No evidence for nuclear introgression despite complete mtDNA replacement in the Carpathian newt (Lissotriton montandoni)

Patterns of interspecific introgression may vary geographically, and the distribution of introgressed variants can yield insight into the historical dynamics of genetic interactions between hybridizing species. Urodele amphibians, often characterized by limited mobility, deep intraspecific genetic structuring and vulnerability to climatic changes, constitute suitable models for such historical inferences. Here, we combine an extensive survey of the mitochondrial (mtDNA) and nuclear (15 microsatellites) genomes in the Carpathian newt, Lissotriton montandoni (Lm) with species distribution modelling (SDM). Populations of the smooth newt, L. vulgaris (Lv) from the areas surrounding the Lm range were also sampled to test whether gene flow between these hybridizing species extends beyond the area of strict syntopy. The extent of introgression differs dramatically between the mitochondrial genome and the nuclear genome. While multiple, spatially and temporally distinct introgression events from Lv resulted in complete mtDNA replacement in Lm, there was little evidence of recent interspecific nuclear gene flow in the assayed markers. Microsatellite differentiation within Lm defines three units, probably derived from separate glacial refugia, located in the northern, eastern and southern part of the Carpathians. In situ survival and range fragmentation of Lm are supported by SDM, corroborating the role of the Carpathians as a major refugial area. Our results, in combination with previous reports of extensive introgression of the major histocompatibility complex (MHC) genes, emphasize the complexity of historical gene exchange between Lm and Lv.     

Characterization of microsatellite loci in two closely related Lissotriton newt species

We have developed eight di- and tetranucleotide Lissotriton microsatellite markers. Eight loci were polymorphic in the palmate newt Lissotriton helveticus and six were polymorphic in the smooth newt L. vulgaris. Polymorphism detected in 33 and 37 individuals per species ranged from 3 to 15 alleles. These markers are suitable for the investigation of population structure, genetic variation and taxonomic identification in the two focal species, and may also be of use in other Lissotriton–Triturus species.     

Gene flow rates in Yugoslavian populations of the smooth newt Triturus vulgaris

Allozymic variation in 22 loci in several Yugoslavian populations of four subspecies of the smooth newt Triturus vulgaris, has been analyzed. The frequency of private alleles and the coefficient of genetic differentiation, F_ST, give very different indirect estimates of the effective number of migrants per generation, Nm. However, such Nm estimates, in most cases higher than 1, imply that gene flow between populations is large enough as to prevent differentiation by random drift. In the case of T.v. vulgaris, of which sixteen populations amply distributed through Yugoslavia were sampled, there is evidence that frequent extinction and recolonization processes might be responsible for the observed genetic structure. This conclusion has been reached after testing the correlation between genetic, environmental and geographical matrices.     

Molecular and morphological evidence of hybridization between newts Triturus vulgaris and T. montandoni (Caudata: Salamandridae) in Slovakia

Natural hybridization and introgression is recorded between the newt species Triturus vulgaris and T. montandoni in Slovakia. To confirm a hybrid status of two putative hybrids, morphological and Randomly Amplified Polymorphic DNA (RAPD) markers specific for T. vulgaris and T. montandoni were used. The individuals found in Zbojská (Veporské vrchy Mts, Slovakia; analyzed morphologically and genetically) and Zubrohlava (Oravská kotlina basin, Slovakia; analyzed only morphologically) possessed markers of both species. Segregation of RAPD markers together with a model-based Bayesian analysis revealed that the specimen from Zbojská belonged to later generation hybrids.     

Massive introgression of major histocompatibility complex (MHC) genes in newt hybrid zones

Variation in the vertebrate major histocompatibility complex (MHC) genes is crucial for fighting pathogen assault. Because new alleles confer a selective advantage, MHC should readily introgress between species, even under limited hybridization. Using replicated transects through two hybrid zones between strongly reproductively isolated European newts, Lissotriton montandoni and L. vulgaris, we demonstrated recent and ongoing MHC class I and II introgression in the Carpathian region. The extent of introgression correlated with the age of contact. In the older zone, MHC similarity between species within transects exceeded similarity between transects within species, implying pervasive introgression ‐ a massive exchange of MHC genes, not limited to specific variants. In simulations, the observed pattern emerged under the combined action of balancing selection and hybridization, but not when these processes acted separately. Thus, massive introgression at advanced stages of divergence can introduce novel and restore previously lost MHC variation, boosting the adaptive potential of hybridizing taxa. In consequence, MHC genes may be the last to stop introgressing between incipient species.     

