Morphology and genetics of Lythrum salicaria from latitudinal gradients of the Northern Hemisphere grown in cold and hot common gardens.

The aim of this project was to compare the phenotypic responses of global populations of Lythrum salicaria in cold/dry and hot/humid environments to determine if phenotypic plasticity varied between the native and invasive ranges, and secondarily if this variation was linked to genetic diversity. Common garden studies were conducted in Trebon, Czech Republic, and Lafayette, Louisiana, USA (cold/dry vs. hot/humid garden, respectively), using populations from latitudinal gradients in Eurasia and North America. Lythrum salicaria seeds collected from the same maternal plants across these latitudinal gradients were germinated and grown in Trebon and Lafayette. Tissue masses (above-, below-ground, inflorescence and total) of these individuals were assessed at the end of each growing season (2006-2008). Worldwide field measurements of L. salicaria height were made by volunteers from 2004-2016. Biomass and height data were analyzed using the General Linear Model framework and multivariate techniques. Molecular markers (amplified fragment length polymorphisms) of individuals used in the common garden study were analyzed using traditional genetic diversity metrics and Bayesian clustering algorithms in STRUCTURE. Reaction norms were developed from differences in maternal plant responses in Trebon versus Lafayette. In the common garden studies, stem/leaf, root and total biomass generally were highest for individuals grown from seeds collected in the southern part of the range in the cold garden, particularly by the third year of the study. In contrast, inflorescence biomass in the cold garden was higher by the third year in individuals from mid-latitude populations. As measured by volunteers, plants were taller in Eurasia than in North America moving from north to south with the pattern switching southward of 40°N latitude. Genetic diversity was similar between native and non-native invasive populations regardless of geographical origin of the seed and was not significantly different in the GLM Select model (p > 0.05). Reaction norm slopes showed that Eurasia had larger values than North America for reaction norms for above-ground and total biomass. Plants from the seeds of mother plants from Turkey had wide variation in total biomass when grown in Trebon versus Lafayette; this variation in response within certain populations may have contributed to the lack of population-level differences in plasticity. These results indicate no loss of genetic diversity for L. salicaria during its North American invasion, nor reduction in plastic tissue allocation responses to a varying environment, which may help explain some of its invasive qualities and which could be of adaptive value under changing future environments.

Evidence that a herbivore tolerance response affects selection on floral traits and inflorescence architecture in purple loosestrife (Lythrum salicaria).

Background and Aims: The study of the evolution of floral traits has generally focused on pollination as the primary driver of selection. However, herbivores can also impose selection on floral traits through a variety of mechanisms, including florivory and parasitism. Less well understood is whether floral and inflorescence architecture traits that influence a plant's tolerance to herbivory, such as compensatory regrowth, alter pollinator-mediated selection. Methods: Because herbivore damage to Lythrum salicaria meristems typically leads to an increase in the number of inflorescences and the size of the floral display, an experiment was conducted to test whether simulated herbivory (i.e. clipping the developing meristem) could alter the magnitude or direction of pollinator-mediated selection on a suite of floral and inflorescence architecture traits. Using a pollen supplementation protocol, pollen limitation was compared in the presence and absence of meristem damage in order to quantify any interaction between pollinator and herbivore-mediated selection on floral traits. Key Results: Surprisingly, in spite of an obvious impact on floral display and architecture, with clipped plants producing more inflorescences and more flowers, there was no difference in pollen limitation between clipped and unclipped plants. Correspondingly, there was no evidence that imposing herbivore damage altered pollinator-mediated selection in this system. Rather, the herbivory treatment alone was found to alter direct selection on floral display, with clipped plants experiencing greater selection for earlier flowering and weaker selection for number of inflorescences when compared with unclipped plants. Conclusions: These findings imply that herbivory on its own can drive selection on plant floral traits and inflorescence architecture in this species, even more so than pollinators. Specifically, herbivory can impose selection on floral traits if such traits influence a plant's tolerance to herbivory, such as through the timing of flowering and/or the compensatory regrowth response.

Phenological shifts of native and invasive species under climate change: insights from the Boechera-Lythrum model.

