Unforeseen plant phenotypic diversity in a dry and grazed world.

Earth harbours an extraordinary plant phenotypic diversity1 that is at risk from ongoing global changes2,3. However, it remains unknown how increasing aridity and livestock grazing pressure-two major drivers of global change4-6-shape the trait covariation that underlies plant phenotypic diversity1,7. Here we assessed how covariation among 20 chemical and morphological traits responds to aridity and grazing pressure within global drylands. Our analysis involved 133,769 trait measurements spanning 1,347 observations of 301 perennial plant species surveyed across 326 plots from 6 continents. Crossing an aridity threshold of approximately 0.7 (close to the transition between semi-arid and arid zones) led to an unexpected 88% increase in trait diversity. This threshold appeared in the presence of grazers, and moved toward lower aridity levels with increasing grazing pressure. Moreover, 57% of observed trait diversity occurred only in the most arid and grazed drylands, highlighting the phenotypic uniqueness of these extreme environments. Our work indicates that drylands act as a global reservoir of plant phenotypic diversity and challenge the pervasive view that harsh environmental conditions reduce plant trait diversity8-10. They also highlight that many alternative strategies may enable plants to cope with increases in environmental stress induced by climate change and land-use intensification.

The added value of the long-term ecological research network to upscale restoration in Europe.

Achieving global restoration targets poses challenges including the need for long-term research and effective monitoring of success, fostering collaborations across diverse fields and actors, ensuring the availability of suitable reference ecosystems, and securing sustained funding. Yet, these conditions are often lacking, limiting the effectiveness of restoration. We provide an overview of ecological restoration practices in the pan-European region of the Long-term Ecological Research Network (eLTER) and demonstrate the importance of eLTER and its potential contributions to support the implementation of the EU Nature Restoration Law. We developed an online questionnaire to collect information on eLTER restoration experts and restoration projects details including the use of eLTER contributions (e.g. infrastructure, data and knowledge), between November 2021 and March 2022. We identified 62 restoration experts and 42 restoration projects from 18 countries. Our results show that eLTER restoration expertise covers most of the European habitats, diverse degradation states and restoration techniques. Most restoration projects (78%) involved long-term monitoring exceeding the average project lifespan, which has proven necessary to achieve restoration success. No common protocol was used for monitoring and evaluation or cost-benefit estimates, but respondents reported effective projects, mostly financed from national funds, and benefits in five ecosystem services on average covered per project. Key eLTER contributions included providing reference ecosystems, biotic and abiotic background data, and interdisciplinary discussion or stakeholder management. Ecological restoration is time intensive and requires long-term research and monitoring standardization to fully understand the restoration process and to ensure comparability across ecosystems. The eLTER network can help address these challenges providing added-value contributions through its infrastructure, long-term datasets, diversity of expertise and strategies that can help identify best restoration practices and support the EU Nature Restoration Law. Finally, additional and long-term funding from the EU and the private sector is needed to achieve global larger-scale restoration targets.

Strategic roadmap to assess forest vulnerability under air pollution and climate change.

Although it is an integral part of global change, most of the research addressing the effects of climate change on forests have overlooked the role of environmental pollution. Similarly, most studies investigating the effects of air pollutants on forests have generally neglected the impacts of climate change. We review the current knowledge on combined air pollution and climate change effects on global forest ecosystems and identify several key research priorities as a roadmap for the future. Specifically, we recommend (1) the establishment of much denser array of monitoring sites, particularly in the South Hemisphere; (2) further integration of ground and satellite monitoring; (3) generation of flux-based standards and critical levels taking into account the sensitivity of dominant forest tree species; (4) long-term monitoring of N, S, P cycles and base cations deposition together at global scale; (5) intensification of experimental studies, addressing the combined effects of different abiotic factors on forests by assuring a better representation of taxonomic and functional diversity across the ~73,000 tree species on Earth; (6) more experimental focus on phenomics and genomics; (7) improved knowledge on key processes regulating the dynamics of radionuclides in forest systems; and (8) development of models integrating air pollution and climate change data from long-term monitoring programs.

