Gene expression and drought response in an invasive thistle

Though rapid phenotypic evolution has been observed in many invasive plant species, less is known about the associated genetic mechanisms. Some hypotheses invoke the evolution of trade-offs in resource allocation to explain phenotypic differences between the native and invaded ranges of a species. Alternately, invasive species may benefit from a generalist strategy and perform well in many environments. Identification of the molecular changes associated with successful invasions can offer clues regarding the mechanistic basis of such hypotheses, even in non-model organisms. To complement studies of phenotypic variation, we investigate gene expression during drought response that might underlie variation in drought tolerance between native and introduced populations of diffuse knapweed (Centaurea diffusa), and possibly contribute to invasion success. Using species-specific microarrays and tissue sampled under drought and control conditions at three time points, we identified genes whose expression either varied constitutively or responded to drought stress differently between ranges. Further, we functionally investigate these genes. Based on these data, invasive populations have constitutively higher levels of expression relating to energy production and lower levels of signal transduction expression relative to native populations. Under drought conditions, invasive populations may maintain energy production and react less strongly to drought than native populations, which may allow them to maintain fitness across moderate environmental variation. This supports the expectation of invasive populations consisting of generalist genotypes capable of fitness homeostasis, which may have facilitated the successful invasion of many environments in North America.     

Contrasting levels of evolutionary potential in populations of the invasive plant Polygonum cespitosum

The amount of quantitative genetic variation within an invasive species influences its ability to adapt to conditions in the new range and its long-term persistence. Consequently, this aspect of genetic diversity (or evolutionary potential) can be a key factor in the success of species invasions. Previous studies have compared the evolutionary potential of populations in introduced versus native ranges of invasive species, but to date no study has examined differences among introduced-range populations of such species in levels of quantitative genetic variation expressed in ecologically relevant environments. We assessed quantitative variation of fitness, life-history, and functional traits in six geographically separate introduced-range populations of the invasive annual Polygonum cespitosum, by comparing norms of reaction for a large sample of genotypes (16–19 per population) expressed in response to two glasshouse environments simulating contrasting habitats in this new range. Patterns of reaction norm diversity varied considerably among the 6 populations studied. Two populations showed very little quantitative genetic variation in both environments. In contrast, two other populations contained significant genetic variation for fitness and life-history traits in the form of genotypes with low performance in both habitats. Finally, two populations showed significant norm of reaction diversity in the form of cross-over interaction: genotypes that performed relatively well in one environment did poorly in the other. Differences among populations in potential selective response are likely to affect the dynamics and future spread of P. cespitosum, since specific populations will likely contribute differently to the invasion process. More generally, our results suggest that the evolutionary component of long-term invasion success may depend on population rather than on species-level processes.     

Lack of adaptation from standing genetic variation despite the presence of putatively adaptive alleles in introduced sweet vernal grass (Anthoxanthum odoratum)

Population genetic theory predicts that the availability of appropriate standing genetic variation should facilitate rapid evolution when species are introduced to new environments. However, few tests of rapid evolution have been paired with empirical surveys for the presence of previously identified adaptive genetic variants in natural populations. In this study, we examined local adaptation to soil Al toxicity in the introduced range of sweet vernal grass (Anthoxanthum odoratum), and we genotyped populations for the presence of Al tolerance alleles previously identified at the long‐term ecological Park Grass Experiment (PGE, Harpenden, UK) in the species native range. We found that markers associated with Al tolerance at the PGE were present at appreciable frequency in introduced populations. Despite this, there was no strong evidence of local adaptation to soil Al toxicity among populations. Populations demonstrated significantly different intrinsic root growth rates in the absence of Al. This suggests that selection on correlated root growth traits may constrain the ability of populations to evolve significantly different root growth responses to Al. Our results demonstrate that genotype–phenotype associations may differ substantially between the native and introduced parts of a species range and that adaptive alleles from a native species range may not necessarily promote phenotypic differentiation in the introduced range.     