Warmer and drier climates have shifted phenologies of many species. However, the magnitude and direction of phenological shifts vary widely among taxa, and it is often unclear when shifts are adaptive or how they affect long-term viability. Here, we model evolution of flowering phenology based on our long-term research of two species exhibiting opposite shifts in floral phenology: Lythrum salicaria, which is invasive in North America, and the sparse Rocky Mountain native Boechera stricta Genetic constraints are similar in both species, but differences in the timing of environmental conditions that favour growth lead to opposite phenological shifts under climate change. As temperatures increase, selection is predicted to favour earlier flowering in native B. stricta while reducing population viability, even if populations adapt rapidly to changing environmental conditions. By contrast, warming is predicted to favour delayed flowering in both native and introduced L. salicaria populations while increasing long-term viability. Relaxed selection from natural enemies in invasive L. salicaria is predicted to have little effect on flowering time but a large effect on reproductive fitness. Our approach highlights the importance of understanding ecological and genetic constraints to predict the ecological consequences of evolutionary responses to climate change on contemporary timescales.This article is part of the themed issue 'Human influences on evolution, and the ecological and societal consequences'.

Variation in style morph frequencies in tristylous Lythrum salicaria in the Iberian Peninsula: the role of geographical and demographic factors.

BACKGROUND AND AIMS: The balance between stochastic forces and negative frequency-dependent selection largely determines style morph frequencies in heterostylous populations. Investigation of morph frequencies at geographical range limits can provide insights into the forces maintaining the floral polymorphism, and the factors causing biased morph ratios. Here, we investigate style morph frequencies in populations at the south-western European range limit of tristylous Lythrum salicaria, to explore the role of demographic and geographical factors influencing morph ratios in its native range. METHODS: We measured morph composition and evenness, and the size of 96 populations, along a north to south latitudinal transect from Galicia to Andalucia, Iberian Peninsula, traversing a steep climatic gradient. To examine the potential influence of morph-specific fitness components on morph ratios, we examined reproductive traits in 19 populations. KEY RESULTS: Most populations of L. salicaria were trimorphic (94·79 %), the majority exhibiting 1 : 1 : 1 morph ratios (68·75 %). Populations with biased morph ratios had a deficiency of the short-styled morph. Population size and morph evenness were positively associated with latitude, with smaller populations and those with less even morph ratios occurring towards the south. Greater variance in morph evenness was evident at the southern range margin. There were no consistent differences in components of reproductive fitness among style morphs, but southern populations produced less fruit and seed than more northerly populations. CONCLUSIONS: Our results demonstrate the influence of finite population size on morph frequencies in L. salicaria. However, they also illustrate the resilience of Iberian populations to the factors causing deviations from isoplethy and morph loss, especially at the southern range limit where populations are smaller. The maintenance of tristyly in small populations of L. salicaria may be aided by the genetic connectivity of populations in agricultural landscapes resulting from gene flow through pollen and seed dispersal.

Rapid adaptation to climate facilitates range expansion of an invasive plant.

Adaptation to climate, evolving over contemporary time scales, could facilitate rapid range expansion across environmental gradients. Here, we examine local adaptation along a climatic gradient in the North American invasive plant Lythrum salicaria. We show that the evolution of earlier flowering is adaptive at the northern invasion front where it increases fitness as much as, or more than, the effects of enemy release and the evolution of increased competitive ability. However, early flowering decreases investment in vegetative growth, which reduces fitness by a factor of 3 in southern environments where the North American invasion commenced. Our results demonstrate that local adaptation can evolve quickly during range expansion, overcoming environmental constraints on propagule production.

Response to enemies in the invasive plant Lythrum salicaria is genetically determined.