Using gradient Forest to predict climate response and adaptation in Cork oak.

Climate change is impacting locally adapted species such as the keystone tree species cork oak (Quercus suber L.). Quantifying the importance of environmental variables in explaining the species distribution can help build resilient populations in restoration projects and design forest management strategies. Using landscape genomics, we investigated the population structure and ecological adaptation of this tree species across the Mediterranean Basin. We applied genotyping by sequencing and derived 2,583 single nucleotide polymorphism markers genotyped from 81 individuals across 17 sites in the studied region. We implemented an approach based on the nearest neighbour haplotype 'coancestry' and uncovered a weak population structure along an east-west climatic gradient across the Mediterranean region. We identified genomic regions potentially involved in local adaptation and predicted differences in the genetic composition across the landscape under current and future climates. Variants associated with temperature and precipitation variables were detected, and we applied a nonlinear multivariate association method, gradient forest, to project these gene-environment relationships across space. The model allowed the identification of geographic areas within the western Mediterranean region most sensitive to climate change: south-western Iberia and northern Morocco. Our findings provide a preliminary assessment towards a potential management strategy for the conservation of cork oak in the Mediterranean Basin.

Bottom-up cascading effects of quarry revegetation deplete bird-mediated seed dispersal services.

Quarrying activities cause profound modifications on ecosystems, such as removal of vegetation cover, biodiversity loss and depletion of ecosystem services. Ecological restoration stands as a solution to revert such effects. Concomitantly, awareness is currently being given on ecosystem services and ecological processes to evaluate restoration efficiency. The objective of the study was to assess restoration success in a quarry subjected to restoration practices for the last 40 years involving the plantation of native Mediterranean vegetation and the non-native Aleppo pine Pinus halepensis. The study was carried out by assessing the effectiveness of seed dispersal service provided by birds in the restored quarry by comparing this service to neighbouring natural (shrubland) and other semi-natural areas (oak-pine mixed open and Aleppo pine forest) present at the landscape. For this purpose, we explored bird composition structure and seed dispersal networks using point counts and faecal samples of mist-netted birds. We also collected vegetation structure data and explored its effect on bird community composition. Our results showed that bird abundance in the restored quarry was significantly lower, and its bird community was compositionally different than natural shrubland and semi-natural areas. For instance, seed-dispersing birds, woody and shrub/ground foragers and partially migrators were the most affected groups at the restored area. Bird community composition and their traits were likely driven by vegetation characteristics, with higher native vegetation cover and fruit richness promoting higher bird abundance and Aleppo pine cover negatively influencing seed-dispersing birds. Concurrently, seed dispersal network in the restored quarry was less complex than in other areas. Seed dispersal services in the restored quarry were below the reported values for neighbouring natural and semi-natural areas and are likely driven by the low abundance of seed-dispersing birds. We consider that the causes affecting this group's low abundance can be related to revegetation measures favouring Aleppo pine, combined with a shallow soil depth and poor soil quality, which may have constrained native vegetation development. We conclude that seed dispersal services at the quarry are depleted, which may suggest a low restoration success concerning ecosystem functioning. Our results strengthen that quarry revegetation with non-native species must be avoided, since it alters bird community composition, and consequently, affects seed dispersal service provided by birds.

Green spaces are not all the same for the provision of air purification and climate regulation services: The case of urban parks.

