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Some plants abandoned photosynthesis and developed full dependency on fungi for nutrition. Most of the so-called mycoheterotrophic plants exhibit high specificity towards their fungal partners. We tested whether natural rarity of mycoheterotrophic plants and usual small and fluctuating population size make their populations more prone to genetic differentiation caused by restricted gene flow and/or genetic drift. We also tested whether these genetic characteristics might in turn shape divergent fungal preferences. We studied the mycoheterotrophic orchid Epipogium aphyllum, addressing the joint issues of genetic structure of its populations over Europe and possible consequences for mycorrhizal specificity within the associated fungal taxa. Out of 27 sampled E. aphyllum populations, nine were included for genetic diversity assessment using nine nuclear microsatellites and plastid DNA. Population genetic structure was inferred based on the total number of populations. Individuals from 17 locations were included into analysis of genetic identity of mycorrhizal fungi of E. aphyllum based on barcoding by nuclear ribosomal DNA. Epipogium aphyllum populations revealed high genetic diversity (uHe = 0.562) and low genetic differentiation over vast distances (FST = 0.106 for nuclear microsatellites and FST = 0.156 for plastid DNA). Bayesian clustering analyses identified only two genetic clusters, with a high degree of admixture. Epipogium aphyllum genets arise from panmixia and display locally variable, but relatively high production of ramets, as shown by a low value of rarefied genotypic richness (Rr = 0.265). Epipogium aphyllum genotype control over partner selection was negligible as (1) we found ramets from a single genetic individual associated with up to 68% of the known Inocybe spp. associating with the plant species, (2) and partner identity did not show any geographic structure. The absence of mosaicism in the mycorrhizal specificity over Europe may be linked to preferential allogamous habit of E. aphyllum and significant gene flow, which tend to promote host generalism.

International audience; The Asian knotweed species complex gathers some of the world’s most successful plant invaders including the Japanese knotweed (Reynoutria japonica), the giant knotweed (R. sachalinensis) and the hybrid of these two species, the Bohemian knotweed (R. × bohemica). Hybrid species often present higher competitive abilities compared to their parent species. While several studies have focused on the effects of knotweed invasion on plant communities, few have simultaneously considered (i) effects of the three taxa on native plant communities and (ii) effects on litter and soil faunal components. In this study, we compared the differential effects of three Asian knotweeds on vegetation and soil macroinvertebrates communities across seven sites on a regional scale in North Western France. All three knotweed species displayed similar negative effects on local plant species richness, while promoting the taxonomic richness of litter-dwelling macroinvertebrates. Belowground macroinvertebrate taxonomic richness appeared strongly reduced by the presence of the hybrid R. × bohemica, significantly more so than those of sites colonized by R. japonica or R. sachalinensis. These changes of belowground communities were correlated to associated changes of composition and richness within plant communities. This study provides new insight into the consequences of ecosystem invasion by these species, especially revealing the even further strength of impacts of the hybrid Bohemian knotweed on local vegetation and belowground macroinvertebrates than those of other Asian knotweeds, which points to the need for monitoring the spatial spread of R. × bohemica and describing further its effects on ecosystem properties.

International audience; Detecting the molecular basis of local adaptation and identifying selective drivers is still challenging in nonmodel species. The use of purely population genetic approaches is limited by some characteristics of genetic systems, such as pleiotropy and polygenic control, and parallel evidence from phenotypic‐based experimental comparisons is required. In long‐lived organisms, the detection of selective pressures might also be precluded by evolutionary lag times in response to the environment. Here, we used the English yew to showcase an example of a multiscale integrative approach in a nonmodel species with limited plant and genomic resources. We combined information from two independent sources, phenotypes in a common environment and genomic data in natural populations, to investigate the signature of selection. Growth differences among populations in a common environment, and phenological patterns of both shoot elongation and male strobili maturation, were associated with climate clines, providing evidence for local adaptation and guiding us in the selection of populations for genomic analyses. We used information on over 25,000 SNPs from c. 1,200 genes to infer the demographic history and to test for molecular signatures of selection at different levels: SNP, gene, and biological pathway. Our results confirmed an overall demographic history of population decline, but we also found evidence for putative local adaptation at the molecular level. We identified or confirmed several candidate genes for positive and negative selection in forest trees, including the pseudo‐response regulator 7 (PRR7), an essential component of the circadian clock in plants. In addition, we successfully tested an approach to detect polygenic adaptation in biological pathways, allowing us to identify the flavonoid biosynthesis pathway as a candidate stress‐response pathway that deserves further attention in other plants. Finally, our study contributes to the emerging view that explaining contemporary standing genetic variation requires considering adaptation to past climates, especially for long‐lived trees.

