Campanula Thyrsoides Research Articles

Glacial History Affected Phenotypic Differentiation in the Alpine Plant, Campanula thyrsoides

Numerous widespread Alpine plant species show molecular differentiation among populations from distinct regions. This has been explained as the result of genetic drift during glacial survival in isolated refugia along the border of the European Alps. Since genetic drift may affect molecular markers and phenotypic traits alike, we asked whether phenotypic differentiation mirrors molecular patterns among Alpine plant populations from different regions. Phenotypic traits can be under selection, so we additionally investigated whether part of the phenotypic differentiation can be explained by past selection and/or current adaptation. Using the monocarpic Campanula thyrsoides as our study species, a common garden experiment with plants from 21 populations from four phylogeographic groups located in regions across the Alps and the Jura Mountains was performed to test for differentiation in morphological and phenological traits. Past selection was investigated by comparing phenotypic differentiation among and within regions with molecular differentiation among and within regions. The common garden results indicated regional differentiation among populations for all investigated phenotypic traits, particularly in phenology. Delayed flowering in plants from the South-eastern Alps suggested adaptation to long sub-mediterranean summers and contrasted with earlier flowering of plants experiencing shorter growing seasons in regions with higher elevation to the West. Comparisons between molecular and phenotypic differentiation revealed diversifying selection among regions in height and biomass, which is consistent with adaptation to environmental conditions in glacial refugia. Within regions, past selection acted against strong diversification for most phenotypic traits, causing restricted postglacial adaptation. Evidence consistent with post-glacial adaptation was also given by negative correlation coefficients between several phenotypic traits and elevation of the population's origin. In conclusion, our study suggests that, irrespective of adaptation of plants to their current environment, glacial history can have a strong and long-lasting influence on the phenotypic evolution of Alpine plants.

Flowering phenology and reproductive fitness along a mountain slope: maladaptive responses to transplantation to a warmer climate in Campanula thyrsoides

In many biomes, global warming has resulted in advanced and longer growing seasons, which has often led to earlier flowering in plant taxa. Elevational gradients are ideal to study the effects of global warming as they allow transplantation of plants from their original cooler higher elevations down to elevations with a prospective climate. We transplanted plants from ten populations of the European alpine monocarpic herb species Campanula thyrsoides L. to three sites along a steep mountain slope (600, 1,235 and 1,850 m above sea level) in the Swiss Alps and asked whether reproductive phenology adjusts plastically to elevation and if these responses were adaptive, i.e. increased the fitness of plants. We further assessed current genetic differentiation in phenotypic traits and whether any such origin effects were due to adaptation to climatic conditions of origin. Our results showed that transplantation to lower elevations caused strong shifts in phenology, with plants starting growth and flowering earlier than plants placed at higher elevations. However, compared to flower production at high elevation, number of flowers per plant decreased 21 % at mid- and 61 % at low elevation. The shift in phenology thus came with a high cost in fitness, and we suggest that phenology is maladaptive when C. thyrsoides faces temperature conditions deviating from its natural amplitude. We conclude that the frequently reported phenological shift in plant species as a response to global warming may include heavy fitness costs that may hamper species survival.

Pollen dispersal and gene flow within and into a population of the alpine monocarpic plant Campanula thyrsoides

Gene flow by seed and pollen largely shapes the genetic structure within and among plant populations. Seed dispersal is often strongly spatially restricted, making gene flow primarily dependent on pollen dispersal within and into populations. To understand distance-dependent pollination success, pollen dispersal and gene flow were studied within and into a population of the alpine monocarpic perennial Campanula thyrsoides. A paternity analysis was performed on sampled seed families using microsatellites, genotyping 22 flowering adults and 331 germinated offspring to estimate gene flow, and pollen analogues were used to estimate pollen dispersal. The focal population was situated among 23 genetically differentiated populations on a subalpine mountain plateau (<10 km(2)) in central Switzerland. Paternity analysis assigned 110 offspring (33·2 %) to a specific pollen donor (i.e. 'father') in the focal population. Mean pollination distance was 17·4 m for these offspring, and the pollen dispersal curve based on positive LOD scores of all 331 offspring was strongly decreasing with distance. The paternal contribution from 20-35 offspring (6·0-10·5 %) originated outside the population, probably from nearby populations on the plateau. Multiple potential fathers were assigned to each of 186 offspring (56·2 %). The pollination distance to 'mother' plants was negatively affected by the mothers' degree of spatial isolation in the population. Variability in male mating success was not related to the degree of isolation of father plants. Pollen dispersal patterns within the C. thyrsoides population are affected by spatial positioning of flowering individuals and pollen dispersal may therefore contribute to the course of evolution of populations of this species. Pollen dispersal into the population was high but apparently not strong enough to prevent the previously described substantial among-population differentiation on the plateau, which may be due to the monocarpic perenniality of this species.