The growing human population concentrated in urban areas lead to the increase of road traffic and artificial areas, consequently enhancing air pollution and urban heat island effects, among others. These environmental changes affect citizen's health, causing a high number of premature deaths, with considerable social and economic costs. Nature-based solutions are essential to ameliorate those impacts in urban areas. While the mere presence of urban green spaces is pointed as an overarching solution, the relative importance of specific vegetation structure, composition and management to improve the ecosystem services of air purification and climate regulation are overlooked. This avoids the establishment of optimized planning and management procedures for urban green spaces with high spatial resolution and detail. Our aim was to understand the relative contribution of vegetation structure, composition and management for the provision of ecosystem services of air purification and climate regulation in urban green spaces, in particular the case of urban parks. This work was done in a large urban park with different types of vegetation surrounded by urban areas. As indicators of microclimatic effects and of air pollution levels we selected different metrics: lichen diversity and pollutants accumulation in lichens. Among lichen diversity, functional traits related to nutrient and water requirements were used as surrogates of the capacity of vegetation to filter air pollution and to regulate climate, and provide air purification and climate regulation ecosystem services, respectively. This was also obtained with very high spatial resolution which allows detailed spatial planning for optimization of ecosystem services. We found that vegetation type characterized by a more complex structure (trees, shrubs and herbaceous layers) and by the absence of management (pruning, irrigation and fertilization) had a higher capacity to provide the ecosystems services of air purification and climate regulation. By contrast, lawns, which have a less complex structure and are highly managed, were associated to a lower capacity to provide these services. Tree plantations showed an intermediate effect between the other two types of vegetation. Thus, vegetation structure, composition and management are important to optimize green spaces capacity to purify air and regulate climate. Taking this into account green spaces can be managed at high spatial resolutions to optimize these ecosystem services in urban areas and contribute to improve human well-being.

Ecosystem services: Urban parks under a magnifying glass.

Urban areas' population has grown during the last century and it is expected that over 60% of the world population will live in cities by 2050. Urban parks provide several ecosystem services that are valuable to the well-being of city-dwellers and they are also considered a nature-based solution to tackle multiple environmental problems in cities. However, the type and amount of ecosystem services provided will vary with each park vegetation type, even within same the park. Our main goal was to quantify the trade-offs in ecosystem services associated to different vegetation types, using a spatially detailed approach. Rather than relying solely on general vegetation typologies, we took a more ecologically oriented approach, by explicitly considering different units of vegetation structure and composition. This was demonstrated in a large park (44ha) located in the city of Almada (Lisbon metropolitan area, Portugal), where six vegetation units were mapped in detail and six ecosystem services were evaluated: carbon sequestration, seed dispersal, erosion prevention, water purification, air purification and habitat quality. The results showed that, when looking at the park in detail, some ecosystem services varied greatly with vegetation type. Carbon sequestration was positively influenced by tree density, independently of species composition. Seed dispersal potential was higher in lawns, and mixed forest provided the highest amount of habitat quality. Air purification service was slightly higher in mixed forest, but was high in all vegetation types, probably due to low background pollution, and both water purification and erosion prevention were high in all vegetation types. Knowing the type, location, and amount of ecosystem services provided by each vegetation type can help to improve management options based on ecosystem services trade-offs and looking for win-win situations. The trade-offs are, for example, very clear for carbon: tree planting will boost carbon sequestration regardless of species, but may not be enough to increase habitat quality. Moreover, it may also negatively influence seed dispersal service. Informed practitioners can use this ecological knowledge to promote the role of urban parks as a nature-based solution to provide multiple ecosystem services, and ultimately improve the design and management of the green infrastructure. This will also improve the science of Ecosystem Services, acknowledging that the type of vegetation matters for the provision of ecosystem services and trade-offs analysis.

Suppression of Plant Defenses by Herbivorous Mites Is Not Associated with Adaptation to Host Plants.

Some herbivores suppress plant defenses, which may be viewed as a result of the coevolutionary arms race between plants and herbivores. However, this ability is usually studied in a one-herbivore-one-plant system, which hampers comparative studies that could corroborate this hypothesis. Here, we extend this paradigm and ask whether the herbivorous spider-mite Tetranychus evansi, which suppresses the jasmonic-acid pathway in tomato plants, is also able to suppress defenses in other host plants at different phylogenetic distances from tomatoes. We test this using different plants from the Solanales order, namely tomato, jimsonweed, tobacco, and morning glory (three Solanaceae and one Convolvulaceae), and bean plants (Fabales). First, we compare the performance of T. evansi to that of the other two most-commonly found species of the same genus, T. urticae and T. ludeni, on several plants. We found that the performance of T. evansi is higher than that of the other species only on tomato plants. We then showed, by measuring trypsin inhibitor activity and life history traits of conspecific mites on either clean or pre-infested plants, that T. evansi can suppress plant defenses on all plants except tobacco. This study suggests that the suppression of plant defenses may occur on host plants other than those to which herbivores are adapted.