Quantifying natural seasonal variation in mutation parameters with mutation accumulation lines

Mutations create novel genetic variants, but their contribution to variation in fitness and other phenotypes may depend on environmental conditions. Furthermore, natural environments may be highly heterogeneous. We assessed phenotypes associated with survival and reproductive success in over 30,000 plants representing 100 mutation accumulation lines of Arabidopsis thaliana across four temporal environments at a single field site. In each of the four assays, environmental variance was substantially larger than mutational variance. For some traits, whether mutational variance was significantly varied between seasons. The founder genotype had mean trait values near the mean of the distribution of the mutation accumulation lines in all field experiments. New mutations also contributed more phenotypic variation than would be predicted, given phenotypic and sequence‐level divergence among natural populations of A. thaliana. The combination of large environmental variance with a mean effect of mutation near zero suggests that mutations could contribute substantially to standing genetic variation.     

SRAP analysis of genetic diversity of nine native populations of wild sugarcane, Saccharum spontaneum, from Sichuan, China

Saccharum spontaneum is a wild sugarcane species that is native to and widely distributed in China. It has been extensively used in sugarcane breeding programs, and is being tested for the development of bioenergy cultivars. In order to provide basic information for the exploitation of this species, we analyzed genetic variation among and within native S. spontaneum populations collected from Sichuan, China. Eighty plants from nine native populations were sampled. Twenty-one sequence-related amplified polymorphism primer pairs generated 235 clearly scorable bands, of which 185 were polymorphic (78.7%). Nei's genetic diversity was 0.2801 and Shannon's information index was 0.4155 across the populations. Genetic diversity parameters, G(ST) value (0.2088) and N(m) value (1.8944), showed that the genetic variation within populations was greater than that among populations. In the cluster analysis, one major grouping was formed by populations from Ya'an and another one by populations from Sichuan basin; a population from Baoxing formed a single cluster. In order to fully comprehend the genetic diversity of cold-tolerant local germplasm in this species, germplasm should be collected from the heterogeneous environments along the northern regions of this species' distribution. The germplasm that we collected should be a valuable resource for Saccharum breeding.     

Genome size reduction can trigger rapid phenotypic evolution in invasive plants

Background and Aims

The study of rapid evolution in invasive species has highlighted the fundamental role played by founder events, emergence of genetic novelties through recombination and rapid response to new selective pressures. However, whether rapid adaptation of introduced species can be driven by punctual changes in genome organization has received little attention. In plants, variation in genome size, i.e. variation in the amount of DNA per monoploid set of chromosomes through loss or gain of repeated DNA sequences, is known to influence a number of physiological, phenological and life-history features. The present study investigated whether change in genome size has contributed to the evolution of greater potential of vegetative growth in invasive populations of an introduced grass.

Methods

The study was based on the recent demonstration that invasive genotypes of reed canarygrass (Phalaris arundinacea) occurring in North America have emerged from recombination between introduced European strains. The genome sizes of more than 200 invasive and native genotypes were measured and their genome size was related to their phenotypic traits measured in a common glasshouse environment. Population genetics data were used to infer phylogeographical relationships between study populations, and the evolutionary history of genome size within the study species was inferred.

Key Results

Invasive genotypes had a smaller genome than European native genotypes from which they are derived. This smaller genome size had phenotypic effects that increased the species'' invasive potential, including a higher early growth rate, due to a negative relationship between genome size and rate of stem elongation. Based on inferred phylogeographical relationships of invasive and native populations, evolutionary models were consistent with a scenario of genome reduction by natural selection during the invasion process, rather than a scenario of stochastic change.

Conclusions

Punctual reduction in genome size could cause rapid changes in key phenotypic traits that enhance invasive ability. Although the generality of genome size variation leading to phenotypic evolution and the specific genomic mechanisms involved are not known, change in genome size may constitute an important but previously under-appreciated mechanism of rapid evolutionary change that may promote evolutionary novelties over short time scales.Key words: Biological invasion, evolutionary models, genome size, Phalaris arundinacea, quantile regression, relative growth rate, rapid evolution     

The lack of genetic bottleneck in invasive Tansy ragwort populations suggests multiple source populations.

