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Recent plant genomic studies provide fine-grained details on the evolutionary consequences of adaptive introgression during crop domestication. Modern genomic approaches and analytical methods now make it possible to better separate the introgression signal from the demographic signal thus providing a more comprehensive and complex picture of the role of introgression in local adaptation. Adaptive introgression has been fundamental for crop expansion and has involved complex patterns of gene flow. In addition to providing new and more favorable alleles of large effect, introgression during the early stages of domestication also increased allelic diversity at adaptive loci. Previous studies have largely underestimated the effect of such increased diversity following introgression. Recent genomic studies in wheat, potato, maize, grapevine, and ryegrass show that introgression of multiple genes, of as yet unknown effect, increased the effectiveness of purifying selection, and promoted disruptive or fluctuating selection in early cultivars and landraces. Historical selection processes associated with introgression from crop wild relatives provide an instructive analog for adaptation to current climate change and offer new avenues for crop breeding research that are expected to be instrumental for strengthening food security in the coming years.

Sedges (Cyperaceae) are commonly pollinated by wind, but some transitions from wind to insect pollination have been reported in tropical sedges. Despite being a temperate sedge, the pollination strategy of Rhynchospora alba may partly depend on insects, as suggested by its white-colored inflorescences. Here we quantify the contribution of insects to the reproductive fitness of R. alba as an exploration of the success of entomophily in temperate sedges. Honeybees and hoverflies seem to be the primary visitors of R. alba in the study site, and pollen of R. alba was recovered from the mouthparts and legs of the insects. Treatments excluding insects led to a lower reproductive fitness (regarding fruit weight and fruit viability) than those not excluding them. Therefore, insects seem to contribute significantly to the pollination of R. alba despite moderate morphological adaptation. We highlight the role of entomophily as an overlooked reproductive mechanism in cold-temperate sedges.

Background and Aims The persistence of a plant population under a specific local climatic regime requires phenotypic adaptation with underlying particular combinations of alleles at adaptive loci. The level of allele diversity at adaptive loci within a natural plant population conditions its potential to evolve, notably towards adaptation to a change in climate. Investigating the environmental factors that contribute to the maintenance of adaptive diversity in populations is thus worthwhile. Within-population allele diversity at adaptive loci can be partly driven by the mean climate at the population site but also by its temporal variability. Methods The effects of climate temporal mean and variability on within-population allele diversity at putatively adaptive quantitative trait loci (QTLs) were evaluated using 385 natural populations of Lolium perenne (perennial ryegrass) collected right across Europe. For seven adaptive traits related to reproductive phenology and vegetative potential growth seasonality, the average within-population allele diversity at major QTLs (HeA) was computed. Key Results Significant relationships were found between HeA of these traits and the temporal mean and variability of the local climate. These relationships were consistent with functional ecology theory. Conclusions Results indicated that temporal variability of local climate has likely led to fluctuating directional selection, which has contributed to the maintenance of allele diversity at adaptive loci and thus potential for further adaptation.

International audience; Human induced environmental change may require rapid adaptation of plant populations and crops, but the genomic basis of environmental adaptation remain poorly understood. We analysed polymorphic loci from the perennial crop Medicago sativa (alfalfa or lucerne) and the annual legume model species M. truncatula to search for a common set of candidate genes that might contribute to adaptation to abiotic stress in both annual and perennial Medicago species. We identified a set of candidate genes of adaptation associated with environmental gradients along the distribution of the two Medicago species. Candidate genes for each species were detected in homologous genomic linkage blocks using genome-environment (GEA) and genome-phenotype association analyses. Hundreds of GEA candidate genes were species-specific, of these, 13.4% (M. sativa) and 24% (M. truncatula) were also significantly associated with phenotypic traits. A set of 168 GEA candidates were shared by both species, which was 25.4% more than expected by chance. When combined, they explained a high proportion of variance for certain phenotypic traits associated with adaptation. Genes with highly conserved functions dominated among the shared candidates and were enriched in gene ontology terms that have shown to play a central role in drought avoidance and tolerance mechanisms by means of cellular shape modifications and other functions associated with cell homeostasis. Our results point to the existence of a molecular basis of adaptation to abiotic stress in Medicago determined by highly conserved genes and gene functions. We discuss these results in light of the recently proposed omnigenic model of complex traits.