The cost of surviving nitrogen excess: energy and protein demand in the lichen Cladonia portentosa as revealed by proteomic analysis.

MAIN CONCLUSION: Different nitrogen forms affect different metabolic pathways in lichens. In particular, the most relevant changes in protein expression were observed in the fungal partner, with NO 3- mostly affecting the energetic metabolism and NH 4+ affecting transport and regulation of proteins and the energetic metabolism much more than NO 3- did. Excess deposition of reactive nitrogen is a well-known agent of stress for lichens, but which symbiont is most affected and how, remains a mystery. Using proteomics can expand our understanding of stress effects on lichens. We investigated the effects of different doses and forms of reactive nitrogen, with and without supplementary phosphorus and potassium, on the proteome of the lichen Cladonia portentosa growing in a 'real-world' simulation of nitrogen deposition. Protein expression changed with the nitrogen treatments but mostly in the fungal partner, with NO3- mainly affecting the energetic metabolism and NH4+ also affecting the protein synthesis machinery. The photobiont mainly responded overexpressing proteins involved in energy production. This suggests that in response to nitrogen stress, the photobiont mainly supports the defensive mechanisms initiated by the mycobiont with an increased energy production. Such surplus energy is then used by the cell to maintain functionality in the presence of NO3-, while a futile cycle of protein production can be hypothesized to be induced by NH4+ excess. External supply of potassium and phosphorus influenced differently the responses of particular enzymes, likely reflecting the many processes in which potassium exerts a regulatory function.

Dehydration rate determines the degree of membrane damage and desiccation tolerance in bryophytes.

Desiccation tolerant (DT) organisms are able to withstand an extended loss of body water and rapidly resume metabolism upon rehydration. This ability, however, is strongly dependent on a slow dehydration rate. Fast dehydration affects membrane integrity leading to intracellular solute leakage upon rehydration and thereby impairs metabolism recovery. We test the hypothesis that the increased cell membrane damage and membrane permeability observed under fast dehydration, compared with slow dehydration, is related to an increase in lipid peroxidation. Our results reject this hypothesis because following rehydration lipid peroxidation remains unaltered, a fact that could be due to the high increase of NO upon rehydration. However, in fast-dried samples we found a strong signal of red autofluorescence upon rehydration, which correlates with an increase in ROS production and with membrane leakage, particularly the case of phenolics. This could be used as a bioindicator of oxidative stress and membrane damage.

Mapping Exposure to Multi-Pollutants Using Environmental Biomonitors-A Multi-Exposure Index.

Atmosphere is a major pathway for transport and deposition of pollutants in the environment. In industrial areas, organic compounds are released or formed as by-products, such as polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F's). Inorganic chemical elements, including lead and arsenic, are also part of the pollutants mixture, and even in low concentrations may potentially be toxic and carcinogenic. However, assessing the spatial pattern of their deposition is difficult due to high spatial and temporal heterogeneity. Lichens have been used as biomonitors of atmospheric deposition, because these organisms encompass greater spatial detail than air monitoring stations and provide an integration of overall pollution. Based upon the ability of lichens to concentrate pollutants such as PCDD/F and chemical elements, the main objectives of this study were to develop a new semi-quantitative multi-pollutant toxicity exposure index (TEQ-like), derived from risk estimates, in an attempt to correlate several atmospheric pollutants to human exposure levels. The actual pollutant concentrations were measured in the environment, from biomonitors (organisms that integrate multi-pollutants), enabling interpolation and mapping of contaminant deposition within the region. Thus, the TEQ-like index provides a spatial representation not from absolute accumulation of the different pollutants, but from the accumulation weighted by their relative risk. The assessment of environmental human exposure to multi-pollutants through atmospheric deposition may be applied to industries to improve mitigation processes or to health stakeholders to target populations for a comprehensive risk assessment, epidemiological studies, and health recommendations.

