Carol Eunmi Lee

Evolutionary genetics of invasive species

The evolutionary genetics of invasive species has been relatively unexplored, but could offer insights into mechanisms of invasions. Recent studies suggest that the invasion success of many species might depend more heavily on their ability to respond to natural selection than on broad physiological tolerance or plasticity. Thus, these studies stress the importance of genetic architecture, selection upon which could result in evolutionary adaptations and possibly speciation. For instance, epistatic interactions and the action of a few genes could facilitate invasion success. These findings emphasize the utility of genomic approaches for determining invasion mechanisms, through analysis of gene expression, gene interactions, and genomic rearrangements that are associated with invasion events.

GLOBAL PHYLOGEOGRAPHY OF A CRYPTIC COPEPOD SPECIES COMPLEX AND REPRODUCTIVE ISOLATION BETWEEN GENETICALLY PROXIMATE “POPULATIONS”

The copepod Eurytemora affinis has a broad geographic range within the Northern Hemisphere, inhabiting coastal regions of North America, Asia, and Europe. A phylogenetic approach was used to determine levels of genetic differentiation among populations of this species, and interpopulation crosses were performed to determine reproductive compatibility. DNA sequences from two mitochondrial genes, large subunit (16S) rRNA (450 bp) and cytochrome oxidase I (COI, 652 bp), were obtained from 38 populations spanning most of the species range and from two congeneric species, E. americana and E. herdmani. Phylogenetic analysis revealed a polytomy of highly divergent clades with maximum sequence divergences of 10% in 16S rRNA and 19% in COI. A power test (difference of a proportion) revealed that amount of sequence data collected was sufficient for resolving speciation events occurring at intervals greater than 300,000 years, but insufficient for determining whether speciation events were approximately simultaneous. Geographic and genetic distances were not correlated (Mantels test; r= 0.023, P= 0.25), suggesting that populations had not differentiated through gradual isolation by distance. At finer spatial scales, there was almost no sharing of mtDNA haplotypes among proximate populations, indicating little genetic exchange even between nearby sites. Interpopulation crosses demonstrated reproductive incompatibility among genetically distinct populations, including those that were sympatric. Most notably, two geographically distant (4000 km) but genetically proximate (0.96% 16S, 0.15% COI) populations exhibited asymmetric reproductive isolation at the F2 generation. Large genetic divergences and reproductive isolation indicate that the morphologically conservative E. affinis constitutes a sibling species complex. Reproductive isolation between genetically proximate populations underscores the importance of using multiple measures to examine patterns of speciation.

RAPID AND REPEATED INVASIONS OF FRESH WATER BY THE COPEPOD EURYTEMORA AFFINIS

Invasions of fresh water by marine organisms have been of great interest to evolutionary biologists and paleontologists because they typically constitute major evolutionary transitions. Recent (< 200 years) invasions of fresh water by brackish or marine species offer an opportunity to understand mechanisms underlying these events, but pathways of invasion from salt water have not been confirmed using genetic data. This study employed mitochondrial DNA sequences (652 base pairs from the cytochrome oxidase I (COI) gene) to reconstruct the geographic and evolutionary history of freshwater invasion by the common estuarine and saltmarsh crustacean Eurytemora affinis (Copepoda; Poppe 1880). Phylogenetic analysis of populations from North America, Europe, and Asia revealed at least eight independent invasions of fresh water from genetically distinct lineages. At least five of these freshwater invasions most likely arose independently in different river drainages, recently from saltwater sources within each river drainage. An analysis of molecular variance (AMOVA) was performed at three geographic scales (among continents, among drainages, and within drainages) to assess the hierarchical distribution of genetic variance. Results indicated that 52% of the genetic variance was explained by differences among drainages, 43% by differences among continents, but only 5% by differences within drainages, thus supporting geographic patterns of invasions inferred from the phylogeny. Physiological experiments were performed to determine whether adults and larvae from saltwater populations could tolerate freshwater conditions. Transfer to zero salinity resulted in high mortalities, but with some survival to the second generation in one population. This study provides genetic evidence and physiological support for rapid transitions from a saline life history into fresh water, with repeated invasions on a global scale.

