The shape of root systems in a mountain meadow: plastic responses or species-specific architectural blueprints?

The efficient uptake of nutrients depends on the ability of roots to respond to gradients of these resources. Although pot experiments have shown that species differ in their ability to proliferate their roots in nutrient-rich patches, the role of such differences in determining root shapes in the field is unclear. We used fine-scale quantitative (q)PCR-based species-specific mapping of roots in a grassland community to reconstruct species-specific root system shapes. We linked them with data from pot experiments on the ability of these species to proliferate in nutrient-rich patches and their rooting depth. We found remarkable diversity in root system shapes, from cylindrical to conical. Interspecific differences in rooting depths in pots were the main determinant of rooting depths in the field, whereas differences in foraging ability played only a minor role. Although some species with strong foraging ability did place their roots into nutrient-rich soil layers, it was not a universal pattern. The results imply that although the vertical differentiation of grassland species is pronounced, it is primarily not driven by the differential plastic response of species to soil nutrient gradients. This may constrain the coexistence of species with similar rooting depths and may instead favour coexistence of species differing in their architectural blueprints.

The importance of ecophysiological traits in response of Festuca rubra to changing climate.

Knowledge of the ability of plants to respond to climate change via phenotypic plasticity or genetic adaptation in ecophysiological traits and of the link of these traits to fitness is still limited. We studied the clonal grass Festuca rubra from 11 localities representing factorially crossed gradients of temperature and precipitation and cultivated them in growth chambers simulating temperature and moisture regime in the four extreme localities. We measured net photosynthetic rate, Fv /Fm , specific leaf area, osmotic potential and stomatal density and length and tested their relationship to proxies of fitness. We found strong phenotypic plasticity in photosynthetic traits and genetic differentiation in stomatal traits. The effects of temperature and moisture interacted (either as conditions of origin or growth chambers), as were effects of growth and origin. The relationships between the ecophysiological and fitness-related traits were significant but weak. Phenotypic plasticity and genetic differentiation of the species indicate the potential ability of F. rubra to adapt to novel climatic conditions. The most important challenge for the plants seems to be increasing moisture exposing plants to hypoxia. However, the plants have the potential to respond to increased moisture by changes in stomatal size and density and adjustments of osmotic potential. Changes in ecophysiological traits translate into variation in plant fitness, but the selection on the traits is relatively weak and depends on actual conditions. Despite the selection, the plants do not show strong local adaptation and local adaptation is thus likely not restricting species ability to adjust to novel conditions.

Maternal effects strengthen interactions of temperature and precipitation, determining seed germination of dominant alpine grass species.

PREMISE: Despite the existence of many studies on the responses of plant species to climate change, there is a knowledge gap on how specific climatic factors and their interactions regulate seed germination in alpine species. This understanding is complicated by the interplay between responses of seeds to the environment experienced during germination, the environment experienced by the maternal plant during seed development and genetic adaptations of the maternal plant to its environment of origin. METHODS: The study species (Anthoxanthum alpinum, A. odoratum) originated from localities with factorial combinations of temperature and precipitation. Seed germination was tested in conditions simulating the extreme ends of the current field conditions and a climate change scenario. We compared the performance of field-collected seeds with that of garden-collected seeds. RESULTS: A change to warmer and wetter conditions resulted in the highest germination of A. alpinum, while A. odoratum germinated the most in colder temperature and with home moisture. The maternal environment did have an impact on plant performance of the study species. Field-collected seeds of A. alpinum tolerated warmer conditions better than those from the experimental garden. CONCLUSIONS: The results demonstrate how knowledge of responses to climate change can increase our ability to understand and predict the fate of alpine species. Studies that aim to understand the germination requirements of seeds under future climates should use experimental designs allowing the separation of genetic differentiation, plasticity and maternal effects and their interactions, since all these mechanisms play an important role in driving species' germination patterns.

Evolutionary potential of a widespread clonal grass under changing climate.

