Evaluating the hologenome concept by analyzing the root-endosphere microbiota of chimeric plants.

The hologenome concept considers the entity formed by a host and its microbiota, the holobiont, as new level of hierarchical organization subject to neutral and selective forces. We used grafted plants to formally evaluate the hologenome concept. We analyzed the root-endosphere microbiota of two independent watermelon and grapevine plant systems, including ungrafted and reciprocal-grafting combinations. Grafted and ungrafted hosts harbor markedly different microbiota compositions. Furthermore, the results indicate a non-random assembly of bacterial communities inhabiting the root endosphere of chimeric plants with interactive effect of both the rootstock and scion on the recruitment of microorganisms. Because chimeric plants did not have a random microbiota, the null hypothesis that holobionts assemble randomly and hologenome concept is an intellectual construction only can be rejected. The study supports the relevance of hologenome as biological level of organization and opens new avenues for a better fundamental understanding of plants as holobionts.

Ecological corridors homogenize plant root endospheric mycobiota.

Ecological corridors promote species coexistence in fragmented habitats where dispersal limits species fluxes. The corridor concept was developed and investigated with macroorganisms in mind, while microorganisms, the invisible majority of biodiversity, were disregarded. We analyzed the effect of corridors on the dynamics of endospheric fungal assemblages associated with plant roots at the scale of 1 m over 2 years (i.e. at five time points) by combining an experimental corridor-mesocosm with high-throughput amplicon sequencing. We showed that plant root endospheric mycobiota were sensitive to corridor effects when the corridors were set up at a small spatial scale. The endospheric mycobiota of connected plants had higher species richness, lower beta-diversity, and more deterministic assembly than the mycobiota of isolated plants. These effects became more pronounced with the development of host plants. Biotic corridors composed of host plants may thus play a key role in the spatial dynamics of microbial communities and may influence microbial diversity and related ecological functions.

Plant microbiota dysbiosis and the Anna Karenina Principle.

Microorganisms are associated with all plants, recently leading to the hologenome concept. We reviewed the assembly processes of plant microbiota and analyzed its structure during the emergence of dysbioses. In particular, we discussed the Anna Karenina Principle (AKP) based on Leo Tolstoy's assertion applied to plant microbiota: 'All healthy microbiota are alike; each disease-associated microbiota is sick in its own way.' We propose the AKP to explain how stochastic processes in plant microbiota assembly due to several external stressors could lead to plant diseases. Finally, we propose the AKP to conceptualize plant dysbioses as a transitory loss of host capacity to regulate its microbiota, implying a loss of function that leads to a reduction of the host's fitness.

Organic agriculture and field edges uphold endospheric wheat microbiota at field and landscape scale.

Agricultural intensification has been demonstrated to induce a loss of biodiversity. Despite the key role of symbiotic microorganisms in plant nutrition and protection, the impact of agricultural intensification on these microorganisms is not fully understood. Organic farming and field edges (as semi-natural elements) may promote a higher microbial diversity thanks to lower anthropic disturbance and higher plant diversity. We sampled wheat individuals in pairs of wheat fields (one organic and one conventional) along a distance gradient to the edges (hedgerow vs. grassy), in 20 landscape windows selected along an uncorrelated gradient of organic farming and hedgerow density. We demonstrated that organic farming shaped microbial composition and increased fungal and bacterial richness, while hedgerows had a neutral or negative effect on richness depending on the microbial phyla considered. In contrast to bacteria, fungal communities were heterogeneously distributed within fields, having a higher diversity for some phyla close to field edges. Overall we highlighted that fungi responded more to the field scale while bacteria were more affected by landscape scale. The effect of agricultural intensification on plant microbiota and therefore on the functions provided by microorganisms to the plants has to be considered at a multiple spatial scale-from field to landscape.

How does interspecific competition modify the response of grass plants against herbicide treatment? A hierarchical concentration-response approach.

