Responses of rhizosphere fungi to the root economics space in grassland monocultures of different age.

Recent studies on root traits have shown that there are two axes explaining trait variation belowground: the collaboration axis with mycorrhizal partners and the conservation ('fast - slow') axis. However, it is yet unknown whether these trait axes affect the assembly of soilborne fungi. We expect saprotrophic fungi to link to the conservation axis of root traits, whereas pathogenic and arbuscular mycorrhizal fungi link to the collaboration axis, but in opposite directions, as arbuscular mycorrhizal fungi might provide pathogen protection. To test these hypotheses, we sequenced rhizosphere fungal communities and measured root traits in monocultures of 25 grassland plant species, differing in age. Within the fungal guilds, we evaluated fungal species richness, relative abundance and community composition. Contrary to our hypotheses, fungal diversity and relative abundance were not strongly related to the root trait axes. However, saprotrophic fungal community composition was affected by the conservation gradient and pathogenic community composition by the collaboration gradient. The rhizosphere AMF community composition did not change along the collaboration gradient, even though the root trait axis was in line with the root mycorrhizal colonization rate. Overall, our results indicate that in the long term, the root trait axes are linked with fungal community composition.

The structure of root-associated fungal communities is related to the long-term effects of plant diversity on productivity.

Root-associated fungi could play a role in determining both the positive relationship between plant diversity and productivity in experimental grasslands, and its strengthening over time. This hypothesis assumes that specialized pathogenic and mutualistic fungal communities gradually assemble over time, enhancing plant growth more in species-rich than in species-poor plots. To test this hypothesis, we used high-throughput amplicon sequencing to characterize root-associated fungal communities in experimental grasslands of 1 and 15 years of age with varying levels of plant species richness. Specifically, we tested whether the relationship between fungal communities and plant richness and productivity becomes stronger with the age of the experimental plots. Our results showed that fungal diversity increased with plant diversity, but this relationship weakened rather than strengthened over the two time points. Contrastingly, fungal community composition showed increasing associations with plant diversity over time, suggesting a gradual build-up of specific fungal assemblages. Analyses of different fungal guilds showed that these changes were particularly marked in pathogenic fungi, whose shifts in relative abundance are consistent with the pathogen dilution hypothesis in diverse plant communities. Our results suggest that root-associated fungal pathogens play more specific roles in determining the diversity-productivity relationship than other root-associated plant symbionts.

An integrated framework of plant form and function: the belowground perspective.

Plant trait variation drives plant function, community composition and ecosystem processes. However, our current understanding of trait variation disproportionately relies on aboveground observations. Here we integrate root traits into the global framework of plant form and function. We developed and tested an overarching conceptual framework that integrates two recently identified root trait gradients with a well-established aboveground plant trait framework. We confronted our novel framework with published relationships between above- and belowground trait analogues and with multivariate analyses of above- and belowground traits of 2510 species. Our traits represent the leaf and root conservation gradients (specific leaf area, leaf and root nitrogen concentration, and root tissue density), the root collaboration gradient (root diameter and specific root length) and the plant size gradient (plant height and rooting depth). We found that an integrated, whole-plant trait space required as much as four axes. The two main axes represented the fast-slow 'conservation' gradient on which leaf and fine-root traits were well aligned, and the 'collaboration' gradient in roots. The two additional axes were separate, orthogonal plant size axes for height and rooting depth. This perspective on the multidimensional nature of plant trait variation better encompasses plant function and influence on the surrounding environment.

Root traits as drivers of plant and ecosystem functioning: current understanding, pitfalls and future research needs.

The effects of plants on the biosphere, atmosphere and geosphere are key determinants of terrestrial ecosystem functioning. However, despite substantial progress made regarding plant belowground components, we are still only beginning to explore the complex relationships between root traits and functions. Drawing on the literature in plant physiology, ecophysiology, ecology, agronomy and soil science, we reviewed 24 aspects of plant and ecosystem functioning and their relationships with a number of root system traits, including aspects of architecture, physiology, morphology, anatomy, chemistry, biomechanics and biotic interactions. Based on this assessment, we critically evaluated the current strengths and gaps in our knowledge, and identify future research challenges in the field of root ecology. Most importantly, we found that belowground traits with the broadest importance in plant and ecosystem functioning are not those most commonly measured. Also, the estimation of trait relative importance for functioning requires us to consider a more comprehensive range of functionally relevant traits from a diverse range of species, across environments and over time series. We also advocate that establishing causal hierarchical links among root traits will provide a hypothesis-based framework to identify the most parsimonious sets of traits with the strongest links on functions, and to link genotypes to plant and ecosystem functioning.

