Differential patterns of within- and between-population genetically based trait variation in Lupinus angustifolius.

BACKGROUND AND AIMS: Within-population genetic and phenotypic variation play a key role in the development of adaptive responses to environmental change. Between-population variation is also an essential element in assessing the evolutionary potential of species in response to changes in environmental conditions. In this context, common garden experiments are a useful tool to separate the genetic and environmental components of phenotypic variation. We aimed to assess within- and between-population phenotypic variation of Lupinus angustifolius L. in terms of its evolutionary potential to adapt to ongoing climate change. METHODS: We evaluated populations' phenotypic variation of foliar, phenological and reproductive traits with a common garden experiment. Patterns of functional trait variation were assessed with (1) mixed model analyses and coefficients of variation (CVs) with confidence intervals, (2) principal component analyses (PCAs) and (3) correlations between pairs of traits. Analyses were performed at the population level (four populations) and at the latitude level (grouping pairs of populations located in two latitudinal ranges). KEY RESULTS: Phenotypic variation had a significant genetic component associated with a latitudinal pattern. (1) Mixed models found lower specific leaf area, advanced flowering phenology and lower seed production of heavier seeds in southern populations, whereas CV analyses showed lower within-latitude variation especially in phenological and reproductive traits in southern populations. (2) PCAs showed a clearer differentiation of phenotypic variation between latitudes than between populations. (3) Correlation analyses showed a greater number of significant correlations between traits in southern populations (25 vs. 13). CONCLUSIONS: Between-population phenotypic variation was determined by contrasting temperature and drought at different latitude and elevation. Southern populations had differential trait values compatible with adaptations to high temperatures and drought. Moreover, they had lower within-population variation and a greater number of trait correlations probably as a result of these limiting conditions, making them more vulnerable to climate change.

Modeling the continua in the outcomes of biotic interactions.

The outcome of many ecological interactions lies somewhere along a continuum between pure positive and pure negative effects. Although the popularity of this idea has notoriously risen in the last decades, with the occurrence of continua in interaction outcomes invoked for a wide variety of interactions, the absence of a precise theoretical treatment has led to considerable inaccuracy and ambiguity in its treatment. We develop here a consumer-resource model to explore the occurrence of continua. This model is based on the assumption that the distribution of individual interaction events includes both negative and positive immediate outcomes, with variable frequencies, for at least one of the interacting species. Our study shows that continua in interaction outcomes happen just by varying the sign and impact of individual events. The exact shape of the continua depends on the proportion of positive versus negative events and the relative magnitude of per-capita interaction strengths. Our model shows that continua in interaction outcomes are a key property of most pairwise interactions and are originated from the variable roles played by the interacting partners. It constitutes a step forward in the paradigm change from discrete categorization of ecological interactions to a new perspective over a continuous space.

Population origin determines the adaptive potential for the advancement of flowering onset in Lupinus angustifolius L. (Fabaceae).

In the present framework of global warming, it is unclear whether evolutionary adaptation can happen quick enough to preserve the persistence of many species. Specifically, we lack knowledge about the adaptive potential of the different populations in relation to the various constraints that may hamper particular adaptations. There is evidence indicating that early flowering often provides an adaptive advantage to plants in temperate zones in response to global warming. Thus, the objective of this study was to assess the adaptive potential for advancing flowering onset in Lupinus angustifolius L. (Fabaceae). Seeds from four populations from two contrasting latitudes in Spain were collected and sown in a common garden environment. Selecting the 25% of the individuals that flowered earlier in the first generation, over three generations, three different early flowering selection lines were established, involving both self-crosses and outcrosses. All artificial selection lines advanced their flowering significantly with respect to the control line in the northernmost populations, but not in the southern ones. Selection lines obtained from outcrossing had a greater advancement in flowering than those from self-crossing. No differences were found in the number or weight of the seeds produced between control and artificial selection lines, probably because plants in the common garden were drip irrigated. These results suggest that northern populations may have a greater adaptive potential and that southern populations may be more vulnerable in the context of climate warming. However, earlier flowering was also associated with changes in other traits (height, biomass, shoot growth, specific leaflet area, and leaflet dry matter content), and the effects of these changes varied greatly depending on the latitude of the population and selection line. Assessments of the ability of populations to cope with climate change through this and other approaches are essential to manage species and populations in a more efficient way.