Ecological interactions are rarely taken into account in environmental risk assessment. The objective of this work was to assess how interspecific competition affects the way plant species react to herbicides and more specifically how it modifies the concentration-response curves that can be built using ecotoxicological bioassays. To do this, we relied on the results of ecotoxicological bioassays on six herbaceous species exposed to isoproturon under two conditions: in presence and in absence of a competitor. At the end of the experiments, eleven endpoints were measured. We modelled these data using a hierarchical modelling framework designed to assess the effects of competition on each of the four parameters of the concentration response curves (e.g. the level of response at the control or the concentration at the inflection point of the curve) simultaneously for the six species. The modelled effects could be of three types, 1) competition had no effect on the parameter, 2) competition had the same effect on the parameter for all species and 3) competition had a different effect on the parameter for each species. Our main hypothesis was that different species would react differently to competition. Results showed that about a half of the estimated parameters showed a modification under competition pressure among which only a fourth showed a species-specific effect, the three other fourth showing the same effect between the different species. Our initial hypothesis was thus not supported as species tended to react in the same way to competition. The competition effect on plants was mainly negative, thus showing that they were more affected by isoproturon under competition pressure. This study therefore establishes how competition modifies plant responses to chemical stress and how this interaction varies from one species to the other.

Past landscape structure drives the functional assemblages of plants and birds.

Landscape structure is a major driver of biodiversity in agricultural landscapes. However, the response of biodiversity can be delayed after landscape changes. This study aimed to determine the effect of current and past landscape structure on plant and bird assemblages. We used a trait-based approach to understand their responses to landscape simplification and habitat fragmentation. We quantified landscape structure at three different years (1963, 1985, 2000) and sampled current plant and bird assemblages in twenty 1 km2 landscape windows located along the Seine Valley (France). For each window, we calculated plant and bird species richness, Community Weighted Variance (CWV), and Community Weighted Mean (CWM) of five functional traits related to dispersal capacity, reproduction, and life-cycle. We detected non-random patterns of traits for both taxa. Plant and bird species richness was lower in simple landscapes. The functional variance of plant traits was higher in landscapes simple in configuration. Both plant and bird assemblages strongly responded to past landscapes, especially their traits related to reproduction and life-cycle. It suggests that landscapes of the Seine valley will face a functional extinction debt. Further research is needed to better predict the delayed response of biodiversity expected to occur after landscape structure changes.

Differentiation of endospheric microbiota in ancient and modern wheat cultivar roots.

Modern plant breeding and agrosystems artificialization could have altered plants' ability to filter and recruit beneficial microorganisms in its microbiota. Thus, compared to modern cultivars, we hypothesized that root-endosphere microbiota in modern wheat cultivars are less resistant to colonization by fungi and bacteria and thus more susceptible to also recruit more pathogens. We used an in-field experimental design including six wheat varieties (three ancient vs. three modern) grown in monoculture and in mixture (three replicates each). Endospheric microbiota of wheat roots were analyzed on four individuals sampled randomly in each plot. Composition-based clustering of sequences was then characterized from amplicon mass-sequencing. We show that the bacterial and fungal microbiota composition in wheat roots differed between ancient and modern wheat cultivar categories. However, the responses observed varied with the group considered. Modern cultivars harbored higher richness of bacterial and fungal pathogens than ancient cultivars. Both cultivar types displayed specific indicator species. A synergistic effect was identified in mixtures of modern cultivars with a higher root endospheric mycobiota richness than expected from a null model. The present study shows the effect of plant breeding on the microbiota associated plant roots. The results call for making a diagnosis of the cultivar's endospheric-microbiota composition. These new results also suggest the importance of a holobiont-vision while considering plant selection in crops and call for better integration of symbiosis in the development of next-generation agricultural practices.

A Landscape of Opportunities for Microbial Ecology Research.