The structure and function of the soil microbiome of urban greenspaces remain largely undetermined. We conducted a global field survey in urban greenspaces and neighboring natural ecosystems across 56 cities from six continents, and found that urban soils are important hotspots for soil bacterial, protist and functional gene diversity, but support highly homogenized microbial communities worldwide. Urban greenspaces had a greater proportion of fast-growing bacteria, algae, amoebae, and fungal pathogens, but a lower proportion of ectomycorrhizal fungi than natural ecosystems. These urban ecosystems also showed higher proportions of genes associated with human pathogens, greenhouse gas emissions, faster nutrient cycling, and more intense abiotic stress than natural environments. City affluence, management practices, and climate were fundamental drivers of urban soil communities. Our work paves the way toward a more comprehensive global-scale perspective on urban greenspaces, which is integral to managing the health of these ecosystems and the well-being of human populations., M.D.-B. and this project were supported by a 2019 Leonardo Grant for Researchers and Cultural Creators, BBVA Foundation (URBANFUN) and by the BES grant agreement no. LRB17\1019 (MUSGONET). M.D.-B. is also supported by a Ramón y Cajal grant from the Spanish Ministry of Science and Innovation (RYC2018-025483-I). N.F. was supported by grants from the U.S. National Science Foundation (DEB1556090 and DEB1542653). L.T. acknowledges support from Norway-Baltic collaboration grant EMP442 and Estonian Science Foundation grant PRG632. B.K.S. acknowledges a research award by the Humboldt Foundation and funding from the Australian Research Council (DP190103714). F.A. is supported by ANID FONDECYT 11180538 and 1170995. S.A. is funded by ANID FONDECYT 1170995 and ANID ANILLO ACT192027. F.B. and J.L.M. acknowledge support from the Spanish Ministry and FEDER funds for the project AGL2017-85755-R, the i-LINK+ 2018 (LINKA20069) from CSIC, as well as funds from “Fundación Séneca” from Murcia Province (19896/GERM/15). C.P. acknowledges support from the Spanish State Plan for Scientific and Technical Research and Innovation (2013–2016), award reference AGL2016-75762-R (AEI/FEDER, UE). M.B. acknowledges support from a Juan de la Cierva Formación grant from the Spanish Ministry of Economy and Competitiveness (FJCI-2018-036520-I). T.P.M. would like to acknowledge contributions from the National Research Foundation of South Africa and cities involved in the South African survey. Slovenian coauthors were supported by the research project J4-1766 “Methodology approaches in genome-based diversity and ecological plasticity study of truffles from their natural distribution areas” and the Research Program in Forest Biology, Ecology, and Technology (P4-0107) of the Slovenian Research Agency. J.D. and A. Rey acknowledge support from the FCT (IF/00950/2014 and SFRH/BDP/108913/2015, respectively), as well as from the MCTES, FSE, UE, and the CFE (UIDB/04004/2020) research unit financed by FCT/MCTES through national funds (PIDDAC). J.P.V. acknowledges financial support from SERB (Science and Engineering Research Board) (EEQ/2017/000775) India. J.-Z.H. and H.-W.H. are financially supported by Australian Research Council (DP170101628).

More severe seasonal stresses resulting from climate change affect the survival of perennial plant species. The growth–survival trade‐off exemplified in dormant species is a key issue to understand adaptation. As the validity of this trade‐off has yet to be tested in non‐dormant species, it was assessed by exploring the intraspecific variability of strategies to face drought and frost within perennial ryegrass. Three common gardens compared 385 European perennial ryegrass populations along a latitudinal environmental gradient over 3‐years. Persistence, productivity and physiological traits were recorded under contrasting seasonal environments. Decoupling plant responses, that is, growth under favourable summers/winters and plant survival under harsh summers/winters, showed a general trade‐off between growth potential and dehydration survival. Three groups of perennial ryegrass populations were identified according to their contrasting strategies: (a) year‐round productive but stress sensitive populations from wet areas; (b) drought‐tolerant populations with low summer growth potential from drought‐prone areas and (c) frost‐tolerant populations with low winter growth potential from frost‐prone areas. Overall, the populations surviving drought best were more resource conservative, whereas populations of the other groups were more resource acquisitive. However, such overall functional patterns were less meaningful than seasonal variations of resource acquisition potentials. The predicted potential biogeographical distribution of these groups suggests shifts of areas of suitability under climate change over the next decades in Europe. Dehydration escape and dehydration tolerance through reduction of growth potential in summer may become the strategies best adapted to an increasingly large area of Europe. The large intraspecific variability of phenological adaptations within perennial ryegrass reveals that the seasonal modulation of growth potential is crucial to plant adaptation under severe chronic abiotic stresses. The global plant economics spectrum cannot account for contrasting seasonal trade‐offs, which points out the importance of integrating phenological traits as key components of plant strategies. The identification of the trade‐off between growth potential and frost or drought stress survival in this non‐dormant species provides key knowledge to understand the future regional distribution of this major species for grassland ecosystem services. A free Plain Language Summary can be found within the Supporting Information of this article.