International audience; Global trade increases plant introductions, but joint introduction of associated microbes is overlooked. We analyzed the ectomycorrhizal fungi of a Caribbean beach tree, seagrape (Coccoloba uvifera, Polygonacaeae), introduced pantropically to stabilize coastal soils and produce edible fruits. Seagrape displays a limited symbiont diversity in the Caribbean. In five regions of introduction (Brazil, Japan, Malaysia, Reunion and Senegal), molecular barcoding showed that seagrape mostly or exclusively associates with Scleroderma species (Basidiomycota) that were hitherto only known from Caribbean seagrape stands. An unknown Scleroderma species dominates in Brazil, Japan and Malaysia, while Scleroderma bermudense exclusively occurs in Reunion and Senegal. Population genetics analysis of S. bermudense did not detect any demographic bottleneck associated with a possible founder effect, but fungal populations from regions where seagrape is introduced are little differentiated from the Caribbean ones, separated by thousands of kilometers, consistently with relatively recent introduction. Moreover, dry seagrape fruits carry Scleroderma spores, probably because, when drying on beach sand, they aggregate spores from the spore bank accumulated by semi-hypogeous Scleroderma sporocarps. Aggregated spores inoculate seedlings, and their abundance may limit the founder effect after seagrape introduction. This rare pseudo-vertical transmission of mycorrhizal fungi likely contributed to efficient and repeated seagrape/Scleroderma co-introductions.

(IF 3.13; Q1); Numerous studies predict a short-term important decrease in fossil resources and stress the need to develop alternative renewable energies, thus European countries aim at increasing biomass production for energetic purposes.One such source of bioenergy could be obtained from forest biomass pools by exporting logging residues, yet this practice would have strong impacts on forest ecosystems (e.g. disturbance of soil biodiversity and chemical properties). Most studies on biomass removal effects focused on boreal forests but responses in temperate forests are still scarcely studied. Soil macrofauna is involved in forest ecosystem functioning through numerous chemical, physical and biological processes and multiple interactions with other organisms. A disturbance of the soil macrofauna community can thus lead to a response of the forest ecosystem as a whole.Experimental plots were set up in 6 northern France forests (either dominated by oak or beech), with a treatment corresponding to Whole Tree Harvesting (WTH) practices compared to control, in order to characterize the response of soil macrofaunal communities to organic matter (OM) export. Our study showed that OM export leads to a short-term loss of abundance in macrofaunal communities in temperate deciduous forest soils that can also lead to an alteration of soil OM cycle, and revealed that the breadth of this negative impact is modulated by tree species and faunal trophic groups. Furthermore, some results of community responses were directly applicable to forest management as marked responses of several taxa revealed four bioindicators of OM disturbance.Therefore, the level of disturbance related to new forest management practices, such as WTH to increase forest biofuel yields, (i) could be characterized by surveying either total soil macrofauna or using tools such as bioindicator taxa and (ii) should be assessed regarding tree stand identity.