Microbes encompass tremendous biodiversity, provide support to all living forms, including humans, and play an important role in many ecosystem services. The rules that govern microorganism community assembly are increasingly revealed due to key advances in molecular and analytical methods but their understanding remain a key challenge in microbial ecology. The existence of biogeographic patterns within microbial communities has been established and explained in relation to landscape-scale processes, including selection, drift, dispersal and mutation. The effect of habitat patchiness on microorganisms' assembly rules remains though incompletely understood. Here, we review how landscape ecology principles can be adapted to explore new perspectives on the mechanisms that determine microbial community structure. To provide a general overview, we characterize microbial landscapes, the spatial and temporal scales of the mechanisms that drive microbial assembly and the feedback between microorganisms and landscape structure. We provide evidence for the effects of landscape heterogeneity, landscape fragmentation and landscape dynamics on microbial community structure, and show that predictions made for macro-organisms at least partly also apply to microorganisms. We explain why emerging metacommunity approaches in microbial ecology should include explicit characterization of landscape structure in their development and interpretation. We also explain how biotic interactions, such as competition, prey-predator or mutualist relations may influence the microbial landscape and may be involved in the above-mentioned feedback process. However, we argue that the application of landscape ecology to the microbial world cannot simply involve transposing existing theoretical frameworks. This is due to the particularity of these organisms, in terms of size, generation time, and for some of them, tight interaction with hosts. These characteristics imply dealing with unusual and dependent space and time scales of effect. Evolutionary processes have also a strong importance in microorganisms' response to their landscapes. Lastly, microorganisms' activity and distribution induce feedback effects on the landscape that have to be taken into account. The transposition of the landscape ecology framework to microorganisms provides many challenging research directions for microbial ecology.

Local-scale dynamics of plant-pesticide interactions in a northern Brittany agricultural landscape.

Soil pollution by anthropogenic chemicals is a major concern for sustainability of crop production and of ecosystem functions mediated by natural plant biodiversity. Understanding the complex effects of soil pollution requires multi-level and multi-scale approaches. Non-target and agri-environmental plant communities of field margins and vegetative filter strips are confronted with agricultural xenobiotics through soil contamination, drift, run-off and leaching events that result from chemical applications. Plant-pesticide dynamics in vegetative filter strips was studied at field scale in the agricultural landscape of a long-term ecological research network in northern Brittany (France). Vegetative filter strips effected significant pesticide abatement between the field and riparian compartments. However, comparison of pesticide usage modalities and soil chemical analysis revealed the extent and complexity of pesticide persistence in fields and vegetative filter strips, and suggested the contribution of multiple sources (yearly carry-over, interannual persistence, landscape-scale contamination). In order to determine the impact of such persistence, plant dynamics was followed in experimentally-designed vegetative filter strips of identical initial composition (Agrostis stolonifera, Anthemis tinctoria/Cota tinctoria, Centaurea cyanus, Fagopyrum esculentum, Festuca rubra, Lolium perenne, Lotus corniculatus, Phleum pratense, Trifolium pratense). After homogeneous vegetation establishment, experimental vegetative filter strips underwent rapid changes within the following two years, with Agrostis stolonifera, Festuca rubra, Lolium perenne and Phleum pratense becoming dominant and with the establishment of spontaneous vegetation. Co-inertia analysis showed that plant dynamics and soil residual pesticides could be significantly correlated, with the triazole fungicide epoxiconazole, the imidazole fungicide prochloraz and the neonicotinoid insecticide thiamethoxam as strong drivers of the correlation. However, the correlation was vegetative-filter-strip-specific, thus showing that correlation between plant dynamics and soil pesticides likely involved additional factors, such as threshold levels of residual pesticides. This situation of complex interactions between plants and soil contamination is further discussed in terms of agronomical, environmental and health issues.

Effect of interspecific competition on species sensitivity distribution models: Analysis of plant responses to chemical stress.

Species Sensitivity Distributions (SSD) are widely used in environmental risk assessment to predict the concentration of a contaminant that is hazardous for 5% of species (HC5). They are based on monospecific bioassays conducted in the laboratory and thus do not directly take into account ecological interactions. This point, among others, is accounted for in environmental risk assessment through an assessment factor (AF) that is applied to compensate for the lack of environmental representativity. In this study, we aimed to assess the effects of interspecific competition on the responses towards isoproturon of plant species representative of a vegetated filter strip community, and to assess its impact on the derived SSD and HC5 values. To do so, we realized bioassays confronting six herbaceous species to a gradient of isoproturon exposure in presence and absence of a competitor. Several modelling approaches were applied to see how they affected the results, using different critical effect concentrations and investigating different ways to handle multiple endpoints in SSD. At the species level, there was a strong trend toward organisms being more sensitive to isoproturon in presence of a competitor than in its absence. At the community level, this trend was also observed in the SSDs and HC5 values were always lower in presence of a competitor (1.12-11.13 times lower, depending on the modelling approach). Our discussion questions the relevance of SSD and AF as currently applied in environmental risk assessment.