Prevalence of mutualism in a simple model of microbial coevolution.

Evolutionary transitions among ecological interactions are widely known, although their detailed dynamics remain absent for most population models. Adaptive dynamics has been used to illustrate how the parameters of population models might shift through evolution, but within an ecological regime. Here we use adaptive dynamics combined with a generalized logistic model of population dynamics to show that transitions of ecological interactions might appear as a consequence of evolution. To this purpose, we introduce a two-microbial toy model in which population parameters are determined by a bookkeeping of resources taken from (and excreted to) the environment, as well as from the byproducts of the other species. Despite its simplicity, this model exhibits all kinds of potential ecological transitions, some of which resemble those found in nature. Overall, the model shows a clear trend toward the emergence of mutualism.

Climate change conditions the selection of rust-resistant candidate wild lentil populations for in situ conservation.

Crop Wild Relatives (CWR) are a valuable source of genetic diversity that can be transferred to commercial crops, so their conservation will become a priority in the face of climate change. Bizarrely, in situ conserved CWR populations and the traits one might wish to preserve in them are themselves vulnerable to climate change. In this study, we used a quantitative machine learning predictive approach to project the resistance of CWR populations of lentils to a common disease, lentil rust, caused by fungus Uromyces viciae-fabae. Resistance is measured through a proxy quantitative value, DSr (Disease Severity relative), quite complex and expensive to get. Therefore, machine learning is a convenient tool to predict this magnitude using a well-curated georeferenced calibration set. Previous works have provided a binary outcome (resistant vs. non-resistant), but that approach is not fine enough to answer three practical questions: which variables are key to predict rust resistance, which CWR populations are resistant to rust under current environmental conditions, and which of them are likely to keep this trait under different climate change scenarios. We first predict rust resistance in present time for crop wild relatives that grow up inside protected areas. Then, we use the same models under future climate IPCC (Intergovernmental Panel on Climate Change) scenarios to predict future DSr values. Populations that are rust-resistant by now and under future conditions are optimal candidates for further evaluation and in situ conservation of this valuable trait. We have found that rust-resistance variation as a result of climate change is not uniform across the geographic scope of the study (the Mediterranean basin), and that candidate populations share some interesting common environmental conditions.

Effects of parental drought on offspring fitness vary among populations of a crop wild relative.

Transgenerational plasticity is a form of non-genetic inheritance that can reduce or enhance offspring fitness depending on parental stress. Yet, the adaptive value of such parental environmental effects and whether their expression varies among populations remain largely unknown. We used self-fertilized lines from climatically distinct populations of the crop wild relative Lupinus angustifolius. In the parental generation, full-siblings were grown in two contrasting watering environments. Then, to robustly separate the within-generation and transgenerational response to drought, we reciprocally assigned the offspring of parents to the same experimental treatments. We measured key functional traits and assessed lifetime reproductive fitness. Offspring of drought-stressed parents produced less reproductive biomass, but a similar number of lighter seeds, in dry soil compared to offspring of genetically identical, well-watered parents, an effect not mediated by differences in seed provisioning. Importantly, while the offspring of parents grown in the favourable environment responded to drought by slightly increasing individual seed mass, the pattern of plasticity of the offspring of drought-grown parents showed the opposite direction, and the negative effects of parental drought on seed mass were more pronounced in populations from cooler and moist habitats. Overall, our results show that parental effects may override immediate adaptive responses to drought and provide evidence of population-level variation in the expression of transgenerational plasticity.

Seventeen 'extinct' plant species back to conservation attention in Europe.

Seventeen European endemic plant species were considered extinct, but improved taxonomic and distribution knowledge as well as ex situ collecting activities brought them out of the extinct status. These species have now been reported into a conservation framework that may promote legal protection and in situ and ex situ conservation.

Geography and Environment Shape Landscape Genetics of Mediterranean Alpine Species Silene ciliata Poiret. (Caryophyllaceae).

