Epigenetic and Phenotypic Responses to Experimental Climate Change of Native and Invasive Carpobrotus edulis.

Despite the recent discoveries on how DNA methylation could help plants to adapt to changing environments, the relationship between epigenetics and climate change or invasion in new areas is still poorly known. Here, we investigated, through a field experiment, how the new expected climate scenarios for Southern Europe, i.e., increased temperature and decreased rainfall, might affect global DNA methylation in relation to phenotypic variation in individuals of clonal plant, Carpobrotus edulis, from its native (Southern African) and invaded (northwestern Iberian Peninsula) area. Our results showed that changes in temperature and rainfall induced phenotypic but not global DNA methylation differences among plants, and the climatic effects were similar for plants coming from the native or invaded areas. The individuals from the Iberian Peninsula showed higher levels of global methylation than their native counterparts from South Africa. We also observed differences between natives and invasive phenotypes in traits related to the pattern of biomass partitioning and to the strategies for water uptake and use and found an epigenetic contribution to phenotypic changes in some leaf traits, especially on the nitrogen isotopic composition. We conclude that the increased temperature and decreased rainfall projected for Southern Europe during the course of the twenty-first century may foster phenotypic changes in C. edulis, possibly endowing this species with a higher ability to successful cope the rapid environmental shifts. The epigenetic and phenotypic divergence that we observed between native and invasive plants suggests an intraspecific functional variation during the process of invasion. This result could indicate that phenotypic plasticity and global DNA methylation are related to the colonization of new habitats. Our findings reinforce the importance of epigenetic plasticity on rapid adaptation of invasive clonal plants.

Functional responses to climate change may increase invasive potential of Carpobrotus edulis.

PREMISE: Biological invasions and climate change are major threats to biodiversity. It is therefore important to anticipate how the climate changes projected for Southern Europe would affect the ecophysiological performance of the invasive South African plant, Carpobrotus edulis (ice plant or sour fig), and its capacity to undergo rapid adaptive evolution. METHODS: We manipulated the climate conditions in a field plot located on the island of Sálvora (northwest of the Iberian Peninsula) to establish a full factorial experiment with C. edulis plants transplanted from four native (southern African) and four invasive (northwestern Iberian Peninsula) populations. Throughout 14 months we measured growth and functional traits of this species under two temperatures (control vs. increased), and two rainfall levels (control vs. reduced). RESULTS: Temperature increased photochemical efficiency and relative growth rate of C. edulis. Rainfall modulated some of the effects of temperature on C and N isotopic composition, and pigment contents. Invasive populations showed lower root mass allocation and higher survival rates, as well as increased water use efficiency, lipid peroxidation, chlorophyll, and xanthophyll cycle pigment contents than native populations. CONCLUSIONS: The increased growth and physiological performances observed under our experimental conditions suggest that the expected climate changes would further promote the invasion of C. edulis. Differences between native and invasive genotypes in survival and functional traits revealed that populations have diverged during the process of invasion, what gives support to the invasiveness hypothesis. Our findings highlight the importance of analyzing intraspecific variability in functional responses to better predict how invasive species will respond to environmental changes.

A test of native plant adaptation more than one century after introduction of the invasive Carpobrotus edulis to the NW Iberian Peninsula.

BACKGROUND: Although the immediate consequences of biological invasions on ecosystems and conservation have been widely studied, the long-term effects remain unclear. Invaders can either cause the extinction of native species or become integrated in the new ecosystems, thus increasing the diversity of these ecosystems and the services that they provide. The final balance of invasions will depend on how the invaders and native plants co-evolve. For a better understanding of such co-evolution, case studies that consider the changes that occur in both invasive and native species long after the introduction of the invader are especially valuable. In this work, we studied the ecological consequences of the more than one century old invasion of NW Iberia by the African plant Carpobrotus edulis. We conducted a common garden experiment to compare the reciprocal effects of competition between Carpobrotus plants from the invaded area or from the native African range and two native Iberian plant species (Artemisia crithmifolia and Helichrysum picardii) from populations exposed or unexposed to the invader. RESULTS: Exposure of H. picardii populations to C. edulis increased their capacity to repress the growth of Carpobrotus. The repression specifically affected the Carpobrotus from the invader populations, not those from the African native area. No effects of exposition were detected in the case of A. crithmifolia. C. edulis plants from the invader populations had higher growth than plants from the species' African area of origin. CONCLUSIONS: We found that adaptive responses of natives to invaders can occur in the long term, but we only found evidence for adaptive responses in one of the two species studied. This might be explained by known differences between the two species in the structure of genetic variance and gene flow between subpopulations. The overall changes observed in the invader Carpobrotus are consistent with adaptation after invasion.

