Basic and Applied Ecology

O_LIFloral nectar plays a critical role in shaping the ecology and evolution of plant-pollinator interactions. Effective and efficient methods that allow for broad-scale sampling of nectar volume and sugar concentration across a diversity of taxa are needed to improve our understanding of many dimensions of mutualistic plant-pollinator interactions--including their basic ecology and evolution, their responses to environmental change, and their conservation and restoration. C_LIO_LIDespite the key importance of nectar for mediating plant-pollinator interactions, quantifying floral nectar in the field from many different plant species is challenging because there is often no one-size-fits-all sampling method that is effective across a diversity of floral structures and nectar traits. Different methods require different preparation, and sampling from many species involves a variety of logistical challenges. C_LIO_LIHere we provide a methodological roadmap for sampling floral nectar in the field from many different plant species. We describe our nectar collection methods in detail, including necessary equipment, calculations, and approaches appropriate for different floral morphologies. We also provide a troubleshooting guide for common problems encountered while collecting nectar in the field. To demonstrate the utility and effectiveness of our methods for collecting nectar from many different species, we present results on nectar trait variation from 53 species in an ecosystem. C_LIO_LIOur method illustrates that nectar traits vary considerably within and among plant species, indicating that large-scale nectar sampling projects are an important consideration for many basic and applied questions in pollination ecology and evolution. We hope that across many plant communities and ecosystems, our paper provides a practical roadmap for how to navigate the complexities of quantifying floral nectar traits. C_LI

Artificial light at night (ALAN) can alter the movement of nocturnal pollinators such as moths, with potential consequences for pollination services. However, most studies have focused on the presence or properties of lights, while the role of spatial lighting configuration remains poorly understood. We conducted a small-scale field experiment to test how different lighting configurations affect pollination success in moth-pollinated plants, using low-intensity LED garden lights. Potted phytometer plants of three moth-pollinated species were exposed to one of three treatments: no-light control, isolated LED point lights, or 25 m linear arrays of multiple LED lights. We quantified pollination success as both the probability of seed set and reproductive output through seed and capsule production. Experimental lighting affected plant reproductive success, with light arrays increasing seed-set probability, seed and seed capsule number relative to unlit controls, whereas point lights showed no clear effect. These results provide preliminary evidence that the spatial configuration of artificial lights may influence nocturnal pollination outcomes. Future work combining phytometer assays with direct tracking of moth movement is needed to assess whether light arrays facilitate or redirect the dispersal of nocturnal pollinators.

Repeated grazing and trampling can reduce forb populations in South African mesic grasslands by limiting photosynthesis and depleting carbohydrate reserves stored in underground organs. We examined the effects of repeated defoliation on growth, bulb starch reserves and water status in the geophyte Eucomis autumnalis over 468 days. Four intense summer and autumn defoliations reduced above-ground production, bulb mass, starch reserves and bulb water pools, with the strongest impacts occurring in late summer and autumn when plants normally replenish reserves. Repeated defoliation disrupted seasonal source sink dynamics, forcing resources into regrowth and driving bulbs towards energetic bankruptcy. These results highlight the importance of avoiding heavy autumn grazing and providing periodic full-year rests to conserve geophytic forbs in mesic grasslands.

A majority of stream restoration efforts in central Europe focus on streams that are less than five meters wide. Restoration aims to increase structural complexity, thereby enhancing habitat heterogeneity, promoting biodiversity, and reestablishing aquatic-terrestrial linkages that can drive responses in adjacent terrestrial communities. However, the effects of small stream restoration on terrestrial biodiversity remain poorly understood because research focuses mainly on large rivers. Here, we investigated the effects of restoration on the structural complexity of the stream channel as well as the terrestrial habitat and plant diversity on a local and landscape scale across 55 small streams in an agricultural landscape. We compared restored stream sections with non-restored ones that were similar to the conditions before restoration. We also assessed how restored sections changed over time since restoration. Restored stream sections showed a higher stream structural complexity and habitat diversity, both of which are targets of active restoration measures. Restoration also increased riparian plant diversity, both directly and indirectly through structural complexity and habitat diversity. Although time since restoration did not influence structural complexity, it drove successional changes in plant communities that became increasingly associated with wetland habitat conditions. Our results demonstrate that small stream restoration effectively increases floodplain habitat and plant diversity in agricultural landscapes, primarily by enhancing water availability in the floodplain. Restoration actions on small streams support biodiversity if they improve stream channel complexity, connect the stream with its floodplain, and create floodplain habitats.