Artificial Intelligence Accidentally Learned Ecology through Video Games.

An advanced artificial intelligence (AI) system defeated the best human players in StarCraft II, a popular real-time strategy game. In a virtual ecosystem, players compete for habitats and resources, unintentionally reproducing many ecological phenomena. We propose to repurpose this AI to test ecological hypotheses that have been intractable using traditional approaches.

The influence of host-plant connectivity on fungal assemblages in the root microbiota of Brachypodium pinnatum.

Dispersal limitation may drive the structure of fungal microbiota of plant roots at small spatial scales. Fungal root microorganisms disperse through the plant rooting systems from hosts to hosts. Due to a pronounced host-preference effect, the composition of endophytic root microbiota may follow plant distribution. A given plant community may hence include a matrix of host-plant species that represent various habitat permeabilities to fungal dispersal in the floristic landscape. We experimentally tested the effect of host-plant isolation on endophytic fungal assemblages (Ascomycota, Basidiomycota, Glomeromycotina) inhabiting Brachypodium pinnatum roots. We calculated host-plant isolation using Euclidean distance (distance-based dispersal limitation) and resistance distance (functional-based dispersal limitation), based on host presences. All fungal groups were more influenced by the resistance distance between B. pinnatum than by the Euclidean distance. Fungal dispersal was hence strongly related to the spatial distribution of the host plants. The fungal groups displayed however different responses (in richness, abundance, and composition) to host isolation. Additionally, fungal assemblages were more strongly controlled by the degree of connectivity between host plants during the prior year than by current connectivity. This discrepancy may be due to changes in plant species coverage in a year and/or to the delay of dispersal response of fungi. This study it the first to demonstrate how small-scale host-plant distributions mediate connectivity in microorganisms. The consequences of plant distributions for the permeability of the floristic landscape to fungi dispersal appear to control fungal assemblages, but with possibly different mechanisms for the different fungal groups.

Effect of floristic composition and configuration on plant root mycobiota: a landscape transposition at a small scale.

Fungal communities in the root endosphere are heterogeneous at fine scale. The passenger hypothesis assumes that this heterogeneity is driven by host plant distribution. Plant composition and host plant configuration should then influence root fungal assemblages. We used a large-scale experimental design of 25 mixtures of grassland plants. We sampled Brachypodium pinnatum in each mesocosm, and used amplicon mass-sequencing to analyze the endospheric mycobiota. We used plant distribution maps to assess plant species richness and evenness (heterogeneity of composition), and patch size and the degree of isolation of B. pinnatum (heterogeneity of configuration) on fungal community assembly. The Glomeromycotina community in B. pinnatum roots was not related to either floristic heterogeneity or productivity. For Ascomycota, the composition of operational taxonomic units (OTUs) was driven by plant evenness while OTU richness decreased with plant richness. For Basidiomycota, richness increased with host plant aggregation and connectivity. Plant productivity influenced Ascomycota, inducing a shift in OTU composition and decreasing evenness. Plant heterogeneity modified root mycobiota, with potential direct (i.e. host preference) and indirect (i.e. adaptations to abiotic conditions driven by plant occurrence over time) effects. Plant communities can be envisioned as microlandscapes consisting of a variety of fungal niches.

Additive effects of connectivity provided by different habitat types drive plant assembly.

