Crop-to-wild gene flow in wild coffee species: the case of Coffea canephora in the Democratic Republic of the Congo.

BACKGROUND AND AIMS: Plant breeders are increasingly turning to crop wild relatives (CWRs) to ensure food security in a rapidly changing environment. However, CWR populations are confronted with various human-induced threats, including hybridization with their nearby cultivated crops. This might be a particular problem for wild coffee species, which often occur near coffee cultivation areas. Here, we briefly review the evidence for wild Coffea arabica (cultivated as Arabica coffee) and Coffea canephora (cultivated as Robusta coffee) and then focused on C. canephora in the Yangambi region in the Democratic Republic of the Congo. There, we examined the geographical distribution of cultivated C. canephora and the incidence of hybridization between cultivated and wild individuals within the rainforest. METHODS: We collected 71 C. canephora individuals from home gardens and 12 C. canephora individuals from the tropical rainforest in the Yangambi region and genotyped them using genotyping-by-sequencing (GBS). We compared the fingerprints with existing GBS data from 388 C. canephora individuals from natural tropical rainforests and the INERA Coffee Collection, a Robusta coffee field gene bank and the most probable source of cultivated genotypes in the area. We then established robust diagnostic fingerprints that genetically differentiate cultivated from wild coffee, identified cultivated-wild hybrids and mapped their geographical position in the rainforest. KEY RESULTS: We identified cultivated genotypes and cultivated-wild hybrids in zones with clear anthropogenic activity, and where cultivated C. canephora in home gardens may serve as a source for crop-to-wild gene flow. We found relatively few hybrids and backcrosses in the rainforests. CONCLUSIONS: The cultivation of C. canephora in close proximity to its wild gene pool has led to cultivated genotypes and cultivated-wild hybrids appearing within the natural habitats of C. canephora. Yet, given the high genetic similarity between the cultivated and wild gene pool, together with the relatively low incidence of hybridization, our results indicate that the overall impact in terms of risk of introgression remains limited so far.

Genetic diversity and structure of the coffee leaf rust fungus Hemileia vastatrix across different coffee management systems in Ethiopia.

Although coffee leaf rust (CLR), caused by Hemileia vastatrix, poses an increasing threat to coffee production in Ethiopia, little is known regarding its genetic diversity and structure and how these are affected by coffee management. Here, we used genetic fingerprinting based on sequence-related amplified polymorphism (SRAP) markers to genotype H. vastatrix samples from different coffee shrubs, across 40 sites, covering four coffee production systems (forest coffee, semi plantation coffee, home garden coffee, and plantation coffee) and different altitudes in Ethiopia. In total, 96 H. vastatrix samples were successfully genotyped with three primer combinations, producing a total of 79 scorable bands. We found 35.44% of amplified bands to be polymorphic, and the polymorphic information content (PIC) was 0.45, suggesting high genetic diversity among our CLR isolates. We also found significant isolation-by-distance across the samples investigated and detected significant differences in fungal genetic composition among plantation coffee and home garden coffee and a marginally significant difference among plantation coffee and forest coffee. Furthermore, we found a significant effect of altitude on CLR genetic composition in the forest coffee and plantation systems. Our results suggest that both spore dispersal and different selection pressures in the different coffee management systems are likely responsible for the observed high genetic diversity and genetic structure of CLR isolates in Ethiopia. When selecting Ethiopian coffee genotypes for crop improvement, it is important that these genotypes carry some resistance against CLR. Because our study shows large variation in genetic composition across relatively short geographical distances, a broad selection of rust isolates must be used for coffee resistance screening.

Genetic composition and diversity of Arabica coffee in the crop's centre of origin and its impact on four major fungal diseases.

Conventional wisdom states that genetic variation reduces disease levels in plant populations. Nevertheless, crop species have been subject to a gradual loss of genetic variation through selection for specific traits during breeding, thereby increasing their vulnerability to biotic stresses such as pathogens. We explored how genetic variation in Arabica coffee sites in southwestern Ethiopia was related to the incidence of four major fungal diseases. Sixty sites were selected along a gradient of management intensity, ranging from nearly wild to intensively managed coffee stands. We used genotyping-by-sequencing of pooled leaf samples (pool-GBS) derived from 16 individual coffee shrubs in each of the 60 sites to assess the variation in genetic composition (multivariate: reference allele frequency) and genetic diversity (univariate: mean expected heterozygosity) between sites. We found that genetic composition had a clear spatial pattern and that genetic diversity was higher in less managed sites. The incidence of the four fungal diseases was related to the genetic composition of the coffee stands, but in a specific way for each disease. In contrast, genetic diversity was only related to the within-site variation of coffee berry disease, but not to the mean incidence of any of the four diseases across sites. Given that fungal diseases are major challenges of Arabica coffee in its native range, our findings that genetic composition of coffee sites impacted the major fungal diseases may serve as baseline information to study the molecular basis of disease resistance in coffee. Overall, our study illustrates the need to consider both host genetic composition and genetic diversity when investigating the genetic basis for variation in disease levels.