Urban green spaces are increasingly recognised as important refuges for biodiversity, yet their ecological value depends strongly on design and management. Here, we investigate how fine-scale structural and microhabitat components shape urban ant assemblages, using ants as indicators of broader arthropod responses to urbanisation. Ant communities were sampled in twelve urban green spaces in Cordoba (southern Spain) over a ten-year period (2004 to 2013) using pitfall traps, alongside detailed characterisation of vegetation structure and ground-layer microhabitats. In total, 38 species and 25,578 individuals were recorded. Microhabitat variables explained 58% of the variation in species occurrence. Community differences among microhabitats were driven primarily by nestedness, with dense herbaceous cover acting as a core habitat and edge-related components contributing disproportionately to beta diversity. Tree abundance showed a unimodal relationship with species richness, with maximum diversity at intermediate densities, while shrub and lawn cover had weak or inconsistent effects. Fine-scale elements such as leaf litter, stones, woody debris, and small bare-ground patches strongly influenced species occurrence by providing thermal refugia, nesting substrates, and foraging opportunities. The invasive Argentine ant (Linepithema humile) exhibited strong but spatially restricted dominance and species-specific negative effects on native ants, emphasising the role of habitat context in mediating invasion impacts. Our results demonstrate that urban biodiversity is maximised by enhancing fine-scale habitat heterogeneity rather than increasing green cover alone. We highlight practical design principles for urban green infrastructure that prioritise structural diversity and ground-layer complexity to support resilient arthropod communities.

Surges in food availability create localized but intense foraging opportunities, often attracting multi-species consumer groups. Agricultural practices can trigger these surges, prompting bird associations. However, the strength and duration of the association, as well as its drivers, remain unclear. This study examines waterbird association with harvesting and ploughing events in rice fields. The duration and magnitude of these associations were determined and three hypotheses addressed to explain them: (1) increased food availability, (2) enhanced foraging success and (3) reduced time allocated to vigilance. Waterbird counts and GPS tracking revealed strong associations with management events. Bird numbers spiked during events but declined within one to two days. Food availability (and soil penetrability) increased significantly during events - crayfish and rice during harvesting, worms and soil penetrability during ploughing - supporting Hypothesis 1. However, this did not improve foraging performance (intake rate, foraging success), rejecting Hypothesis 2. Higher competition, interference or kleptoparasitism in these large mixed-species flocks may offset increased food availability benefits. Alternatively, functional responses of target species may limit prey intake due to physiological or behavioural constraints. Hypothesis 3 was also unsupported, as birds did not reduce vigilance. It is plausible that birds may be drawn to events by the perception of a feast, not actual benefits. Gregariousness and foraging behaviour by local enhancement may explain such associations. Results highlight the complexity of bird responses to food surges while suggesting waterbirds in rice fields maintain stable foraging performance during agricultural management events and otherwise, indicating resilience to agricultural timing shifts.

Climate change is expected to alter plant populations not only through direct environmental shifts but also via changes in biotic interactions, such as with herbivores and pathogens. As plant species are also expected to differ in their responses to both climate and antagonists, plant responses to both factors are expected to be variable and species-specific. To assess whether interactive effects of climate and antagonists on plant population dynamics are common and whether the strength and direction of plant responses vary across species, we conducted a multi-year field experiment that manipulated realistic climate change and experimentally reduced insect herbivores and fungal pathogens. We measured responses of plant vital rates, such as survivorship, growth, and reproduction across six grassland species. Using Integral Projection Models (IPMs) and Life Table Response Experiments (LTREs), we quantified changes in population growth rate across experimental treatments and the contribution of each vital rate to that observed change. Two of the study species declined so drastically over the course of the experiment that demographic quantification of population growth rates was not possible. From the remaining species, Bromus erectus and Plantago lanceolata show significant interactive responses of climate and antagonist reduction on population growth rates. In contrast, Dianthus carthusianorum and Tragopogon orientalis showed limited responses to experimental treatments. Notably, our results indicate that in some species biotic interactions may amplify the effects of climate change: the presence of plant antagonists exacerbates the negative effects of the future climate treatment on plant population dynamics. Our findings highlight the complexity in predicting plant population responses to climate change and provide insights for grassland management under future environmental conditions.