International audience; Disentangling the origin of species-genetic diversity correlations (SGDCs) is a challenging task that provides insight into the way that neutral and adaptive processes influence diversity at multiple levels. Genetic and species diversity are comprised by components that respond differently to the same ecological processes. Thus, it can be useful to partition species and genetic diversity into their different components to infer the mechanisms behind SGDCs. In this study, we applied such an approach using a high-elevation Andean wetland system, where previous evidence identified neutral processes as major determinants of the strong and positive covariation between plant species richness and AFLP genetic diversity of the common sedge Carex gayana. To tease apart putative neutral and non-neutral genetic variation of C. gayana, we identified loci putatively under selection from a dataset of 1,709 SNPs produced using restriction site-associated DNA sequencing (RAD-seq). Significant and positive relationships between local estimates of genetic and species diversities (alpha-SGDCs) were only found with the putatively neutral loci datasets and with species richness, confirming that neutral processes were primarily driving the correlations and that the involved processes differentially influenced local species diversity components (i.e., richness and evenness). In contrast, SGDCs based on genetic and community dissimilarities (beta-SGDCs) were only significant with the putative non-neutral datasets. This suggests that selective processes influencing C. gayana genetic diversity were involved in the detected correlations. Together, our results demonstrate that analyzing distinct components of genetic and species diversity simultaneously is useful to determine the mechanisms behind species-genetic diversity relationships.

The growing demand for timber and the boom in massive tree-planting programs could mean the spreading of mismanaged tree plantations worldwide. Here, we apply the concept of ecological intensification to forestry systems as a viable biodiversity-focused strategy that could be critical to develop productive, yet sustainable, tree plantations. Tree plantations can be highly productive if tree species are properly combined to complement their ecological functions. Simultaneously considering soil biodiversity and animal-mediated biocontrol will be critical to minimize the reliance on external inputs. Integrating genetic, functional, and demographic diversity across heterogeneous landscapes should improve resilience under climate change. Designing ecologically intensified plantations will mean breaking the timber productivity versus conservation dichotomy and assuring the maintenance of key ecosystem services at safe levels.

The natural genetic diversity of agricultural species is an essential genetic resource for breeding programs aiming to improve their ecosystem and production services. A large natural ecotype diversity is usually available for most grassland species. This could be used to recombine natural climatic adaptations and agronomic value to create improved populations of grassland species adapted to future regional climates. However describing natural genetic resources can be long and costly. Molecular markers may provide useful information to help this task. This opportunity was investigated for Lolium perenne L., using a set of 385 accessions from the natural diversity of this species collected right across Europe and provided by genebanks of several countries. For each of these populations, genotyping provided the allele frequencies of 189,781 SNP markers. GWAS were implemented for over 30 agronomic and/or putatively adaptive traits recorded in three climatically contrasted locations (France, Belgium, Germany). Significant associations were detected for hundreds of markers despite a strong confounding effect of the genetic background; most of them pertained to phenology traits. It is likely that genetic variability in these traits has had an important contribution to environmental adaptation and ecotype differentiation. Genomic prediction models calibrated using natural diversity were found to be highly effective to describe natural populations for almost all traits as well as commercial synthetic populations for some important traits such as disease resistance, spring growth or phenological traits. These results will certainly be valuable information to help the use of natural genetic resources of other species.