Adaptive responses are probably the most effective long-term responses of populations to climate change, but they require sufficient evolutionary potential upon which selection can act. This requires high genetic variance for the traits under selection and low antagonizing genetic covariances between the different traits. Evolutionary potential estimates are still scarce for long-lived, clonal plants, although these species are predicted to dominate the landscape with climate change. We studied the evolutionary potential of a perennial grass, Festuca rubra, in western Norway, in two controlled environments corresponding to extreme environments in natural populations: cold-dry and warm-wet, the latter being consistent with the climatic predictions for the country. We estimated genetic variances, covariances, selection gradients and response to selection for a wide range of growth, resource acquisition and physiological traits, and compared their estimates between the environments. We showed that the evolutionary potential of F. rubra is high in both environments, and genetic covariances define one main direction along which selection can act with relatively few constraints to selection. The observed response to selection at present is not sufficient to produce genotypes adapted to the predicted climate change under a simple, space for time substitution model. However, the current populations contain genotypes which are pre-adapted to the new climate, especially for growth and resource acquisition traits. Overall, these results suggest that the present populations of the long-lived clonal plant may have sufficient evolutionary potential to withstand long-term climate changes through adaptive responses.

Recent similarity in distribution ranges does not mean a similar postglacial history: a phylogeographical study of the boreal tree species Alnus incana based on microsatellite and chloroplast DNA variation.

We reconstructed the historical pattern of postglacial biogeographic range expansion of the boreal tree species Alnus incana in Europe. To assess population genetic structure and diversity, we performed a combined analysis of nuclear microsatellite loci and chloroplast DNA sequences (65 populations, 1004 individuals). Analysis of haplotype and microsatellite diversity revealed that southeastern refugial populations situated in the Carpathians and the Balkan Peninsula did not spread north and cannot be considered as important source populations for postglacial recolonization of Europe; populations in Eastern Europe did not establish Fennoscandian populations; populations in Fennoscandia and Eastern Europe have no unique genetic cluster, but represent a mix with a predominant cluster typical for Central Europe; and that colonization of Fennoscandia and Eastern Europe took place from Central Europe. Our findings highlight the importance of an effective refugium in Central Europe located outside classical southern refugia confirming the existence of northern refugia for boreal trees in Europe. The postglacial range expansion of A. incana did not follow the model established for Picea abies. Fennoscandian populations are not derived from Eastern European ones, but from Central European ones.

Flow cytometry, microsatellites and niche models reveal the origins and geographical structure of Alnus glutinosa populations in Europe.

BACKGROUND AND AIMS: Polyploidy in plants has been studied extensively. In many groups, two or more cytotypes represent separate biological entities with distinct distributions, histories and ecology. This study examines the distribution and origins of cytotypes of Alnus glutinosa in Europe, North Africa and western Asia. METHODS: A combined approach was used involving flow cytometry and microsatellite analysis of 12 loci in 2200 plants from 209 populations combined with species distribution modelling using MIROC and CCSM climatic models, in order to analyse (1) ploidy and genetic variation, (2) the origin of tetraploid A. glutinosa, considering A. incana as a putative parent, and (3) past distributions of the species. KEY RESULTS: The occurrence of tetraploid populations of A. glutinosa in Europe is determined for the first time. The distribution of tetraploids is far from random, forming two geographically well-delimited clusters located in the Iberian Peninsula and the Dinaric Alps. Based on microsatellite analysis, both tetraploid clusters are probably of autopolyploid origin, with no indication that A. incana was involved in their evolutionary history. A projection of the MIROC distribution model into the Last Glacial Maximum (LGM) showed that (1) populations occurring in the Iberian Peninsula and North Africa were probably interconnected during the LGM and (2) populations occurring in the Dinaric Alps did not exist throughout the last glacial periods, having retreated southwards into lowland areas of the Balkan Peninsula. CONCLUSIONS: Newly discovered tetraploid populations are situated in the putative main glacial refugia, and neither of them was likely to have been involved in the colonization of central and northern Europe after glacial withdrawal. This could mean that neither the Iberian Peninsula nor the western part of the Balkan Peninsula served as effective refugial areas for northward post-glacial expansion of A. glutinosa.