How connectivity affects plant assemblages is a central issue in landscape ecology. So far, empirical studies have produced contradictory results, possibly because studies: (1) inaccurately assess connectivity by prioritizing the respective effect of the type of habitat on plant assemblages and (2) omit the range of possible plant responses to connectivity depending on dispersal vectors. We focused on three dominant habitat types in agricultural landscapes (woodland, grassland and cropland), and analysed the effect of connectivity on herbaceous plant assemblage similarity for three primary dispersal modes (animal-dispersed, wind-dispersed and unassisted). Using circuit theory, we measured connectivity provided by woodland, grassland and cropland habitats independently. The similarity of plant assemblages was evaluated relative to the random expectation based on the regional pool. Overall, plant assemblage similarity in woodlands and temporary grasslands was dependent on connectivity, but not in wheat croplands. Only animal-dispersed species responded to connectivity. The similarity of animal-dispersed assemblages in woodlands was increased by the connectivity provided by woodland habitats, but was reduced by cropland habitats, whereas in temporary grasslands, similarity was increased by the connectivity provided by cropland habitats. Our results suggest that animal-dispersed species supplement their dispersal pathways, thus improving our knowledge of plant assembly rules in fragmented landscapes.

Clonal Plants as Meta-Holobionts.

The holobiont concept defines a given organism and its associated symbionts as a potential level of selection over evolutionary time. In clonal plants, recent experiments demonstrated vertical transmission of part of the microbiota from one ramet (i.e., potentially autonomous individual) to another within the clonal network (i.e., connections by modified stems present in ∼35% of all plants). Because of this heritability, and potentially reciprocal exchange of microbes between generations of ramets, we propose to extend the existing holobiont framework to the concept of meta-holobiont. A meta-holobiont is a network of holobionts that can exchange biomolecules and microbiota across generations, thus impacting the fitness of both biological scales: holobionts and meta-holobionts. Specifically, meta-holobiont dynamics can result in sharing, specialization, and division of labor across plant clonal generations. This paper, which coins the meta-holobiont concept, is expected to stimulate discussion and to be applied beyond the context of networked clonal plants (e.g., to social insects).

A microorganisms' journey between plant generations.

BACKGROUND: Plants are colonized by a great diversity of microorganisms which form a microbiota and perform additional functions for their host. This microbiota can thus be considered a toolbox enabling plants to buffer local environmental changes, with a positive influence on plant fitness. In this context, the transmission of the microbiota to the progeny represent a way to ensure the presence of beneficial symbionts within the habitat. Examples of such transmission have been mainly described for seed transmission and concern a few pathogenic microorganisms. We investigated the transmission of symbiotic partners to plant progeny within clonal plant network. METHODS: We used the clonal plant Glechoma hederacea as plant model and forced newly emitted clonal progeny to root in separated pots while controlling the presence of microorganisms. We used an amplicon sequencing approach of 16S and 18S rRNA targeting bacteria/archaea and fungi respectively to describe the root microbiota of mother and clonal-plant offspring. RESULTS: We demonstrated the vertical transmission of a significant proportion of the mother plants' symbiotic bacteria and fungi to the daughters. Interestingly, archaea were not transmitted to the daughter plants. Transmitted communities had lower richness, suggesting a filtration during transmission. We found that the transmitted pool of microorganisms was similar among daughters, constituting the heritability of a specific cohort of microorganisms, opening a new understanding of the plant holobiont. We also found significant effects of distance to the mother plant and of growth time on the richness of the microbiota transmitted. CONCLUSIONS: In this clonal plant, microorganisms are transmitted between individuals through connections, thereby ensuring the availability of microbe partners for the newborn plants as well as the dispersion between hosts for the microorganisms. This previously undescribed ecological process allows the dispersal of microorganisms in space and across plant generations. As the vast majority of plants are clonal, this process might be therefore a strong driver of ecosystem functioning and assembly of plant and microorganism communities in a wide range of ecosystems.

Functionally dissimilar neighbors accelerate litter decomposition in two grass species.