Ecophysiological differentiation between two invasive species of Carpobrotus competing under different nutrient conditions.

PREMISE: Hybridization between the South African invasive species Carpobrotus edulis and C. acinaciformis in Europe has led to the formation of highly aggressive morphotypes referred to in the scientific literature as the new large "hybrid swarm" C. aff. acinaciformis. In the present study, we aimed to determine whether the taxonomic differentiation between taxa coincides with ecophysiological differentiation. With this aim, we tested for differences between both morphotypes in functional traits related to competitive ability and resource-use efficiency. Assuming that the complex hybrid C. aff. acinaciformis is more vigorous, depends more strongly on vegetative reproduction, and invests less in sexual reproduction than C. edulis, we predicted that the hybrid would show higher competitive ability and better physiological performance compared with the species. METHODS: We used a comparative ecophysiological approach to assess the extent to which two Carpobrotus morphotypes coexisting in northwestern Spain differ in physiological, reproductive, and growth traits when competing under different soil nutrients in controlled greenhouse conditions. RESULTS: C. aff. acinaciformis had a greater relative growth rate and water-use and photochemical efficiencies compared to C. edulis. However, C. edulis appeared to be more responsive to incremental change in soil nutrients than C. aff. acinaciformis. They also differed in the amount of resources invested in reproduction. CONCLUSIONS: The study findings demonstrate that the taxonomic differentiation between taxa corresponds to ecophysiological differentiation, warranting a detailed examination of all existing trades-offs to predict the long-term outcomes of the interaction between these taxa.

Evergreen or deciduous trees for capturing PAHs from ambient air? A case study.

Tree canopies play a key role in the cycling of polycyclic aromatic hydrocarbons (PAHs) in terrestrial ecosystems, as leaves can capture PAHs from the air. In this study, accumulation of PAHs was compared in an evergreen species, P. pinaster, and in a deciduous species, Q. robur, in relation to some physio-morphological characteristics. For this purpose, pine needles and oak leaves collected from different sites across Galicia (NW Spain) were analysed to determine PAH contents, specific leaf area, stomatal density and conductance. Leaves and needles contained similar total amounts of PAHs. The major contribution of particle-bound PAHs in oak (the concentrations of 4- and 5-ring PAHs were two times higher, and those of 6-ring PAHs five times higher in oak than in pine) may be related to the higher specific leaf area (13 and 4 cm2 g-1 dry mass in respectively oak and pine). However, the major contribution of vapor-phase PAHs in pines may be affected by the stomatal conductance (two times higher in pine than in oak). Moreover, an increase in the diameter at breast height of trees led to an increase in accumulation of PAHs, with pine capturing higher amounts of low and medium molecular weight PAHs. The study findings underline the potential role of trees in improving air quality, taking into account the canopy biomass and life cycle.

Division of Labor Brings Greater Benefits to Clones of Carpobrotus edulis in the Non-native Range: Evidence for Rapid Adaptive Evolution.

Why some species become invasive while others do not is a central research request in biological invasions. Clonality has been suggested as an attribute that could contribute to plant invasiveness. Division of labor is an important advantage of clonal growth, and it seems reasonable to anticipate that clonal plants may intensify this clonal attribute in an invaded range because of positive selection on beneficial traits. To test this hypothesis, we collected clones of Carpobrotus edulis from native and invasive populations, grew pairs of connected and severed ramets in a common garden and under negative spatial covariance of nutrients and light to induce division of labor, and measured biomass allocation ratios, final biomass, and photochemical efficiency. Our results showed that both clones from the native and invaded range develop a division of labor at morphological and physiological level. However, the benefit from the division of labor was significantly higher in apical ramets from the invaded range than in ramets from the native area. This is a novel and outstanding result because it provides the first evidence that the benefit of a key clonal trait such as division of labor may have been subjected to evolutionary adaptation in the invaded range. The division of labor can therefore be considered an important trait in the invasiveness of C. edulis. An appropriate assessment of the influence of clonal traits in plant invasions seems key for understanding the underlying mechanisms behind biological invasions of new environments.