A starting guide to root ecology: strengthening ecological concepts and standardising root classification, sampling, processing and trait measurements.

In the context of a recent massive increase in research on plant root functions and their impact on the environment, root ecologists currently face many important challenges to keep on generating cutting-edge, meaningful and integrated knowledge. Consideration of the below-ground components in plant and ecosystem studies has been consistently called for in recent decades, but methodology is disparate and sometimes inappropriate. This handbook, based on the collective effort of a large team of experts, will improve trait comparisons across studies and integration of information across databases by providing standardised methods and controlled vocabularies. It is meant to be used not only as starting point by students and scientists who desire working on below-ground ecosystems, but also by experts for consolidating and broadening their views on multiple aspects of root ecology. Beyond the classical compilation of measurement protocols, we have synthesised recommendations from the literature to provide key background knowledge useful for: (1) defining below-ground plant entities and giving keys for their meaningful dissection, classification and naming beyond the classical fine-root vs coarse-root approach; (2) considering the specificity of root research to produce sound laboratory and field data; (3) describing typical, but overlooked steps for studying roots (e.g. root handling, cleaning and storage); and (4) gathering metadata necessary for the interpretation of results and their reuse. Most importantly, all root traits have been introduced with some degree of ecological context that will be a foundation for understanding their ecological meaning, their typical use and uncertainties, and some methodological and conceptual perspectives for future research. Considering all of this, we urge readers not to solely extract protocol recommendations for trait measurements from this work, but to take a moment to read and reflect on the extensive information contained in this broader guide to root ecology, including sections I-VII and the many introductions to each section and root trait description. Finally, it is critical to understand that a major aim of this guide is to help break down barriers between the many subdisciplines of root ecology and ecophysiology, broaden researchers' views on the multiple aspects of root study and create favourable conditions for the inception of comprehensive experiments on the role of roots in plant and ecosystem functioning.

Biodiversity increases the resistance of ecosystem productivity to climate extremes.

It remains unclear whether biodiversity buffers ecosystems against climate extremes, which are becoming increasingly frequent worldwide. Early results suggested that the ecosystem productivity of diverse grassland plant communities was more resistant, changing less during drought, and more resilient, recovering more quickly after drought, than that of depauperate communities. However, subsequent experimental tests produced mixed results. Here we use data from 46 experiments that manipulated grassland plant diversity to test whether biodiversity provides resistance during and resilience after climate events. We show that biodiversity increased ecosystem resistance for a broad range of climate events, including wet or dry, moderate or extreme, and brief or prolonged events. Across all studies and climate events, the productivity of low-diversity communities with one or two species changed by approximately 50% during climate events, whereas that of high-diversity communities with 16-32 species was more resistant, changing by only approximately 25%. By a year after each climate event, ecosystem productivity had often fully recovered, or overshot, normal levels of productivity in both high- and low-diversity communities, leading to no detectable dependence of ecosystem resilience on biodiversity. Our results suggest that biodiversity mainly stabilizes ecosystem productivity, and productivity-dependent ecosystem services, by increasing resistance to climate events. Anthropogenic environmental changes that drive biodiversity loss thus seem likely to decrease ecosystem stability, and restoration of biodiversity to increase it, mainly by changing the resistance of ecosystem productivity to climate events.

Leaf area and pubescence drive sedimentation on leaf surfaces during flooding.

Worldwide, stream water is increasingly loaded with sediments and nutrients, due to processes such as accelerated soil erosion and overfertilization caused by agricultural intensification. This leads to increases in eutrophication and silting up of bottom sediments. Floodplains can play an important role in mitigating these problems, by removing sediment from rivers via water filtration and retention. Fine sediment is accumulated on the soil in between plants as well as on plant surfaces. However, it is still poorly understood how plant species facilitate leaf surface sedimentation via their leaf traits. In a flume experiment, we investigated to what extent the leaf traits (area, length, perimeter, pinnation, pubescence, surface roughness, flexibility and wettability) influence leaf surface sedimentation. We exposed leaves of 30 plant species to an artificial flood, and measured the fine sediment load the leaves captured after 24 h. Our results show that leaf traits overall explain 65% of the variation of fine sedimentation on leaves. Especially adaxial pubescence and leaf area strongly drove sedimentation. Hairy leaves accumulate more sediment per leaf area, presumably, because hairs create a buffer zone of reduced flow velocity which enhances sedimentation between the hairs. Additionally, for leaves with no or few hairs, sedimentation decreased with increasing leaf area, because most likely the more turbulent boundary layer of larger leaves allows less sediment to settle. Our results provide a first understanding of how plants can be selected based on their leaf traits for maximizing the sediment retention on floodplains, thereby providing a key ecosystem service.