Jacobaea vulgaris (Asteraceae) is a species of Eurasian origin that has become a serious non-indigenous weed in Australia, New Zealand, and North America. We used neutral molecular markers to (1) test for genetic bottlenecks in invasive populations and (2) to investigate the invasion pathways. It is for the first time that molecular markers were used to unravel the process of introduction in this species.The genetic variation of 15 native populations from Europe and 16 invasive populations from Australia, New Zealand and North America were compared using the amplified fragment length polymorphisms (AFLP's). An analysis of molecular variance showed that a significant part (10%) of the total genetic variations between all individuals could be explained by native or invasive origin.Significant among-population differentiation was detected only in the native range, whereas populations from the invasive areas did not significantly differ from each other; nor did the Australian, New Zealand and North American regions differ within the invasive range. The result that native populations differed significantly from each other and that the amount of genetic variation, measured as the number of polymorphic bands, did not differ between the native and invasive area, strongly suggests that introductions from multiple source populations have occurred. The lack of differentiation between invasive regions suggests that either introductions may have occurred from the same native sources in all invasive regions or subsequent introductions took place from one into another invasive region and the same mix of genotypes was subsequently introduced into all invasive regions.An assignment test showed that European populations from Ireland, the Netherlands and the United Kingdom most resembled the invasive populations.     

Effects of competition with Bromus tectorum on early establishment of Poa secunda accessions: can seed source impact restoration success?

When landscapes are heavily impacted by biological invasion, local populations of native plant species may no longer be adapted to altered environmental conditions. In these cases, it is useful to investigate alternative sources of germplasm, such as cultivated varieties, for planting at restoration sites. This study compared cultivated and wild (local) varieties of the native perennial bunchgrass, Poa secunda J. Presl, grown with and without the exotic, invasive Bromus tectorum L. in a greenhouse setting. While P. secunda cultivars emerged and grew more rapidly than wild seed sources, this advantage declined in the presence of B. tectorum and cultivated germplasm did not outperform wild accessions in the presence of an invasive species. Given the novel genetic background of cultivars and their potential to alter patterns of dominance in native plant communities, we recommend the use of local or regional wild seed sources when possible to conserve regional patterns of genetic diversity and adaptation. Use of multiple seed sources may increase the potential for capturing vigorous genotypes in the restoration seed mix. In cases where sites are heavily impacted by exotic, invasive species, other control measures will be necessary to improve establishment of native species in grassland restoration programs.     

Locally adapted traits maintained in the face of high gene flow

Gene flow between phenotypically divergent populations can disrupt local adaptation or, alternatively, may stimulate adaptive evolution by increasing genetic variation. We capitalised on historical Trinidadian guppy transplant experiments to test the phenotypic effects of increased gene flow caused by replicated introductions of adaptively divergent guppies, which were translocated from high‐ to low‐predation environments. We sampled two native populations prior to the onset of gene flow, six historic introduction sites, introduction sources and multiple downstream points in each basin. Extensive gene flow from introductions occurred in all streams, yet adaptive phenotypic divergence across a gradient in predation level was maintained. Descendants of guppies from a high‐predation source site showed high phenotypic similarity with native low‐predation guppies in as few as ~12 generations after gene flow, likely through a combination of adaptive evolution and phenotypic plasticity. Our results demonstrate that locally adapted phenotypes can be maintained despite extensive gene flow from divergent populations.     

Patterns of introduction and adaptation during the invasion of Aegilops triuncialis (Poaceae) into Californian serpentine soils

Multiple introductions can play a prominent role in explaining the success of biological invasions. One often cited mechanism is that multiple introductions of invasive species prevent genetic bottlenecks by parallel introductions of several distinct genotypes that, in turn, provide heritable variation necessary for local adaptation. Here, we show that the invasion of Aegilops triuncialis into California, USA, involved multiple introductions that may have facilitated invasion into serpentine habitats. Using microsatellite markers, we compared the polymorphism and genetic structure of populations of Ae. triuncialis invading serpentine soils in California to that of accessions from its native range. In a glasshouse study, we also compared phenotypic variation in phenological and fitness traits between invasive and native populations grown on loam soil and under serpentine edaphic conditions. Molecular analysis of invasive populations revealed that Californian populations cluster into three independent introductions (i.e. invasive lineages). Our glasshouse common garden experiment found that all Californian populations exhibited higher fitness under serpentine conditions. However, the three invasive lineages appear to represent independent pathways of adaptation to serpentine soil. Our results suggest that the rapid invasion of serpentine habitats in California may have been facilitated by the existence of colonizing Eurasian genotypes pre‐adapted to serpentine soils.     