Grasslands are vulnerable to increasing drought with global warming, but process-based models lack the mechanistic knowledge required to predict the magnitude of drought impacts. While a plant hydraulics framework has been successful in advancing process understanding of drought responses in trees, and how drought responses vary across rainfall gradients, similar approaches have rarely been applied to grasses. Here, we quantified the progression of key drought response processes in sixteen dominant perennial grasses (seven C3 and nine C4) with differing climatic origins across eastern Australia. We found that stomatal closure, hydraulic impairment and leaf browning occurred concurrently, in contrast to the progressive sequence typically observed in trees. We also found that drought response traits were not correlated with species climate of origin. The early impairment of leaf hydraulic conductance and leaf browning along with the lack of correlation with climate of origin suggest that grasses may employ fundamentally different strategies to adapt to low water availability than trees. These results highlight the need for grass-specific parameterization of drought responses in process-based models.

Agroecosystems are dynamic ecosystems, constituted by patches of vegetation and agricultural use, where biodiversity is shaped by spatial and temporal variability. While most studies have focused on spatial composition and configuration, the role of temporal variability remains poorly understood. Yet, temporal dynamics can strongly modify species composition, abundance, and persistence in ecological communities. Temporal variability is particularly relevant in agroecosystems with rainfed agriculture where environmental conditions shift dramatically between rainy and dry seasons. In this paper, we assess the role of temporal variability on biodiversity maintenance in an agricultural matrix using a metacommunity model that simulates an agricultural landscape under rainfed conditions, that is, with abrupt seasonal changes in the agricultural patches. This model couples a local community network dynamic with a migration dynamic and is based on empirically documented features of rainfed agricultural matrices. Our results show that temporal variability provides new opportunities for species to recover from low densities. However, the effect of temporal variability is not straightforward. It depends on the initial and final conditions, the migration and mortality rates and the intensity of temporal variability. Overall, our findings highlight the need to further investigate temporal variability to better understand its role in shaping biodiversity in agricultural landscapes.

Plants form holobionts by associating with diverse microbiota. Self-organization gives rise to emergent properties of the holobiont, such as increased resistance to pathogens. However, the local factors contributing to the self-organization are not well understood. We hypothesized that nematode communities and their associated microbiota govern the rhizobiome of the model plant holobiont tomato in terms of its suppression of root invasion by the parasite Meloidogyne hapla, and that the soil legacy influences the suppressive potential mediated by these biota. In pot experiments, a resistant tomato holobiont was favored by assembly in the presence of a nematode community conditioned by tomato plants, compared to oilseed rape or fallow soil. Nematode communities conditioned by tagetes could enhance resistance even better than tomato. Microbiota from crushed tomato-conditioned nematode communities increased resistance of the tomato holobiont, compared to microbiota of nematode communities conditioned by maize, or heat-inactivated microbiota. The 0.2 micrometre filtered microbiota from crushed nematodes had the same effect, suggesting a role of nematode-associated bacteriophages in holobiont assembly. The results indicate that soil nematodes and their associated microbiota play a role in the local organization and stabilization of plant holobionts. They can influence the resistance of plants that subsequently grow in the same soil. From an applied perspective, crop rotation schemes that alter nematode-microbiota communities could be harnessed to engineer crop holobionts.

Plant roots are broadly colonized by endophytic fungi with saprotrophic capabilities, but our understanding of whether they function in ways that are beneficial or detrimental to the host remains limited to model organisms. We hypothesized that endophytic fungi broadly affect plant access to soil nutrients, particularly organic forms that are typically not directly available to the plant. To address this, we paired 41 fungal endophytes with switchgrass (Panicum virgatum L.) and provided either inorganic or organic forms of nitrogen (N) and phosphorus (P). We evaluated how the fungi affected plant tissue N and P as well as plant growth. We also examined if these outcomes could be predicted from fungal phylogenetic relationships, in vitro traits of the fungi, or characteristics of the habitat from which fungi were isolated. There was substantial variation in both plant N (0.05-0.63%) and P (0.02-0.10%) acquisition that depended on the interaction of fungus and nutrient treatment. More fungi were beneficial for plant N than for P and shoot nutrients generally increased more than root nutrients from fungal associations. However, fungal effects on plant nutrients were not predicted by fungal traits, habitat traits, or fungal phylogenetic relationships. This unpredictability highlights a key challenge for incorporating endophytes into nutrient management strategies. Improving our ability to predict endophyte impacts on host nutrient acquisition will require identifying the mechanisms underlying observed beneficial effects and scaling up to realistic, diverse root microbial communities.