Spatial genetic structure of Campanula thyrsoides across the European Alps: Indications for glaciation-driven allopatric subspeciation

Abstract Quaternary climate change had profound impacts on the geographical distribution and genetic structure of plant species which is hypothesized to have triggered allopatric speciation due to spatial isolation. However, evidence is scarce despite recent advances that indicate glacial history and substrate requirements as main drivers of spatial genetic structures. Here we built upon these studies to test the role of glaciations on the morphological and ecological differentiation within the calcicolous Campanula thyrsoides across its European Alpine distribution range. We collected samples from 51 populations (1173 plants), used five microsatellite markers, estimated diversity ( N a , H E ) and differentiation ( D est , G ST_est , F ST ) and applied Bayesian clustering analysis and tessellation methods. We found support for four genetically distinct groups of populations, arranged from West to East: (i) France and Western Switzerland, (ii) Central and most of Eastern Switzerland, (iii) parts of Eastern Switzerland and Central Austria, and (iv) Southeastern Austria, Slovenia and Northeastern Italy. Among-subspecies variance was 8.4% and each subspecies was highly differentiated ( C.*thyrsoides : D est = 0.47; C.*carniolica : D est = 0.58). Geographic structuring of within-population diversity was not related to refugia outside of previously-glaciated terrain but to subspecies: the more thermophilic C.*carniolica showed significantly lower levels of within-population diversity and higher numbers of private alleles. The location of the genetic break lines between these four groups of populations corresponds to well-known biogeographic barriers. However, the phylogeographic pattern has elements formerly found in both calcicolous and silicicolous species and thus questions the generality of substrate-related patterns. Within C.*thyrsoides , population admixture upon secondary contact may have led to high genetic diversity across the distribution range. Given the geographic and genetic differences of the subspecies we conclude that differentiation between C.*thyrsoides and C.*carniolica represents a case of glaciation-driven allopatric subspeciation reinforced by missing secondary contact due to incomplete post-glacial recolonization of potential habitats.

Differentiation in morphology and flowering phenology between two Campanula thyrsoides L. subspecies

Subspecies are usually characterised by sets of morphological discontinuities. By means of common garden experiments, we investigated genetic differentiation in morphological and phenological traits in two geographically disjunct subspecies of Campanula thyrsoides L., i.e. subsp. thyrsoides (=C.* thyrsoides) occurring in the European Alps and Jura Mountains, and subsp. carniolica (=C.* carniolica) occurring in the Southeastern Alps and the Dinaric Arc. Nine out of 16 investigated traits were significantly different between C.* thyrsoides and C.* carniolica. For C.* carniolica inflorescence length was 1.4×, and above-ground biomass 2.7× higher, while flower density was significantly lower. Campanula* carniolica also showed delayed flowering and flower development from bottom to top as compared to C.* thyrsoides which flowered from top to bottom. The inflorescence growth was indeterminate and flowering took several weeks in C.* carniolica, whereas C.* thyrsoides showed determinate flowering, rapidly opening all flowers within a few days. This differentiation in flowering phenology is likely to be adaptive. The submediterranean climate favours indeterminate flowering in C.* carniolica, allowing ongoing growth of the inflorescence throughout the long summer until environmental conditions worsen, whereas determinate and early flowering in C.* thyrsoides is favourable in the short growing season in the high Alps where seed production must be secured before temperature drops. Glacial survival in refugia with different climates (alpine vs. submediterranean) may have caused this regional differentiation.

Genotypic and environmental variation in specific leaf area in a widespread Alpine plant after transplantation to different altitudes

Specific leaf area (SLA) is an important plant functional trait as it is an indicator of ecophysiological characteristics like relative growth rate, stress tolerance and leaf longevity. Substantial intraspecific variation in SLA is common and usually correlates with environmental conditions. For instance, SLA decreases with increasing altitude, which is understood as adjustment to temperature. It is generally assumed that intraspecific variation is mostly the result of environmentally induced phenotypic plasticity, but genetic effects may also be present, due to local adaptation or genetic drift. In this study, genotypic and environmental effects on SLA were experimentally separated for the widespread Alpine bell flower Campanula thyrsoides by transplanting plants to three common gardens at contrasting altitudes (600, 1,235 and 1,850 m a.s.l.). Seeds were sampled from 18 populations in four phylogeographic regions within the European Alps. A strong plastic response was observed: SLA decreased with increasing altitude of the common gardens (22.0% of variation). The phylogeographic regions were differentiated in SLA in the common gardens (10.1% of variation), indicating that SLA is at least partly genetically determined. Plants from the six easternmost populations experienced a submediterranean climate and showed decreased SLA values in the three common gardens compared to populations to the west, which may be explained as adaptation to drought. Within these submediterranean populations, SLA decreased with altitude of origin in two out of three common gardens. Concluding, SLA shows strong phenotypic plasticity as well as substantial genetic effects, the latter probably being the result of adaptation to local conditions rather than genetic drift.

