Extensive pollen-mediated gene flow across intensively managed landscapes in an insect-pollinated shrub native to semiarid habitats.

Our knowledge of the impact of landscape fragmentation on gene flow patterns is mainly drawn from tropical and temperate ecosystems, where landscape features, such as the distance of a tree to the forest edge, drive connectivity and mating patterns. Yet, the structure of arid and semiarid plant communities - with open canopies and a scattered distribution of trees - differs greatly from those that are well-characterized in the literature. As a result, we ignore whether the documented consequences of landscape fragmentation on plant mating and gene flow patterns also hold for native plant communities in arid and semiarid regions. We investigated the relative contribution of plant traits, pollinator activity, and individual neighbourhood in explaining variation in mating and gene flow patterns of an insect-pollinated semiarid arborescent shrub, Ziziphus lotus, at three sites embedded in highly altered agriculture landscapes. We used 14 SSRs, seed paternity analyses, and individual mixed effect mating models (MEMMi) to estimate the individual mating variables and the pollen dispersal kernel at each site. Individual spatial location, flower density, and floral visitation rate explained most of the variation of mating variables. Unexpectedly, individual correlated paternity was very low and shrubs surrounded by the most degraded matrix exhibited an increased fraction of pollen immigration and a high effective number of pollen donors per mother shrub. Overall, our results reveal that an active pollinator assemblage ensures highly efficient mating, and maintains pollen-mediated gene flow and notable connectivity levels, even in highly altered landscapes, potentially halting genetic isolation within and between distant sites.

Plant species abundance and phylogeny explain the structure of recruitment networks.

Established plants can affect the recruitment of young plants, filtering out some and allowing the recruitment of others, with profound effects on plant community dynamics. Recruitment networks (RNs) depict which species recruit under which others. We investigated whether species abundance and phylogenetic distance explain the structure of RNs across communities. We estimated the frequency of canopy-recruit interactions among woody plants in 10 forest assemblages to describe their RNs. For each RN, we determined the functional form (linear, power or exponential) best describing the relationship of interaction frequency with three predictors: canopy species abundance, recruit species abundance and phylogenetic distance. We fitted models with all combinations of predictor variables, from which we simulated RNs. The best functional form of each predictor was the same in most communities (linear for canopy species abundance, power for recruit species abundance and exponential for phylogenetic distance). The model including all predictor variables was consistently the best in explaining interaction frequency and showed the best performance in predicting RN structure. Our results suggest that mechanisms related to species abundance are necessary but insufficient to explain the assembly of RNs. Evolutionary processes affecting phylogenetic divergence are critical determinants of RN structure.

Variation in functional responses to water stress and differentiation between natural allopolyploid populations in the Brachypodium distachyon species complex.

Background and Aims: Some polyploid species show enhanced physiological tolerance to drought compared with their progenitors. However, very few studies have examined the consistency of physiological drought response between genetically differentiated natural polyploid populations, which is key to evaluation of the importance of adaptive evolution after polyploidization in those systems where drought exerts a selective pressure. Methods: A comparative functional approach was used to investigate differentiation of drought-tolerance-related traits in the Brachypodium species complex, a model system for grass polyploid adaptive speciation and functional genomics that comprises three closely related annual species: the two diploid parents, B. distachyon and B. stacei, and the allotetraploid derived from them, B. hybridum. Differentiation of drought-tolerance-related traits between ten genetically distinct B. hybridum populations and its ecological correlates was further analysed. Key Results: The functional drought response is overall well differentiated between Brachypodium species. Brachypodium hybridum allotetraploids showed a transgressive expression pattern in leaf phytohormone content in response to drought. In contrast, other B. hybridum physiological traits correlated to B. stacei ones. Particularly, proline and water content were the traits that best discriminated these species from B. distachyon under drought. Conclusions: After polyploid formation and/or colonization, B. hybridum populations have adaptively diverged physiologically and genetically in response to variations in aridity.