Plant litter decomposition is a key regulator of nutrient recycling. In a given environment, decomposition of litter from a focal species depends on its litter quality and on the efficiency of local decomposers. Both may be strongly modified by functional traits of neighboring species, but the consequences for decomposition of litter from the focal species remain unknown. We tested whether decomposition of a focal plant's litter is influenced by the functional-trait dissimilarity to the neighboring plants. We cultivated two grass species (Brachypodium pinnatum and Elytrigia repens) in experimental mesocosms with functionally similar and dissimilar neighborhoods, and reciprocally transplanted litter. For both species, litter quality increased in functionally dissimilar neighborhoods, partly as a result of changes in functional traits involved in plant-plant interactions. Furthermore, functional dissimilarity increased overall decomposer efficiency in one species, probably via complementarity effects. Our results suggest a novel mechanism of biodiversity effects on ecosystem functioning in grasslands: interspecific functional diversity within plant communities can enhance intraspecific contributions to litter decomposition. Thus, plant species might better perform in diverse communities by benefiting from higher remineralization rates of their own litter.

AM fungi patchiness and the clonal growth of Glechoma hederacea in heterogeneous environments.

The effect of AM fungi spatial distribution on individual plant development may determine the dynamics of the whole plant community. We investigated whether clonal plants display, like for other resources, a foraging or a specialization response, to adapt to the distribution of AM fungi. Two separate experiments were done to investigate the response of Glechoma hederacea to a heterogeneous distribution of a mixture of 3 AM fungi species, and the effects of each species on colonization and allocation traits. No specialization and a limited foraging response to the heterogeneous distribution of AM fungi was observed. An effect of the AM fungal species on plant mass allocation and ramet production, but not on spacer length, was detected. Two possible explanations are proposed: (i) the plant's responses are buffered by differences in individual effects of the fungal species or their root colonization intensity. (ii) the initial heterogeneous distribution of AM fungi is perceived as homogeneous by the plant either by reduced physiological integration or due to the transfer of AM fungi propagules through the stolons. Microscopic and DNA sequencing analyses provided evidence of this transfer, thus demonstrating the role of stolons as dispersal vectors of AM fungi within the plant clonal network.

Abiotic stressors and stress responses: What commonalities appear between species across biological organization levels?

Organisms are regularly subjected to abiotic stressors related to increasing anthropogenic activities, including chemicals and climatic changes that induce major stresses. Based on various key taxa involved in ecosystem functioning (photosynthetic microorganisms, plants, invertebrates), we review how organisms respond and adapt to chemical- and temperature-induced stresses from molecular to population level. Using field-realistic studies, our integrative analysis aims to compare i) how molecular and physiological mechanisms related to protection, repair and energy allocation can impact life history traits of stressed organisms, and ii) to what extent trait responses influence individual and population responses. Common response mechanisms are evident at molecular and cellular scales but become rather difficult to define at higher levels due to evolutionary distance and environmental complexity. We provide new insights into the understanding of the impact of molecular and cellular responses on individual and population dynamics and assess the potential related effects on communities and ecosystem functioning.

Plant traits respond to the competitive neighbourhood at different spatial and temporal scales.

BACKGROUND AND AIMS: Clonal plants can plastically modify their traits in response to competition, but little is known regarding the spatio-temporal scale at which a competitive neighbourhood determines the variability in species traits. This study tests the hypothesis that the local neighbourhood can be expected to influence the processes that are involved in competition tolerance and avoidance, and that this effect depends on organ lifespan. METHODS: Fragments of the rhizomatous Elytrigia repens (Poaceae) were sampled in 2012 in experimental plant communities that varied in species identity and abundance. These communities had been cultivated since 2009 in mesocosms in a common garden. Fragment performance, shoot and clonal traits were measured, and the effects of past and present local neighbourhoods of five different radius sizes (5-25 cm) were examined. Past and present local neighbourhood compositions were assessed in 2011 and 2012, respectively. KEY RESULTS: Most of the measured traits of E. repens responded to the local neighbourhood (5-10 cm radius), with an additional effect of the larger neighbourhood (20-25 cm radius) on ramet height, leaf dry matter content, maximal internode length and specific rhizome mass. Contrary to the expectation of the hypothesis, the temporal influence was not due to the organ lifespan. Indeed, five of the eight traits studied responded to both the past and present neighbourhoods. With the exception of specific rhizome mass, all trait responses were explained by the abundance of specific species. CONCLUSIONS: This study demonstrates that the traits of a single clonal individual can respond to different competitive environments in space and time. The results thus contribute to the understanding of competition mechanisms.