MULTIVARIATE ADAPTATION BUT NO INCREASE IN COMPETITIVE ABILITY IN INVASIVE GERANIUM CAROLINIANUM L. (GERANIACEAE)

Adaptive evolution can affect the successful establishment of invasive species, but changes in selective pressures, loss of genetic variation in relevant traits, and/or altered trait correlations can make adaptation difficult to predict. We used a common‐garden experiment to assess trait correlations and patterns of adaptation in the invasive plant, Geranium carolinianum, sampled across 20 populations in its native (United States) and invasive (China) ranges. We used multivariate Q_ST – F_ST tests to determine if phenotypic differences between countries are attributable to adaptation. We also compared population‐level variation within each country to assess whether local adaptation resulted in similar multivariate phenotypes in the United States and China. Between countries, most phenotypic differences are indistinguishable from genetic drift, although we detected a signature of adaptation to the colder, drier winters in China. There was no evidence for increases in invasive traits in China. Within countries, strong multivariate adaptation appears to be driven by latitudinal climatic variation in the United States, but not in China. Additionally, adaptive trait combinations as well as their underlying correlations differ between the two countries, indicating that adaptation in invasive populations does not parallel patterns in native populations due to differences in selection pressures, genetic constraints, or both.     

Colony social structure in native and invasive populations of the social wasp Vespula pensylvanica

Social insects rank among the most invasive of terrestrial species. The success of invasive social insects stems, in part, from the flexibility derived from their social behaviors. We used genetic markers to investigate if the social system of the invasive wasp, Vespula pensylvanica, differed in its introduced and native habitats in order to better understand variation in social phenotype in invasive social species. We found that (1) nestmate workers showed lower levels of relatedness in introduced populations than native populations, (2) introduced colonies contained workers produced by multiple queens whereas native colonies contained workers produced by only a single queen, (3) queen mate number did not differ significantly between introduced and native colonies, and (4) workers from introduced colonies were frequently produced by queens that originated from foreign nests. Thus, overall, native and introduced colonies differed substantially in social phenotype because introduced colonies more frequently contained workers produced by multiple, foreign queens. In addition, the similarity in levels of genetic variation in introduced and native habitats, as well as observed variation in colony social phenotype in native populations, suggest that colony structure in invasive populations may be partially associated with social plasticity. Overall, the differences in social structure observed in invasive V. pensylvanica parallel those in other, distantly related invasive social insects, suggesting that insect societies often develop similar social phenotypes upon introduction into new habitats.     

Genetic diversity and structure of the globally invasive tree, <Emphasis Type="Italic">Paraserianthes lophantha</Emphasis> subspecies <Emphasis Type="Italic">lophantha</Emphasis>, suggest an introduction history characterised by varying propagule pressure

An emerging insight in invasion biology is that intra-specific genetic variation, human usage, and introduction histories interact to shape genetic diversity and its distribution in populations of invasive species. We explore these aspects for the tree species Paraserianthes lophantha subsp. lophantha, a close relative of Australian wattles (genus Acacia). This species is native to Western Australia and is invasive in a number of regions globally. Using microsatellite genotype and DNA sequencing data, we show that native Western Australian populations of P. lophantha subsp. lophantha are geographically structured and are more diverse than introduced populations in Australia (New South Wales, South Australia, and Victoria), the Hawaiian Islands, Portugal, and South Africa. Introduced populations varied greatly in the amount of genetic diversity contained within them, from being low (e.g. Portugal) to high (e.g. Maui, Hawaiian Islands). Irrespective of provenance (native or introduced), all populations appeared to be highly inbred (F _IS ranging from 0.55 to 0.8), probably due to selfing. Although introduced populations generally had lower genetic diversity than native populations, Bayesian clustering of microsatellites and phylogenetic diversity indicated that introduced populations comprise a diverse array of genotypes, most of which were also identified in Western Australia. The dissimilarity in the distribution and number of genotypes in introduced regions suggests that non-native populations originated from different native sources and that introduction events differed in propagule pressure.     