Evolutionary origins of invasive populations

What factors shape the evolution of invasive populations? Recent theoretical and empirical studies suggest that an evolutionary history of disturbance might be an important factor. This perspective presents hypotheses regarding the impact of disturbance on the evolution of invasive populations, based on a synthesis of the existing literature. Disturbance might select for life‐history traits that are favorable for colonizing novel habitats, such as rapid population growth and persistence. Theoretical results suggest that disturbance in the form of fluctuating environments might select for organismal flexibility, or alternatively, the evolution of evolvability. Rapidly fluctuating environments might favor organismal flexibility, such as broad tolerance or plasticity. Alternatively, longer fluctuations or environmental stress might lead to the evolution of evolvability by acting on features of the mutation matrix. Once genetic variance is generated via mutations, temporally fluctuating selection across generations might promote the accumulation and maintenance of genetic variation. Deeper insights into how disturbance in native habitats affects evolutionary and physiological responses of populations would give us greater capacity to predict the populations that are most likely to tolerate or adapt to novel environments during habitat invasions. Moreover, we would gain fundamental insights into the evolutionary origins of invasive populations.

Morphological stasis in the Eurytemora affinis species complex (Copepoda: Temoridae)

Morphological stasis has long been regarded as one of the most challenging problems in evolutionary biology. This study focused on the copepod species complex, Eurytemora affinis, as a model system to determine pattern and degree of morphological stasis. This study revealed discordant rates of morphological differentiation, molecular evolution, and reproductive isolation, where speciation was accompanied by lack of morphological differentiation in secondary sex characters. Comparisons were made among phylogenies based on morphometrics, nuclear (allozyme) loci, and mitochondrial DNA (mtDNA) sequences from cytochrome oxidase I, for a total of 43 populations within the complex. These systematic relationships were also compared to patterns of reproductive isolation. In addition, genetic subdivision of nuclear molecular (allozyme) markers (GST) and quantitative (morphological) characters (QST) were determined to infer evolutionary forces driving morphological differentiation. The morphometric phylogeny revealed that all clades, excluding the European clade, were morphologically undifferentiated and formed a polytomy (multifurcation). Morphometric distances were not correlated with mtDNA distances, or with patterns of reproductive isolation. In contrast, nuclear and mtDNA phylogenies were mostly congruent. Reproductive isolation proved to be the most sensitive indicator of speciation, given that two genetically and morphologically proximate populations showed evidence of hybrid breakdown. Quantitative genetic (morphological) subdivision (QST = 0.162) was lower than nuclear genetic subdivision (GST = 0.617) for four laboratory-reared North American populations, indicating retarded evolution of morphological characters. This result contrasts with most other species, where QST typically exceeds GST as a result of directional selection. Thus, in all but the European populations, evolution of the secondary sex characters was marked by morphological stasis, even between reproductively-isolated populations.

Evolution of Physiological Tolerance and Performance During Freshwater Invasions

Abstract Invasive species that penetrate habitat boundaries are likely to experience strong selection and rapid evolution. This study documents evolutionary shifts in tolerance and performance following the invasion of fresh water by the predominantly estuarine and salt marsh copepod Eurytemora affinis. Common-garden experiments were performed on freshwater-invading (Lake Michigan) and ancestral saline (St. Lawrence marsh) populations to measure shifts in adult survival (at 0, 5, and 25 PSU), and survival during development and development time (both using full-sib clutches split across 0, 5, 15, and 25 PSU). Results showed clear evidence of heritable shifts in tolerance and performance associated with freshwater invasions. The freshwater population exhibited a gain in low-salinity tolerance and a reduction in high-salinity tolerance relative to the saline population, suggesting tradeoffs. These tradeoffs were supported by negative genetic correlations between survival at fresh (0 PSU) versus higher salinities. Mortality in response to salinity occurred primarily before metamorphosis, suggesting that selection in response to salinity had acted primarily on the early life-history stages. The freshwater population exhibited curious patterns of life-history evolution across salinities, relative to the saline population, of retarded development to metamorphosis but accelerated development from metamorphosis to adulthood. This pattern might reflect tradeoffs between development rate and survival in fresh water at the early life-history stages, but some other selective force acting on later life-history stages. Significant effects of clutch (genotype) and clutch-by-salinity interaction (G × E) on survival and development time in both populations indicated ample genetic variation as substrate for natural selection. Variation for high-salinity tolerance was present in the freshwater population despite negative genetic correlations between high- and low-salinity tolerance. Results implicate the importance of natural selection and document the evolution of reaction norms during freshwater invasions.

Anthropogenically induced adaptation to invade (AIAI): contemporary adaptation to human-altered habitats within the native range can promote invasions.

Adaptive evolution is currently accepted as playing a significant role in biological invasions. Adaptations relevant to invasions are typically thought to occur either recently within the introduced range, as an evolutionary response to novel selection regimes, or within the native range, because of long‐term adaptation to the local environment. We propose that recent adaptation within the native range, in particular adaptations to human‐altered habitat, could also contribute to the evolution of invasive populations. Populations adapted to human‐altered habitats in the native range are likely to increase in abundance within areas frequented by humans and associated with human transport mechanisms, thus enhancing the likelihood of transport to a novel range. Given that habitats are altered by humans in similar ways worldwide, as evidenced by global environmental homogenization, propagules from populations adapted to human‐altered habitats in the native range should perform well within similarly human‐altered habitats in the novel range. We label this scenario ‘Anthropogenically Induced Adaptation to Invade’. We illustrate how it differs from other evolutionary processes that may occur during invasions, and how it can help explain accelerating rates of invasions.