Genetic diversity and structure in wild Robusta coffee (Coffea canephora A. Froehner) populations in Yangambi (DR Congo) and their relation to forest disturbance.

Degradation and regeneration of tropical forests can strongly affect gene flow in understorey species, resulting in genetic erosion and changes in genetic structure. Yet, these processes remain poorly studied in tropical Africa. Coffea canephora is an economically important species, found in the understorey of tropical rainforests of Central and West Africa, and the genetic diversity harboured in its wild populations is vital for sustainable coffee production worldwide. Here, we aimed to quantify genetic diversity, genetic structure, and pedigree relations in wild C. canephora populations, and we investigated associations between these descriptors and forest disturbance and regeneration. Therefore, we sampled 256 C. canephora individuals within 24 plots across three forest categories in Yangambi (DR Congo), and used genotyping-by-sequencing to identify 18,894 SNPs. Overall, we found high genetic diversity, and no evidence of genetic erosion in C. canephora in disturbed old-growth forest, as compared to undisturbed old-growth forest. In addition, an overall heterozygosity excess was found in all populations, which was expected for a self-incompatible species. Genetic structure was mainly a result of isolation-by-distance, reflecting geographical location, with low to moderate relatedness at finer scales. Populations in regrowth forest had lower allelic richness than populations in old-growth forest and were characterised by a lower inter-individual relatedness and a lack of isolation-by-distance, suggesting that they originated from different neighbouring populations and were subject to founder effects. Wild Robusta coffee populations in the study area still harbour high levels of genetic diversity, yet careful monitoring of their response to ongoing forest degradation remains required.

Multiple taxa inoculants of arbuscular mycorrhizal fungi enhanced colonization frequency, biomass production, and water use efficiency of cassava (Manihot esculenta).

Increasing water use efficiency (WUE) in crops is critical to maintaining agricultural production under climate change-exacerbated drought. One of these approaches may consist of leveraging on the beneficial interactions between crops and arbuscular mycorrhizal fungi (AMF). In this study, we investigated how inoculation with AMF from three different taxa (Claroideoglomus etunicatum (T1), Gigaspora margarita (T2), and Rhizophagus irregularis (T3)) and their combination (T123) and a non-inoculated "control" treatment in a greenhouse could achieve increased biomass production and water use efficiency in cassava under three levels of water availability (100% PC, 60%-moderate stress, and 30%-severe stress). Whereas T1 and T2 resulted in a lower growth rate for the plants than the control, T123 enhanced cassava height and the number of petioles and leaves. T123 and T3 increased the total plant dry biomass in comparison with uninoculated plants by 30% and 26%, respectively. The T123 and plants inoculated with T3 significantly increased cassava above-ground biomass by 19% as compared to T1 (8.68 ± 2.44 g) and T2 (8.68 ± 2.44 g) inoculated plants. T123 resulted in higher WUE, which was validated by the leaf carbon (δ13C) isotopic signature, significantly outperforming cassava with T1 and T2, yet there was no difference between the control and T3. Overall, this study demonstrated that the use of multiple AMF from different taxa can increase cassava growth and WUE under greenhouse conditions.

The impact of habitat loss on molecular signatures of coevolution between an iconic butterfly (Alcon blue) and its host plant (Marsh gentian).