The study of the drivers that shape spatial genetic structure across heterogeneous landscapes is one of the main approaches used to understand population dynamics and responses in changing environments. While the Isolation-by-Distance model (IBD) assumes that genetic differentiation increases among populations with geographical distance, the Isolation-by-Resistance model (IBR) also considers geographical barriers and other landscape features that impede gene flow. On the other hand, the Isolation-by-Environment model (IBE) explains genetic differentiation through environmental differences between populations. Although spatial genetic studies have increased significantly in recent years, plants from alpine ecosystems are highly underrepresented, even though they are great suitable systems to disentangle the role of the different factors that structure genetic variation across environmental gradients. Here, we studied the spatial genetic structure of the Mediterranean alpine specialist Silene ciliata across its southernmost distribution limit. We sampled three populations across an altitudinal gradient from 1850 to 2400 m, and we replicated this sample over three mountain ranges aligned across an E-W axis in the central part of the Iberian Peninsula. We genotyped 20 individuals per population based on eight microsatellite markers and used different landscape genetic tools to infer the role of topographic and environmental factors in shaping observed patterns along the altitudinal gradient. We found a significant genetic structure among the studied Silene ciliata populations which was related to the orography and E-W configuration of the mountain ranges. IBD pattern arose as the main factor shaping population genetic differentiation. Geographical barriers between mountain ranges also affected the spatial genetic structure (IBR pattern). Although environmental variables had a significant effect on population genetic diversity parameters, no IBE pattern was found on genetic structure. Our study reveals that IBD was the driver that best explained the genetic structure, whereas environmental factors also played a role in determining genetic diversity values of this dominant plant of Mediterranean alpine environments.

Identification of ecogeographical gaps in the Spanish Aegilops collections with potential tolerance to drought and salinity.

Drought, one of the most important abiotic stress factors limiting biomass, significantly reduces crop productivity. Salinization also affects the productivity of both irrigated and rain-fed wheat crops. Species of genus Aegilops can be considered crop wild relatives (CWR) of wheat and have been widely used as gene sources in wheat breeding, especially in providing resistance to pests and diseases. Five species (Ae. biuncialis, Ae. geniculata, Ae. neglecta, Ae. triuncialis and Ae. ventricosa) are included in the Spanish National Inventory of CWRs. This study aimed to identify ecogeographic gaps in the Spanish Network on Plant Genetic Resources for Food and Agriculture (PGRFA) with potential tolerance to drought and salinity. Data on the Spanish populations of the target species collected and conserved in genebanks of the Spanish Network on PGRFA and data on other population occurrences in Spain were compiled and assessed for their geo-referencing quality. The records with the best geo-referencing quality values were used to identify the ecogeographical variables that might be important for Aegilops distribution in Spain. These variables were then used to produce ecogeographic land characterization maps for each species, allowing us to identify populations from low and non-represented ecogeographical categories in ex situ collections. Predictive characterization strategy was used to identify 45 Aegilops populations in these ecogeographical gaps with potential tolerance to drought and salinity conditions. Further efforts are being made to collect and evaluate these populations.

Ranking of critical species to preserve the functionality of mutualistic networks using the k-core decomposition.

BACKGROUND: Network analysis has become a relevant approach to analyze cascading species extinctions resulting from perturbations on mutualistic interactions as a result of environmental change. In this context, it is essential to be able to point out key species, whose stability would prevent cascading extinctions, and the consequent loss of ecosystem function. In this study, we aim to explain how the k-core decomposition sheds light on the understanding the robustness of bipartite mutualistic networks. METHODS: We defined three k-magnitudes based on the k-core decomposition: k-radius, k-degree, and k-risk. The first one, k-radius, quantifies the distance from a node to the innermost shell of the partner guild, while k-degree provides a measure of centrality in the k-shell based decomposition. k-risk is a way to measure the vulnerability of a network to the loss of a particular species. Using these magnitudes we analyzed 89 mutualistic networks involving plant pollinators or seed dispersers. Two static extinction procedures were implemented in which k-degree and k-risk were compared against other commonly used ranking indexes, as for example MusRank, explained in detail in Material and Methods. RESULTS: When extinctions take place in both guilds, k-risk is the best ranking index if the goal is to identify the key species to preserve the giant component. When species are removed only in the primary class and cascading extinctions are measured in the secondary class, the most effective ranking index to identify the key species to preserve the giant component is k-degree. However, MusRank index was more effective when the goal is to identify the key species to preserve the greatest species richness in the second class. DISCUSSION: The k-core decomposition offers a new topological view of the structure of mutualistic networks. The new k-radius, k-degree and k-risk magnitudes take advantage of its properties and provide new insight into the structure of mutualistic networks. The k-risk and k-degree ranking indexes are especially effective approaches to identify key species to preserve when conservation practitioners focus on the preservation of ecosystem functionality over species richness.

