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Increasing environmental changes primarily due to anthropogenic impacts, are affecting organisms all over the planet. In general, scientists distinguish between three different ways in which organisms can respond to environmental changes in their habitat: extinction, dispersal and adaptation. An example of organisms which are highly adaptable and can easily cope with new and changing environments are invasive species which are able to colonize new habitats with only few individuals. To successfully survive in their new environment, invasive species adapt fast to novel abiotic and biotic parameters, such as different temperature regimes. Phenotypic plasticity which enables organisms to quickly modify their phenotype to new environmental conditions, explains the success in adaptation of invasive species.
While underlying mechanisms of phenotypic plasticity are not fully understood, one possible “motor” of phenotypic plasticity is epigenetics. Especially DNA methylation could explain the fast changes of the organism’s phenotype due to plasticity when experiencing changing environments, as invasive species do. DNA methylation could even contribute to the adaptation of invasive species via phenotypic plasticity, especially with clonally reproducing species. Methods such as common garden experiments with clonally reproducing species are a useful tool to differentiate between phenotypic plasticity and genetic adaptation because the confusing effects of genetic variation are lowered in clonally reproducing species.
Our overall goal was to evaluate the genetic adaptive potential of New Zealand mud snail (Potamopyrgus antipodarum) populations from Europe since they went through an extreme bottleneck after colonizing Europe only 180-360 generations ago. Seemingly, two different clonal lineages colonized Europe because two 16 s rRNA and cytochrome b haplotypes were found across different European countries, haplotypes t and z. The NZMS is a highly successful invasive species that is nowadays nearly globally distributed. The shells of the NZMS show a habitat-dependent high variability and are a fitness-relevant trait. The high variability in shell morphology is due to both genetic variation and phenotypic plasticity. To disentangle genetic from environmental effects on the shell morphology NZMS, we conducted a common garden experiment. We kept asexually reproducing females from eleven European populations in climate cabinets with three different temperatures to produce offspring. We compared shell size and shape across three generations using the geometric morphometrics approach. Furthermore, we estimated reaction norms, maternal effects, broad-sense heritability, the coefficient of genetic variation (CVA) and evolvability (IA) in shell size and shape across different temperature conditions. Additionally, we investigated the reproductive rate of the parental generation.
Results showed that the shell morphology of the parental generation differed across populations. In contrast, the shell morphology of offspring generations became more similar. The reaction norms of the F1 generation were rather variable across the three temperatures. However, we were able to observe a haplotype-dependent pattern across the reaction norms suggesting a restricted genetic differentiation among NZMS in Europe. We detected high heritability values in size indicating a high genetic influence. Heritability values for shape were lower than in size. Generally, heritability varied slightly depending on temperature. Size seemed to have a higher evolvability than shape. However, the values of all our calculations were very low which indicates that the European NZMS populations are genetically diminished. The reproductive rate of the parental generation was rather haplotype than temperature dependent. In summary, we were able to display that the NZMS is capable to plastically adapt its shell morphology to different temperatures showing significant differences between the two haplotypes. Nevertheless, the low evolvability values indicate that little genetic variation has formed since the arrival of the NZMS in Europe and therefore, European NZMS seem to have a reduced ability to react to selection.
These results implied that phenotypic plasticity is important for the adaptation to different environmental conditions in the NZMS and maybe other molluscan species. Since classical experimental approaches can only describe the resulting phenotypes, we also intended to shed more light on the mechanistic side of environmentally induced phenotypic modifications using DNA methylation analysis. Although molluscs represent one of the most diverse taxa within the metazoan and are found in many different habitats, our knowledge of the DNA methylation in molluscs is scarce. Therefore, we aimed at deepening and summarizing our understanding about DNA methylation in molluscs. Publicly available molluscan genomic and transcriptomic data of all eight mollusc classes was downloaded to search for DNA methyltransferases (DNMTs 1-3) responsible for DNA methylation. Additionally, we estimated the normalized CpG dinucleotide content (CpG o/e) indicating the presence/absence and the frequency of DNA methylation in the genome. The CpG o/e ratio refers to the level of DNA methylation in the genome. Based on the sensitivity of methylated cytosines to mutate into thymine residues, species having a high germline methylation in genomic regions over evolutionary time, also have a lower CpG content, which is called CpG depletion. In contrary, species with a limited germline methylation in genomic regions over evolutionary time, show a higher CpG content and lack CpG depletion. The presence or absence of CpG depletion can be calculated with the CpG o/e ratio. Ultimately, the goal of our analyses was to gain insight into the evolution of methylation in molluscs.
We detected DNMTs in all eight mollusc classes and in most of the species. It is therefore plausible that the last common ancestor of molluscs has already had the enzymatic machinery which is needed for DNA methylation. However, various species did not possess the complete DNMT toolkit indicating evolutionary modification in DNA methylation. In general, we found a wide distribution of the bimodal CpG o/e pattern in six mollusc classes, resulting from CpG depletion. The genes in these groups seem to be divided into genes with a high degree of methylation and genes with a lower degree of methylation. This implies that DNA methylation seems to be rather common in molluscs. Species of Solenogastres and Monoplacophora were not or only sparsely methylated. It seems that those mollusc groups have undergone a reduction in DNA methylation. We hope that our investigations will demonstrate the lacking knowledge in epigenetics of molluscs and encourage scientist to execute and continue genetic studies on molluscs.