Habitat loss is threatening natural communities worldwide. Small and isolated populations suffer from inbreeding and genetic drift, which jeopardize their long-term survival and adaptive capacities. However, the consequences of habitat loss for reciprocal coevolutionary interactions remain poorly studied. In this study, we investigated the effects of decreasing habitat patch size and connectivity associated with habitat loss on molecular signatures of coevolution in the Alcon blue butterfly (Phengaris alcon) and its most limited host, the marsh gentian (Gentiana pneumonanthe). Because reciprocal coevolution is characterized by negative frequency-dependent selection as a particular type of balancing selection, we investigated how signatures of balancing selection vary along a gradient of patch size and connectivity, using single nucleotide polymorphisms (SNPs). We found that signatures of coevolution were unaffected by patch characteristics in the host plants. On the other hand, more pronounced signatures of coevolution were observed in both spatially isolated and in large Alcon populations, together with pronounced spatial variation in SNPs that are putatively involved in coevolution. These findings suggest that habitat loss can facilitate coevolution in large butterfly populations through limiting swamping of locally beneficial alleles by maladaptive ones. We also found that allelic richness (Ar) of the coevolutionary SNPs is decoupled from neutral Ar in the butterfly, indicating that habitat loss has different effects on coevolutionary as compared with neutral processes. We conclude that this specialized coevolutionary system requires particular conservation interventions aiming at generating a spatial mosaic of both connected and of isolated habitat to maintain coevolutionary dynamics.

Could green infrastructure supplement ecosystem service provision from semi-natural grasslands?

Ancient semi-natural grasslands in Europe are important for ecosystem service (ES) provision. Often, the surrounding matrix contains 'Grassland Green Infrastructure' (GGI) that contain grassland species which have the potential to supplement grassland ES provision across the landscape. Here we investigate the potential for GGI to deliver a set of complementary ES, driven by plant composition.We surveyed 36 landscapes across three European countries comprising core grasslands and their surrounding GGI. We calculated community-level values of plant species characteristics to provide indicators for four ES: nature conservation value, pollination, carbon storage and aesthetic appeal.Inferred ES delivery for GGI was substantially lower than in core grasslands for conservation, pollination and aesthetic appeal indicators, but not for carbon storage. These differences were driven by the GGI having 17% fewer plant species, and compositional differences, with 61% of species unique to the core grasslands. In addition, connectivity to the core, the amount of GGI and inferred seed dispersal distances by livestock, were strongly positively correlated with conservation value, pollination and aesthetic indicators. All ES indicators showed similar responses to the GGI spatial structure and distance to the core, suggesting robust effects of these drivers on ES. We projected that improved landscape-wide delivery of nature conservation value and pollination could be achieved through targeted GGI management. Reductions in the distances seeds would need to disperse, more GGI, along with a diversification of the GGI elements, were predicted to enhance service credits.We conclude that for vegetation-related ES, species surveys can be employed to assess potential ES delivery. Creating and enhancing GGI is a useful landscape management strategy to supplement the ES delivered by ancient grasslands.

Signatures of polygenic adaptation align with genome-wide methylation patterns in wild strawberry plants.

Epigenetic inheritance can drive adaptive evolution independently of DNA sequence variation. However, to what extent epigenetic variation represents an autonomous evolutionary force remains largely elusive. Through gene ontology and comparative analyses of genomic and epigenomic variation of wild strawberry plants raised in distinct drought settings, we characterised genome-wide covariation between single nucleotide polymorphisms (SNPs) and differentially methylated cytosines (DMCs). Covariation between SNPs and DMCs was independent of genomic proximity, but instead associated with fitness-related processes such as stress responses, genome regulation and reproduction. We expected this functional SNP-DMC covariation to be driven by adaptive evolution canalising SNP and DMC variation, but instead observed significantly lower covariation with DMCs for adaptive rather than for neutral SNPs. Drought-induced DMCs frequently co-varied with tens of SNPs, suggesting high genomic redundancy as a broad potential basis for polygenic adaptation of gene expression. Our findings suggest that stress-responsive DMCs initially co-vary with many SNPs under increased environmental stress, and that natural selection acting upon several of these SNPs subsequently reduces standing covariation with stress-responsive DMCs. Our study supports DNA methylation profiles that represent complex quantitative traits rather than autonomous evolutionary forces. We provide a conceptual framework for polygenic regulation and adaptation shaping genome-wide methylation patterns in plants.

Spatial variability and environmental drivers of cassava-arbuscular mycorrhiza fungi (AMF) associations across Southern Nigeria.