A glacial survivor of the alpine Mediterranean region: phylogenetic and phylogeographic insights into Silene ciliata Pourr. (Caryophyllaceae).

Silene ciliata Pourr. (Caryophyllaceae) is a species with a highly disjunct distribution which inhabits the alpine mountains of the Mediterranean Basin. We investigated the phylogeny and phylogeography of the species to (a) clarify the long-suggested division of S. ciliata into two subspecies, (b) evaluate its phylogenetic origin and (c) assess whether the species' diversification patterns were affected by the Mediterranean relief. For this purpose, we collected DNA from 25 populations of the species that inhabit the mountains of Portugal, Spain, France, Italy, former Yugoslav Republic of Macedonia, Bulgaria and Greece and studied the plastid regions rbcL, rps16 and trnL. Major intraspecific variation was supported by all analyses, while the possibility of the existence of more varieties or subspecies was not favoured. Plastid DNA (cpDNA) evidence was in accordance with the division of S. ciliata into the two subspecies, one spreading west (Iberian Peninsula and Central Massif) and the other east of the Alps region (Italian and Balkan Peninsula). This study proposes that the species' geographically disconnected distribution has probably derived from vicariance processes and from the Alps acting as a barrier to the species' dispersal. The monophyletic origin of the species is highly supported. cpDNA patterns were shown independent of the chromosome evolution in the populations and could have resulted from a combination of geographic factors providing links and barriers, climatic adversities and evolutionary processes that took place during Quaternary glaciations.

Rethinking the logistic approach for population dynamics of mutualistic interactions.

Mutualistic communities have an internal structure that makes them resilient to external perturbations. Late research has focused on their stability and the topology of the relations between the different organisms to explain the reasons of the system robustness. Much less attention has been invested in analyzing the systems dynamics. The main population models in use are modifications of the r-K formulation of logistic equation with additional terms to account for the benefits produced by the interspecific interactions. These models have shortcomings as the so-called r-K formulation diverges under some conditions. In this work, we introduce a model for population dynamics under mutualism that preserves the original logistic formulation. It is mathematically simpler than the widely used type II models, although it shows similar complexity in terms of fixed points and stability of the dynamics. We perform an analytical stability analysis and numerical simulations to study the model behavior in general interaction scenarios including tests of the resilience of its dynamics under external perturbations. Despite its simplicity, our results indicate that the model dynamics shows an important richness that can be used to gain further insights in the dynamics of mutualistic communities.

URJC GB dataset: Community-based seed bank of Mediterranean high-mountain and semi-arid plant species at Universidad Rey Juan Carlos (Spain).