There is an increasingly urgent need to understand and predict how organisms will cope with the environmental consequences of global climate change. Adaptation in any form can be mediated by genetic adaptation and/or by phenotypic plasticity. Disentangling these two adaptive processes is critical in understanding and predicting adaptive responses to environmental change. Usually, disentangling genetic adaptation from phenotypic plasticity requires common garden experiments conducted under controlled laboratory conditions. While these experiments are powerful, it is often difficult to translate the results into natural populations and extrapolate to naturally occurring phenotypic variation. One solution to this problem is provided by the many examples of invasive species that exhibit wide phenotypic variation and that reproduce asexually. Besides selecting the appropriate in situ model, one must carefully choose a relevant trait to investigate. Ecomorphology has been a central theme in evolutionary biology because it reflects how organisms can adapt to their environment through their morphology. Intraspecific ecomorphological studies are especially well suited to identify adaptive pressures and provide insights into the microevolutionary mechanisms leading to the phenotypic differentiation.
One excellent candidate for an intraspecific ecomorphological study aiming to understand adaptation through genetic adaptation and phenotypic plasticity is the invasive New Zealand mudsnail Potamopyrgus antipodarum Gray (1853). This ovoviviparous snail features high variability in shell morphology and has successfully invaded a wide range of fresh- and brackish water habitats around the world. The evolutionary and ecological situations in this species’ native and invasive ranges is drastically different. In New Zealand, P. antipodarum’s native range, sexual and asexual individuals coexist and experience selective pressure by sterilizing endoparasites. By contrast, only a few asexual lineages have been established in invaded regions around the globe, where parasite infection is extremely rare. Here, we took advantage of the low genetic diversity among asexually reproducing European individuals in an attempt to characterize the relative contribution of genetic variation and phenotypic plasticity to the wide variation in shell morphology of this snail.
Analysing the ecomorphology of 425 European P. antipodarum in a geometric-morphometric framework, using brood size as proxy for fecundity, and mtDNA and nuclear SNPs to account for relatedness and identify reproductive mode, we hypothesized that 1) shell variation in the invasive range should be adaptive with respect to colonization of novel habitats, and 2) at least some of the variation might be caused by phenotypic plasticity. We then expanded our ecomorphological scope by analysing 996 native specimens, expecting 1) genetic and morphological diversity to be higher in the native range compared to invaded regions; 2) morphological diversity to be higher in sexual compared to asexual individuals according to the frozen niche hypothesis; and 3) shell morphology to be habitat specific, hence adaptative. In a last part, we used computational fluid dynamics simulations to calculate relative drag and lift forces of three shell morphologies (globular, intermediate, and slender). Here, we tested the overall hypothesis that shell morphology in gastropods is an adaptation against dislodgement through lift rather than drag forces, which would explain the counterintuitive presence of wider shells with shorter spires in lotic environments. With a final flow tank experiment, we tested the specific hypothesis that the dislocation velocity of living snails is positively linked to foot size, and that the latter can be predicted by shell morphology, in particular the aperture area as assumed by several authors.
As expected, we found genetic and morphological diversity to be higher in native than in invasive snails, but surprisingly no higher morphological diversity in sexual versus asexual individuals. The relationships between shell morphology, habitat, and fecundity were complex. Shape variation was primarily linked to genetic relatedness, but specific environmental factors including flow rate induced similar shell shapes. By contrast, shell size was largely explained by environmental factors. Fecundity was correlated with size, but showed trade-offs with shape in increasingly extreme conditions. With increasing flow and in smaller habitats such as springs, the trend of shell shape becoming wider was reversed, i.e. snails with slender shells were brooding more embryos. This increase in fitness was explained by our CFD simulations: in lotic habitats, slender shells experience less drag and lift forces compared to globular shells. We found no correlation between foot size and shell shape or aperture area showing that the assumed aperture/foot area correlation should be used with caution and cannot be generalized for all aquatic gastropod species. Finally, shell morphology and foot size were not related to dislodgement speed in our flow tank experiment. We concluded that the relationship of shell morphology and flow velocity is more complex than assumed. Hence, other traits must play a major role in decreasing dislodgement risk in stream gastropods, e.g. specific behaviours or pedal mucus stickiness. Although we did not find that globular shells are adaptations decreasing dislodgement risk, we cannot rule out that they are still flow related adaptations. For instance, globular shells are more crush-resistant and might therefore represent a flow adaptation in terms of diminishing damage caused by tumbling after dislodgement or against lotic specific crush-type predators.
At this point, we can conclude that shell morphology in P. antipodarum varies at least in part as an adaptation to specific environmental factors. This study shows how essential it is to reveal how plastic, genetically as well as phenotypically, adaptive traits in species can be and to identify the causal factors and how these adaptations affect the fitness in order to better predict how organisms will cope with changing environmental conditions.