Cassava, forming starch-rich, tuberous roots, is an important staple crop in smallholder farming systems in sub-Saharan Africa. Its relatively good tolerance to drought and nutrient-poor soils may be partly attributed to the crop's association with arbuscular mycorrhiza fungi (AMF). Yet insights into AMF-community composition and richness of cassava, and knowledge of its environmental drivers are still limited. Here, we sampled 60 cassava fields across three major cassava-growing agro-ecological zones in Nigeria and used a DNA meta-barcoding approach to quantify large-scale spatial variation and evaluate the effects of soil characteristics and common agricultural practices on AMF community composition, richness and Shannon diversity. We identified 515 AMF operational taxonomic units (OTUs), dominated by Glomus, with large variation across agro-ecological zones, and with soil pH explaining most of the variation in AMF community composition. High levels of soil available phosphorus reduced OTU richness without affecting Shannon diversity. Long fallow periods (> 5 years) reduced AMF richness compared with short fallows, whereas both zero tillage and tractor tillage reduced AMF diversity compared with hoe tillage. This study reveals that the symbiotic relationship between cassava and AMF is strongly influenced by soil characteristics and agricultural management and that it is possible to adjust cassava cultivation practices to modify AMF diversity and community structure.

Both organic and integrated pest management of apple orchards maintain soil health as compared to a semi-natural reference system.

Growing concerns about the negative environmental impacts of agriculture have resulted in the increasing adoption of farming systems that try to reconcile crop production with environmental sustainability, such as organic farming. As organic farming refrains from using synthetic inputs, it heavenly relies on maintaining soil health. However, it is still poorly understood how organic management performs in terms of maintaining soil health in real commercial and heterogeneous farm settings as compared to conventional management, and especially as compared to a natural reference system. Here, we compared a set of soil health indicators among 24 commercial apple orchards that were either managed organically or conventionally using Integrated Pest Management (IPM) practices. In addition, we quantified the same indicators in 12 semi-natural grasslands as a benchmark to assess to what extent soil processes and functions have been degraded due to agricultural practices. As soil heath indicators, we quantified soil bulk density, organic matter content, organic carbon content, organic carbon stock, total nitrogen (N), potential heterotrophic respiration, potential net N mineralization, litter decomposition and litter stabilization, and we added the diversity of the herbaceous vegetation and the soil microbiome as covariates in our models. We found no differences between organic and IPM orchards, and neither of the farming systems showed evidence of impaired soil health compared to the semi-natural benchmark, with the exception of higher decomposition rates measured in both orchard types. We observed, however, high spatial variation in soil health between drive and crop rows within the orchards. Especially in the IPM orchards, crop rows showed impaired soil health compared to the adjacent drive rows, indicating that there is still opportunity to improve soil management in the IPM system. In addition, our results show that a considerable part of the variation in soil characteristics can be attributed to the study site, suggesting that both natural heterogeneity and personal management preferences by individual farmers are more important than the management system. Overall, and at least in terms of the soil variables measured in this study, our results suggest that perennial crop systems can be managed in a sustainable way, without jeopardizing soil health.

Improved genotypes and fertilizers, not fallow duration, increase cassava yields without compromising arbuscular mycorrhizal fungus richness or diversity.

Arbuscular mycorrhizal fungi (AMF) are ubiquitous in agroecosystems, but their role in mediating agricultural yield remains contested. Field experiments testing effects of realistic agronomic practices of intensification on AM fungus composition and yields are scarce, especially in the low-input systems of sub-Saharan Africa. A large, full-factorial field experiment was conducted in South-Kivu (DR Congo), testing effects of fallow duration (6 vs. 12 months), genotype (landrace vs. improved), and fertilizer management (control vs. five combinations omitting N, P, K, and/or secondary macro- and micronutrients) on yields of cassava, an important staple crop strongly colonized by AMF. Furthermore, we used DNA-metabarcoding to evaluate effects of these agronomic practices on the AM fungal communities on the roots. The shorter fallow duration strongly increased diversity and richness of AMF, but this did not correspond with increased yields. Cassava yield was mainly determined by genotype, being largest for the improved genotype, which coincided with a significantly higher sum of AM fungal sequences. Effects of fertilizer or genotype on community composition were minor to absent. We found no evidence that increased AMF richness and diversity enhanced cassava yields. In contrast, the use of the improved genotype and mineral fertilizers strongly benefitted yields, without compromising richness or diversity of AMF. Cassava-AMF associations in this work appear robust to fertilizer amendments and modern genotype improvement.

Diversity and community structure of ericoid mycorrhizal fungi in European bogs and heathlands across a gradient of nitrogen deposition.