Scaling of Gelatinous Clutches: Effects of Siblings' Competition for Oxygen on Clutch Size and Parental Investment per Offspring

Theories on the evolution of clutch size are primarily influenced by examples from terrestrial animals, yet most animal phyla occur exclusively in water. Oxygen has a lower diffusion coefficient and lower solubility in water than in air, and siblings in aquatic clutches often compete for oxygen. Mitigating this competition could affect allocation of resources to offspring. Gelatinous clutches are common in aquatic habitats and have evolved multiple times in many phyla. We hypothesized that spacing of embryos by gel enhances delivery of oxygen but that gel is organically costly. A model of diffusion predicts that clutch thickness should scale inversely with the square root of embryo concentration, indicating a need to reduce embryo concentration (and increase gel volume) disproportionately with increasing clutch thickness. For embryos in artificial clutches constructed with agarose gel, development was faster in clutches with more gel per embryo, as predicted. For natural gelatinous clutches of gastropods, thick clutches had disproportionately larger volumes of gel and disproportionately more organic material invested in gel relative to embryos. Thus, for aquatic gelatinous clutches, requirements for oxygen supply can affect trade‐offs involving clutch thickness and parental investment per offspring: resources are diverted to gel, and the proportion diverted increases with clutch thickness.

PUMPING IONS: RAPID PARALLEL EVOLUTION OF IONIC REGULATION FOLLOWING HABITAT INVASIONS

Marine to freshwater colonizations constitute among the most dramatic evolutionary transitions in the history of life. This study examined evolution of ionic regulation following saline‐to‐freshwater transitions in an invasive species. In recent years, the copepod Eurytemora affinis has invaded freshwater habitats multiple times independently. We found parallel evolutionary shifts in ion‐motive enzyme activity (V‐type H+ ATPase, Na+/K+‐ATPase) across independent invasions and in replicate laboratory selection experiments. Freshwater populations exhibited increased V‐type H+ ATPase activity in fresh water (0 PSU) and declines at higher salinity (15 PSU) relative to saline populations. This shift represented marked evolutionary increases in plasticity. In contrast, freshwater populations displayed reduced Na+/K+‐ATPase activity across all salinities. Most notably, modifying salinity alone during laboratory selection experiments recapitulated the evolutionary shifts in V‐type H+ ATPase activity observed in nature. Maternal and embryonic acclimation could not account for the observed shifts in enzyme activity. V‐type H+ ATPase function has been hypothesized to be critical for freshwater and terrestrial adaptations, but evolution of this enzyme function had not been previously demonstrated in the context of habitat transitions. Moreover, the speed of these evolutionary shifts was remarkable, within a few generations in the laboratory and a few decades in the wild.

Molecular ecology of zebra mussel invasions

The invasion of the zebra mussel, Dreissena polymorpha, into North American waters has resulted in profound ecological disturbances and large monetary losses. This study examined the invasion history and patterns of genetic diversity among endemic and invading populations of zebra mussels using DNA sequences from the mitochondrial cytochrome oxidase I (COI) gene. Patterns of haplotype frequency indicate that all invasive populations of zebra mussels from North America and Europe originated from the Ponto‐Caspian Sea region. The distribution of haplotypes was consistent with invasive populations arising from the Black Sea drainage, but could not exclude the possibility of an origin from the Caspian Sea drainage. Similar haplotype frequencies among North American populations of D. polymorpha suggest colonization by a single founding population. There was no evidence of invasive populations arising from tectonic lakes in Turkey, while lakes in Greece and Macedonia contained only Dreissena stankovici. Populations in Turkey might be members of a sibling species complex of D. polymorpha. Ponto‐Caspian derived populations of D. polymorpha (θ = 0.0011) and Dreissena bugensis (one haplotype) exhibited low levels of genetic diversity at the COI gene, perhaps as a result of repeated population bottlenecks. In contrast, geographically isolated tectonic lake populations exhibited relatively high levels of genetic diversity (θ = 0.0032 to 0.0134). It is possible that the fluctuating environment of the Ponto‐Caspian basin facilitated the colonizing habit of invasive populations of D. polymorpha and D. bugensis. Our findings were concordant with the general trend of destructive freshwater invaders in the Great Lakes arising from the Ponto‐Caspian Sea basin.