The interplay between aridity and competition determines colonization ability, exclusion and ecological segregation in the heteroploid Brachypodium distachyon species complex.

A higher competitive advantage of polyploid plants compared with their parental diploids is frequently invoked to explain their establishment success, colonization of novel environments and cytotypic ecological segregation, yet there is scarce experimental evidence supporting such hypotheses. Here, we investigated whether differential competitive ability of species of the Brachypodium distachyon (Poaceae) species complex, a model system for genomic, ecological and evolutionary studies of temperate grasses, contributes to explaining their ecological segregation as well as their coexistence in diploid/allotetraploid contact zones. We conducted two field experiments in dry and humid localities to evaluate the tolerance to competition of diploids and allotetraploids in densely occupied environments, and to parameterize models of intra- and intercytotype competition as a mechanism for species exclusion/coexistence. We provide experimental evidence supporting the hypothesis that, under natural field conditions, allotetraploids have superior ecological success compared with one of their parental diploids in terms of both colonizing competitive habitats and intercytotypic competition, with the balance of intra/intercytotype competition favoring polyploid population establishment. These findings, together with previous data on ecogeographic segregation and adaptive response to water stress, suggest that the interplay between aridity and competitive outcome determines the ability to colonize competitive environments, the exclusion of diploids, especially in arid localities, and species geographic segregation.

Facilitation contributes to Mediterranean woody plant diversity but does not shape the diversity-productivity relationship along aridity gradients.

The diversity-productivity relationship (humped-back model (HBM)) and the stress-gradient (SGH) hypotheses may be connected when productivity is limited primarily by aridity. We analytically connect both hypotheses and assess the contribution of facilitation to woody plant richness along the aridity gradient of the Western Mediterranean floristic region. We monitored regeneration niches of woody plants, obtaining rarefied species richness and plant relative interaction indices in 54 forests and scrublands in a 1750-km geographical range across Spain, Morocco and the Canary Islands. We verified the monotonic increase in facilitation with aridity postulated by SGH and the humped-shape pattern of species richness expected from HBM, which became manifest after expanding the aridity gradient or crossing vegetation types. Along the gradient, interaction balance turned into facilitation earlier in forest than in scrublands. The effects of aridity and interaction balance on species diversity were additive rather than interdependent. Facilitation is an important driver of woody species richness at macroecological scales because it added up to diversity in most sites, with enhanced contribution with increased stress. The HBM was not shaped by species interactions. Results suggest that facilitation may act in Mediterranean vegetation buffering against critical transitions between states allowing woody plant communities to cope with the rise in aridity expected with global warming.

Environmental niche variation and evolutionary diversification of the Brachypodium distachyon grass complex species in their native circum-Mediterranean range.

UNLABELLED: • PREMISE OF THE STUDY: We conducted environmental niche modeling (ENM) of the Brachypodium distachyon s.l. complex, a model group of two diploid annual grasses (B. distachyon, B. stacei) and their derived allotetraploid (B. hybridum), native to the circum-Mediterranean region. We (1) investigated the ENMs of the three species in their native range based on present and past climate data; (2) identified potential overlapping niches of the diploids and their hybrid across four Quaternary windows; (3) tested whether speciation was associated with niche divergence/conservatism in the complex species; and (4) tested for the potential of the polyploid outperforming the diploids in the native range.• METHODS: Geo-referenced data, altitude, and 19 climatic variables were used to construct the ENMs. We used paleoclimate niche models to trace the potential existence of ancestral gene flow among the hybridizing species of the complex.• KEY RESULTS: Brachypodium distachyon grows in higher, cooler, and wetter places, B. stacei in lower, warmer, and drier places, and B. hybridum in places with intermediate climatic features. Brachypodium hybridum had the largest niche overlap with its parent niches, but a similar distribution range and niche breadth.• CONCLUSIONS: Each species had a unique environmental niche though there were multiple niche overlapping areas for the diploids across time, suggesting the potential existence of several hybrid zones during the Pleistocene and the Holocene. No evidence of niche divergence was found, suggesting that species diversification was not driven by ecological speciation but by evolutionary history, though it could be associated to distinct environmental adaptations.