International audience; Germplasm from perennial ryegrass (Lolium perenne L.) natural populations is useful for breeding because of its adaptation to a wide range of climates. Climate-adaptive genes can be detected from associations between genotype, phenotype and climate but an integrated framework for the analysis of these three sources of information is lacking. We used two approaches to identify adaptive loci in perennial ryegrass and their effect on phenotypic traits. First, we combined Genome-Environment Association (GEA) and GWAS analyses. Then, we implemented a new test based on a Canonical Correlation Analysis (CANCOR) to detect adaptive loci. Furthermore, we improved the previous perennial ryegrass gene set by de novo gene prediction and functional annotation of 39,967 genes. GEA-GWAS revealed eight outlier loci associated with both environmental variables and phenotypic traits. CANCOR retrieved 633 outlier loci associated with two climatic gradients, characterized by cold-dry winter versus mild-wet winter and long rainy season versus long summer, and pointed out traits putatively conferring adaptation at the extremes of these gradients. Our CANCOR test also revealed the presence of both polygenic and oligogenic climatic adaptations. Our gene annotation revealed that 374 of the CANCOR outlier loci were positioned within or close to a gene. Co-association networks of outlier loci revealed a potential utility of CANCOR for investigating the interaction of genes involved in polygenic adaptations. The CANCOR test provides an integrated framework to analyse adaptive genomic diversity and phenotypic responses to environmental selection pressures that could be used to facilitate the adaptation of plant species to climate change.

Although hybridisation through genome duplication is well known, hybridisation without genome duplication (homoploid hybrid speciation, HHS) is not. Few well-documented cases have been reported. A possible instance of HHS in Medicago prostrata Jacq. was suggested previously, based on only two genes and one individual. We tested whether this species was formed through HHS by sampling eight nuclear loci and 22 individuals, with additional individuals from related species, using gene capture and Illumina sequencing. Phylogenetic inference and coalescent simulations were performed to infer the causes of gene tree incongruence. We found no evidence that phylogenetic differences among M. prostrata individuals were the result of HHS. Instead, an autopolyploid origin of tetraploids with introgression from tetraploids of the M. sativa complex is likely. We argue that tetraploid M. prostrata individuals constitute a new species, characterised by a partially non-overlapping distribution and distinctive alleles (from the M. sativa complex). No gene flow from tetraploid to diploid M. prostrata is apparent, suggesting partial reproductive isolation. Thus, speciation via autopolyploidy appears to have been reinforced by introgression. This raises the intriguing possibility that introgressed alleles may be responsible for the increased range exploited by tetraploid M. prostrata with respect to that of the diploids.

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International audience; Premise Although hybridization has played an important role in the evolution of many plant species, phylogenetic reconstructions that include hybridizing lineages have been historically constrained by the available models and data. Restriction-site-associated DNA sequencing (RADseq) has been a popular sequencing technique for the reconstruction of hybridization in the next-generation sequencing era. However, the utility of RADseq for the reconstruction of complex evolutionary networks has not been thoroughly investigated. Conflicting phylogenetic relationships in the genus Medicago have been mainly attributed to hybridization, but the specific hybrid origins of taxa have not been yet clarified. Methods We obtained new molecular data from diploid species of Medicago section Medicago using single-digest RADseq to reconstruct evolutionary networks from gene trees, an approach that is computationally tractable with data sets that include several species and complex hybridization patterns. Results Our analyses revealed that assembly filters to exclusively select a small set of loci with high phylogenetic information led to the most-divergent network topologies. Conversely, alternative clustering thresholds or filters on the number of samples per locus had a lower impact on networks. A strong hybridization signal was detected for M. carstiensis and M. cretacea, while signals were less clear for M. rugosa, M. rhodopea, M. suffruticosa, M. marina, M. scutellata, and M. sativa. Conclusions Complex network reconstructions from RADseq gene trees were not robust under variations of the assembly parameters and filters. But when the most-divergent networks were discarded, all remaining analyses consistently supported a hybrid origin for M. carstiensis and M. cretacea.

Floral nectar spurs are widely considered a key innovation promoting diversification in angiosperms by means of pollinator shifts. We investigated the macroevolutionary dynamics of nectar spurs in the tribe Antirrhineae (Plantaginaceae), which contains 29 genera and 300–400 species (70–80% spurred). The effect of nectar spurs on diversification was tested, with special focus on Linaria, the genus with the highest number of species. We generated the most comprehensive phylogeny of Antirrhineae to date and reconstructed the evolution of nectar spurs. Diversification rate heterogeneity was investigated using trait-dependent and trait-independent methods, and accounting for taxonomic uncertainty. The association between changes in spur length and speciation was examined within Linaria using model testing and ancestral state reconstructions. We inferred four independent acquisitions of nectar spurs. Diversification analyses revealed that nectar spurs are loosely associated with increased diversification rates. Detected rate shifts were delayed by 5–15 Myr with respect to the acquisition of the trait. Active evolution of spur length, fitting a speciational model, was inferred in Linaria, which is consistent with a scenario of pollinator shifts driving diversification. Nectar spurs played a role in diversification of the Antirrhineae, but diversification dynamics can only be fully explained by the complex interaction of multiple biotic and abiotic factors., This work was supported by the Marie Curie Intra‐European Fellowship LINARIA‐SPECIATION (FP7‐PEOPLE‐2013‐IEF, reference 624396), an Isaac Newton Trust Research Grant (Trinity College, Cambridge), a Juan de la Cierva fellowship to MF‐M (Spanish Ministry of Economy and Competitivity, reference IJCI‐2015‐23459) and a Generalitat Valenciana postdoctoral grant to AJ (Ministry of Education, Culture and Sport, reference BEST/2014/264).