Genetic diversity, phenotypic variation and local adaptation in the alpine landscape: case studies with alpine plant species

Plant survival in alpine landscapes is constantly challenged by the harsh and often unpredictable environmental conditions. Steep environmental gradients and patchy distribution of habitats lead to small size and spatial isolation of populations and restrict gene flow. Agricultural land use has further increased the diversity of habitats below and above the treeline. We studied the consequences of the highly structured alpine landscape for evolutionary processes in four study plants: Epilobium fleischeri, Geum reptans, Campanula thyrsoides and Poa alpina. The main questions were: (1) How is genetic diversity distributed within and among populations and is it affected by altitude, population size or land use? (2) Do reproductive traits such as allocation to sexual or vegetative reproduction vary with altitude or land use? Furthermore, we studied if seed weight increases with altitude. Within-population genetic diversity of the four species was high and mostly not related to altitude and population size. Nevertheless, genetic differentiation among populations was pronounced and strongly increasing with distance. In Poa alpina genetic diversity was affected by land use. Results suggest considerable genetic drift among populations of alpine plants. Reproductive allocation was affected by altitude and land use in Poa alpina and by succession in Geum reptans. Seed weight was usually higher in alpine species than in related lowland species. We conclude that the evolutionary potential to respond to global change is mostly intact in alpine plants, even at high altitude. Phenotypic variability is shaped by adaptive as well as by random evolutionary processes; moreover plastic responses to growth conditions seem to be crucial for survival of plants in the alpine landscape.

Evolutionary demography of long‐lived monocarpic perennials: a time‐lagged integral projection model

Summary The evolution of flowering strategies (when and at what size to flower) in monocarpic perennials is determined by balancing current reproduction with expected future reproduction, and these are largely determined by size‐specific patterns of growth and survival. However, because of the difficulty in following long‐lived individuals throughout their lives, this theory has largely been tested using short‐lived species (&lt; 5 years). Here, we tested this theory using the long‐lived monocarpic perennial Campanula thyrsoides which can live up to 16 years. We used a novel approach that combined permanent plot and herb chronology data from a 3‐year field study to parameterize and validate integral projection models (IPMs). Similar to other monocarpic species, the rosette leaves of C. thyrsoides wither over winter and so size cannot be measured in the year of flowering. We therefore extended the existing IPM framework to incorporate an additional time delay that arises because flowering demography must be predicted from rosette size in the year before flowering. We found that all main demographic functions (growth, survival probability, flowering probability and fecundity) were strongly size‐dependent and there was a pronounced threshold size of flowering. There was good agreement between the predicted distribution of flowering ages obtained from the IPMs and that estimated in the field. Mostly, there was good agreement between the IPM predictions and the direct quantitative field measurements regarding the demographic parameters λ, R0 and T. We therefore conclude that the model captures the main demographic features of the field populations. Elasticity analysis indicated that changes in the survival and growth function had the largest effect (c. 80%) on λ and this was considerably larger than in short‐lived monocarps. We found only weak selection pressure operating on the observed flowering strategy which was close to the predicted evolutionary stable strategy. Synthesis. The extended IPM accurately described the demography of a long‐lived monocarpic perennial using data collected over a relatively short period. We could show that the evolution of flowering strategies in short‐ and long‐lived monocarps seem to follow the same general rules but with a longevity‐related emphasis on survival over fecundity.