Male attractiveness is influenced by UV wavelengths in a newt species but not in its close relative

Background

Functional communication in the UV range has been reported in Invertebrates and all major groups of Vertebrates but Amphibians. Although perception in this wavelength range has been shown in a few species, UV signalling has not been demonstrated in this group. One reason may be that in lentic freshwater habitats, litter decomposition generates dissolved organic carbon that absorbs UV radiation and thus hinders its use for visual signalling. We tested the effect of male UV characteristics on female sexual preference in two newt species that experience contrasting levels of UV water transmission when breeding.

Methodology/Principal Findings

We analysed water spectral characteristics of a sample of breeding ponds in both species. We quantified male ventral coloration and measured male attractiveness under two lighting conditions (UV present, UV absent) using a no-choice female preference design. UV transmission was higher in Lissotriton vulgaris breeding sites. Male UV patterns also differed between experimental males of the two species. We observed a first common peak around 333 nm, higher in L. vulgaris, and a second peak around 397 nm, more frequent and higher in L. helveticus. Male attractiveness was significantly reduced in L. vulgaris when UV was not available but not in L. helveticus. Male attractiveness depended on the hue of the first UV peak in L. vulgaris.

Conclusion/Significance

Our study is the first report of functional UV-based communication in Amphibians. Interestingly, male spectral characteristics and female preferences were consistent with the differences in habitat observed between the two species as L. helveticus often breeds in ponds containing more UV blocking compounds. We discuss the three hypotheses proposed so far for UV signalling in animals (enhanced signal detectability, private communication channel, indicator of individual quality).     

Lags in the response of plant assemblages to global warming depends on temperature-change velocity

Aim

Current global warming is driving changes in biological assemblages by increasing the number of thermophilic species while reducing the number of cold-adapted species, leading to thermophilization of these assemblages. However, there is increasing evidence that thermophilization might not keep pace with global warming, resulting in thermal lags. Here, we quantify the magnitude of thermal lags of plant assemblages in Norway during the last century and assess how their spatio-temporal variation is related to variables associated with temperature-change velocity, topographic heterogeneity, and habitat type.

Location

Norway.

Time period

1905–2007.

Major taxa studied

Vascular plants.

Methods

We inferred floristic temperature from 16,351 plant assemblages and calculated the floristic temperature anomaly (difference between floristic temperature and baseline temperature) and thermal lag index (difference between reconstructed floristic temperature and observed climatic temperature) from 1905 until 2007. Using generalized least squares models, we analysed how the variation in observed lags since 1980 is related to temperature-change velocity (measured as magnitude, rate of temperature change, and distance to past analogous thermal conditions), topographic heterogeneity, and habitat type (forest versus non-forest), after accounting for the baseline temperature.

Results

The floristic temperature anomaly increases overall during the study period. However, thermophilization falls behind temperature change, causing a constantly increasing lag for the same period. The thermal lag index increases most strongly in the period after 1980, when it is best explained by variables related to temperature-change velocity. We also find a higher lag in non-forested areas, while no relationship is detected between the degree of thermal lag and fine-scale topographic heterogeneity.

Main conclusions

The thermal lag of plant assemblages has increased as global warming outpaces thermophilization responses. The current lag is associated with different dimensions of temperature-change velocity at a broad landscape scale, suggesting specifically that limited migration is an important contributor to the observed lags.     

Differential introgression across newt hybrid zones: Evidence from replicated transects

Genomic heterogeneity of divergence between hybridizing species may reflect heterogeneity of introgression, but also processes unrelated to hybridization. Heterogeneous introgression and its repeatability can be directly tested in natural hybrid zones by examining multiple transects. Here, we studied hybrid zones between the European newts Lissotriton montandoni and two lineages of Lissotriton vulgaris, with replicate transects within each zone. Over 1,000 nuclear genes located on a linkage map and mitochondrial DNA were investigated using geographical and genomic clines. Overall, the five transects were all similar, showing hallmarks of strong reproductive isolation: bimodal distribution of genotypes in central populations and narrow allele frequency clines. However, the extent of introgression differed between the zones, possibly as a consequence of their different ages, as suggested by the analysis of heterozygosity runs in diagnostic markers. In three transects genomic signatures of small‐scale (~2 km) zone movements were detected. We found limited overlap of cline outliers between transects, and only weak evidence of stronger differentiation of introgression between zones than between transects within zones. Introgression was heterogeneous across linkage groups, with patterns of heterogeneity similar between transects and zones. Predefined candidates for increased or reduced introgression exhibited only a subtle tendency in the expected direction, suggesting that interspecific differentiation is not a reliable indicator for the strength of introgression. These hierarchically sampled hybrid zones of apparently different ages show how introgression unfolds with time and offer an excellent opportunity to dissect the dynamics of hybridization and architecture of reproductive isolation at advanced stages of speciation.     