International audience; Aim: Grasslands have been pivotal in the development of herbivore breeding since the Neolithic and still represent the most widespread agricultural land use across Europe. However, it remains unclear whether the current large‐scale genetic variation of plant species found in natural grasslands of Europe is the result of human activities or natural processes. Location: Europe. Taxon: Lolium perenne L. (perennial ryegrass). Methods: We reconstructed the phylogeographic history of L. perenne, a dominant grassland species, using 481 natural populations, including 11 populations of closely related taxa. We combined Genotyping‐by‐Sequencing (GBS) and pool‐Sequencing (pool‐Seq) to obtain high‐quality allele frequency calls of ~500 k SNP loci. We performed genetic structure analyses and demographic reconstructions based on the site frequency spectrum (SFS). We additionally used the same genotyping protocol to assess the genomic diversity of a set of 32 cultivars representative of the L. perenne cultivars widely used for forage purposes. Results: Expansion across Europe took place during the Würm glaciation (12–110 kya), a cooling period that decreased the dominance of trees in favour of grasses. Splits and admixtures in L. perenne fit historical climate changes in the Mediterranean basin. The development of agriculture in Europe (7–3.5 kya), that caused an increase in the abundance of grasslands, did not have an effect on the demographic patterns of L. perenne. We found that most modern cultivars are closely related to natural diversity from north-western Europe. Thus, modern cultivars do not represent the wide genetic variation found in natural populations. Main conclusions: Demographic events in L. perenne can be explained by the changing climatic conditions during the Pleistocene. Natural populations maintain a wide genomic variability at continental scale that has been minimally exploited by recent breeding activities. This variability constitutes valuable standing genetic variation for future adaptation of grasslands to climate change, safeguarding the agricultural services they provide.

Although hybridization has played an important role in the evolution of many species, phylogenetic reconstructions that include hybridizing lineages have been historically constrained by the available models and data. Recently, the combined development of high-throughput sequencing and evolutionary network models offer new opportunities for phylogenetic inference under complex patterns of hybridization in the context of incomplete lineage sorting. Restriction site associated DNA sequencing (RADseq) has been a popular sequencing technique for evolutionary reconstructions of close relatives in the Next Generation Sequencing (NGS) era. However, the utility of RADseq data for the reconstruction of complex evolutionary networks has not been thoroughly discussed. Here, we used new molecular data collected from diploid perennialMedicagospecies using single-digest RADseq to reconstruct evolutionary networks from gene trees, an approach that is computationally tractable with datasets that include several species and complex patterns of hybridization. Our analyses revealed that complex network reconstructions from RADseq-derived gene trees were not robust under variations of the assembly parameters and filters. Filters to exclusively select loci with high phylogenetic information created datasets that retrieved the most anomalous topologies. Conversely, alternative clustering thresholds or filters on the number of samplesperlocus affected the level of missing data but had a lower impact on networks. When most anomalous networks were discarded, all remaining network analyses consistently supported a hybrid origin forM. carstiensisandM. cretacea.