Despite the ecological significance of ericoid mycorrhizal fungi, little is known about the abiotic and biotic factors driving their diversity and community composition. To determine the relative importance of abiotic and biotic filtering in structuring ericoid mycorrhizal fungal communities, we established 156 sampling plots in two highly contrasting environments but dominated by the same Ericaceae plant species: waterlogged bogs and dry heathlands. Plots were located across 25 bogs and 27 dry heathlands in seven European countries covering a gradient in nitrogen deposition and phosphorus availability. Putatively ericoid mycorrhizal fungal communities in the roots of 10 different Ericaceae species were characterized using high-throughput amplicon sequencing. Variation in ericoid mycorrhizal fungal communities was attributed to both habitat and soil variables on the one hand and host plant identity on the other. Communities differed significantly between bogs and heathlands and, in a given habitat, communities differed significantly among host plant species. Fungal richness was negatively related to nitrogen deposition in bogs and phosphorus availability in bogs and heathlands. Our results demonstrate that both abiotic and biotic filtering shapes ericoid mycorrhizal fungal communities and advocate an environmental policy minimizing excess nutrient input in these nutrient-poor ecosystems to avoid loss of ericoid mycorrhizal fungal taxa.

Effects of single and multiple species inocula of arbuscular mycorrhizal fungi on the salinity tolerance of a Bangladeshi rice (Oryza sativa L.) cultivar.

Soil salinization due to sea level rise and groundwater irrigation has become an important agronomic problem in many parts of the world. Symbiosis between crop species and arbuscular mycorrhizal fungi (AMF) may alleviate salt stress-induced detrimental effects on crop growth and yield, for example, through helping the host plant to selectively absorb potassium while avoiding uptake of excessive sodium. Here, we performed a greenhouse experiment to evaluate growth, grain yield, and salt tolerance of a Bangladeshi rice cultivar under three levels of salt stress (0, 75, and 120 mM) after inoculation with three different AMF species from three different genera (Funnelliformis mosseae (BEG12), Acaulospora laevis (BEG13), and Gigaspora margarita (BEG34)), singly and in combination. We found that under salt stress, AMF inoculation enhanced total chlorophyll concentration, shoot K+/Na+ ratio, and lowered shoot Na+/root Na+ ratio, accompanied by increased root biomass, spikelet fertility, and grain yield compared with the non-inoculated control plants. Specifically, we found that the combination of BEG13 and BEG34 increased rice yield by 125 and 143% as compared with the non-inoculated controls, at the 75 and 120mM salt levels, respectively. In general, the low AMF diversity treatments (one species or a combination of two AMF species) were found to be the most effective in mediating salt stress tolerance for the majority of the measured crop performance variables. Overall, our results indicate that specific AMF species can promote the salt tolerance and productivity of rice, likely by increasing photosynthetic efficiency and restricting Na+ uptake and transport from root to shoot in AMF-inoculated plants.

Local abiotic conditions are more important than landscape context for structuring arbuscular mycorrhizal fungal communities in the roots of a forest herb.

Due to human influence, large tracts of natural vegetation have been cleared and replaced by other types of land use, resulting in highly fragmented landscapes consisting of small fragments of well-conserved habitat scattered within a matrix of intensively managed land. Changes in land use and associated fragmentation have important consequences for biodiversity in the remaining fragments. Most studies so far have investigated the impact of land use change on macro-organisms, but little is known about how landscape fragmentation affects microbial communities. Here, we studied how changes in land use and abiotic conditions affected the arbuscular mycorrhizal fungal (AMF) communities in the roots of the forest herb Stachys sylvatica. Root samples were collected from 40 populations occurring in fragmented forest patches of varying age and size embedded within an agricultural landscape. Our results showed that forest age and isolation did not affect AMF diversity or community composition, suggesting that AMF disperse easily throughout the landscape and that AMF communities reassemble fast in recently established forest patches. On the other hand, AMF richness increased with increasing forest area, indicating that small forest sizes limit AMF richness. Additionally, AMF richness increased with increasing soil pH and decreased with soil nitrate content, while AMF community composition was affected plant-available phosphorus. Overall, these results show that landscape context is less important than local abiotic conditions for structuring AMF communities. However, the significant area effect indicates that further reductions in forest area will lead to impoverished AMF communities, potentially affecting long-term plant fitness and community structure.

Biodiversity and human health: mechanisms and evidence of the positive health effects of diversity in nature and green spaces.