Higher genetic diversity in recolonized areas than in refugia of Alnus glutinosa triggered by continent-wide lineage admixture.

Genetic admixture is supposed to be an important trigger of species expansions because it can create the potential for selection of genotypes suitable for new climatic conditions. Up until now, however, no continent-wide population genetic study has performed a detailed reconstruction of admixture events during natural species expansions. To fill this gap, we analysed the postglacial history of Alnus glutinosa, a keystone species of European swamp habitats, across its entire distribution range using two molecular markers, cpDNA and nuclear microsatellites. CpDNA revealed multiple southern refugia located in the Iberian, Apennine, Balkan and Anatolian Peninsulas, Corsica and North Africa. Analysis of microsatellites variation revealed three main directions of postglacial expansion: (i) from the northern part of the Iberian Peninsula to Western and Central Europe and subsequently to the British Isles, (ii) from the Apennine Peninsula to the Alps and (iii) from the eastern part of the Balkan Peninsula to the Carpathians followed by expansion towards the Northern European plains. This challenges the classical paradigm that most European populations originated from refugial areas in the Carpathians. It has been shown that colonizing lineages have met several times and formed secondary contact zones with unexpectedly high population genetic diversity in Central Europe and Scandinavia. On the contrary, limited genetic admixture in southern refugial areas of A. glutinosa renders rear-edge populations in the Mediterranean region more vulnerable to extinction due to climate change.

Effect of DNA methylation, modified by 5-azaC, on ecophysiological responses of a clonal plant to changing climate.

Epigenetic regulation of gene expression is expected to be an important mechanism behind phenotypic plasticity. Whether epigenetic regulation affects species ecophysiological adaptations to changing climate remains largely unexplored. We compared ecophysiological traits between individuals treated with 5-azaC, assumed to lead to DNA demethylation, with control individuals of a clonal grass originating from and grown under different climates, simulating different directions and magnitudes of climate change. We linked the ecophysiological data to proxies of fitness. Main effects of plant origin and cultivating conditions predicted variation in plant traits, but 5-azaC did not. Effects of 5-azaC interacted with conditions of cultivation and plant origin. The direction of the 5-azaC effects suggests that DNA methylation does not reflect species long-term adaptations to climate of origin and species likely epigenetically adjusted to the conditions experienced during experiment set-up. Ecophysiology translated to proxies of fitness, but the intensity and direction of the relationships were context dependent and affected by 5-azaC. The study suggests that effects of DNA methylation depend on conditions of plant origin and current climate. Direction of 5-azaC effects suggests limited role of epigenetic modifications in long-term adaptation of plants. It rather facilitates fast adaptations to temporal fluctuations of the environment.

Ericaceous dwarf shrubs affect ectomycorrhizal fungal community of the invasive Pinus strobus and native Pinus sylvestris in a pot experiment.

This study aimed to elucidate the relationship between ericaceous understorey shrubs and the diversity and abundance of ectomycorrhizal fungi (EcMF) associated with the invasive Pinus strobus and native Pinus sylvestris. Seedlings of both pines were grown in mesocosms and subjected to three treatments simulating different forest microhabitats: (a) grown in isolation and grown with (b) Vaccinium myrtillus or (c) Vaccinium vitis-idaea. Ericaceous plants did not act as a species pool of pine mycobionts and inhibited the ability of the potentially shared species Meliniomyces bicolor to form ectomycorrhizae. Similarly, Ericaceae significantly reduced the formation of Thelephora terrestris ectomycorrhizae in P. sylvestris. EcMF species composition in the mesocosms was strongly affected by both the host species and the presence of an ericaceous neighbour. When grown in isolation, P. strobus root tips were predominantly colonised by Wilcoxina mikolae, whereas those of P. sylvestris were more commonly colonised by Suillus and Rhizopogon spp. Interestingly, these differences were less evident (Suillus + Rhizopogon spp.) or absent (W. mikolae) when the pines were grown with Ericaceae. P. strobus exclusively associated with Rhizopogon salebrosus s.l., suggesting the presence of host specificity at the intrageneric level. Ericaceous plants had a positive effect on colonisation of P. strobus root tips by R. salebrosus s.l. This study demonstrates that the interaction of selective factors such as host species and presence of ericaceous plants may affect the realised niche of the ectomycorrhizal fungi.