AimGrasslands have been pivotal in the development of herbivore breeding since the Neolithic and are still nowadays the most widespread agricultural land-use across Europe. However, it remains unclear whether the current large-scale genetic variation of plant species found in natural grasslands of Europe is the result of human activities or natural processes.LocationEurope.TaxonLolium perenne L (perennial ryegrass).MethodsWe reconstructed the phylogeographic history of L. perenne, a dominant grassland species, using 481 natural populations including 11 populations from closely related taxa. We combined the Genotyping-by-Sequencing (GBS) and Pool-sequencing (Pool-seq) methods to obtain high-quality allele frequency calls of ~ 500 k SNP loci. We performed genetic structure analyses and demographic reconstructions based on the site frequency spectrum (SFS). We additionally used the same genotyping protocol to assess the genomic diversity of a set of 32 cultivars representative of the L. perenne cultivars widely used for forage purposes.ResultsExpansion across Europe took place during the Würm glaciation (12-110 kya), a cooling period that decreased the dominance of trees in favour of grasses. Splits and admixtures in L. perenne fit historical sea level changes in the Mediterranean basin. The development of agriculture in Europe (7-3.5 kya), that caused an increase in the abundance of grasslands, did not have an effect on the demographic patterns of L. perenne. We found little differentiation between modern cultivars and certain natural variants. However, modern cultivars do not represent the wide genetic variation found in natural populations.Main conclusionsDemographic events in L. perenne can be explained by the changing climatic conditions during the Pleistocene. Natural populations maintain a wide genomic variability at continental scale that has been underused by recent breeding activities. This variability constitutes valuable standing genetic variation for future adaptation of grasslands to climate change, safeguarding the agricultural services they provide.

International audience; Allele frequency fingerprinting of heterogeneous plant populations of outbreeding species can be used for variety identification, association mapping, genomic selection or characterization of genetic resources. In the FACCE-JPI GrassLandscape project, we empirically validated a pool-GBS method for Genome-Wide Allele Frequency Fingerprinting (GWAFF). As pool-GBS cannot be targeted to predefined loci such as candidate genes, we integrated it with targeted resequencing using a highly multiplexed amplicon sequencing strategy to measure allele frequencies (pool-HiPlex). A pool-HiPlex assay was designed that amplifies 185 amplicons in 41 L. perenne genes in just two parallel PCRreactions. We validated pool-GBS and pool-HiPlex using pools of 48 individuals, chosen to represent a wide range of genetic diversity in L. perenne, and addressed completeness, reproducibility and accuracy of allele frequencies with >1000 HiPlex SNPs and >150.000 GBS SNPs. We consistently found high correlations between allele frequencies obtained by genotyping individual plants and pool genotyping on leaf tissue pools and DNA extract pools. We also analyzed the error introduced at various steps of the protocol such as weighing, DNA-quantification, pooling, ligation and/or PCRamplification. Applying a minor allele frequency threshold of 5% or 3% effectively removed nonreproducible SNPs in pool-GBS and pool-HiPlex, respectively. Allele frequency spectra could be obtained for single SNPs as well as for haplotypes spanning neighboring SNPs using read-backed phasing. Application of this methodology to a set of 470 natural populations of L. perenne sampled across Europe and the fertile Crescent revealed a geographical pattern of genetic differentiation in this species.

International audience; Grasslands have been pivotal in the development of domestic herbivore breeding since the Neolithic and are nowadays the most widespread agricultural land-use over Europe. We have demonstrated that the genealogy of natural Lolium perenne (perennial ryegrass), from permanent grasslands across Europe and the Fertile Crescent, can be traced back to the pre-Holocene epoch (> 12 kya). Demographic events in this species, as reconstructed from genetic data, can be explained by the changing climatic conditions during Quaternary glaciations in Europe. Specific population splits and admixtures fit historical sea level changes across the Mediterranean basin. According to our estimations, population expansion across continental Europe took place during the Würm glaciation (12-110 kya), a cooling period that decreased the dominance of trees in favor of herbs, and notably grasses. The development of agriculture and herbivore breeding that started in the late Holocene (3.5 kya) and caused an increase in the abundance of grasses across Europe did not impact the genetic structure of Lolium perenne at continental scale. Furthermore, our results reveal little differentiation between cultivars released by contemporary plant breeding and natural strains of L. perenne, pointing to the very recent start of intensive human directional selection in herbage grasses. We expose that natural grasslands harbor wide genetic variability at continental scale that has enabled grasses to survive extreme climatic changes across millennia. This genetic variability has been underused by recent breeding activities and constitute valuable standing genetic variation for future adaptation of grasslands to Climate Change, safeguarding the agricultural services they provide.