Due to climate change, extreme weather events such as droughts are becoming more frequent and intense. This has a profound impact on plant performance and ecological interactions, including those involving herbivorous insects. The combined impact of drought stress and insect herbivory on plant metabolism has rarely been studied, particularly in woody plants. In this study, we investigated the influence of varying degrees of drought, both alone and in combination with herbivory by the leaf beetle Chrysomela tremulae, on the morphological and chemical characteristics of black poplar (Populus nigra) trees using a full factorial experimental design. We quantified morphological traits, volatile organic compound (VOC) emissions, phytohormone and amino acid concentrations, and phenolic profiles. Drought conditions increased the concentrations of salicylic acid (SA) and abscisic acid (ABA), while feeding induced ABA and SA. Amino acid profiles shifted significantly under drought conditions, particularly in beetle-infested plants. In contrast, salicinoids, which are the most important phenolic defense compounds in poplars, remained relatively stable. We also observed significant compound-specific effects on both constitutive and herbivore-induced VOC emissions. Our results demonstrate that drought and insect herbivory exert a joint influence on the chemical responses of P. nigra across multiple metabolic pathways. These findings highlight how the interaction between abiotic and biotic stresses can influence the defense chemistry of trees, which will consequently affect ecological interactions in forest ecosystems in the face of climate change.

1. Stomata and leaf traits are key regulators of plant water use efficiency and are expected to have changed in response to rising atmospheric CO2 concentrations and climate warming over the past centuries. However, long-term data documenting such changes are rare. 2. We leveraged herbarium collections to track changes in stomatal characteristics and leaf traits in 656 individuals of invasive Japanese knotweed and its hybrid Bohemian knotweed collected across their European range and spanning 160 years of invasive spread. 3. We found that several functional traits including stomatal density and maximum anatomical stomatal conductance did not show significant changes over time but that plants adjusted their stomatal size and shape over time, and that these changes were associated with increased atmospheric CO2 levels. Interestingly, Reynoutria japonica showed increases in stomatal size and stomatal elongation, while the hybrid R. x bohemica showed a reduction in stomatal size. Traits also varied systematically with climates of origin. Plants from warmer origins with higher evaporative demands during the growing season had thicker leaves, lower SLA, smaller stomata and higher stomatal density, indicating more conservative water-use strategies. Stomatal density and gas exchange capacity co-varied with leaf structural traits, and there was a trade-off between stomatal size and number. Overall, fast leaf economic traits were associated with slow physiological traits. 4. Our results suggest that stomatal anatomical plasticity may enhance climate resilience by maintaining a stable maximum gas exchange capacity across environmental gradients. Herbarium collections provide a unique resource for reconstructing plant responses to historical environmental changes and understanding intraspecific trait variation.

Understanding the relationships between soil macrofauna and decomposition is crucial for predicting how land-use change impacts ecosystem function in fragmented systems. This is because soil macrofauna affect decomposition and also respond to the changes in abiotic conditions across habitat gradients. This study investigates edge effects on the macrofauna contributions to decomposition across forest-field ecotones. We used bait lamina assay to quantify aboveground and belowground feeding activity of soil macrofauna in Autumn 2025 in three deciduous forest-old field ecotones and one coniferous forest-old field ecotone, in Southwestern Ontario, Canada. Vegetation diversity and composition, LAI and soil characteristics (i.e., soil organic matter, pH, temperature and moisture) were measured at each plot along the ecotone. Pitfall trap data collected in Summer 2025 at the same sites were used to characterize macrofauna communities. We used generalized linear mixed effects models to estimate the effect of distance to edge, site, and depth into the soil on bait lamina consumption and soil macrofauna, with transect nested within site as random effects. Consumption activity increased with distance into the forest from the field, with the edge representing an intermediate; and, decreased with increasing depth into the soil. In contrast, soil macrofauna abundance, especially isopods, decrease with distance into the forest from the field. These trends varied significantly across sites, so that consumption activity and abundance sometimes remained constant across the ecotone (i.e., site x distance interaction). The results demonstrate that macrofaunal contributions to bait consumption varied along the ecotone, shaped by interacting environmental gradients and shifts in community composition unique to each site.