The Germplasm Bank of Universidad Rey Juan Carlos was created in 2008 and currently holds 235 accessions and 96 species. This bank focuses on the conservation of wild-plant communities and aims to conserve ex situ a representative sample of the plant biodiversity present in a habitat, emphasizing priority ecosystems identified by the Habitats Directive. It is also used to store plant material for research and teaching purposes. The collection consists of three subcollections, two representative of typical habitats in the center of the Iberian Peninsula: high-mountain pastures (psicroxerophylous pastures) and semi-arid habitats (gypsophylic steppes), and a third representative of the genus Lupinus. The high-mountain subcollection currently holds 153 accessions (63 species), the semi-arid subcollection has 76 accessions (29 species,) and the Lupinus subcollection has 6 accessions (4 species). All accessions are stored in a freezer at -18 °C in Kilner jars with silica gel. The Germplasm Bank of Universidad Rey Juan Carlos follows a quality control protocol which describes the workflow performed with seeds from seed collection to storage. All collectors are members of research groups with great experience in species identification. Herbarium specimens associated with seed accessions are preserved and 63% of the records have been georreferenced with GPS and radio points. The dataset provides unique information concerning the location of populations of plant species that form part of the psicroxerophylous pastures and gypsophylic steppes of Central Spain as well as populations of genus Lupinus in the Iberian Peninsula. It also provides relevant information concerning mean seed weight and seed germination values under specific incubation conditions. This dataset has already been used by researchers of the Area of Biodiversity and Conservation of URJC as a source of information for the design and implementation of experimental designs in these plant communities. Since they are all active subcollections in continuous growth, data is updated regularly every six months and the latest version can be accessed through the GBIF data portal at http://www.gbif.es:8080/ipt/resource.do?r=germoplasma-urjc. This paper describes the URJC Germplasm Bank and its associated dataset with the aim of disseminating the dataset and explaining how it was derived.

Unravelling genetics at the top: mountain islands or isolated belts?

BACKGROUND AND AIMS: In mountain plant populations, local adaptation has been described as one of the main responses to climate warming, allowing plants to persist under stressful conditions. This is especially the case for marginal populations at their lowest elevation, as they are highly vulnerable. Adequate levels of genetic diversity are required for selection to take place, while high levels of altitudinal gene flow are seen as a major limiting factor potentially precluding local adaptation processes. Thus, a compromise between genetic diversity and gene flow seems necessary to guarantee persistence under oncoming conditions. It is therefore critical to determine if gene flow occurs preferentially between mountains at similar altitudinal belts, promoting local adaptation at the lowest populations, or conversely along altitude within each mountain. METHODS: Microsatellite markers were used to unravel genetic diversity and population structure, inbreeding and gene flow of populations at two nearby altitudinal gradients of Silene ciliata, a Mediterranean high-mountain cushion plant. KEY RESULTS: Genetic diversity and inbreeding coefficients were similar in all populations. Substantial gene flow was found both along altitudinal gradients and horizontally within each elevation belt, although greater values were obtained along altitudinal gradients. Gene flow may be responsible for the homogeneous levels of genetic diversity found among populations. Bayesian cluster analyses also suggested that shifts along altitudinal gradients are the most plausible scenario. CONCLUSIONS: Past population shifts associated with glaciations and interglacial periods in temperate mountains may partially explain current distributions of genetic diversity and population structure. In spite of the predominance of gene flow along the altitudinal gradients, local genetic differentiation of one of the lower populations together with the detection of one outlier locus might support the existence of different selection forces at low altitudes.

Characterization of microsatellites in the mountain plant Armeria caespitosa (Plumbaginaceae) and transferability to congeners.

PREMISE OF THE STUDY: The focus of this study is to develop microsatellite markers in Armeria caespitosa, a narrow endemic of central Spain. Microsatellite loci are sought to clarify population structure and estimate gene flux among populations. METHODS AND RESULTS: Enriched microsatellite genomic libraries were used for microsatellite isolation. Sixteen microsatellite loci were characterized, eight of which can be used for successful genotyping. Allele number ranged from two to seven per locus. Observed and expected heterozygosity ranged from 0.300 to 0.800 and from 0.296 to 0.733, respectively. Cross-amplification of seven and six loci was successful for A. maritima and A. cantabrica, respectively. CONCLUSIONS: These microsatellites are suitable in the study of population genetics and gene flow among A. caespitosa populations. The information provided by these markers may be useful in the study of this plant's response to global warming.

Seedling dynamics at elevation limits: Complex interactions beyond seed and microsite limitations.