Micro-habitat use within a guild of newt larvae (Trituridae) in an Alpine lake

Population and ecological parameters such as numbers of larvae, microhabitat use, niche breadth and niche overlap of three species of syntopic larval newts (Alpine newt Triturus alpestris, Italian crested newt T. carnifex, and common newt T. vulgaris) were studied for two years in a small pond at 1160 m a.s.l. in NE Slovenia. Differences in microhabitat partitioning among larval newts were small. The largest niche breadth was estimated for larval T. alpestris, and the narrowest estimate was for larval T. carnifex in both years. Ecological differences seem to be very small and quite variable among sites and years. It appears that the developmental stage and size of newt larvae are more important in explaining resource partitioning than the characteristics of each species. Because of the absence of potential invertebrate predators and adult newts in the second half of the breeding season, the injuries could only be caused by intra-and interspecific predation attempts.     

Agricultural intensification alters marbled newt genetic diversity and gene flow through density and dispersal reduction

Recent agricultural intensification threatens global biodiversity with amphibians being one of the most impacted groups. Because of their biphasic life cycle, amphibians are particularly vulnerable to habitat loss and fragmentation that often result in small, isolated populations and loss of genetic diversity. Here, we studied how landscape heterogeneity affects genetic diversity, gene flow and demographic parameters in the marbled newt, Triturus marmoratus, over a hedgerow network landscape in Western France. While the northern part of the study area consists of preserved hedged farmland, the southern part was more profoundly converted for intensive arable crops production after WWII. Based on 67 sampled ponds and 10 microsatellite loci, we characterized regional population genetic structure and evaluated the correlation between landscape variables and (i) local genetic diversity using mixed models and (ii) genetic distance using multiple regression methods and commonality analysis. We identified a single genetic population characterized by a spatially heterogeneous isolation-by-distance pattern. Pond density in the surrounding landscape positively affected local genetic diversity while arable crop land cover negatively affected gene flow and connectivity. We used demographic inferences to quantitatively assess differences in effective population density and dispersal between the contrasted landscapes characterizing the northern and southern parts of the study area. Altogether, results suggest recent land conversion affected T. marmoratus through reduction in both effective population density and dispersal due to habitat loss and reduced connectivity.     

A comparative study of mitotic and meiotic chromosomes of the Montandon's newt,Triturus montandoni (Urodela: Salamandridae) from Poland and Rumania

A karyological analysis was carried out on two populations ofTriturus montandoni, one from Poland and another from Rumania. For both samples, morphometric characteristics and C-banding pattern of mitotic chromosomes are provided. Only slight differences between specimens from two geographic localities were found. Data on chiasma frequency and distribution are presented for male meiosis. No sex-related heteromorphism was found and for none of the chromosomal arms was a consistent absence of chiasmata recorded. There was a relatively high proportion of spermatocyte metaphases I with chiasmata on both arms of all chromosomes in all specimens studied. It is concluded that there are no well-defined sex chromosomes in the chromosomal complement of the maleT. montandoni. The findings are compared with previous studies on chromosome morphology, C-banding pattern, and meiosis in closely related species,T. vulgaris andT. helveticus.     