Glandular trichomes as an inflorescence defence mechanism against insect herbivores in Iberian columbines.

Glandular trichomes play a defensive role against herbivores in the leaves of many plant species. However, their functional role in inflorescences has not been studied, even though theory suggests that tissues with a higher fitness value, such as inflorescences, should be better defended. Using manipulative experiments, we analysed the defensive role of glandular trichomes against herbivorous insects in the inflorescence of Iberian columbines (genus Aquilegia), and its inter-population and inter-taxa variation in relation to herbivore abundance and potential selective pressure. The experiments were conducted in eight populations belonging to four subspecies of two columbines (Aquilegia vulgaris and Aquilegia pyrenaica). For each population, we estimated the density of glandular trichomes in the inflorescences, the abundance of insects stuck in the inflorescences, the abundance of small herbivorous insects, the incidence of damage on flowers and fruits, and the fruit set. The density of glandular trichomes on the inflorescence of A. vulgaris and A. pyrenaica was higher in regions of higher herbivore abundance. We also found that when the plants lose the protection of glandular trichomes, small insects have better access to flowers and fruits, causing more damage and reducing plant fitness. This study concludes that glandular trichomes are part of an adaptive response against phytophagous insect herbivory. The observed variation in herbivore pressure between taxa, likely caused by habitat differentiation, might have played a role in trait differentiation through divergent selection. This result adds evidence to the differentiation of the Iberian columbines through habitat specialization.

Linking topological structure and dynamics in ecological networks.

Interaction networks are basic descriptions of ecological communities and are at the core of community dynamics models. Knowledge of their structure should enable us to understand dynamical properties of ecological communities. However, the relationships between dynamical properties of communities and qualitative descriptors of network structure remain unclear. To improve our understanding of such relationships, we develop a framework based on the concept of strongly connected components, which are key structural components of networks necessary to explain stability properties such as persistence and robustness. We illustrate this framework for the analysis of qualitative empirical food webs and plant-plant interaction networks. Both types of networks exhibit high persistence (on average, 99% and 80% of species, respectively, are expected to persist) and robustness (only 0.2% and 2% of species are expected to disappear following the extinction of a species). Each of the networks is structured as a large group of interconnected species accompanied by much smaller groups that most often consist of a single species. This low-modularity configuration can be explained by a negative modularity-stability relationship. Our results suggest that ecological communities are not typically structured in multispecies compartments and that compartmentalization decreases robustness.

Environmental aridity is associated with cytotype segregation and polyploidy occurrence in Brachypodium distachyon (Poaceae).

• The ecological and adaptive significance of plant polyploidization is not well understood and no clear pattern of association between polyploid frequency and environment has emerged. Climatic factors are expected to predict cytotype distribution. However, the relationship among climate, cytotype distribution and variation of abiotic stress tolerance traits has rarely been examined. • Here, we use flow cytometry and root-tip squashes to examine the cytotype distribution in the temperate annual grass Brachypodium distachyon in 57 natural populations distributed across an aridity gradient in the Iberian Peninsula. We further investigate the link between environmental aridity, ploidy, and variation of drought tolerance and drought avoidance (flowering time) traits. • Distribution of diploids (2n = 10) and allotetraploids (2n = 30) in this species is geographically structured throughout its range in the Iberian Peninsula, and is associated with aridity gradients. Importantly, after controlling for geographic and altitudinal effects, the link between aridity and polyploidization occurrence persisted. Water-use efficiency varied between ploidy levels, with tetraploids being more efficient in the use of water than diploids under water-restricted growing conditions. • Our results indicate that aridity is an important predictor of polyploid occurrence in B. distachyon, suggesting a possible adaptive origin of the cytotype segregation.

