{"paper_id":"02a8eddc-7e8e-4bcf-8c49-6080c06c61c6","body_text":"Effects of soil nutrient enrichment on biomass, herbivores and their predators differ between tree species in the Brazilian Cerrado | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Effects of soil nutrient enrichment on biomass, herbivores and their predators differ between tree species in the Brazilian Cerrado Carla Faleiro Tinoco, Sílvia Castro, Rodrigo Damasco Daud, Vanessa Leonel Falchi, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6968991/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 07 Feb, 2026 Read the published version in Oecologia → Version 1 posted 4 You are reading this latest preprint version Abstract Human-induced changes in nitrogen (N) and phosphorus (P) global cycles, significantly impact plant growth and nutritional composition, thereby affecting ecosystem dynamics. However, research on the effects of increased nutrient availability often focuses on plant community-level effects, overlooking interspecific variability and neglecting impacts on higher trophic levels. Using a controlled fertilization experiment with six tree species that naturally occur in the Cerrado biome (Brazilian savannas) we showed that there is significant interspecific variation in how plants responded to nutrient changes, indicating competitive advantages for certain native species under a scenario of increased soil nutrient availability. Such effects propagated to higher trophic levels (herbivores and their predators), also varying between plant host species. The strength and direction of N input effect depended on P levels and the type of herbivores. Large invertebrate leaf-herbivores were less affected than phytophagous mites. Impacts on higher trophic levels (predatory mites) were less pronounced than on phytophagous mites. Overall, we show that ongoing soil nutrient enrichment has the potential to alter interspecific competition dynamics in plant communities with consequences for ecological interaction partners. These findings have important implications for conservation and ecosystem management, especially in areas highly exposed to soil nutrient enrichment due to farming and industrial activities. plant-herbivore interaction phytophagous mites fertilization effects soil nutrient enrichment trees and shrubs Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Biogeochemical flows, especially those of nitrogen (N) and phosphorous (P), are greatly affected by anthropogenic activity, affecting ecosystems worldwide (Steffen et al., 2015 ). While N is a limiting nutrient for plants in most environments, human-driven increases in its availability are leading to biodiversity losses and changes in ecosystem functioning (Bobbink et al., 2010 ). Most studies that evaluate the impacts of ongoing soil nutrient enrichment on terrestrial communities tend to focus on community-level patterns of plants (Bobbink et al., 2010 ; Stevens et al., 2018 ). However, increased N availability can also lead to a variety of bottom-up effects, influencing trophic interactions through several mechanisms (Chen et al., 2010 ). Such effects on plants and interacting partners likely depend on traits of the plant species that regulate how the plants make use of the existing nutrients (Barbosa et al., 2014 ). Despite this, only a few studies investigate the interspecific variability of these effects (e.g. Barbosa et. al 2014 ) and how such impacts propagate to higher trophic levels at species level (e.g., herbivores and their natural enemies; but see Chen et al., 2010 ; Pöyry et al., 2017 ; Stevens et al., 2018 ; David et al., 2019 for community level analyses or reviews on propagation of impacts of nutrient enrichment). This knowledge gap is particularly pronounced in tropical regions and may limit our ability to predict the impacts of global change and plan appropriate management actions. Savanna ecosystems, such as the Cerrado, have ancient geological formations with advanced soil weathering processes, limiting the development and productivity of its vegetation by the low availability of nutrients in the soil, mainly N and P (oligotrophic soils, Bustamante et al., 2006 ; Haridasan, 2008 ). Many species in these environments have specific and energetically costly strategies that increase access to soil nutrients (e.g., root structures like root dimorphism, resorption of nutrients before leaf senescence and symbioses with microorganisms; Haridasan, 2008 ; Oliveira et al., 2015 ). These strategies require a substantial investment and can, depending on plant traits (Barbosa et al., 2014 ), result in less investment of plants in above-ground growth (Hoffmann & Franco, 2003 ; Lambers et al., 2020 ). Hence, in a scenario of increased nutrient availability, the negative impacts on plants above-ground growth may be more accentuated in these oligotrophic and highly biodiverse