Plant diversity influenced gross nitrogen mineralization, microbial ammonium consumption and gross inorganic N immobilization in a grassland experiment.

Gross rates of nitrogen (N) turnover inform about the total N release and consumption. We investigated how plant diversity affects gross N mineralization, microbial ammonium (NH4+) consumption and gross inorganic N immobilization in grasslands via isotopic pool dilution. The field experiment included 74 plots with 1-16 plant species and 1-4 plant functional groups (legumes, grasses, tall herbs, small herbs). We determined soil pH, shoot height, root, shoot and microbial biomass, and C and N concentrations in soil, microbial biomass, roots and shoots. Structural equation modeling (SEM) showed that increasing plant species richness significantly decreased gross N mineralization and microbial NH4+ consumption rates via increased root C:N ratios. Root C:N ratios increased because of the replacement of legumes (low C:N ratios) by small herbs (high C:N ratios) and an increasing shoot height, which was positively related with root C:N ratios, with increasing species richness. However, in our SEM remained an unexplained direct negative path from species richness to both N turnover rates. The presence of legumes increased gross N mineralization, microbial NH4+ consumption and gross inorganic N immobilization rates likely because of improved N supply by N2 fixation. The positive effect of small herbs on microbial NH4+ consumption and gross inorganic N immobilization could be attributed to their increased rhizodeposition, stimulating microbial growth. Our results demonstrate that increasing root C:N ratios with increasing species richness slow down the N cycle but also that there must be additional, still unidentified processes behind the species richness effect potentially including changed microbial community composition.

Functional groups differ in trait means, but not in trait plasticity to species richness in local grassland communities.

Despite growing interest in incorporating intraspecific variation of functional traits in community-level studies, it remains unclear whether species classified into functional groups based on interspecific trait differences are similar regarding their variation in trait expression in response to varying plant diversity and composition in local communities. In a large biodiversity experiment (Jena Experiment) designed on a trait-based a priori definition of functional groups (grasses, legumes, small herbs, tall herbs), we studied means, extent of variation (coefficient of variation across communities) and plasticity to increased plant diversity (slopes over a logarithmic species richness ranging from 1, 2, 4, 8 and 16 to 60 species) for nine functional traits. Species means and extent of variation in traits related to nitrogen (N) acquisition and N use differed among functional groups and were more similar in phylogenetically closely related species than expected by chance. Species in the same functional group showed a weak phylogenetic signal and varied widely in means and extent of variation in traits related to shoot architecture and to a smaller extent in leaf traits related to carbon acquisition. This indicated that functional groups were less distinguishable in light than in nitrogen acquisition strategies. The direction and degree of trait plasticity to increasing species richness did not show a phylogenetic signal and were not different among functional groups, but varied largely among species within functional groups. Correlation structures in trait means, extent of trait variation and trait plasticity revealed functional tradeoffs in the acquisition of nitrogen and light across species. While correlations between trait means and extent of trait variation varied from trait to trait (positive, negative or unrelated), trait means and trait plasticity were mostly unrelated. Our results suggest that the concept of functional groups is viable, but context-specific trait measurements are required to improve our understanding about the functional significance of intraspecific trait variation and interspecific trait differences in local plant communities.

Origin context of trait data matters for predictions of community performance in a grassland biodiversity experiment.

Plant functional traits may explain the positive relationship between species richness and ecosystem functioning, but species-level trait variation in response to growth conditions is often ignored in trait-based predictions of community performance. In a large grassland biodiversity experiment (Jena Experiment), we measured traits on plants grown as solitary individuals, in monocultures or in mixtures. We calculated two measures of community-level trait composition, i.e., community-weighted mean traits (CWM) and trait diversity (Rao's quadratic entropy; FD) based on different contexts in which traits were measured (trait origins). CWM and FD values of the different measurement origins were then compared regarding their power to predict community biomass production and biodiversity effects quantified with the additive partitioning method. Irrespective of trait origin, models combining CWM and FD values as predictors best explained community biomass and biodiversity effects. CWM values based on monoculture, mixture-mean or community-specific trait data were similarly powerful predictors, but predictions became worse when trait values originated from solitary-grown individuals. FD values based on monoculture traits were the best predictors of community biomass and net biodiversity effects, while FD values based on community-specific traits were the best predictors for complementarity and selection effects. Traits chosen as best CWM predictors were not strongly affected by trait origin but traits chosen as best FD predictors varied strongly dependent on trait origin and altered the predictability of community performance. We conclude that by adjusting their functional traits to species richness and even specific community compositions, plants can change community-level trait compositions, thereby also changing community biomass production and biodiversity effects. Incorporation of these plastic trait adjustments of plants in trait-based ecology can improve its predictive power in explaining biodiversity-ecosystem functioning relationships.