Introduction: Natural environments and green spaces provide ecosystem services that enhance human health and well-being. They improve mental health, mitigate allergies and reduce all-cause, respiratory, cardiovascular and cancer mortality. The presence, accessibility, proximity and greenness of green spaces determine the magnitude of their positive health effects, but the role of biodiversity (including species and ecosystem diversity) within green spaces remains underexplored. This review describes mechanisms and evidence of effects of biodiversity in nature and green spaces on human health. Sources of data: We identified studies listed in PubMed and Web of Science using combinations of keywords including 'biodiversity', 'diversity', 'species richness', 'human health', 'mental health' and 'well-being' with no restrictions on the year of publication. Papers were considered for detailed evaluation if they were written in English and reported data on levels of biodiversity and health outcomes. Areas of agreement: There is evidence for positive associations between species diversity and well-being (psychological and physical) and between ecosystem diversity and immune system regulation. Areas of concern: There is a very limited number of studies that relate measured biodiversity to human health. There is more evidence for self-reported psychological well-being than for well-defined clinical outcomes. High species diversity has been associated with both reduced and increased vector-borne disease risk. Growing points: Biodiversity supports ecosystem services mitigating heat, noise and air pollution, which all mediate the positive health effects of green spaces, but direct and long-term health outcomes of species diversity have been insufficiently studied so far. Areas timely for research: Additional research and newly developed methods are needed to quantify short- and long-term health effects of exposure to perceived and objectively measured species diversity, including health effects of nature-based solutions and exposure to microbiota.

Abiotic rather than biotic filtering shapes the arbuscular mycorrhizal fungal communities of European seminatural grasslands.

Although it is well known that arbuscular mycorrhizal fungi (AMF) play a key role in the functioning of natural ecosystems, the underlying drivers determining the composition of AMF communities remain unclear. In this study, we established 138 sampling plots at 46 grassland sites, consisting of 26 acidic grasslands and 20 calcareous grasslands spread across eight European countries, to assess the relative importance of abiotic and biotic filtering in driving AMF community composition and structure in both the grassland soils and in the roots of 13 grassland plant species. Soil AMF communities differed significantly between acidic and calcareous grasslands. In root AMF communities, most variance was attributable to soil variables while very little variation was explained by host plant identity. Root AMF communities in host plant species occurring in only one grassland type closely resembled the soil AMF communities of that grassland type and the root AMF communities of other host plant species occurring in the same grassland type. The observed AMF-host plants networks were not modular but nested. Our results indicate that abiotic conditions, rather than biotic filtering through host plant specificity, are the most important drivers in shaping AMF communities in European seminatural grasslands.

Retention of gene diversity during the spread of a non-native plant species.

Spatial expansion, which is a crucial stage in the process to successful biological invasion, is anticipated to profoundly affect the magnitude and spatial distribution of genetic diversity in novel colonized areas. Here, we show that, contrasting common expectations, Pyrenean rocket (Sisymbrium austriacum), retained SNP diversity as this introduced plant species descended in the Meuse River Basin. Allele frequencies did not mirror between-population distances along the predominant expansion axis. Reconstruction of invasion history based on the genotypes of historical herbarium specimens indicated no influence of additional introductions or multiple points of entry on this nongradual pattern. Assignment analysis suggested the admixture of distant upstream sources in recently founded downstream populations. River dynamics seem to have facilitated occasional long-distance dispersal which brought diversity to the expansion front and so maintained evolutionary potential. Our findings highlight the merit of a historical framework in interpreting extant patterns of genetic diversity in introduced species and underscore the need to integrate long-distance dispersal events in theoretical work on the genetic consequences of range expansion.

Does the seed bank contribute to the build-up of a genetic extinction debt in the grassland perennial Campanula rotundifolia?