Phylogeography of a widespread species: pre-glacial vicariance, refugia, occasional blocking straits and long-distance migrations.

Phylogeographic studies give us the opportunity to reconstruct the historical migrations of species and link them with climatic and geographic variation. They are, therefore, a key tool to understanding the relationships among biology, geology and history. One of the most interesting biogeographical areas of the world is the Mediterranean region. However, in this area, the description of concordant phylogeographic patterns is quite scarce, which limits the understanding of evolutionary patterns related to climate. Species with one-dimensional distribution ranges, such as the strawberry tree (Arbutus unedo), are particularly useful to unravel these patterns. Here, we describe its phylogeographic structure and check for concordance with patterns seen in other Mediterranean plants: longitudinal/latitudinal clines of diversity, evidence for glacial refugia and the role of sea straits in dispersal. We also identify the most likely source for the disjunct Irish population. With this aim, we sequenced four chloroplast non-coding fragments of A. unedo from 23 populations covering its whole distribution. We determined the genetic diversity, population structure, haplotype genealogy and time to the most recent common ancestor. The genealogy revealed two clades that separated during the last 700 ky but before the last glacial maximum. One clade occupies Atlantic Iberia and North Africa, while the other occurs in the Western Mediterranean. The Eastern Mediterranean is inhabited by newer haplotypes derived from both clades, while the Irish population is closely related to Iberian demes. The straits of Sicily and Gibraltar partially restricted the gene flow. We concluded that a vicariance event during the Late Quaternary in the western end of the species' range followed by eastward migration seems a likely explanation for the observed phylogeographic pattern. The role of straits indicates an occasional communication between Europe and North Africa, suggesting that the latter was a novel refugia. The East-West genetic split in Iberia is consistent with the refugia-within-refugia model. Finally, the strawberry tree possibly reached Ireland from Iberia instead of throughout the maritime fringe of France as previously thought.

Low among-provenance differences in structural and functional plasticity in response to nutrients in saplings of the circum-Mediterranean tree Arbutus unedo L.

The Mediterranean region is an area of special interest for conservation where the incidence of multiple drivers of global change is expected to increase. One of the factors predicted to change is soil-nutrient availability, an essential factor for plant growth. Thus, study of the effects of variation in this parameter is especially relevant in species with a circum-Mediterranean distribution, such as Arbutus unedo L., in which the different provenances grow in different habitats, which must differ in nutritional conditions. We aimed to determine the effect of provenance on plasticity, to establish whether structural and morphological traits differ in the level of plasticity and to assess how nutrients affect the photosynthetic light response. In a common garden experiment, we studied seven provenances from the circum-Mediterranean range of A. unedo and established two nutrient treatments (low and high nutrient availability). We measured physiological and structural traits in 1-year-old sapling and determined a phenotypic plasticity index (PPI) to quantify the level of plasticity, whereas the radiation effects were tested by construction and analysis of light response curves. Interestingly, provenance did not explain a significant amount of variance, but the plasticity was four times higher for the structural traits than for the physiological traits. Therefore, the plasticity to nutrient availability will not favour or prevent the expansion or contraction of the range of any of these provenances of A. unedo. Furthermore, the structural plasticity demonstrated the ability of the strawberry tree to optimize resource allocation, whereas the physiology remained stable, thus avoiding extra expenditure. The study findings also suggest that increased availability of nutrients would improve the performance of the species during the Mediterranean summer, characterized by high irradiance. These abilities will be key to the survival of saplings of the species under the future scenario of changes in nutrient availability.

Population Structure of a Widespread Species under Balancing Selection: The Case of Arbutus unedo L.