Tracking the Spatial Fate of PCDD/F Emissions from a Cement Plant by Using Lichens as Environmental Biomonitors.

In an area with multiple sources of air pollution, it is difficult to evaluate the spatial impact of a minor source. Here, we describe the use of lichens to track minor sources of air pollution. The method was tested by transplanting lichens from a background area to the vicinity of a cement manufacturing plant that uses alternative fuel and is located in a Natural Park in an area surrounded by other important sources of pollution. After 7 months of exposure, the lichens were collected and analyzed for 17 PCDD/F congeners. The PCDD/F profiles of the exposed lichens were dominated by TCDF (50%) and OCDD (38%), which matched the profile of the emissions from the cement plant. The similarity in the profiles was greatest for lichens located northeast of the plant (i.e., in the direction of the prevailing winds during the study period), allowing us to evaluate the spatial impact of this source. The best match was found for sites located on the tops of mountains whose slopes faced the cement plant. Some of the sites with highest influence of the cement plant were the ones with the highest concentrations, whereas others were not. Thus, our newly developed lichen-based method provides a tool for tracking the spatial fate of industrially emitted PCDD/Fs regardless of their concentrations. The results showed that the method can be used to validate deposition models for PCDD/F industrial emissions in sites with several sources and characterized by complex orography.

Evaluating green infrastructure in urban environments using a multi-taxa and functional diversity approach.

Forested areas within cities host a large number of species, responsible for many ecosystem services in urban areas. The biodiversity in these areas is influenced by human disturbances such as atmospheric pollution and urban heat island effect. To ameliorate the effects of these factors, an increase in urban green areas is often considered sufficient. However, this approach assumes that all types of green cover have the same importance for species. Our aim was to show that not all forested green areas are equal in importance for species, but that based on a multi-taxa and functional diversity approach it is possible to value green infrastructure in urban environments. After evaluating the diversity of lichens, butterflies and other-arthropods, birds and mammals in 31 Mediterranean urban forests in south-west Europe (Almada, Portugal), bird and lichen functional groups responsive to urbanization were found. A community shift (tolerant species replacing sensitive ones) along the urbanization gradient was found, and this must be considered when using these groups as indicators of the effect of urbanization. Bird and lichen functional groups were then analyzed together with the characteristics of the forests and their surroundings. Our results showed that, contrary to previous assumptions, vegetation density and more importantly the amount of urban areas around the forest (matrix), are more important for biodiversity than forest quantity alone. This indicated that not all types of forested green areas have the same importance for biodiversity. An index of forest functional diversity was then calculated for all sampled forests of the area. This could help decision-makers to improve the management of urban green infrastructures with the goal of increasing functionality and ultimately ecosystem services in urban areas.

Disentangling natural and anthropogenic sources of atmospheric sulfur in an industrial region using biomonitors.

Despite reductions in atmospheric sulfur (S) concentrations due to abatement policies in some countries, modeling the dispersion of this pollutant and disentangling anthropogenic sources from natural ones is still of great concern. Lichens have been used as biomonitors of the impacts of S for over 40 years, but their potential as source-tracers of specific sources, including natural ones, remains unexplored. In fact, few attempts have been made to try to distinguish and spatially model different sources of S using lichens. We have measured S concentrations and isotopic values in lichens within an industrial coastal region where different sources of S, natural and anthropogenic, interplay. We detected a prevailing influence of natural sea-originated S that mixed with anthropogenic sources of S. We were then able to disentangle the sources of S, by removing the ocean influence on S isotopic values, enabling us to model the impact of different anthropogenic sources on S deposition and highlighting the potential use of lichens to evaluate the weight of different types of anthropogenic sources.