Genetic Structure of Remnant Populations and Cultivars of Switchgrass (Panicum virgatum) in the Context of Prairie Conservation and Restoration

Switchgrass (Panicum virgatum L.) is a dominant, perennial C₄ grass of North American tallgrass prairies with cultivars that are widely used in grassland restoration, pastures, and landscaping. However, these cultivars may be genetically dissimilar to small, remnant populations, raising concerns about altered genetic composition of native populations through gene flow. To address this issue on a local scale in Ohio and Illinois, we used microsatellite markers to characterize genetic diversity and differentiation of 10 remnant prairie populations (5 in each state) and 8 common cultivars. The bulk of genetic variation was found to reside within rather than among wild populations, consistent with the outcrossing breeding system of switchgrass. Genetic diversity was similar among the remnant populations despite large differences in area (approximately 2–2,590 ha), highlighting the importance of small native populations as reservoirs of variation and potential seed sources for prairie restoration. Cultivars generally had similar levels of variation to the wild populations, but we found clear genetic dissimilarity between wild and cultivated gene pools (especially for Kanlow, but also Trailblazer, Blackwell, Dacotah, Summer, and Sunburst cultivars). This suggests that using cultivars in local prairie restoration efforts may alter the genetic composition of wild populations. Whether such changes are deemed as negative depends on the cultivar under consideration and specific conservation goals for preserving native switchgrass populations. Patterns of genetic variation in remnant prairie populations and potential cultivar sources can be used to develop guidelines for restoration as well as future planting of cultivars for biofuels.     

Contrasting levels of genotype by environment interaction for life history and morphological traits in invasive populations of Zaprionus indianus (Diptera: Drosophilidae)

It has been demonstrated that phenotypic plasticity and genotype by environment interaction are important for coping with new and heterogeneous environments during invasions. Zaprionus indianus Gupta (Diptera: Drosophilidae) is an Afrotropical invasive fly species introduced to the South American continent in 1999. This species is generalist and polyphagous, since it develops and feeds in several different fruit species. These characteristics of Z. indianus suggest that phenotypic plasticity and genotype by environment interaction may be important in this species invasion process. In this sense, our aim was to investigate the role of genetic variation for phenotypic plasticity (genotype by environment interaction) in Z. indianus invasion of the South American continent. Specifically, we quantified quantitative genetic variation and genotype by environment interactions of morphological and life history traits in different developmental environments, that is, host fruits. This was done in different populations in the invasive range of Z. indianus in Argentina. Results showed that Z. indianus populations have considerable amounts of quantitative genetic variation. Also, genotype by environment interactions was detected for the different traits analyzed in response to the different developmental environments. Interestingly, the amounts and patterns of these parameters differed between populations. We interpreted these results as the existence of differences in evolutionary potential between populations that have an important role in the short‐ and long‐term success of the Z. indianus invasion process.     

RAPD variation within and among natural populations of outcrossing buffalograss [Buchloë dactyloides (Nutt.) Engelm.]

RAPD markers provide a powerful tool for the investigation of genetic variation in natural and domesticated populations. Recent studies of strain/cultivar identification have shown extensive RAPD divergence among, but little variation within, inbred species or cultivars. In contrast, little is known about the pattern and extent of RAPD variation in heterogeneous, outcrossing species. We describe the population genetic variation of RAPD markers in natural, diploid sources of dioecious buffalograss [Buchloë dactyloides (Nutt.) Engelm.]. Buffalograss is native to the semi-arid regions of the Great Plains of North America, where it is important for rangeland forage, soil conservation, and as turfgrass. Most sources of buffalograss germplasm are polyploid; diploid populations are previously known only from semi-arid Central Mexico. This is the first report of diploids from humid Gulf Coastal Texas. These two diploid sources represent divergent adaptive ecotypes. Seven 10-mer primers produced 98 polymorphic banding sites. Based on the presence/ absence of bands, a genetic distance matrix was calculated. The new Analysis of Molecular Variance (AMOVA) technique was used to apportion the variation among individuals within populations, among populations within adaptive regions, and among regions. There was considerable variation within each of the four populations, and every individual was genetically distinct. Even so, genetic divergence was found among local populations. Within-population variation was larger and among-population variation smaller in Mexico than in Texas. The largest observed genetic differences were those between the two regional ecotypes. These patterns of genetic variation were very different from those reported for inbred species and provide important baseline data for cultivar identification and continuing studies of the evolution of polyploid races in this species.     

Molecular genetic approaches to elucidate the ecological and evolutionary issues associated with biological invasions

Biological invasions may cause serious damage to the native environments and threaten the native biodiversity. Molecular genetic approaches have been found to be powerful tools for investigating the ecological and evolutionary aspects of biological invasions because the genetic structure and level of genetic variation of an invasive species are changed following its invasion. The present article reviews the use of molecular markers in addressing various aspects of invasive species. The application of these techniques has shown that many invasive species are actually "cryptic" species – species whose uniqueness is only recognizable at the genetic level. An estimation of the actual number of invasive species is essential when evaluating its ecological and economic impacts. Molecular genetic approaches have also enabled the source populations of invasive species to be identified. Reconstructions of invasion histories are crucial to preventing future invasions and conserving the native biodiversity, while comparisons of genetic variations between the native and introduced populations provide valuable opportunities to elucidate the mechanisms of rapid adaptation demonstrated by many invasive species.     