ContextArtificial light at night (ALAN) has profound impacts on individual organisms and entire communities. Still, humans tend to underestimate the true biological (spatial) footprint of ALAN, in part because of our limited sensitivity to light compared to other organisms. ObjectivesWe sought to demonstrate how far ALAN can reach into dark spaces at levels that can impact organismal behavior and physiology using a fundamental physical law, the inverse square law. MethodsWe created a spatially explicit model of light spread on real landscapes, parameterized using increasingly available landscape-scale vegetation data to account for attenuation through forests and blocking by topographic relief. ResultsLighting types common in rural areas can produce biologically important effects more than 1 kilometer from the source, and effects of large lights might stretch 3 kilometers or more. The footprint of a light is determined by the complex and multidimensional interaction between characteristics of the light itself and the environment. For example, attenuation through a dense forest might decrease the footprint of a light more than 90% compared to the same light on a grassland. In complex environments, even small changes in light placement and characteristics can lead to large changes in the biological footprint of the light. ConclusionsDesigners and land stewards must account for lighting type, brightness, directionality, and reflected light to create ecologically responsible lighting. Vertical vegetation and topography strongly influence the propagation of biologically detrimental light, and environmental context is vital when planning and installing lights to minimize the biological impacts.

1. Sandy beaches along urbanized coasts are increasingly managed through beach nourishment and hard infrastructure, yet these interventions often constrain natural dune-building processes. Along the Belgian coast, where much of the beach-dune interface is bordered by dikes, promenades and intensive recreation, strandline vegetation may provide an overlooked mechanism for retaining sand and initiating embryo dune development. 2. We assessed the potential for four pioneer dune plant species (Cakile maritima, Calamagrostis arenaria, Elymus farctus and Salsola kali) to establish, develop vegetation cover and contribute to sand accumulation along the Belgian coast. Using field surveys from 2017-2023, LiDAR-derived beach elevation and annual sediment dynamics, we modelled species occurrence and abundance/cover in low-disturbance reference zones and projected these relationships across the wider coastline. 3. Occurrence models identified where abiotic conditions allow plants to establish and persist until the late growing season, whereas zero-inflated abundance/cover models estimated expected vegetation development across environmental gradients. Predicted occurrence was widespread for several species, suggesting that the abiotic gradients modelled here are not the primary constraints on potential establishment across large parts of the coast. In contrast, expected abundance/cover showed stronger species-specific responses, particularly to sand accretion, indicating that sediment dynamics mainly affect post-establishment vegetation development rather than occurrence alone. 4. Independent field measurements of embryo dunes showed positive relationships between vegetation cover and local sand accumulation for all four species. When scaled using spatial predictions of potential abundance/cover, pioneer vegetation could retain substantial volumes of sand, with Cakile maritima contributing the largest share, followed by Salsola kali, Elymus farctus and Calamagrostis arenaria. Estimated volumes depended on assumptions about whether vegetation occurs as dispersed units or aggregated patches. 5. Synthesis and applications. Our results show that, even along a heavily urbanized and nourished coastline, abiotic conditions can support strandline vegetation and embryo dune initiation where disturbance is reduced. Management actions such as limiting trampling, adapting beach cleaning and protecting strandline vegetation could enhance the retention of nourished sand and support nature-based coastal defense. Rather than replacing engineered interventions, strandline vegetation may increase the efficiency with which available sediment is retained within the beach-dune system.

Protected areas cannot halt biodiversity loss in isolation; integrating them with surrounding human-dominated landscapes is critical. However, this integration is challenged by substantial landscape heterogeneity at their borders, hindering our understanding of cross-border changes in ecosystem service provision. We introduce a novel framework for characterizing these dynamics by analyzing ecosystem service gradients along protected area borders. For 16 protected areas in the French Alps, we assessed 12 ecosystem services using a mix of established biophysical models and novel connectivity-based models for mobile species. These were aggregated into three stakeholder-driven domains reflecting respectively rural, cultural, and urban management priorities. Automated polynomial regression analysis classified borders into five gradient types. The most common were 'Decreasing Gradients', representing a decline in ecosystem services outside the protected area, and 'Increasing Gradients', with the opposite pattern. Our analysis reveals these patterns are driven by specific landscape configurations, uncovering frequent trade-offs between the three management priorities, where, for instance, landscapes supporting rural priorities often degrade cultural and urban ones. We also identify key opportunities for synergies, by identifying areas where ecosystem services for all three priority domains increase simultaneously outside the protected area. This spatially explicit typology provides a powerful diagnostic tool for designing targeted interventions, such as prioritizing habitat restoration where ecosystem services decline or managing agricultural landscapes to mitigate conflicts across management priorities, supporting a more effective integration of protected areas into the wider landscape.