No trade‐offs in interspecific interference ability and predation susceptibility in newt larvae

Coexistence of species with similar requirements is allowed, among others, through trade‐offs between competitive ability and other ecological traits. Although interspecific competition is based on two mechanisms, exploitation of resources and physical interference, trade‐off studies largely consider only species’ ability to exploit resources. Using a mesocosm experiment, we examined the trade‐off between interference competition ability and susceptibility to predation in larvae of two newt species, Ichthyosaura alpestris and Lissotriton vulgaris. In the presence of heterospecifics, L. vulgaris larvae slowed somatic growth and developmental rates, and experienced a higher frequency of injuries than in conspecific environments which suggests asymmetrical interspecific interference. During short‐term predation trials, L. vulgaris larvae suffered higher mortality than I. alpestris. Larvae of the smaller species, L. vulgaris, had both lower interference and antipredator performance than the larger I. alpestris, which suggests a lack of trade‐off between interference competition ability and predator susceptibility. We conclude that interference competition may produce a positive rather than negative relationship with predation susceptibility, which may contribute to the elimination of subordinate species from common habitats.     

No distinct barrier effects of highways and a wide river on the genetic structure of the Alpine newt (<Emphasis Type="Italic">Ichthyosaura alpestris</Emphasis>) in densely settled landscapes

Linear landscape elements such as roads, railways and rivers have been shown to act as barriers to dispersal and gene flow, hence impeding functional connectivity and increasing genetic differentiation between individuals or populations on opposite sides of the barrier. Such putative barriers act through a confluence of mechanisms, including crossing mortality, barrier avoidance and modifications to organisms’ effective dispersal patterns. Small, terrestrial animals such as amphibians are predicted to be vulnerable to the effects of such barriers given their limited locomotive performance and their dependence on spatially distinct breeding habitats. Here, we examined the effects of highways and a wide river on Ichthyosaura alpestris in three regions of northern Switzerland by measuring the genetic differentiation between local populations and describing the spatial genetic structure. Moreover, we estimated effective population sizes as an indicator for the susceptibility of populations to random genetic drift. Based on genetic differentiation, we found evidence to suggest that the highways and river acted as barriers to gene flow for the newt in the study regions, but results were inconsistent when ignoring breeding ponds with low samples sizes. Admixture-based genetic clustering suggested the delineation of the genotypes to rough regional clusters, with only weak structure inferred within these clusters. Thus, results suggest that at present, highways and rivers do not substantially affect the genetic structure of I. alpestris within northern Switzerland in a negative manner. Alternatively, the lack of a distinct genetic structure in regional newt populations may be explained by, e.g., large effective population sizes.     

Landscape genetics in a changing world: disentangling historical and contemporary influences and inferring change

Landscape genetics seeks to determine the effect of landscape features on gene flow and genetic structure. Often, such analyses are intended to inform conservation and management. However, depending on the many factors that influence the time to reach equilibrium, genetic structure may more strongly represent past rather than contemporary landscapes. This well‐known lag between current demographic processes and population genetic structure often makes it challenging to interpret how contemporary landscapes and anthropogenic activity shape gene flow. Here, we review the theoretical framework for factors that influence time lags, summarize approaches to address this temporal disconnect in landscape genetic studies, and evaluate ways to make inferences about landscape change and its effects on species using genetic data alone or in combination with other data. Those approaches include comparing correlation of genetic structure with historical versus contemporary landscapes, using molecular markers with different rates of evolution, contrasting metrics of genetic structure and gene flow that reflect population genetic processes operating at different temporal scales, comparing historical and contemporary samples, combining genetic data with contemporary estimates of species distribution or movement, and controlling for phylogeographic history. We recommend using simulated data sets to explore time lags in genetic structure, and argue that time lags should be explicitly considered both when designing and interpreting landscape genetic studies. We conclude that the time lag problem can be exploited to strengthen inferences about recent landscape changes and to establish conservation baselines, particularly when genetic data are combined with other data.     

Conservation genetics of the central newt (Notophthalmus viridescens) in Iowa: the importance of a biogeographic framework

In light of global declines in amphibian populations, genetic data have become increasingly important for understanding population structure and for revealing hidden diversity. At the species level, Notophthalmus viridescens is an IUCN species of “least concern”, but the subspecies N. v. louisianensis (central newt) is listed as “threatened” in Iowa, a state on the western periphery of the species range. Genetic data were collected from 282 N. v. louisianensis from 14 sites in Iowa. Sequences from 1,054 nucleotides of mitochondrial DNA from Iowa newts revealed unexpected diversity in the form of two major haplotype groups that are not sister clades, with southern Iowa N. v. louisianensis being more closely related to N. v. piaropicola (peninsula newt) from Florida than to consubspecifics in Iowa. Sequence differentiation indicates that the two lineages of newts present in Iowa diverged near the beginning of the Pleistocene. Northern and southern Iowa haplotypes were found together at one site, indicating an opportunity for hybridization near Remington’s biogeographic suture zone 1, a hotspot for hybridization in other species. Three microsatellite loci provided additional evidence for distinctness of northern and southern Iowa newt populations. This study highlights the relevance of historical biogeography to conservation, as management strategies for N. v. louisianensis in Iowa must reflect previously unrecognized diversity in this species. Nuclear and mitochondrial data indicate genetic isolation of nearby populations on the same drainage, and field data suggest the decline of one study population, emphasizing the need for identification and protection of newt breeding sites in Iowa.     

