haplotypes of mitochondrial DNA reveal a zone of genetic transition among western Palearctic water frogs extending across northeastern Greece and European Turkey. At the western end of the zone, allozymes characteristic of Central European frogs known as Pelophylax ridibundus predominate, whereas at the eastern end, alleles characteristic of western Anatolian water frogs (P. cf. bedriagae) prevail. The ND3 haplotypes reveal 2 major clades, 1 characteristic of Anatolian frogs, the other of European; the European clade itself has distinct eastern and western subclades. Both the 2 major clades and the 2 subclades overlap within the transition zone. Using Bayesian model selection methods, allozyme data suggest considerable immigration into the Nestos River area from eastern and western populations. In contrast, the ND3 data suggest that migration rates are so high among all locations that they form a single panmictic unit; the best model for allozymes is second best for mitochondrial DNA (mtDNA). Nuclear markers (allozymes), which have roughly 4 times as deep a coalescent history as mtDNA data and thus may reflect patterns over a longer time, indicate that eastern and western refugial populations have expanded since deglaciation (in the last 10,000 years) and have met near the Nestos River, whereas the mtDNA with its smaller effective population size has already lost the signal of partitioning into refugia.

Genetic diversity and distribution patterns of diploid and polyploid hybrid water frog populations (Pelophylax esculentus complex) across Europe

Polyploidization is a rare yet sometimes successful way for animals to rapidly create geno‐ and phenotypes that may colonize new habitats and quickly adapt to environmental changes. In this study, we use water frogs of the Pelophylax esculentus complex, comprising two species (Pelophylax lessonae, genotype LL; Pelophylax ridibundus, RR) and various diploid (LR) and triploid (LLR, LRR) hybrid forms, summarized as P. esculentus, as a model for studying recent hybridization and polyploidization in the context of speciation. Specifically, we compared the geographic distribution and genetic diversity of diploid and triploid hybrids across Europe to understand their origin, maintenance and potential role in hybrid speciation. We found that different hybrid and parental genotypes are not evenly distributed across Europe. Rather, their genetic diversity is structured by latitude and longitude and the presence/absence of parental species but not of triploids. Highest genetic diversity was observed in central and eastern Europe, the lowest in the northwestern parts of Europe. This gradient can be explained by the decrease in genetic diversity during postglacial expansion from southeastern glacial refuge areas. Genealogical relationships calculated on the basis of microsatellite data clearly indicate that hybrids are of multiple origin and include a huge variety of parental genomes. Water frogs in mixed‐ploidy populations without any parental species (i.e. all‐hybrid populations) can be viewed as evolutionary units that may be on their way towards hybrid speciation. Maintenance of such all‐hybrid populations requires a continuous exchange of genomes between diploids and triploids, but scenarios for alternative evolutionary trajectories are discussed.

Batrachochytrium dendrobatidis in Germany: distribution, prevalences, and prediction of high risk areas

In Germany, the pathogenic fungus Batrachochytrium dendrobatidis (Bd) was detected in 11 indigenous frog species, 4 newt species, and 1 salamander species in 64 out of the 181 locations (35%) investigated. Among the 3450 samples collected between 2003 and 2011, 284 (8.2%) were positive for Bd infections. The highest prevalences were observed in Alytes obstetricans (17.8% of individuals, 20% of populations), followed by Ichthyosaura alpestris (14.7%, 22.2%), Bombina variegata (13.9%, 38.5%), and water frogs comprising 2 species, Pelophylax lessonae and P. ridibundus, and their hybrid form P. esculentus (13.5%, 29.0%). Bd is widespread; areas of higher prevalence were detected in eastern, southeastern, western, and southwestern Germany. Our data indicate that drift fencing of amphibians is not a risk factor for the anthropogenic spread of Bd. Although chytridiomycosis outbreaks have never been observed in Germany, it cannot be excluded that Bd infections affect the dynamics of local amphibian populations. Among the questions still to be answered is whether juveniles are more susceptible to Bd infections than adults. Further work, especially long-term observations including capture-mark-recapture studies, is required to clarify the impact Bd has on amphibians in Germany and Central Europe.

New records of Batrachochytrium dendrobatidis in Chilean frogs.

We used molecular techniques to examine 11 species of frogs in 6 localities in southern Chile to ascertain the incidence of the chytrid fungus Batrachochytrium dendrobatidis (Bd). We detected the fungus in 2 localities (Coñaripe and Raúl Marín Balmaceda) in 3 species: Batrachyla leptopus, Pleurodema thaul and Rhinoderma darwinii. Our findings expand the list of Bd hosts to include B. leptopus and P. thaul and extend the spatial distribution in Chile to include the southernmost Bd record at Raúl Marín Balmaceda.

Is premeiotic genome elimination an exclusive mechanism for hemiclonal reproduction in hybrid males of the genus Pelophylax

BackgroundThe ability to eliminate a parental genome from a eukaryotic germ cell is a phenomenon observed mostly in hybrid organisms displaying an alternative propagation to sexual reproduction. For most taxa, the underlying cellular pathways and timing of the elimination process is only poorly understood. In the water frog hybrid Pelophylax esculentus (parental taxa are P. ridibundus and P. lessonae) the only described mechanism assumes that one parental genome is excluded from the germline during metamorphosis and prior to meiosis, while only second genome enters meiosis after endoreduplication. Our study of hybrids from a P. ridibundus—P. esculentus-male populations known for its production of more types of gametes shows that hybridogenetic mechanism of genome elimination is not uniform.ResultsUsing comparative genomic hybridization (CGH) on mitotic and meiotic cell stages, we identified at least two pathways of meiotic mechanisms. One type of Pelophylax esculentus males provides supporting evidence of a premeiotic elimination of one parental genome. In several other males we record the presence of both parental genomes in the late phases of meiotic prophase I (diplotene) and metaphase I.ConclusionSome P. esculentus males have no genome elimination from the germ line prior to meiosis. Considering previous cytological and experimental evidence for a formation of both ridibundus and lessonae sperm within a single P. esculentus individual, we propose a hypothesis that genome elimination from the germline can either be postponed to the meiotic stages or absent altogether in these hybrids.