Plants are less negatively affected by flooding when growing in species-rich plant communities.

Flooding is expected to increase in frequency and severity in the future. The ecological consequences of flooding are the combined result of species-specific plant traits and ecological context. However, the majority of past flooding research has focused on individual model species under highly controlled conditions. An early summer flooding event in a grassland biodiversity experiment in Jena, Germany, provided the opportunity to assess flooding responses of 60 grassland species in monocultures and 16-species mixtures. We examined plant biomass, species-specific traits (plant height, specific leaf area (SLA), root aerenchyma, starch content) and soil porosity. We found that, on average, plant species were less negatively affected by the flood when grown in higher-diversity plots in July 2013. By September 2013, grasses were unaffected by the flood regardless of plant diversity, and legumes were severely negatively affected regardless of plant diversity. Plants with greater SLA and more root aerenchyma performed better in September. Soil porosity was higher in higher-diversity plots and had a positive effect on plant performance. As floods become more frequent and severe in the future, growing flood-sensitive plants in higher-diversity communities and in soil with greater soil aeration may attenuate the most negative effects of flooding.

Plant diversity maintains long-term ecosystem productivity under frequent drought by increasing short-term variation.

Increasing frequency of extreme climatic events can disrupt ecosystem processes and destabilize ecosystem functioning. Biodiversity may dampen these negative effects of environmental perturbations to provide greater ecosystem stability. We assessed the effects of plant diversity on the resistance, recovery and stability of experimental grassland ecosystems in response to recurring summer drought over 7 yr. Plant biomass production was reduced during the summer drought treatment compared with control plots. However, the negative effect of drought was relatively less pronounced at high than at low plant diversity, demonstrating that biodiversity increased ecosystem resistance to environmental perturbation. Furthermore, more diverse plant communities compensated for the reduced productivity during drought by increasing spring productivity compared to control plots. The drought-induced compensatory recovery led to increased short-term variations in productivity across growing seasons in more diverse communities that stabilized the longer-term productivity across years. Our findings show that short-term variation between seasons in the face of environmental perturbation can lead to longer-term stability of annual productivity in diverse ecosystems compared to less diverse ecosystems.

Root chemistry and soil fauna, but not soil abiotic conditions explain the effects of plant diversity on root decomposition.

Plant diversity influences many ecosystem functions including root decomposition. However, due to the presence of multiple pathways via which plant diversity may affect root decomposition, our mechanistic understanding of their relationships is limited. In a grassland biodiversity experiment, we simultaneously assessed the effects of three pathways-root litter quality, soil biota, and soil abiotic conditions-on the relationships between plant diversity (in terms of species richness and the presence/absence of grasses and legumes) and root decomposition using structural equation modeling. Our final structural equation model explained 70% of the variation in root mass loss. However, different measures of plant diversity included in our model operated via different pathways to alter root mass loss. Plant species richness had a negative effect on root mass loss. This was partially due to increased Oribatida abundance, but was weakened by enhanced root potassium (K) concentration in more diverse mixtures. Equally, grass presence negatively affected root mass loss. This effect of grasses was mostly mediated via increased root lignin concentration and supported via increased Oribatida abundance and decreased root K concentration. In contrast, legume presence showed a net positive effect on root mass loss via decreased root lignin concentration and increased root magnesium concentration, both of which led to enhanced root mass loss. Overall, the different measures of plant diversity had contrasting effects on root decomposition. Furthermore, we found that root chemistry and soil biota but not root morphology or soil abiotic conditions mediated these effects of plant diversity on root decomposition.

High plant diversity is needed to maintain ecosystem services.