tropical ecosystems (e.g., Lambers et al., 2020 ). The change in plant nutritional quality, induced by increased N availability, may be more pronounced in soils co-limited by N and P and can affect consumers. Indeed, increased N availability tends to increase P limitation both in plants and their consumers (Vogels et al., 2023 ). Understanding the diversity of responses of plant trees in oligotrophic soils (e.g., Cerrado) to changes in nutrient soil levels is essential to better predict impacts of global environmental changes and to define adequate conservation and restoration practices. Effects of increased N availability can vary between plant species depending on their nutritional requirements. In a scenario of increased nutrient availability, plant species adapted to N rich soils (nitrophilous) can better take advantage of such increase than species that are adapted to oligotrophic soils (nitrophobous) (Bobbink et al., 2010 ; Stevens et al., 2018 ). Such, N-driven changes in plant species composition (i.e., abundance, richness, evenness) will hence affect the availability of resources for primary consumers, such as abundance of leaves (Throop & Lerdau, 2004 ; Stevens et al., 2018 ). Another important pathway through which soil nutrient enrichment can affect plant consumers is through changes in the nutritional quality of plant resources. Plant species naturally differ in their primary (e.g., protein and carbohydrate; Wilson et al., 2019 ) and secondary (e.g., alkaloids; Kessler & Kalske, 2018 ) compound content, and such variability is known to influence primary consumers performance and fitness (Throop & Lerdau, 2004 ; Nijssen et al., 2017 ; Stevens et al., 2018 ). The concentration of such compounds in leaves is affected by changes in soil N availability, especially when P is not a factor limiting N fixation (Throop & Lerdau, 2004 , Vitousek et al., 2010 ), changing how herbivores perceive the quality of the plant as a food resource. Indeed, several amino acids necessary for herbivore growth and reproduction are solely obtained through diet (i.e., are essential amino acids), thus, an increase in N soil levels can change the quality and palatability of plants as a food resource (Throop & Lerdau, 2004 ; Li et al., 2016 ; Stevens et al., 2018 ). Furthermore, increasing N supply, can also significantly decrease concentrations of secondary C-based compounds (e.g., polyphenolics and phenolics), which act as herbivory defenses, reducing plant resistance to herbivory (Chen & Ni, 2011 ; Sun et al., 2020 ). Consequently, such N-driven effects are likely to make the plant more susceptible to herbivory (Throop & Lerdau, 2004 ; Li et al., 2016 ; Stevens et al., 2018 ). The effects of changes of soil N content in plants and herbivores can also propagate to higher trophic levels, e.g., changing the susceptibility of herbivorous insects to their natural enemies (i.e., predators, parasitoids, and pathogens; Throop & Lerdau, 2004 ) or changing the production of plant’s volatile defense compounds that act as foraging signals for natural enemies (Chen & Ni, 2011 ). However, there is still little information on how changes in soil nutrient levels propagate from plants to herbivores and predators, particularly in tropical ecosystems. In this work, we used a controlled fertilization experiment to investigate the impacts of soil nutrient changes on plant biomass (number of leaves), herbivory, and predation in six tree species that naturally occur in Cerrado (but also occur in other biomes). Since the experimental plants were not exposed to competition with other plants, we expected that, moderated N increases would lead to a positive effect on number of leaves, these effects being more accentuated when P is not a limiting factor (expectation 1). We also expected herbivores to prefer plants grown in soils with moderate levels of N added than plants grown in low levels of N, these effects cascading to herbivore natural enemies (expectation 2). On the other hand, above certain thresholds of N and P levels, the plant may suffer a nutritional imbalance or start to invest more in toxic compounds (Throop & Lerdau, 2004 ; Chen et al., 2010 ), reducing the herbivores and their natural enemies (expectation 3). Materials and methods This study was carried out in five study sites within the Campus (Samambaia) of Federal University of Goiás (Table S1 ), Goiânia, Goiás, Brazil. The study sites were at least 500 m apart. Due to the limited dispersal ability of mites (e.g., Jung, 2005 ) and insect herbivores (Ricketts et al., 2008 ; Zurbuchen et al., 2010 ) this distance ensured that plants in each site were exposed to independent communities of these guilds. The sites are within a region of Brazilian Cerrado, with climate classified as Aw (Köppen), where rainfall ranges from 1600 to 1900 mm year − 1 , with an average annual temperature