PREMISE OF THE STUDY: Limited availability of seeds and microsites are important constraints for seedling emergence and survival. However, little is known about population-level feedbacks between seed and microsite limitation and how these relationships vary in contrasting conditions. Focusing on Armeria caespitosa, a high-mountain endemic, we asked whether seedling establishment was simultaneously limited by seed and microsite availability, whether the balance between seed and microsite limitations varied with contrasting environments, and whether seed and microsite limitations interacted with each other. • METHODS: We studied seedling emergence and survival at the edges of the elevation range of A. caespitosa in central Spain over 4 years. We used mixed linear models and structural equation modeling. • KEY RESULTS: Our results showed that A. caespitosa recruits similar numbers of new seedlings per plot in contrasting environments. Seedling emergence and survival in A. caespitosa were both limited by seed and microsite availability. Habitat suitability (the surrogate of microsite limitation) varied between populations and indirectly affected fertility (the surrogate of seed limitation). • CONCLUSIONS: Seedling emergence and survival are both seed and microsite limited, there are interactions between seed and microsite limitations, and both factors, which control seedling establishment, and their interconnections, vary among local populations.

Inherited variability in multiple traits determines fitness in populations of an annual legume from contrasting latitudinal origins.

BACKGROUND AND AIMS: Variation in fitness depends on corresponding variation in multiple traits which have both genetically controlled and plastic components. These traits are subjected to varying degrees of local adaptation in specific populations and, consequently, are genetically controlled to different extents. In this study it is hypothesized that modulation of different traits would have contrasting relevance for the fitness of populations of diverse origins. Specifically, assuming that environmental pressures vary across a latitudinal gradient, it is suggested that inherited variation in traits differentially determines fitness in annual Lupinus angustifolius populations from contrasting latitudinal origins in western Spain. METHODS: Seeds of L. angustifolius from three contrasting origins were grown in a common garden. Traits related to more plastic vegetative growth and more genetically conserved phenology were measured, together with estimates of reproductive success. Fitness was estimated by the number of viable seeds per plant. Structural Equation Models were used to infer causal relationships among multiple traits and fitness, separating the direct and indirect effects of morphological, phenological and reproductive traits. KEY RESULTS: Phenological, vegetative and reproductive traits accounted for most of the fitness variation. Fitness was highest in plants of southernmost origin, mainly due to earlier flowering. Fitness within each seed origin was controlled by variation in different traits. Southern origin plants that grew to a larger size achieved higher fitness. However, plant size in plants of northernmost origin was irrelevant, but early flowering promoted higher fitness. Variation in fruit and seed set had a greater effect on the fitness of plants of central origin than phenological and size variation. CONCLUSIONS: It is concluded that modulation of a functional trait can be relevant to fitness in a given population (i.e. affecting intensity and direction), but irrelevant in other populations. This points to the need to consider integrated phenotypes when trying to unravel local adaptation effects over single traits.

Genetic fingerprinting of germplasm accessions as an aid for species conservation: a case study with Borderea chouardii (Dioscoreaceae), one of the most critically endangered Iberian plants.

BACKGROUND AND AIMS: Molecular markers have changed previous expectations about germplasm collections of endangered plants, as new perspectives aim at holding a significant representation of all the genetic diversity in the studied species to accomplish further conservation initiatives successfully. Borderea chouardii is a critically endangered allotetraploid dioecious member of Dioscoreaceae, known from a single population in the Iberian pre-Pyrenees. This population was reported to be highly structured into two genetically distinct groups of individuals corresponding to their spatial separation along the vertical cliff where it grows. In 1999, the Spanish Government of Aragón launched the first conservation programme for the ex situ preservation of this species, and since then a seed collection has been conserved at the Germplasm Bank of the Universidad Politécnica de Madrid. However, as some seed samples had not been labelled clearly at the time of collection, their origin was uncertain. METHODS: Genetic variation in germplasm accessions of B. chouardii was investigated using microsatellite (simple sequence repeat; SSR) markers. KEY RESULTS: The 17 primer pairs used detected 62 SSR alleles in the 46 samples analysed from five different germplasm stocks. Eight alleles scored from the wild population were not detected in the germplasm samples analysed. The relatedness of the germplasm samples to the wild subpopulations through neighbour-joining clustering, principal coordinates analysis (PCO) and assignment tests revealed a biased higher representation of the genetic diversity of the lower cliff (43 samples) subpopulation than that of the upper cliff (three samples). CONCLUSIONS: The collection of additional samples from the upper cliff is recommended to achieve a better representation of the genetic diversity of this subpopulation. It is also recommended that these stocks should be managed separately according to their distinct microspatial origin in order to preserve the genetic substructuring of the wild population.