Global climate change has increased the frequency and intensity of drought events, making it urgent to understand how native species respond to water deficit (WD). In biodiverse environments such as tropical forests, simple methods are needed to study multiple species simultaneously. This can help predict how natural environments will respond to climate change and guide the strategic selection of drought-resistant species for reforestation. This study aimed to: (1) adapt an existing simple and inexpensive method to apply a controlled WD on tree seedlings from tropical species commonly produced in nurseries for restoration projects, suitable for greenhouse experiments; and (2) evaluate the effectiveness of this method in generating ecophysiological responses to WD that allow the estimation of species' drought resistance. Ten native tree species from the Semideciduous Seasonal Forest (SSF), a phytophysiognomy of the Atlantic Forest, were selected. An existing method was adapted to implement capillary irrigation, in which the bases of the seedling tubes were placed in floral foam blocks positioned inside 15 L plastic containers filled with water. A gradual and severe WD was applied to five seedlings of each species by removing all water from the containers, leaving only the water retained in the saturated floral foam available for plant uptake. The remaining seedlings were maintained well-watered (containers full and foam saturated) as the control group. Stomatal conductance (gs) was measured daily for all seedlings until they reached 50% or less of their initial gs (igs); at this point, stem water potential ({Psi}w) was measured. Both gs and {Psi}w differed significantly among treatments and species (p < 0.01). Ficus guaranitica and Heliocarpus popayanensis were the only species that did not show significant {Psi}w differences between treatments, indicating higher drought resistance. In contrast, Campomanesia xanthocarpa and Eugenia uniflora had the lowest {Psi}w values under WD, suggesting lower drought resistance. The remaining species were distributed along a gradient of responses to WD. Additionally, no correlation was found between {Psi}w and gs at 50% igs in the WD group (rho = 0.16, p = 0.26). The method proved effective in inducing controlled WD and generating measurable ecophysiological responses, offering a useful tool for screening native species for drought resistance.

O_LIEuropean beech (Fagus sylvatica L.) is a widely distributed, ecologically and economically important deciduous tree species in European forests, but is increasingly threatened by drought stress. Volatile organic compounds (VOCs) are ubiquitous plant metabolites that may serve as non-invasive biomarkers of drought stress, yet they have rarely been studied in European beech. C_LIO_LIIn this study, we examined VOC responses of European beech to experimental drought across diverse genetic backgrounds in a common garden. The 72 four-year-old beech saplings represented three genetic clusters, seven provenances (geographic seed sources), and 12 maternal seed families. Half of the saplings were assigned to the drought treatment and received no water for 14 days, while the remaining saplings served as controls and were watered as required. VOC profiles, quantified as peak heights of mass spectral features, were measured for all individuals during pre-drought, drought, and rewatering periods. C_LIO_LIWe found that pre-drought VOC profiles, in particular monoterpenes, varied significantly among genetic backgrounds. Experimental drought significantly altered VOC profiles, characterized by increased green leaf volatiles and decreased monoterpenes, oxidized terpenoid derivatives, and other fatty acid derivatives. Reductions in monoterpenes persisted after rewatering, indicating a drought legacy effect. Drought responses were largely conserved across genetic backgrounds, with significant seed family-specific responses detected for only three VOC features. C_LIO_LIOur findings suggest that VOC profiles are genetically structured yet highly plastic under drought and highlight their potential as non-invasive biomarkers for monitoring drought stress in European beech under climate change. C_LI

While mutualistic symbioses with nitrogen-fixing bacteria enable plants to access fixed nitrogen, they also require substantial carbon investment. Under carbon limitation, such as shading, shifts in biomass allocation can decouple symbiotic investment from leaf and root growth, potentially compromising plant nitrogen status. Because shading shifts biomass allocation toward light acquisition, it could influence nitrogen fixing symbiosis in two opposing ways. If nodulation remains coupled to leaves rather than roots, nitrogen status should be maintained despite reduced root growth. Alternatively, if root growth constrains nodulation, nitrogen status should decline. We tested these hypotheses by manipulating light availability (full sunlight vs. 50% shade) and quantifying biomass allocation and symbiotic nodulation. Under shading, plants allocated proportionally more biomass to shoots than to roots and invested less biomass in root nodules. Relationships between nodulation and leaf or root biomass differed between treatments but converged with increasing plant size, although shaded plants never attained the root biomass observed in full sunlight. Leaf nitrogen concentration was maintained under shading because nodulation remained coupled to leaf investment despite reduced root allocation. These findings highlight that, under carbon limitation, maintaining leaf and nodule coupling enables plants to reduce nodule investment without compromising the nitrogen benefits of symbiosis.