Studies on ectomycorrhizal fungi (EcMF) populations, their genetic diversity, dynamics and the ecological drivers of their structure have long struggled with characterisation of fungal infraspecific variability, until development of molecular tools. Population dynamics have enhanced understanding of basic biological features of EcMF at population level, such as mating systems, local dispersal patterns and relationships between soil mycelial colonisation and fructification. Investigating population structure and dynamics also allowed the transfer of ecological colonisation strategies derived from plant ecology to EcMF species, although local environmental drivers and individual variations can shape populations beyond such ecological strategies. Characterisation of genetic diversity of EcMF populations from contrasted habitats has then been used to explore the role of environmental drivers, for instance soil parameters, in shaping the genetic structure and adaptive responses of populations. Detection of genetic structure of populations also proved relevant to explore host specialisation versus generalism in the ectomycorrhizal symbiosis. Extended up to the breadth of species’ range, investigation through the prism of landscape genetics and demographic reconstruction helped deciphering ancient and modern environmental drivers of population diversity, indicating the value of EcMF for testing biogeographic hypotheses. Present development of high-throughput sequencing methods is now allowing to explore the evolutionary mechanisms and traits beyond EcMF population history and response to environmental variation.

Biogeographical patterns and large-scale genetic structure have been little studied in ectomycorrhizal (EM) fungi, despite the ecological and economic importance of EM symbioses. We coupled population genetics and phylogenetic approaches to understand spatial structure in fungal populations on a continental scale. Using nine microsatellite markers, we characterized gene flow among 16 populations of the widespread EM basidiomycete Laccaria amethystina over Europe (i.e. over 2900 km). We also widened our scope to two additional populations from Japan (10 4 km away) and compared them with European populations through microsatellite markers and multilocus phylogenies, using three nuclear genes (NAR, G6PD and ribosomal DNA) and two mitochondrial ribosomal genes. European L. amethystina populations displayed limited differentiation (average FST = 0.041) and very weak isolation by distance (IBD). This panmictic European pattern may result from effective aerial dispersal of spores, high genetic diversity in populations and mutualistic interactions with multiple hosts that all facilitate migration. The multilocus phylogeny based on nuclear genes confirmed that Japanese and European specimens were closely related but clustered on a geographical basis. By using microsatellite markers, we found that Japanese populations were strongly differentiated from the European populations (FST = 0.416), more than expected by extrapolating the European pattern of IBD. Population structure analyses clearly separated the populations into two clusters, i.e. European and Japanese clusters. We discuss the possibility of IBD in a continuous population (considering some evidence for a ring species over the Northern Hemisphere) vs. an allopatric speciation over Eurasia, making L. amethystina a promising model of intercontinental species for future studies.

International audience; Stem canker of oilseed rape (canola, Brassica napus) is associated with a species complex of two closely related fungal species, Leptosphaeria maculans and L. biglobosa. Of these, L. maculans is the most damaging and develops gene-for-gene relationships with the host. Here, a wide scale analysis of the L. maculans - L. biglobosa species complex was performed throughout the American continent (23 locations from Chile to Canada) plus several locations inWestern Australia for comparison purposes, based on a collection of 1132 isolates from infected tissues of a susceptible cultivar. Fungal species were discriminated on the basis of morphological, phytopathological and molecular criteria and showed that L. biglobosa was closely associated with L. maculans in most of the locations. Multiple gene phylogeny using sequences of ITS, actin and b-tubulin confirmed the prevalence of the L. biglobosa ‘canadensis’ sub-clade in Canada, whereas up to three different sub-clades of L. biglobosa were found in Georgia (USA). Race structure of L. maculans was investigated using a combination of pathogenicity tests and PCR amplification of avirulence alleles AvrLm1, AvrLm4 and AvrLm6. Three contrasting situations were observed: (i) race structure in Ontario, Chile and Georgia was related to that of European and Western Australian populations, with a low race diversity; (ii) only one race was found in Mexico, and not found outside of this country; (iii) a large diversity of races was observed in central Canada (Manitoba, Alberta and Saskatchewan) with very specific features including maintenance of avirulence alleles absent from Europe, absence of the AvrLm7 allele common in Europe (or eastern Canada) and wide location-to-location variability.