BACKGROUND AND AIMS: The enemy release hypothesis assumes that invasive plants lose their co-evolved natural enemies during introduction into the new range. This study tested, as proposed by the evolution of increased competitive ability (EICA) hypothesis, whether escape from enemies results in a decrease in defence ability in plants from the invaded range. Two straightforward aspects of the EICA are examined: (1) if invasives have lost their enemies and their defence, they should be more negatively affected by their full natural pre-invasion herbivore spectrum than their native conspecifics; and (2) the genetic basis of evolutionary change in response to enemy release in the invasive range has not been taken sufficiently into account. METHODS: Lythrum salicaria (purple loosestrife) from several populations in its native (Europe) and invasive range (North America) was exposed to all above-ground herbivores in replicated natural populations in the native range. The experiment was performed both with plants raised from field-collected seeds as well as with offspring of these where maternal effects were removed. KEY RESULTS: Absolute and relative leaf damage was higher for introduced than for native plants. Despite having smaller height growth rate, invasive plants attained a much larger final size than natives irrespective of damage, indicating large tolerance rather than effective defence. Origin effects on response to herbivory and growth were stronger in second-generation plants, suggesting that invasive potential through enemy release has a genetic basis. CONCLUSIONS: The findings support two predictions of the EICA hypothesis - a genetically determined difference between native and invasive plants in plant vigour and response to enemies - and point to the importance of experiments that control for maternal effects and include the entire spectrum of native range enemies.

Population divergence along lines of genetic variance and covariance in the invasive plant Lythrum salicaria in eastern North America.

Evolution during biological invasion may occur over contemporary timescales, but the rate of evolutionary change may be inhibited by a lack of standing genetic variation for ecologically relevant traits and by fitness trade-offs among them. The extent to which these genetic constraints limit the evolution of local adaptation during biological invasion has rarely been examined. To investigate genetic constraints on life-history traits, we measured standing genetic variance and covariance in 20 populations of the invasive plant purple loosestrife (Lythrum salicaria) sampled along a latitudinal climatic gradient in eastern North America and grown under uniform conditions in a glasshouse. Genetic variances within and among populations were significant for all traits; however, strong intercorrelations among measurements of seedling growth rate, time to reproductive maturity and adult size suggested that fitness trade-offs have constrained population divergence. Evidence to support this hypothesis was obtained from the genetic variance-covariance matrix (G) and the matrix of (co)variance among population means (D), which were 79.8% (95% C.I. 77.7-82.9%) similar. These results suggest that population divergence during invasive spread of L. salicaria in eastern North America has been constrained by strong genetic correlations among life-history traits, despite large amounts of standing genetic variation for individual traits.

Evolutionary constraints on adaptive evolution during range expansion in an invasive plant.

Biological invasions may expose populations to strong selection for local adaptation along geographical gradients in climate. However, evolution during contemporary timescales can be constrained by low standing genetic variation and genetic correlations among life-history traits. We examined limits to local adaptation associated with northern migration of the invasive wetland plant purple loosestrife (Lythrum salicaria) using a selection model incorporating a trade-off between flowering time and size at reproduction, and common garden experiments of populations sampled along a latitudinal transect of approximately 1200 km in eastern North America. A strong trade-off between flowering time and size at reproduction caused early-flowering plants to be smaller with reduced seed production in northern populations. Northward spread was associated with a decline in genetic variance within populations and an increase in genetic skew for flowering time and size, with limited genetic variation for small, early-flowering genotypes. These patterns were predicted by our selection model of local adaptation to shorter growing seasons and were not consistent with expectations from non-adaptive processes. Reduced fecundity may limit population growth and rates of spread in northern populations. Identifying genetic constraints on key life-history traits can provide novel insights into invasion dynamics and the causes of range limits in introduced species.

Comparison of quantitative and molecular genetic variation of native vs. invasive populations of purple loosestrife (Lythrum salicaria L., Lythraceae).

Study of adaptive evolutionary changes in populations of invasive species can be advanced through the joint application of quantitative and population genetic methods. Using purple loosestrife as a model system, we investigated the relative roles of natural selection, genetic drift and gene flow in the invasive process by contrasting phenotypical and neutral genetic differentiation among native European and invasive North American populations (Q(ST) - F(ST) analysis). Our results indicate that invasive and native populations harbour comparable levels of amplified fragment length polymorphism variation, a pattern consistent with multiple independent introductions from a diverse European gene pool. However, it was observed that the genetic variation reduced during subsequent invasion, perhaps by founder effects and genetic drift. Comparison of genetically based quantitative trait differentiation (Q(ST)) with its expectation under neutrality (F(ST)) revealed no evidence of disruptive selection (Q(ST) > F(ST)) or stabilizing selection (Q(ST) < F(ST)). One exception was found for only one trait (the number of stems) showing significant sign of stabilizing selection across all populations. This suggests that there are difficulties in distinguishing the effects of nonadaptive population processes and natural selection. Multiple introductions of purple loosestrife may have created a genetic mixture from diverse source populations and increased population genetic diversity, but its link to the adaptive differentiation of invasive North American populations needs further research.