Aim The biogeographical congruence hypothesis, that similar spatiotemporal patterns of geographical distribution exist across lineages, is revisited in this study, and biogeographical processes in presumed Madrean–Tethyan plants are investigated by employing phylogenetic analyses. Location Mediterranean and Californian floristic regions. Methods The snapdragons (tribe Antirrhineae, Plantaginaceae) are one of the plant groups that best illustrate disjunctions between the New World (14 genera) and the Old World (15 genera). A time-calibrated phylogeny (ndhF sequences) and ancestral-area reconstructions were used to test the hypothesis of biogeographical congruence. We estimated support for sister-group relationships together with the probability of temporal congruence of snapdragons and five additional angiosperm groups using a biogeographical approach based on Bayesian inference, parsimony and maximum-likelihood methods. Results Synchronous divergences of four phylogenetically independent Mediterranean/Californian lineages within Antirrhineae were inferred for the Miocene. This result constitutes the first example of high biogeographical congruence within the same plant group. Analyses of five additional angiosperm groups previously considered exemplars of Madrean–Tethyan disjunctions revealed a total of 10 Mediterranean/Californian sister-group lineages, mostly with Miocene divergence times. In particular, our contrasting biogeographical analysis favoured a prevalent colonization process post-dating the separation of America and Eurasia (Eocene) for at least eight angiosperm lineages. Main conclusions Explicit testing of the Madrean–Tethyan hypothesis did not support predominant vicariance for Mediterranean/Californian sister groups as previously proposed. Instead, eight Mediterranean/Californian sister-group lineages displayed a Miocene divergence, including considerable biogeographical congruence within Antirrhineae (four independent lineages) and Cistaceae (two lineages).

The evolutionary patterns of the Mediterranean flora during the Quaternary have been relatively well documented based on phylogenetic and biogeographic analyses, but few studies have addressed the evolutionary traits that determined diversification and range expansion success during this period. We analysed previously published and newly generated sequences of three plastid noncoding regions (rpl32-trnL(UAG) , trnS-trnG and trnL-trnF), the nuclear ribosomal internal transcribed spacer (ITS) and a low-copy nuclear gene intron (AGT1) of Linaria sect. Supinae, a group of angiosperms that diversified in the Quaternary. The origin and recent colonization dynamics of closely related lineages were inferred by biogeographic reconstruction and phylogeographic analyses, while breeding system experiments coupled with ecological and morphological data were used to test association with range expansion and diversification. A combination of traits, including selfing, short lifespan and the ability to tolerate a wide variety of substrates, were key factors underlying range expansion after long-distance dispersal throughout the Mediterranean basin. By contrast, self-incompatibility may have promoted higher diversification rates in narrow ranges of the Iberian Peninsula. We argue that a few traits contributed to the adoption of two contrasting strategies that may have been predominant in the evolution of Mediterranean angiosperms.

Anthropogenic global climate change is expected to cause severe range contractions among alpine plants. Alpine areas in the Mediterranean region are of special concern because of the high abundance of endemic species with narrow ranges. This study combined species distribution models, population structure analyses and Bayesian skyline plots to trace the past and future distribution and diversity of Linaria glacialis, an endangered narrow endemic species that inhabits summits of Sierra Nevada (Spain). The results showed that: (i) the habitat of this alpine-Mediterranean species in Sierra Nevada suffered little changes during glacial and interglacial stages of late Quaternary; (ii) climatic oscillations in the last millennium (Medieval Warm Period and Little Ice Age) moderately affected the demographic trends of L. glacialis; (iii) future warming conditions will cause severe range contractions; and (iv) genetic diversity will not diminish at the same pace as the distribution range. As a consequence of the low population structure of this species, genetic impoverishment in the alpine zones of Sierra Nevada should be limited during range contraction. We conclude that maintenance of large effective population sizes via high mutation rates and high levels of gene flow may promote the resilience of alpine plant species when confronted with global warming.

Premise of research. Toadflaxes (Linaria Mill., ∼150 spp. from the Palearctic region) constitute the largest genus of the snapdragon lineage (tribe Antirrhineae). Here we provide the first extensive phylogenetic testing of systematic and evolutionary hypotheses about toadflaxes.Methodology. Internal transcribed spacer (ITS) sequences were obtained for 94 species representing all sections of Linaria recognized by recent taxonomic treatments, as well as three species of the morphologically related American genus Nuttallanthus. In addition, 71 sequences representing the remaining 26 genera of Antirrhineae were gathered to test the monophyly of toadflaxes. Phylogenetic analyses were conducted using Bayesian inference and maximum likelihood. Evaluation of alternative topologies was assessed by means of Bayes factor analyses.Pivotal results. Linaria and Nuttallanthus constituted a monophyletic group within the Antirrhineae. Linaria was recovered as a paraphyletic group with Nuttallanthus nested within it. Six m...