Intra-population comparison of vegetative and floral trait heritabilities estimated from molecular markers in wild Aquilegia populations.

Measuring heritable genetic variation is important for understanding patterns of trait evolution in wild populations, and yet studies of quantitative genetic parameters estimated directly in the field are limited by logistic constraints, such as the difficulties of inferring relatedness among individuals in the wild. Marker-based approaches have received attention because they can potentially be applied directly to wild populations. For long-lived, self-compatible plant species where pedigrees are inadequate, the regression-based method proposed by Ritland has the appeal of estimating heritabilities from marker-based estimates of relatedness. The method has been difficult to implement in some plant populations, however, because it requires significant variance in relatedness across the population. Here, we show that the method can be readily applied to compare the ability of different traits to respond to selection, within populations. For several taxa of the perennial herb genus Aquilegia, we estimated heritabilities of floral and vegetative traits and, combined with estimates of natural selection, compared the ability to respond to selection of both types of traits under current conditions. The intra-population comparisons showed that vegetative traits have a higher potential for evolution, because although they are as heritable as floral traits, selection on them is stronger. These patterns of potential evolution are consistent with macroevolutionary trends in the European lineage of the genus.

Assessing ecological specialization of an ant-seed dispersal mutualism through a wide geographic range.

Specialization in species interactions is of central importance for understanding the ecological structure and evolution of plant-animal mutualisms. Most plant-animal mutualisms are facultative and strongly asymmetric. In particular, myrmecochory (seed dispersal by ants) has been regarded as a very generalized interaction. Although some recent studies have suggested that only a few ant species are really important for dispersal, no rigorous measurement of the specialization in ant-seed dispersal mutualisms has been performed. Here, we use individual plants as basic units for replication to investigate the generalization-specialization of the herb Helleborus foetidus on its ant dispersers over a considerable part of its geographical range. We define generalization in terms of diversity components (species richness and evenness) of the ant visitor that realizes dispersal by removing diaspores. We obtain truly comparable values of ant visitor diversity, distinguishing among different functional groups of visitors and identifying incidental visitors and real ant dispersers. Using null model approaches, we test the null hypothesis that ant-mediated dispersal is a generalized mutualism. At least two premises should be confirmed to validate the hypothesis: (1) diaspores are dispersed by multiple ant-visitor species, and (2) diaspore dispersal is significantly equitable. Though up to 37 ant species visited diaspores across 10 populations, only two large formicines, Camponotus cruentatus and Formica lugubris, were responsible for the vast majority of visits resulting in dispersal in most populations and years, which strongly suggests that ant seed dispersal in H. foetidus is ecologically specialized. Interestingly, specialization degree was unrelated to dispersal success across populations. Our study offers new insights into the spatiotemporal dynamics of myrmecochory. We propose the existence of an alternative scenario to extensive generalization. In this new scenario, generalization is replaced by ecological specialization, which is determined by the intrinsic traits of the plant species rather than by the ecological context in which the interaction takes place.

Climate change decreases suitable areas for rapeseed cultivation in Europe but provides new opportunities for white mustard as an alternative oilseed for biofuel production.