Arbutus unedo L. is an evergreen shrub with a circum-Mediterranean distribution that also reaches the Eurosiberian region in northern Iberia, Atlantic France, and a disjunct population in southern Ireland. Due to the variety of climatic conditions across its distribution range, the populations of A. unedo were expected to display local adaptation. Conversely, common garden experiments revealed that diverse genotypes from a range of provenances produce similar phenotypes through adaptive plasticity, suggesting the action of stabilizing selection across its climatically heterogeneous range. Nonetheless, since a uniform response might also result from extensive gene flow, we have inferred the population structure of A. unedo and assessed whether its extended and largely one-dimensional range influences gene flow with the help of AFLP genotypes for 491 individuals from 19 populations covering the whole range of the species. As we had anticipated, gene flow is restricted in A. unedo, providing further support to the hypothesis that stabilizing selection is the most likely explanation for the homogeneous phenotypes along the range. The Euro-Siberian populations were not particularly isolated from the Mediterranean. Instead, there was a distinct genetic divide between the populations around the Mediterranean Sea and those sampled along Atlantic coasts from northern Africa up to Ireland. This genetic structure suggests the action of historic rather than biogeographic factors as it seems consistent with a scenario of independent glacial refugia in the Atlantic and Mediterranean portions of the range of A. unedo. Genetic exchange was likewise restricted within each set of populations. Nevertheless, isolation-by-distance (IBD) was stronger, and F ST increased faster with distance, along the Atlantic, suggesting that gene flow might be larger among Mediterranean populations. Genetic diversity was significantly lower in NW Iberia and Ireland than in other populations whereas Ireland was more closely related to populations in NW Iberia than to a population in Atlantic France, suggesting a postglacial stepping-stone colonization of the Atlantic coast. Altogether, our results show that stabilizing selection is able to homogenize the phenotypic response even when population structure is strong, gene flow is constrained, and the phylogeographic past is complex.

Adaptive plasticity to heterogeneous environments increases capacity for division of labor in the clonal invader Carpobrotus edulis (Aizoaceae).

UNLABELLED: • PREMISE OF THE STUDY: Clonality has been proposed as an important mechanism favoring plant invasions, but few studies have been conducted to determine the role of clonal traits on successful invaders. An interesting trait associated with clonality is the capacity for division of labor. Division of labor requires a negative spatial correlation between the availabilities of two essential resources and ramet specialization for locally abundant resources to increase the overall performance of the clone. We hypothesized that the capacity for division of labor in the clonal invader Carpobrotus edulis will be selected in those clones from patchy environments where this trait could be an advantage.• METHODS: Morphological and physiological division of labor was compared between clones from coastal sand dunes (where nutrients and light show a negative spatial covariance) and from rocky coasts (where nutrients and light are homogenously distributed).• KEY RESULTS: Clones from coastal sand dunes showed a greater capacity than clones from rocky coasts for division of labor. Specialization for abundance was found at the morphological (biomass allocated to roots) and the physiological (photochemical efficiency) level.• CONCLUSIONS: The greater ability for division of labor in the patchy environment where the presence of this trait would be more beneficial demonstrates the existence of local adaptation and suggests that rapid evolution in clonal traits could be contributing to the success of the invader C. edulis. This study is one of the few showing that division of labor is under selection and is the first reporting adaptive division of labor of an aggressive invader.

Together but different: co-occurring dune plant species differ in their water- and nitrogen-use strategies.

Stress factors may severely constrain the range of plant physiological responses in harsh environments. Convergence of traits is expected in coastal dunes because of environmental filtering imposed by severe abiotic factors. However, the wide range of morphological and phenological traits exhibited by coexisting dune species suggests considerable variation in functional traits. We hypothesized that the constraints imposed by structural traits ought to translate into physiological differences. Five dominant species with different morphological traits, but coexisting in a homogeneous dune area in Northwest Spain, were selected for study. Soil characteristics and leaf functional traits were measured in April, June and November 2008. Integrated water-use efficiency (assessed by C isotope discrimination) and N acquisition and use strategies (estimated by N isotope composition) varied significantly among species and the differences changed over time. Species differences in specific leaf area, relative water content, leaf N and C:N ratio, also varied over time. The species differed in stomatal density but not in soil characteristics, with the exception of pH. Species differences in functional traits related to the use of resources suggest species niche segregation. Species-specific temporal effects on the use of these resources support temporal niche differentiation. Somewhat in contrast to the findings of previous studies on harsh environments, this study revealed a considerable level of functional diversity and complexity, suggesting that dune plant species have evolved species-specific strategies to survive by partitioning growth-limiting resources.

Soil water content and patterns of allocation to below- and above-ground biomass in the sexes of the subdioecious plant Honckenya peploides.