Spatial isolation and genetic differentiation in naturally fragmented plant populations of the Swiss Alps

Aims The effect Of anthropogenic landscape fragmentation on the genetic diversity and adaptive potential of plant populations is a major issue in conservation biology. However, little is known about the partitioning of genetic diversity in alpine species, which occur in naturally fragmented habitats. Here, we, investigate molecular patterns of three alpine plants (Epilobium fleischeri, Geum reptans and Campanula thyrsoides) across Switzerland and ask whether Spatial isolation has led to high levels of populations differentiation, increasing over distance, and a decrease of within-population variability. We further hypothesize that file contrasting potential for long-distance dispersal (LDD) of Seed in these Species will considerably influence and explain diversity partitioning. Methods For each study species, we Sampled 20-23 individuals from each of 20-32 populations across entire Switzerland. We applied Random Amplified Polymorphic Dimorphism markers to assess genetic diversity within (Nei's expected heterozygosity, H-e; percentage of polymorphic hands, P-P) and among (analysis of molecular variance, Phi(st)) populations and correlated population size and altitude with within-populalion diversity. Spatial patterns of genetic relatedness were investigated using Mantel tests and standardized major axis regression as well as unweighted pair group method with arithmetic mean cluster analyses and Monmonier's algorithm. To avoid known biases, We standardized the numbers of populations, individuals and markers using multiple random reductions. We modelled LDD with a high alpine wind data set using the terminal velocity and height of seed release as key parameters. Additionally, we assessed a number of important life-history traits and factors that potentially influence genetic diversity partitioning (e.g. breeding system, longevity and population size). Important findings For all three species, We found a significant isolation-by-distance relationship but only a moderately high differentiation among populations (Phi(st): 22.7, 48 and 16.8%, for E. fleischeri, G. reptans and C. thyrsoides, respectively). Within-population diversity (H-c: 0.19-0.21, P-p: 62-75%) was not reduced in comparison to known results from lowland species and even small populations with < 50 reproductive individuals contained high levels of genetic diversity. We further found no indication that a high long-distance seed dispersal potential enhances genetic connectivity among populations. Gene flow seems to have a strong stochastic component causing large dissimilarity between population pairs irrespective of the spatial distance. Our results suggest that other life-history traits, especially the breeding System, may play an important role in genetic diversity partitioning. We conclude that spatial isolation in the alpine environment has a strong influence on population relatedness but that a number of factors can considerably influence the strength of this relationship.

The biological flora of Central Europe: Campanula thyrsoides L.

Campanula thyrsoides is one of the few monocarpic perennials of temperate alpine mountain ecosystems and native to the European Alps and adjacent mountain ranges. Individuals are rare but locally abundant and the species is protected in most of the Alpine countries. The exceptionally yellow-flowering member of the Campanulaceae grows preferably in alpine meadows with a moderate disturbance regime on limestone or carbonate bearing schists. Traditionally regarded as a rosette-forming ‘‘biennial’’, individuals usually flower after 8 years and grow considerably older at higher altitudes. This article reviews the taxonomy, morphology, distribution, ecology, life cycle, population biology, and genetics of this species as well as its status in the European countries.

No inbreeding depression in an outcrossing alpine species: The breeding system of Campanula thyrsoides

Plants that live in fragmented landscapes, where populations are isolated from each other and in which long-distance dispersal is essential for colonization of empty sites, reproduction should be favoured by self-compatibility (Baker's law). Nevertheless, outcrossing mechanisms, such as self-incompatibility and dichogamy, are common in many species and are often maintained by inbreeding depression in the fitness of selfed progeny. Here, we studied the breeding system and the consequences of selfing and sister mating in Campanula thyrsoides, a short-lived perennial monocarp, which is found in the naturally fragmented landscape of the Alps. An experiment with controlled pollinations was set up in the common garden with plants grown from seeds originating from 14 seed families, collected in the siliceous Central Alps, where this plant is found on isolated carbonate bearing outcrops. Our results indicate that C. thyrsoides has a strong self-incompatibility system (SI) with no or low seed set in selfed flowers compared to outcrossed and sister-crossed flowers. Moreover, the SI system in C. thyrsoides did not break down with flower age as in some other Campanula species. Surprisingly, there was no significant difference in seed set, seed weight, germination percentage, seedling survival and size between outcrossed and sister-crossed offspring, which indicates no inbreeding depression. We suggest that the absence of inbreeding depression in this outcrossing species might be a result of frequent bottlenecks during colonization of the isolated habitats in the alpine landscape.

Development and characterization of microsatellite DNA markers for the Alpine plant species Campanula thyrsoides

AbstractWe isolated and characterized eight polymorphic microsatellite markers for the Alpine plant species Campanula thyrsoides (Campanulaceae). Number of alleles per locus ranged from six to 12 and the observed heterozygosity was between 0.529 and 0.900. Observed vs. expected heterozygote deficits were significantly different in one out of eight loci following Bonferroni's correction for multiple tests. We did not find evidence for linkage disequilibrium between locus pairs. The microsatellite markers are being used for the study of genetic variation and gene flow within and among populations of C. thyrsoides in the Swiss Alps.