Evolutionary Rescue as a Mechanism Allowing a Clonal Grass to Adapt to Novel Climates.

Filing gaps in our understanding of species' abilities to adapt to novel climates is a key challenge for predicting future range shifts and biodiversity loss. Key knowledge gaps are related to the potential for evolutionary rescue in response to climate, especially in long-lived clonally reproducing species. We illustrate a novel approach to assess the potential for evolutionary rescue using a combination of reciprocal transplant experiment in the field to assess performance under a changing climate and independent growth chamber assays to assess growth- and physiology-related plant trait maxima and plasticities of the same clones. We use a clonal grass, Festuca rubra, as a model species. We propagated individual clones and used them in a transplant experiment across broad-scale temperature and precipitation gradients, simulating the projected direction of climate change in the region. Independent information on trait maxima and plasticities of the same clones was obtained by cultivating them in four growth chambers representing climate extremes. Plant survival was affected by interaction between plant traits and climate change, with both trait plasticities and maxima being important for adaptation to novel climates. Key traits include plasticity in extravaginal ramets, aboveground biomass, and osmotic potential. The direction of selection in response to a given climatic change detected in this study mostly contradicted the natural trait clines indicating that short-term selection pressure as identified here does not match long-term selection outcomes. Long-lived clonal species exposed to different climatic changes are subjected to consistent selection pressures on key traits, a necessary condition for adaptation to novel conditions. This points to evolutionary rescue as an important mechanism for dealing with climate change in these species. Our experimental approach may be applied also in other model systems broadening our understanding of evolutionary rescue. Such knowledge cannot be easily deduced from observing the existing field clines.

Adaptive differentiation of Festuca rubra along a climate gradient revealed by molecular markers and quantitative traits.

Species response to climate change is influenced by predictable (selective) and unpredictable (random) evolutionary processes. To understand how climate change will affect present-day species, it is necessary to assess their adaptive potential and distinguish it from the effects of random processes. This will allow predicting how different genotypes will respond to forecasted environmental change. Space for time substitution experiments are an elegant way to test the response of present day populations to climate variation in real time. Here we assess neutral and putatively adaptive variation in 11 populations of Festuca rubra situated along crossed gradients of temperature and moisture using molecular markers and phenotypic measurements, respectively. By comparing population differentiation in putatively neutral molecular markers and phenotypic traits (QST-FST comparisons), we show the existence of adaptive differentiation in phenotypic traits and their plasticity across the climatic gradient. The observed patterns of differentiation are due to the high genotypic and phenotypic differentiation of the populations from the coldest (and wettest) environment. Finally, we observe statistically significant covariation between markers and phenotypic traits, which is likely caused by isolation by adaptation. These results contribute to a better understanding of the current adaptation and evolutionary potential to face climate change of a widespread species. They can also be extrapolated to understand how the studied populations will adjust to upcoming climate change without going through the lengthy process of phenotyping.

Transgenerational plasticity as an important mechanism affecting response of clonal species to changing climate.

In spite of the increasing number of studies on the importance of transgenerational plasticity for species response to novel environments, its effects on species ability to respond to climate change are still largely unexplored. We study the importance of transgenerational plasticity for response of a clonal species Festuca rubra. Individuals from four natural populations representing two levels of temperature and two levels of precipitation were cultivated in four growth chambers that simulate the temperature and precipitation of origin of the populations (maternal phase). Each population was represented in each growth chamber. After 6 months, single young ramets of these plants were reshuffled among the growth chambers and let to grow for additional 2 months (offspring phase). The results show that transgenerational effects (i.e., maternal phase conditions) significantly modify species response to novel climates, and the direction and intensity of the response depend on the climate of origin of the plants. For traits related to recourse acquisition, the conditions of maternal phase, either alone or in interaction mainly with climate of origin, had stronger effect than the conditions of cultivation. Overall, the maternal climate interacted more intensively with the climate of origin than with the offspring climate. The direction of the effect of the maternal climate was of different directions and intensities depending on plant origin and trait studied. The data demonstrated strong significant effects of conditions during maternal phase on species response to novel climates. These transgenerational affects were, however, not adaptive. Still, transgenerational plasticity may be an important driver of species response to novel conditions across clonal generations. These effects thus need to be carefully considered in future studies exploring species response to novel climates. This will also have strong effects on species performance under increasingly variable climates expected to occur with the climate change.