Re-establishment of clinal variation in flowering time among introduced populations of purple loosestrife (Lythrum salicaria, Lythraceae).

Range expansion during biological invasion requires that invaders adapt to geographical variation in climate, which should yield latitudinal clines in reproductive phenology. We investigated geographic variation in life history among 25 introduced populations of Lythrum salicaria, a widespread European invader of North American wetlands. We detected a strong latitudinal cline in initiation of flowering and size at flowering, which paralleled that reported among native populations. Plants from higher latitudes flowered earlier and at a smaller size than those from lower latitudes, even when raised in a uniform glasshouse. Early flowering was associated with greatly reduced reproductive output, but this was not associated with latitudinal variation in abundance, and probably did not result from a genetic correlation between time to and size at flowering. As introduction to North America c. 200 years ago, L. salicaria has re-established latitudinal clines in life history, probably as an evolutionary response to climatic selection.

Phenotypic plasticity of native vs. invasive purple loosestrife: a two-state multivariate approach.

The differences in phenotypic plasticity between invasive (North American) and native (German) provenances of the invasive plant Lythrum salicaria (purple loosestrife) were examined using a multivariate reaction norm approach testing two important attributes of reaction norms described by multivariate vectors of phenotypic change: the magnitude and direction of mean trait differences between environments. Data were collected for six life history traits from native and invasive plants using a split-plot design with experimentally manipulated water and nutrient levels. We found significant differences between native and invasive plants in multivariate phenotypic plasticity for comparisons between low and high water treatments within low nutrient levels, between low and high nutrient levels within high water treatments, and for comparisons that included both a water and nutrient level change. The significant genotype x environment (G x E) effects support the argument that invasiveness of purple loosestrife is closely associated with the interaction of high levels of soil nutrient and flooding water regime. Our results indicate that native and invasive plants take different strategies for growth and reproduction; native plants flowered earlier and allocated more to flower production, while invasive plants exhibited an extended period of vegetative growth before flowering to increase height and allocation to clonal reproduction, which may contribute to increased fitness and invasiveness in subsequent years.

Evidence of hybridization between Lythrum salicaria (purple loosestrife) and L. alatum (winged loosestrife) in North America.

BACKGROUND AND AIMS: Although Lythrum salicaria (purple loosestrife) was introduced to North America from Europe in the early 1800s, it did not become invasive until the 1930s. Whether hybridization with L. alatum (winged loosestrife) could have played a role in its ultimate spread was tested. METHODS: Six diagnostic morphological traits (flower number per axil, leaf placement, calyx pubescence, style type, plant height and leaf shape) were surveyed in 30 populations of Lythrum across eastern North America. Patterns of AFLP variation were also evaluated using five primer pairs in a 'global screen' of the same North American populations of L. salicaria and L. alatum described above, in L. salicaria from 11 European populations located in Germany, England, Ireland, Austria and Finland, and in six L. salicaria cultivars. KEY RESULTS: All of the North American L. salicaria populations had individuals with alternate leaf placement and 1-2 flowers per leaf axil, which have not been described in Eurasian L. salicaria but predominate in North American L. alatum. In addition, two L. salicaria populations were intermediate in height and leaf ratio between the typical L. salicaria and L. alatum populations in their native fields and when grown in a common greenhouse. In screens of variation patterns using 279 AFLPs, only two fragments were found that clearly supported introgression from L. alatum to L. salicaria. CONCLUSIONS: The evidence indicates that L. salicaria may have hybridized with L. alatum, but if so, only a small fraction of L. alatum genes have been retained in the genome of L. salicaria. This is unlikely to have led to a dramatic adaptive shift unless the introgression of a few key genes into L. salicaria stimulated a genomic reorganization. It is more likely that crossing among genotypes of L. salicaria from multiple introductions provided the necessary variability for new adaptations to arise.