BACKGROUND AND AIMS: Dioecious plants often show sex-specific differences in growth and biomass allocation. These differences have been explained as a consequence of the different reproductive functions performed by the sexes. Empirical evidence strongly supports a greater reproductive investment in females. Sex differences in allocation may determine the performance of each sex in different habitats and therefore might explain the spatial segregation of the sexes described in many dimorphic plants. Here, an investigation was made of the sexual dimorphism in seasonal patterns of biomass allocation in the subdioecious perennial herb Honckenya peploides, a species that grows in embryo dunes (i.e. the youngest coastal dune formation) and displays spatial segregation of the sexes at the studied site. The water content in the soil of the male- and female-plant habitats at different times throughout the season was also examined. METHODS: The seasonal patterns of soil-water availability and biomass allocation were compared in two consecutive years in male and female H. peploides plants by collecting soil and plant samples in natural populations. Vertical profiles of below-ground biomass and water content were studied by sampling soil in male- and female-plant habitats at different soil depths. KEY RESULTS: The sexes of H. peploides differed in their seasonal patterns of biomass allocation to reproduction. Males invested twice as much in reproduction than females early in the season, but sexual differences became reversed as the season progressed. No differences were found in above-ground biomass between the sexes, but the allocation of biomass to below-ground structures varied differently in depth for males and females, with females usually having greater below-ground biomass than males. In addition, male and female plants of H. peploides had different water-content profiles in the soil where they were growing and, when differences existed (usually in the upper layers of the soil), the water content of the soil was higher for the female plants had than for the male plants. CONCLUSIONS: Sex-differential timing of investment in reproduction and differential availability and use of resources from the soil (particularly water) are factors that probably offset the costs of reproduction in the above-ground growth in males and females of H. peploides. The results suggest that the patterns of spatial segregation of the sexes observed in H. peploides may contribute to maximize each sex's growth and reproduction.

Development, photosynthetic activity and habitat selection of the clonal plant Fragaria vesca growing in copper-polluted soil.

The ability of clonal systems to spread by ramet production may expose the clone to spatial heterogeneity. This study explored the physiological and morphological responses in the clonal plant Fragaria vesca L. growing in homogeneous (Cu-contaminated or uncontaminated) or in heterogeneous environments with patches of contrasting quality (Cu-contaminated or uncontaminated). We also investigated the potential of this species to selectively establish ramets within a heterogeneous environment. In heterogeneous environments, plants expanded ramets randomly, but selectively established ramets in the favourable patches. We discuss whether the selective establishment of ramets is a consequence of direct suppression of plant growth due to copper toxicity. The assimilate demand from offspring ramets in unfavourable environments increased the chlorophyll content and photosynthetic efficiency of parents by a feedback regulation process. Integration ameliorated the effects of copper on the photochemical efficiency of the offspring ramets. We did not observe integration costs, in terms of total biomass, for parents supporting ramets in Cu-contaminated environments, although we did detect costs in terms of ramet production. Parents with offspring ramets in Cu-contaminated environments produced 25 times more reproductive biomass than parents with offspring ramets in uncontaminated environments. We interpret this as a strategy for escaping from stressful environments. In this study, we extend the concept of physiological integration in clonal plants to include photochemical responses.

Effects of windspeed on the growth and biomass allocation of white mustard Sinapis alba L.

We examined how different wind speeds and interactions between plant age and wind affect growth and biomass allocation of Sinapis alba L. (white mustard). Physiological and growth measurements were made on individuals of white mustard grown in controlled-environment wind tunnels at windspeeds of 0.3, 2.2 and 6.0 ms-1 for 42 days. Plants were harvested at four different dates. Increasing wind speed slightly increased transpiration and stomatal conductance. We did not observe a significant decline in the photosynthetic rate per unit of leaf area. Number of leaves, stem length, leaf area and dry weights of total biomass and plant parts were significantly lower in plants exposed at high wind speed conditions. There were no significant differences in the unit leaf rate nor relative growth rates, although these were always lower in plants grown at high wind speed. Allocation and architectural parameters were also examined. After 42 days of exposure to wind, plants showed higher leaf area ratio, root and leaf weight ratios and root/shoot ratio than those grown at control treatment. Only specific leaf area declined significantly with wind speed, but stem and reproductive parts also decreased. The responses of plants to each wind speed treatment depended on the age of the plant for most of the variables. It is suggested that wind operates in logarithmic manner, with relatively small or no effect at lower wind speeds and a much greater effect at higher speeds. Since there is no evidence of a significant reduction in photosynthetic rate of Sinapis with increasing wind speed it is suggested that the effect of wind on plant growth was due to mechanical effects leading to changes in allocation and developmental patterns.