The role of pollinators in floral divergence has long attracted the attention of evolutionary biologists. Although abundant studies have reported the effect of pollinators on flower-shape variation and plant speciation, the influence of pollinators on plant species differentiation during rapid radiations and the specific consequences of shifts among similar pollinators are not well understood. Here, we evaluate the association between pollinators and floral morphology in a closely related and recently diversifying clade of Linaria species (sect. Supinae subsect. Supinae). Our approach combined pollinator observations, functional floral morphometric measures and phylogenetic comparative analyses. The fauna visiting Linaria species was determined by extensive surveys and categorized by a modularity algorithm, and the size and shape of flowers were analysed by means of standard and geometric morphometric measures. Standard measures failed to find relationships between the sizes of representative pollinators and flowers. However, discriminant function analyses of geometric morphometric data revealed that pollination niches are finer predictors of flower morphologies in Linaria if compared with phylogenetic relationships. Species with the most restrictive flowers displayed the most slender spurs and were pollinated by bees with larger proboscides. These restrictive flower shapes likely appeared more than once during the evolutionary history of the study group. We show that floral variation can be driven by shifts between pollinators that have been traditionally included in a single functional group, and discuss the consequences of such transitions for plant species differentiation during rapid radiations., This work was supported by the Span-ish Ministry of Education through projects CGL2009-10031 and CGL2012-34736, by the Spanish NationalResearch Council (CSIC) through a CSIC:JAEpre fel-lowship to Jose Luis Blanco Pastor and by the SpanishMinistry of Science and Innovation through a FPI fel-lowship to Isabel M. Liberal.

We examined the phylogenetic history of Linaria with special emphasis on the Mediterranean sect. Supinae (44 species). We revealed extensive highly supported incongruence among two nuclear (ITS, AGT1) and two plastid regions (rpl32-trnL(UAG), trnS-trnG). Coalescent simulations, a hybrid detection test and species tree inference in *BEAST revealed that incomplete lineage sorting and hybridization may both be responsible for the incongruent pattern observed. Additionally, we present a multilabelled *BEAST species tree as an alternative approach that allows the possibility of observing multiple placements in the species tree for the same taxa. That permitted the incorporation of processes such as hybridization within the tree while not violating the assumptions of the *BEAST model. This methodology is presented as a functional tool to disclose the evolutionary history of species complexes that have experienced both hybridization and incomplete lineage sorting. The drastic climatic events that have occurred in the Mediterranean since the late Miocene, including the Quaternary-type climatic oscillations, may have made both processes highly recurrent in the Mediterranean flora.

Background A central aim of island biogeography is to understand the colonization history of insular species using current distributions, fossil records and genetic diversity. Here, we analyze five plastid DNA regions of the endangered Juniperus brevifolia, which is endemic to the Azores archipelago. Methodology/Principal Findings The phylogeny of the section Juniperus and the phylogeographic analyses of J. brevifolia based on the coalescence theory of allele (plastid) diversity suggest that: (1) a single introduction event likely occurred from Europe; (2) genetic diversification and inter-island dispersal postdated the emergence of the oldest island (Santa Maria, 8.12 Ma); (3) the genetic differentiation found in populations on the islands with higher age and smaller distance to the continent is significantly higher than that on the younger, more remote ones; (4) the high number of haplotypes observed (16), and the widespread distribution of the most frequent and ancestral ones across the archipelago, are indicating early diversification, demographic expansion, and recurrent dispersal. In contrast, restriction of six of the seven derived haplotypes to single islands is construed as reflecting significant isolation time prior to colonization. Conclusions/Significance Our phylogeographic reconstruction points to the sequence of island emergence as the key factor to explain the distribution of plastid DNA variation. The reproductive traits of this juniper species (anemophily, ornithochory, multi-seeded cones), together with its broad ecological range, appear to be largely responsible for recurrent inter-island colonization of ancestral haplotypes. In contrast, certain delay in colonization of new haplotypes may reflect intraspecific habitat competition on islands where this juniper was already present., B. Rumeu and J.L. Blanco-Pastor were funded by a Spanish National Research Council grant µ(CSIC: JAE-PRE; http://www.csic.es/web/guest/programa-ja​e). This contribution is framed within the projects CGL2010-18759 and PI2007/053 and financed by the Spanish Ministry of Science and Innovation (http://www.micinn.es), and the Canary Island Government (http://www.gobcan.es), respectively, both of which were partially funded by the European Union. The Organismo Autónomo de Parques Nacionales (http://www.mma.es/secciones/el_ministeri​o/organismos/oapn/) also financed part of this work (project 051/2010). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.