Oilseed crops, including several mustards, are cultivated as biofuel sources worldwide. However, common mustard crops (e.g., the rapeseed Brassica napus) grow naturally in mesic temperate regions, which are expected to be impaired by global warming and increased aridity. In particular, increased aridity is predicted to reduce the oil concentration and seed yield of rapeseed crops. There is therefore an urgent need to identify alternative bioenergy crops that are preadapted to future climatic conditions. An alternative to conventional Brassica species for biodiesel production is the white mustard Sinapis alba, which is native to the circum-Mediterranean region and has a high seed lipid content. S. alba grows spontaneously in olive groves and other widespread Mediterranean crops; accordingly, it could be easily cultivated by companion planting to improve ecosystem function by decreasing soil loss, controlling microbial disease, and assisting in the maintenance of biodiversity. In this study, using species distribution modeling, we predicted climatically suitable areas for the cultivation of S. alba in Western Europe across the Mediterranean Basin under present climatic conditions and several climate change scenarios. We show that current climatically suitable areas for S. alba cultivation do not overlap with those for B. napus. Unlike B. napus, S. alba could be cultivated throughout most of the circum-Mediterranean region. According to our models, increases in aridity and annual mean temperatures will expand the climatically suitable areas for S. alba in the Mediterranean Basin. However, suitable areas for the cultivation of B. napus will decrease significantly in Western Europe. Our results indicate that S. alba is a strong, environmentally safe candidate for biofuel production throughout the Mediterranean Basin and other Western European countries, especially under climate change scenarios that are expected to impair current oilseed crops.

Development and characterization of microsatellite primers in the endangered Mediterranean shrub Ziziphus lotus (Rhamnaceae).

PREMISE OF THE STUDY: Microsatellite primers were developed to characterize and evaluate patterns of genetic diversity and structure in the endangered Mediterranean shrub Ziziphus lotus (Rhamnaceae). METHODS AND RESULTS: Twenty microsatellite primers were developed for Z. lotus, of which 14 were polymorphic. We evaluated microsatellite polymorphism in 97 specimens from 18 Spanish and seven Moroccan populations. Between two and eight alleles were found per locus, and the average number of alleles was 5.54. Observed heterozygosity and expected heterozygosity ranged from 0.08 to 0.90 and from 0.08 to 0.82, respectively. Nine of these primers also amplified microsatellite loci in Z. jujuba. CONCLUSIONS: The microsatellite markers described here will be useful in studies on genetic variation, population genetic structure, and gene flow in the fragmented habitat of this species. These markers are a valuable resource for designing appropriate conservation measures for the species in the Mediterranean range.

Geographic mosaic of plant evolution: extrafloral nectary variation mediated by ant and herbivore assemblages.

Herbivory is an ecological process that is known to generate different patterns of selection on defensive plant traits across populations. Studies on this topic could greatly benefit from the general framework of the Geographic Mosaic Theory of Coevolution (GMT). Here, we hypothesize that herbivory represents a strong pressure for extrafloral nectary (EFN) bearing plants, with differences in herbivore and ant visitor assemblages leading to different evolutionary pressures among localities and ultimately to differences in EFN abundance and function. In this study, we investigate this hypothesis by analyzing 10 populations of Anemopaegma album (30 individuals per population) distributed through ca. 600 km of Neotropical savanna and covering most of the geographic range of this plant species. A common garden experiment revealed a phenotypic differentiation in EFN abundance, in which field and experimental plants showed a similar pattern of EFN variation among populations. We also did not find significant correlations between EFN traits and ant abundance, herbivory and plant performance across localities. Instead, a more complex pattern of ant-EFN variation, a geographic mosaic, emerged throughout the geographical range of A. album. We modeled the functional relationship between EFNs and ant traits across ant species and extended this phenotypic interface to characterize local situations of phenotypic matching and mismatching at the population level. Two distinct types of phenotypic matching emerged throughout populations: (1) a population with smaller ants (Crematogaster crinosa) matched with low abundance of EFNs; and (2) seven populations with bigger ants (Camponotus species) matched with higher EFN abundances. Three matched populations showed the highest plant performance and narrower variance of EFN abundance, representing potential plant evolutionary hotspots. Cases of mismatched and matched populations with the lowest performance were associated with abundant and highly detrimental herbivores. Our findings provide insights on the ecology and evolution of plant-ant guarding systems, and suggest new directions to research on facultative mutualistic interactions at wide geographic scales.

Natural variation, differentiation, and genetic trade-offs of ecophysiological traits in response to water limitation in Brachypodium distachyon and its descendent allotetraploid B. hybridum (Poaceae).