Native drivers of fish life history traits are lost during the invasion process

Rapid adaptation to global change can counter vulnerability of species to population declines and extinction. Theoretically, under such circumstances both genetic variation and phenotypic plasticity can maintain population fitness, but empirical support for this is currently limited. Here, we aim to characterize the role of environmental and genetic diversity, and their prior evolutionary history (via haplogroup profiles) in shaping patterns of life history traits during biological invasion. Data were derived from both genetic and life history traits including a morphological analysis of 29 native and invasive populations of topmouth gudgeon Pseudorasbora parva coupled with climatic variables from each location. General additive models were constructed to explain distribution of somatic growth rate (SGR) data across native and invasive ranges, with model selection performed using Akaike's information criteria. Genetic and environmental drivers that structured the life history of populations in their native range were less influential in their invasive populations. For some vertebrates at least, fitness‐related trait shifts do not seem to be dependent on the level of genetic diversity or haplogroup makeup of the initial introduced propagule, nor of the availability of local environmental conditions being similar to those experienced in their native range. As long as local conditions are not beyond the species physiological threshold, its local establishment and invasive potential are likely to be determined by local drivers, such as density‐dependent effects linked to resource availability or to local biotic resistance.     

Multiple common garden experiments suggest lack of local adaptation in an invasive ornamental plant

Aims Adaptive evolution along geographic gradients of climatic conditions is suggested to facilitate the spread of invasive plant species, leading to clinal variation among populations in the introduced range. We investigated whether adaptation to climate is also involved in the invasive spread of an ornamental shrub, Buddleja davidii, across western and central Europe.Methods We combined a common garden experiment, replicated in three climatically different central European regions, with reciprocal transplantation to quantify genetic differentiation in growth and reproductive traits of 20 invasive B. davidii populations. Additionally, we compared compensatory regrowth among populations after clipping of stems to simulate mechanical damage.Important findings Our results do not provide evidence for clinal variation among invasive B. davidii populations: populations responded similarly to the different environments, and trait values were not correlated to climatic conditions or geographic coordinates of their home sites. Moreover, we did not detect differences in the compensatory ability of populations. We suppose that the invasive spread of B. davidii has been facilitated by phenotypic plasticity rather than by adaptation to climate and that continent-wide shuffling of cultivars due to horticultural trade may have limited local adaptation so far.     

Phenotypic and genetic differentiation between native and introduced plant populations

Plant invasions often involve rapid evolutionary change. Founder effects, hybridization, and adaptation to novel environments cause genetic differentiation between native and introduced populations and may contribute to the success of invaders. An influential idea in this context has been the Evolution of Increased Competitive Ability (EICA) hypothesis. It proposes that after enemy release plants rapidly evolve to be less defended but more competitive, thereby increasing plant vigour in introduced populations. To detect evolutionary change in invaders, comparative studies of native versus introduced populations are needed. Here, we review the current empirical evidence from: (1) comparisons of phenotypic variation in natural populations; (2) comparisons of molecular variation with neutral genetic markers; (3) comparisons of quantitative genetic variation in a common environment; and (4) comparisons of phenotypic plasticity across different environments. Field data suggest that increased vigour and reduced herbivory are common in introduced plant populations. In molecular studies, the genetic diversity of introduced populations was not consistently different from that of native populations. Multiple introductions of invasive plants appear to be the rule rather than the exception. In tests of the EICA hypothesis in a common environment, several found increased growth or decreased resistance in introduced populations. However, few provided a full test of the EICA hypothesis by addressing growth and defence in the same species. Overall, there is reasonable empirical evidence to suggest that genetic differentiation through rapid evolutionary change is important in plant invasions. We discuss conceptual and methodological issues associated with cross-continental comparisons and make recommendations for future research. When testing for EICA, greater emphasis should be put on competitive ability and plant tolerance. Moreover, it is important to address evolutionary change in characteristics other than defence and growth that could play a role in plant invasions.