Biodiversity is rapidly declining worldwide, and there is consensus that this can decrease ecosystem functioning and services. It remains unclear, though, whether few or many of the species in an ecosystem are needed to sustain the provisioning of ecosystem services. It has been hypothesized that most species would promote ecosystem services if many times, places, functions and environmental changes were considered; however, no previous study has considered all of these factors together. Here we show that 84% of the 147 grassland plant species studied in 17 biodiversity experiments promoted ecosystem functioning at least once. Different species promoted ecosystem functioning during different years, at different places, for different functions and under different environmental change scenarios. Furthermore, the species needed to provide one function during multiple years were not the same as those needed to provide multiple functions within one year. Our results indicate that even more species will be needed to maintain ecosystem functioning and services than previously suggested by studies that have either (1) considered only the number of species needed to promote one function under one set of environmental conditions, or (2) separately considered the importance of biodiversity for providing ecosystem functioning across multiple years, places, functions or environmental change scenarios. Therefore, although species may appear functionally redundant when one function is considered under one set of environmental conditions, many species are needed to maintain multiple functions at multiple times and places in a changing world.

Plant diversity and root traits benefit physical properties key to soil function in grasslands.

Plant diversity loss impairs ecosystem functioning, including important effects on soil. Most studies that have explored plant diversity effects belowground, however, have largely focused on biological processes. As such, our understanding of how plant diversity impacts the soil physical environment remains limited, despite the fundamental role soil physical structure plays in ensuring soil function and ecosystem service provision. Here, in both a glasshouse and a long-term field study, we show that high plant diversity in grassland systems increases soil aggregate stability, a vital structural property of soil, and that root traits play a major role in determining diversity effects. We also reveal that the presence of particular plant species within mixed communities affects an even wider range of soil physical processes, including hydrology and soil strength regimes. Our results indicate that alongside well-documented effects on ecosystem functioning, plant diversity and root traits also benefit essential soil physical properties.

Predicting stochastic community dynamics in grasslands under the assumption of competitive symmetry.

Community dynamics is influenced by multiple ecological processes such as environmental spatiotemporal variation, competition between individuals and demographic stochasticity. Quantifying the respective influence of these various processes and making predictions on community dynamics require the use of a dynamical framework encompassing these various components. We here demonstrate how to adapt the framework of stochastic community dynamics to the peculiarities of herbaceous communities, by using a short temporal resolution adapted to the time scale of competition between herbaceous plants, and by taking into account the seasonal drops in plant aerial biomass following winter, harvesting or consumption by herbivores. We develop a hybrid inference method for this novel modelling framework that both uses numerical simulations and likelihood computations. Applying this methodology to empirical data from the Jena biodiversity experiment, we find that environmental stochasticity has a larger effect on community dynamics than demographic stochasticity, and that both effects are generally smaller than observation errors at the plot scale. We further evidence that plant intrinsic growth rates and carrying capacities are moderately predictable from plant vegetative height, specific leaf area and leaf dry matter content. We do not find any trade-off between demographical components, since species with larger intrinsic growth rates tend to also have lower demographic and environmental variances. Finally, we find that our model is able to make relatively good predictions of multi-specific community dynamics based on the assumption of competitive symmetry.

Plant species diversity affects soil-atmosphere fluxes of methane and nitrous oxide.

Plant diversity effects on ecosystem functioning can potentially interact with global climate by altering fluxes of the radiatively active trace gases nitrous oxide (N2O) and methane (CH4). We studied the effects of grassland species richness (1-16) in combination with application of fertilizer (nitrogen:phosphorus:potassium = 100:43.6:83 kg ha(-1) a(-1)) on N2O and CH4 fluxes in a long-term field experiment. Soil N2O emissions, measured over 2 years using static chambers, decreased with species richness unless fertilizer was added. N2O emissions increased with fertilization and the fraction of legumes in plant communities. Soil CH4 uptake, a process driven by methanotrophic bacteria, decreased with plant species numbers, irrespective of fertilization. Using structural equation models, we related trace gas fluxes to soil moisture, soil inorganic N concentrations, nitrifying and denitrifying enzyme activity, and the abundance of ammonia oxidizers, nitrite oxidizers, and denitrifiers (quantified by real-time PCR of gene fragments amplified from microbial DNA in soil). These analyses indicated that plant species richness increased soil moisture, which in turn increased N cycling-related activities. Enhanced N cycling increased N2O emission and soil CH4 uptake, with the latter possibly caused by removal of inhibitory ammonium by nitrification. The moisture-related indirect effects were surpassed by direct, moisture-independent effects opposite in direction. Microbial gene abundances responded positively to fertilizer but not to plant species richness. The response patterns we found were statistically robust and highlight the potential of plant biodiversity to interact with climatic change through mechanisms unrelated to carbon storage and associated carbon dioxide removal.