Background and Aims: Habitat fragmentation threatens global biodiversity. Many plant species persist in habitat fragments via persistent life cycle stages such as seed banks, generating a species extinction debt. Here, seed banks are hypothesized to cause a temporal delay in the expected loss of genetic variation, which can be referred to as a genetic extinction debt, as a possible mechanism behind species extinction debts. Methods: Fragmented grassland populations of Campanula rotundifolia were examined for evidence of a genetic extinction debt, investigating if the seed bank contributed to the extinction debt build-up. The genetic make-up of 15 above- and below-ground populations was analysed in relation to historical and current levels of habitat fragmentation, both separately and combined. Key Results: Genetic diversity was highest in above-ground populations, though below-ground populations contained 8 % of unique alleles that were absent above-ground. Above-ground genetic diversity and composition were related to historical patch size and connectivity, but not current patch characteristics, suggesting the presence of a genetic extinction debt in the above-ground populations. No such relationships were found for the below-ground populations. Genetic diversity measures still showed a response to historical but not present landscape characteristics when combining genetic diversity of the above- and below-ground populations. Conclusions: The fragmented C. rotundifolia populations exhibited a genetic extinction debt. However, the role of the seed banks in the build-up of this extinction debt is probably small, since the limited, unique genetic diversity of the seed bank alone seems unable to counter the detrimental effects of habitat fragmentation on the population genetic structure of C. rotundifolia.

Biogeographical Patterns of Legume-Nodulating Burkholderia spp.: from African Fynbos to Continental Scales.

UNLABELLED: Rhizobia of the genus Burkholderia have large-scale distribution ranges and are usually associated with South African papilionoid and South American mimosoid legumes, yet little is known about their genetic structuring at either local or global geographic scales. To understand variation at different spatial scales, from individual legumes in the fynbos (South Africa) to a global context, we analyzed chromosomal (16S rRNA, recA) and symbiosis (nifH, nodA, nodC) gene sequences. We showed that the global diversity of nodulation genes is generally grouped according to the South African papilionoid or South American mimosoid subfamilies, whereas chromosomal sequence data were unrelated to biogeography. While nodulation genes are structured on a continental scale, a geographic or host-specific distribution pattern was not detected in the fynbos region. In host range experiments, symbiotic promiscuity of Burkholderia tuberum STM678(T) and B phymatum STM815(T) was discovered in selected fynbos species. Finally, a greenhouse experiment was undertaken to assess the ability of mimosoid (Mimosa pudica) and papilionoid (Dipogon lignosus, Indigofera filifolia, Macroptilium atropurpureum, and Podalyria calyptrata) species to nodulate in South African (fynbos) and Malawian (savanna) soils. While the Burkholderia-philous fynbos legumes (D lignosus, I filifolia, and P calyptrata) nodulated only in their native soils, the invasive neotropical species M pudica did not develop nodules in the African soils. The fynbos soil, notably rich in Burkholderia, seems to retain nodulation genes compatible with the local papilionoid legume flora but is incapable of nodulating mimosoid legumes that have their center of diversity in South America. IMPORTANCE: This study is the most comprehensive phylogenetic assessment of root-nodulating Burkholderia and investigated biogeographic and host-related patterns of the legume-rhizobial symbiosis in the South African fynbos biome, as well as at global scales, including native species from the South American Caatinga and Cerrado biomes. While a global investigation of the rhizobial diversity revealed distinct nodulation and nitrogen fixation genes among South African and South American legumes, regionally distributed species in the Cape region were unrelated to geographic and host factors.

Transatlantic invasion routes and adaptive potential in North American populations of the invasive glossy buckthorn, Frangula alnus.

BACKGROUND AND AIMS: Many invasive species severely threaten native biodiversity and ecosystem functioning. One of the most prominent questions in invasion genetics is how invasive populations can overcome genetic founder effects to establish stable populations after colonization of new habitats. High native genetic diversity and multiple introductions are expected to increase genetic diversity and adaptive potential in the invasive range. Our aim was to identify the European source populations of Frangula alnus (glossy buckthorn), an ornamental and highly invasive woody species that was deliberately introduced into North America at the end of the 18th century. A second aim of this study was to assess the adaptive potential as an explanation for the invasion success of this species. METHODS: Using a set of annotated single-nucleotide polymorphisms (SNPs) that were assigned a putative function based on sequence comparison with model species, a total of 38 native European and 21 invasive North American populations were subjected to distance-based structure and assignment analyses combined with population genomic tools. Genetic diversity at SNPs with ecologically relevant functions was considered as a proxy for adaptive potential. KEY RESULTS: Patterns of invasion coincided with early modern transatlantic trading routes. Multiple introductions through transatlantic trade from a limited number of European port regions to American urban areas led to the establishment of bridgehead populations with high allelic richness and expected heterozygosity, allowing continuous secondary migration to natural areas. CONCLUSIONS: Targeted eradication of the urban populations, where the highest genetic diversity and adaptive potential were observed, offers a promising strategy to arrest further invasion of native American prairies and forests.