Consequence of altered nitrogen cycles in the coupled human and ecological system under changing climate: The need for long-term and site-based research.

Anthropogenically derived nitrogen (N) has a central role in global environmental changes, including climate change, biodiversity loss, air pollution, greenhouse gas emission, water pollution, as well as food production and human health. Current understanding of the biogeochemical processes that govern the N cycle in coupled human-ecological systems around the globe is drawn largely from the long-term ecological monitoring and experimental studies. Here, we review spatial and temporal patterns and trends in reactive N emissions, and the interactions between N and other important elements that dictate their delivery from terrestrial to aquatic ecosystems, and the impacts of N on biodiversity and human society. Integrated international and long-term collaborative studies covering research gaps will reduce uncertainties and promote further understanding of the nitrogen cycle in various ecosystems.

Differential proteomics of dehydration and rehydration in bryophytes: evidence towards a common desiccation tolerance mechanism.

All bryophytes evolved desiccation tolerance (DT) mechanisms during the invasion of terrestrial habitats by early land plants. Are these DT mechanisms still present in bryophytes that colonize aquatic habitats? The aquatic bryophyte Fontinalis antipyretica Hedw. was subjected to two drying regimes and alterations in protein profiles and sucrose accumulation during dehydration and rehydration were investigated. Results show that during fast dehydration, there is very little variation in protein profiles, and upon rehydration proteins are leaked. On the other hand, slow dehydration induces changes in both dehydration and rehydration protein profiles, being similar to the protein profiles displayed by the terrestrial bryophytes Physcomitrella patens (Hedw.) Bruch and Schimp. and, to what is comparable with Syntrichia ruralis (Hedw.) F. Weber and D. Mohr. During dehydration there was a reduction in proteins associated with photosynthesis and the cytoskeleton, and an associated accumulation of proteins involved in sugar metabolism and plant defence mechanisms. Upon rehydration, protein accumulation patterns return to control values for both photosynthesis and cytoskeleton whereas proteins associated with sugar metabolism and defence proteins remain high. The current results suggest that bryophytes from different ecological adaptations may share common DT mechanisms.

Where the not-so-wild things are in cities? The influence of social-ecological factors in urban trees at multiple scales.

Green infrastructure plays an essential role in cities due to the ecosystem services it provides. However, these elements are shaped by social and ecological factors that influence their distribution and diversity, affecting ecological functions and human well-being. Here, we analyzed neighborhood tree distribution - trees in pocket parks, squares and along streets - in Lisbon (Portugal) and modelled tree abundance and taxonomic and functional diversity, at the parish and local scales, considering a comprehensive list of social and ecological factors. For the functional analyses, we included functional traits linked to dispersal, resilience to important perturbations in coastal Mediterranean cities, and ecosystem services delivery. Our results show not only that trees are unevenly distributed across the city, but that there is a strong influence of social factors on all biological indices considered. At the parish and local scales, abundance and diversity responded to different factors, with abundance being linked to both social and ecological variables. Although the influence of social factors on urban trees can be expected, by modelling their influence we can quantify how much humans modify urban landscapes at a structural and functional level. These associations can underlie potential biodiversity filters and should be analyzed over time to inform decisions that support long-term ecological resilience, maximize trait functional expression, and increase equity in ecosystem services delivery.

Changes in bryophyte functional composition during post-fire succession.