International audience; Leptosphaeria maculans, a dothideomycete fungus causing stem canker on oilseed rape, develops gene-for-gene interactions with its host plants. It has the ability to rapidly adapt to selection pressure exerted by cultivars harbouring novel resistance genes as exemplified recently by the 3-year evolution towards virulence at the AvrLm1 locus in French populations. The AvrLm1 avirulence gene was recently cloned and shown to be a solo gene within a 269 kb non-coding, heterochromatin-like region. Here we describe the sequencing of the AvrLm1 genomic region in one avirulent and two virulent isolates to investigate the molecular basis of evolution towards virulence at the AvrLm1 locus. For these virulent isolates, the gain of virulence was linked to a 260 kb deletion of a chromosomal segment spanning AvrLm1 and deletion breakpoints were identical or similar. Among the 460 isolates analysed from France, Australia and Mexico, a similar large deletion was apparent in > 90% of the virulent isolates. Deletion breakpoints were also strongly conserved in most of the virulent isolates, which led to the hypothesis that a unique deletion event leading to the avrLm1 virulence has diffused in pathogen populations. These data finally suggest that retrotransposons are key drivers in genome evolution and adaptation to novel selection pressure in L. maculans.

Despite the large body of research devoted to understanding the role of Quaternary glacial cycles in the genetic divergence of European trees, the differential contribution of geographic isolation and/or environmental adaptation in creating population genetic divergence remains unexplored. In this study, we used a long-lived tree (Taxus baccata) as a model species to investigate the impact of Quaternary climatic changes on genetic diversity via neutral (isolation-by-distance) and selective (isolation-by-adaptation) processes. We applied approximate Bayesian computation to genetic data to infer its demographic history, and combined this information with past and present climatic data to assess the role of environment and geography in the observed patterns of genetic structure. We found evidence that yew colonized Europe from the East, and that European samples diverged into two groups (Western, Eastern) at the beginning of the Quaternary glaciations, c. 2.2 Myr before present. Apart from the expected effects of geographical isolation during glacials, we discovered a significant role of environmental adaptation during interglacials at the origin of genetic divergence between both groups. This process may be common in other organisms, providing new research lines to explore the effect of Quaternary climatic factors on present-day patterns of genetic diversity. © 2015 New Phytologist Trust.

Ectomycorrhizal (EM) fungi are major microbial components of boreal, temperate and Mediterranean forests, as well as some tropical forest ecosystems. Nearly two decades of studies have clarified many aspects of their population biology, based on several model species from diverse lineages of fungi where the EM symbiosis evolved, i.e. among Hymenomycetes and, to a lesser extent, among Ascomycetes. In this review, we show how tools for individual recognition have changed, shifting from the use of somatic incompatibility reactions to dominant and non-specific markers (such as random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP)) and, more recently, to co-dominant and specific markers (such as microsatellites and single nucleotide polymorphisms (SNPs)). At the same time, the theoretical focus has also changed. In earlier studies, a major aim was the description of genet size and popul/ation strategy. For example, we show how some studies supported or challenged the simple, classical model of colonization of new forest stands by ruderal (R) species, propagating by spores and forming small genets, progressively replaced in older forests by more competitive (C) species, propagating by mycelial growth and forming larger genets. By contrast, more recent studies give insights into some genetic traits, such as partners’ assortment (allo- versus autogamy), genetic structure of populations and gene flow that turn out to depend both on distance and on whether spores are animal- or wind-dispersed. We discuss the rising awareness that (i) many morphospecies contain cryptic biological species (often sympatric) and (ii) trans- and inter-continental species may often contain several biological species isolated by distance. Finally, we show the emergence of biogeographic approaches and call for some aspects to be developed, such as fine-scale and long-term population monitoring, analyses of subterranean populations of extra-radical mycelia, or more model species from the tropics, as well as from the Ascomycetes (whose genetic idiosyncrasies are discussed). With the rise of the ‘-omics’ sciences, analysis of population structure for non-neutral genes is expected to develop, and forest management and conservation biology will probably profit from published and expected work.