Differences in tolerance to water stress may underlie ecological divergence of closely related ploidy lineages. However, the mechanistic basis of physiological variation governing ecogeographical cytotype segregation is not well understood. Here, using Brachypodium distachyon and its derived allotetraploid B. hybridum as model, we test the hypothesis that, for heteroploid annuals, ecological divergence of polyploids in drier environments is based on trait differentiation enabling drought escape. We demonstrate that under water limitation allotetraploids maintain higher photosynthesis and stomatal conductance and show earlier flowering than diploids, concordant with a drought-escape strategy to cope with water stress. Increased heterozygosity and greater genetic variability and plasticity of polyploids could confer a superior adaptive capability. Consistent with these predictions, we document (1) greater standing within-population genetic variation in water-use efficiency (WUE) and flowering time in allotetraploids, and (2) the existence of (nonlinear) environmental clines in physiology across allotetraploid populations. Increased gas exchange and diminished WUE occurred at the driest end of the gradient, consistent with a drought-escape strategy. Finally, we found that allotetraploids showed weaker genetic correlations than diploids congruous with the expectation of relaxed pleiotropic constraints in polyploids. Our results suggest evolutionary divergence of ecophysiological traits in each ploidy lineage.

Plant traits, environmental factors, and pollinator visitation in winter-flowering Helleborus foetidus (Ranunculaceae).

BACKGROUND AND AIMS: This study examined the effect of plant traits and environmental factors on pollinator visitation in the winter-flowering Helleborus foetidus (Ranunculaceae) in three distant regions in the Iberian Peninsula. METHODS: Geographical variation in floral visitor assemblage, plant traits and environmental factors were analysed during the flowering season. KEY RESULTS: Differences were found in all plant traits measured (number of open flowers, flower size, number of stamens per flower, and number of nectaries) both within and among regions, and differences among regions in all the environmental factors considered (air temperature, exposure to sunlight, canopy cover, and distance to the nearest neighbour). Differences were also found among regions in the probability that plants would be visited by pollinators. CONCLUSIONS: The results show that, although floral display (i.e. number of open flowers on a plant on a given day) consistently explained among-plant differences in visitation rate in all regions, visitation rate was not significantly affected by any other biological or environmental variable. In Helleborus foetidus, then, 'how' the plant is would seem to be more important than 'where' is it.

Interaction of pollinators and herbivores on plant fitness suggests a pathway for correlated evolution of mutualism- and antagonism-related traits.

Different kinds of plant-animal interactions are ordinarily studied in isolation, yet considering the combined fitness effects of mutualistic and antagonistic interactions is essential to understanding plant character evolution. Functional, structural, or phylogenetic associations between attractive and defensive traits may be nonadaptive or result from correlational selection on sets of herbivory- and pollination-linked traits. Nonadditivity of fitness effects of mutualists and antagonists, a requisite for correlational selection, was experimentally tested in the field. We created experimental populations of the insect-pollinated perennial herb, Helleborus foetidus, at 16 different locations distributed among three regions in the Iberian Peninsula. Plants experienced one of four possible selective regimes generated by independently weakening the effects of pollinators and herbivores (flower and fruit predators) according to a two-way fully factorial design. Effects were assessed in terms of number of next-generation offspring recruited per mother plant under natural field conditions. Differences among H. foetidus plants in the strength of their interactions with pollinators and herbivores translated into differential fitness, as measured in terms of recruited offspring, and subsequent changes in plant population densities. A strong, geographically consistent nonadditivity in the fitness consequences of pollinators and herbivores was found also. Plants possessing the particular combination of "traits" simultaneously enhancing pollination and escape from herbivores enjoyed a disproportionate fitness advantage over plants possessing any of the other three possible "trait" combinations. Results suggest a simple, possibly widespread ecological pathway favoring the adaptive correlated evolution of mutualism- and antagonism-related plant traits in pollinator-dependent plants suffering intense flower and fruit herbivory.