Climate and land-use changes are altering fire regimes in many regions around the world. To date, most studies have focused on the effects of altered fire regimes on woody and herbaceous communities, while the mechanisms driving post-fire bryophyte succession remain poorly understood, particularly in Mediterranean-type ecosystems. Here, we examined changes in bryophyte functional composition along a post-fire chronosequence (ranging from 1 to 20+ years) in Pyrenean oak woodlands (northeastern Portugal). To do so, we defined bryophyte functional groups based on seven morphological, reproductive, and life history traits. Then, we fitted linear and structural equation models to disentangle the direct and indirect effects of fire (time since fire and fire intensity), vegetation structure, climate, topography, and edaphic conditions on the abundance of each group. We identified two main functional groups: early colonizers (species with traits associated with strong colonization ability and desiccation tolerance) and perennial stayers (species with high competitive ability, i.e., large perennial mosses). Overall, the abundance of early colonizer species decreased with time since fire and increased with fire intensity, while the opposite was observed for perennial stayers. Thus, successional dynamics reflected a trade-off between species' competitive and colonization abilities, highlighting the role of biotic interactions later in succession. Patterns of functional composition were also consistent with changes in environmental conditions during succession, suggesting that species may experience stressful conditions (i.e., high radiation and low water availability) in early stages of post-fire succession. Our results also indicate that increased fire intensity may alter successional trajectories, leading to long-term changes in bryophyte communities. By understanding the response of bryophyte communities to fire, we were able to identify species with potential use as soil restoration materials.

Shifts in plant functional groups along an aridity gradient in a tropical dry forest.

Increasing aridity associated with climate change may lead to the crossing of critical ecosystem thresholds in drylands, compromising ecosystem services for millions of people. In this context, finding tools to detect at early stages the effects of increasing aridity on ecosystems is extremely urgent to avoid irreversible damage. Here, we assess shifts in plant community functional structure along a spatial aridity gradient in tropical dryland (Brazilian Caatinga), to select the most appropriate plant functional groups as ecological indicators likely useful to predict temporal ecosystem trajectories in response to aridity. We identified seven plant functional groups based on 13 functional traits associated with plant establishment, defense, regeneration, and dispersal, whose relative abundances changed, linearly and non-linearly, with increasing aridity, showing either increasing or decreasing trends. Of particular importance is the increase in abundance of plants with high chemical defense and Crassulacean Acid Metabolism (CAM) photosynthetic pathway, with increasing aridity. We propose the use of these functional groups as early warning indicators to detect aridity impacts on these dryland ecosystems and shifts in ecosystem functioning. This information can also be used in the elaboration of mitigation and ecological restoration measures to prevent and revert current and future climate change impacts on tropical dry forests.

More than trees: Stand management can be used to improve ecosystem diversity, structure and functioning 20 years after forest restoration in drylands.

In Mediterranean drylands, extensive areas have been restored by reforestation over the past decades to improve diversity, soil fertility, and tree natural regeneration, contributing to halting desertification and land degradation. However, evaluating reforestation success usually relies on tree survival, while holistic and long-term evaluations of reforestation success based on ecosystem diversity, structure and functioning are scarce. In this work, we provide the first assessment that combines the evaluation of planted trees and indicators of ecosystem diversity, structure, and functioning in established reforestations with three native Mediterranean species along a climatic gradient. We sampled 43 20-year-old stands with umbrella pine, holm oak and cork oak in Portugal, and tested the effects of tree species composition, stand management (i.e., differences in tree density and shrub cover), and edaphoclimatic conditions, on the size of planted trees, species diversity, structural complexity and indicators of ecosystem functioning related to productivity, soil nutrients and tree natural regeneration. Our results show that, after 20 years of reforestation, stand management was an essential driver of plant diversity and ecosystem functioning. Higher tree density, particularly of oaks, and higher shrub cover improved plant diversity, ecosystem productivity, and oak regeneration. The latter was also improved by structural complexity. Tree composition effects highlighted the importance of pine management to avoid competition. Since we evaluated these reforestations along a climatic gradient, we also conclude that climate influenced pine and holm oak size, ecosystem productivity, and soil C/N. Our research, by being based on assessing the long-term reforestation success in a more holistic way, highlighted the importance of stand management for improving ecosystem diversity and functioning in these restored systems. Practices such as increasing tree density up to ~800 trees/ha and allowing a shrub cover of ca. 30 %, may improve the ecological condition of future and currently reforested areas across the Mediterranean region.