Pyrola rotundifolia (Ericaceae, Pyroleae tribe) is an understorey subshrub that was recently demonstrated to receive considerable amount of carbon from its fungal mycorrhizal associates. So far, little is known of the identity of these fungi and the mycorrhizal anatomy in the Pyroleae. Using 140 mycorrhizal root fragments collected from two Estonian boreal forests already studied in the context of mixotrophic Ericaceae in sequence analysis of the ribosomal DNA internal transcribed spacer region, we recovered 71 sequences that corresponded to 45 putative species in 19 fungal genera. The identified fungi were mainly ectomycorrhizal basidiomycetes, including Tomentella, Cortinarius, Russula, Hebeloma, as well as some ectomycorrhizal and/or endophytic ascomycetes. The P. rotundifolia fungal communities of the two forests did not differ significantly in terms of species richness, diversity and nutritional mode. The relatively high diversity retrieved suggests that P. rotundifolia does not have a strict preference for any fungal taxa. Anatomical analyses showed typical arbutoid mycorrhizae, with variable mantle structures, uniseriate Hartig nets and intracellular hyphal coils in the large epidermal cells. Whenever compared, fungal ultrastructure was congruent with the molecular identification. Similarly to other mixotrophic and autotrophic pyroloids in the same forests, P. rotundifolia shares its mycorrhizal fungal associates with surrounding trees that are likely a carbon source for pyroloids.

It is commonly assumed that ectomycorrhizal (ECM) fungi associated with temperate forest tree roots are not host-specific. Because this assumption relies on species delineations based on fruitbodies morphology or ribosomal DNA sequences, host-specific, cryptic biological species cannot be ruled out. To demonstrate that Laccaria amethystina has true generalist abilities, we sampled 510 fruitbodies on three French sites situated 150-450 km away from each other. At each site, populations from monospecific stands (Abies alba, Castanea europea and Fagus sylvatica) or mixed stands (F. sylvatica + Quercus robur or Q. robur +Carpinus betulus) were sampled. Three different sets of markers were used for genotyping: (i) five microsatellite loci plus the ribosomal DNA intergenic spacer, (ii) the mitochondrial large ribosomal DNA subunit, and (iii) direct amplification of length polymorphism (DALP), a new method for fungi providing dominant markers. Evidence for allogamous populations (with possible inbreeding at local scale) and possibly for biparental mitochondrial inheritance was found. All markers congruently demonstrated that L. amethystina populations show little structure at this geographical scale, indicating high gene flow (as many as 50% of founding spores in all populations being of external origin). Our results also showed that host species contributed even less to population differentiation, and there was no evidence for cryptic biological species. This first in situ demonstration of a true multihost ability in an ECM species is discussed in terms of ecology and evolutionary biology.

International audience; Stem canker of crucifers is caused by an ascomycete species complex comprising of two main species, Leptosphaeria maculans and L. biglobosa. These are composed of at least seven distinct subclades based on biochemical data or on sequences of internal transcribed spacer (ITS), the mating type MAT1-2 or fragments of actin or β-tubulin genes. In the course of a wide-scale characterization of the race structure of L. maculans from Western Australia, a few isolates from two locations failed to amplify specific sequences of L. maculans, i.e., the mating-type or minisatellite alleles. Based on both pathogenicity tests and ITS size, these isolates were classified as belonging to the L. biglobosa species. Parsimony and distance analyses performed on ITS, actin and β-tubulin sequences revealed that these isolates formed a new L. biglobosa subclade, more related to the Canadian L. biglobosa ‘canadensis’ subclade than to the L. biglobosa ‘australensis’ isolates previously described in Australia (Victoria). They are termed here as L. biglobosa ‘occiaustralensis’. These isolates were mainly recovered from resistant oilseed rape cultivars that included the Brassica rapa sp. sylvestris-derived resistance source, but not from the susceptible cv. Westar. The pathogenicity of L. biglobosa ‘occiaustralensis’ to cotyledons of most oilseed rape genotypes was higher than that of L. biglobosa ‘canadensis’ or L. biglobosa ‘australensis’ isolates.