Migration patterns of subgenus Alnus in Europe since the last glacial maximum: a systematic review.

BACKGROUND/AIMS: Recently, new palaeoecological records supported by molecular analyses and palaeodistributional modelling have provided more comprehensive insights into plant behaviour during the last Quaternary cycle. We reviewed the migration history of species of subgenus Alnus during the last 50,000 years in Europe with a focus on (1) a general revision of Alnus history since the Last Glacial Maximum (LGM), (2) evidence of northern refugia of Alnus populations during the LGM and (3) the specific history of Alnus in particular European regions. METHODOLOGY: We determined changes in Alnus distribution on the basis of 811 and 68 radiocarbon-dated pollen and macrofossil sites, respectively. We compiled data from the European Pollen Database, the Czech Quaternary Palynological Database, the Eurasian Macrofossil Database and additional literature. Pollen percentage thresholds indicating expansions or retreats were used to describe patterns of past Alnus occurrence. PRINCIPAL FINDINGS: An expansion of Alnus during the Late Glacial and early Holocene periods supports the presence of alders during the LGM in southern peninsulas and northerly areas in western Europe, the foothills of the Alps, the Carpathians and northeastern Europe. After glaciers withdrew, the ice-free area of Europe was likely colonized from several regional refugia; the deglaciated area of Scandinavia was likely colonized from a single refugium in northeastern Europe. In the more northerly parts of Europe, we found a scale-dependent pattern of Alnus expansion characterised by a synchronous increase of Alnus within individual regions, though with regional differences in the times of the expansion. In southern peninsulas, the Alps and the Carpathians, by contrast, it seems that Alnus expanded differently at individual sites rather than synchronously in whole regions. CONCLUSIONS: Our synthesis supports the idea that northern LGM populations were important sources of postglacial Alnus expansion. The delayed Alnus expansion apparent in some regions was likely a result of environmental limitations.

European invasion of North American Pinus strobus at large and fine scales: high genetic diversity and fine-scale genetic clustering over time in the adventive range.

BACKGROUND: North American Pinus strobus is a highly invasive tree species in Central Europe. Using ten polymorphic microsatellite loci we compared various aspects of the large-scale genetic diversity of individuals from 30 sites in the native distribution range with those from 30 sites in the European adventive distribution range. To investigate the ascertained pattern of genetic diversity of this intercontinental comparison further, we surveyed fine-scale genetic diversity patterns and changes over time within four highly invasive populations in the adventive range. RESULTS: Our data show that at the large scale the genetic diversity found within the relatively small adventive range in Central Europe, surprisingly, equals the diversity found within the sampled area in the native range, which is about thirty times larger. Bayesian assignment grouped individuals into two genetic clusters separating North American native populations from the European, non-native populations, without any strong genetic structure shown over either range. In the case of the fine scale, our comparison of genetic diversity parameters among the localities and age classes yielded no evidence of genetic diversity increase over time. We found that SGS differed across age classes within the populations under study. Old trees in general completely lacked any SGS, which increased over time and reached its maximum in the sapling stage. CONCLUSIONS: Based on (1) the absence of difference in genetic diversity between the native and adventive ranges, together with the lack of structure in the native range, and (2) the lack of any evidence of any temporal increase in genetic diversity at four highly invasive populations in the adventive range, we conclude that population amalgamation probably first happened in the native range, prior to introduction. In such case, there would have been no need for multiple introductions from previously isolated populations, but only several introductions from genetically diverse populations.