Landscape genetics of the Alpine newt (<Emphasis Type="Italic">Mesotriton alpestris</Emphasis>) inferred from a strip-based approach

Habitat destruction and fragmentation are known to strongly affect dispersal by altering the quality of the environment between populations. As a consequence, lower landscape connectivity is expected to enhance extinction risks through a decrease in gene flow and the resulting negative effects of genetic drift, accumulation of deleterious mutations and inbreeding depression. Such phenomena are particularly harmful for amphibian species, characterized by disjunct breeding habitats. The dispersal behaviour of amphibians being poorly understood, it is crucial to develop new tools, allowing us to determine the influence of landscape connectivity on the persistence of populations. In this study, we developed a new landscape genetics approach that aims at identifying land-uses affecting genetic differentiation, without a priori assumptions about associated ecological costs. We surveyed genetic variation at seven microsatellite loci for 19 Alpine newt (Mesotriton alpestris) populations in western Switzerland. Using strips of varying widths that define a dispersal corridor between pairs of populations, we were able to identify land-uses that act as dispersal barriers (i.e. urban areas) and corridors (i.e. forests). Our results suggest that habitat destruction and landscape fragmentation might in the near future affect common species such as M. alpestris. In addition, by identifying relevant landscape variables influencing population structure without unrealistic assumptions about dispersal, our method offers a simple and flexible tool of investigation as an alternative to least-cost models and other approaches.     

Fine scale spatial genetic structure of two syntopic newts across a network of ponds: implications for conservation

In this study we used genetic approaches to assess the influence of landscape features on the dispersal patterns and genetic structure of two newt species (Triturus macedonicus and Lissotriton vulgaris) living syntopically in a network of ponds. Multilocus genotypes were used to detect and measure genetic variation patterns, population genetic structure and levels of gene flow. We interpret results on the basis of the different dispersal properties of the two species and explored the influence of certain landscape features, such as road and channel networks, on population connectivity. We found marked differences in the spatial genetic patterns of the respective species, which can be explained by their different dispersal properties. The road network seems to act as a barrier to dispersal in the overland dispersing L. vulgaris, while the channel network maintains connectivity in the aquatic dispersing T. macedonicus. The simultaneous and comparative consideration of species in a given area offers a much better understanding of the processes that govern population dynamics and persistence, providing valuable knowledge useful in conservation and management design.     

Sexual dimorphism in the skull geometry of newt species of <Emphasis Type="Italic">Ichthyosaura,Triturus</Emphasis> and <Emphasis Type="Italic">Lissotriton</Emphasis> (Salamandridae,Caudata, Amphibia)

In this study, we applied geometric morphometrics to explore variations in the level and pattern of sexual size dimorphism (SSD) and sexual shape dimorphism (SShD) of the ventral cranium in three different Modern Eurasian newt taxa (Ichthyosaura alpestris, Triturus species group and Lissotriton vulgaris). The ventral cranium is the part of the skull that is more directly related to foraging and feeding. Our results indicate that the level and pattern of sexual dimorphism in the ventral cranium differ among Modern Eurasian newt taxa. Regarding sexual dimorphism in skull size, Ichthyosaura alpestris and Triturus species show female-biased patterns (females are larger than males), whereas Lissotriton vulgaris appears to be non-dimorphic in skull size. In I. alpestris and Triturus species, SShD is mostly absent, whereas in L. vulgaris, SShD is more pronounced. A high level of variation between populations in both SSD and SShD indicates that local conditions may have a profound effect on the magnitude and direction of sexual dimorphism. The significant sexual differences in ventral cranium size and shape indicate possible subtle intersexual differences in ecological demands due to diet specialisation, in spite of similar general ecological settings.