between 20 ºC and 22 ºC (Alvares et al., 2013 ). Besides sharing the same climatic regime, the areas exhibit a similar pattern of insolation, rainfall, and winds due to their proximity. Focal plant species The study focused on six tree species that naturally occur in the Cerrado biome, being relatively common. These species also occur in other Brazilian biomes, having different spatial ranges within South America, some being more widespread than others (Table S2). The selected species were chosen considering the availability of individuals in local commercial nurseries and cover a variety of strategies of nutrient acquisition (Table S2). Four of the selected species are fast-growing: Schinus terebinthifolia Raddi (Anacardiaceae) and Solanum lycocarpum A.St.-Hil. (Solanaceae) being pioneer species and Inga vera subsp. affinis (DC.) T.D.Penn. and Psidium guineense Sw. (Myrtaceae) being early secondary successional plants. S. terebinthifola and I. vera are also known to occur in biomes where nutrients are typically not a limitation (Pampas, Pantanal, Amazonia, and Atlantic Forest, Table S2). Two species are slow growing typical of later secondary successional stages, Campomanesia eugenioides (Cambess.) D.Legrand ex Landrum (Myrtaceae), and Eugenia involucrata DC. (Myrtaceae), the last occurring in biomes where nutrients are typically not a limitation (Pampas and Atlantic Forest, Table S2). All S. lycocarpum seedlings were obtained from the sowing of seeds collected from trees in natural areas. Four seeds were added to each pot and, before the addition of fertilizer, only the largest plant was left in each pot. For the other five plant species, saplings were directly acquired from local nurseries. A total of 730 individuals were reared in pots throughout the entire experiment. In October 2018, 350 individuals (70 individuals of S. terebinthifolia , I. vera , E. involucrate , C. eugenioides and S. lycocarpum ) were transferred to 11 L pots. The individuals of I. vera were planted in 18 L pots, due to its growing requirements. Due to high mortality, a further 180 individuals (45 saplings of C. eugenioides , 40 of E. involucrate , 25 of I. vera e 40 of S.terebinthifolia and 30 seeds of S. lycocarpum ) were acquired in February 2019 and 200 more individuals were added in November 2019 (50 saplings of S.terebinthifolia , 30 of I. vera and 60 seeds of S. lycocarpum ), including a new plant species, P. guineense (60 saplings of P. guineense ). Due to lack of availability of seedlings of C. eugenioides and E. involucrata in the nursery, no extra saplings of these species were included in the experiment. The soil used in the experiment was red underground soil (oxisol) commercially purchased, which was impoverished in terms of organic matter (0.75%, i.e., 3 to 6 times less than normal levels in Cerrado having between 2 and 5% of organic matter; Resck et al., 1991 ; Ruggiero et al., 2002 ; Lopes & Guilherme, 2016). Soil P levels are like those found in preserved natural soil from the Cerrado [ca. 1.83 mg/dm 3 , which corresponds to 3.66 kg/ha, normal levels below 2 mg/dm 3 (4 kg/ha), Lopes & Guilherme, 2016]. Experimental set up In each of the five study sites (Table S1 ) in a complete randomized block design arrangement (Figure S1 ), with six treatments with different levels of nutrients (see below). Each treatment contained three pots from each of the plant species (the exception being P. guineense that due to lack of available seedlings only had two pots). The position of each treatment in each study site and the order of the plants in each treatment was randomized. A drip irrigation system has been installed and regulated to provide approximately 750 ml of water per day for each sapling. Treatments combined three different levels of N (N0 = without addition of N, N1 = 60 kg.ha − 1 per application, N2 = 130 kg.ha − 1 per application) and two levels of P (P0 = without addition of P, P1 = 40 kg.ha − 1 per application). The application of N was done in the form of Urea [CO(NH 2 ) 2 ] and for P we used simple superphosphate [Ca(H 2 PO 4 ) 2 ]. The nutrient concentration used aims to replicate the practices recommended for fertilizer use agricultural crops farms. The application of 60 kg.ha − 1 of N and 40 kg.ha − 1 of P corresponds to a recommended level for some of the most common crop species in the study region (i.e., common bean, see Ramos et al., 2018 ), and 130 kg.ha − 1 of N corresponds to an excessive level of N, but is frequently applied by farmers in Cerrado environments (Ramos et al., 2018 ). The amount of fertilizer (N and P) applied in each pot was calculated based on their concentration in the applied elemental form (45% of N in Urea and 21% of P in simple superphosphate for P). The nutrients were applied directly to the soil in each pot approximately every three months since November 2018. Plant vegetative metrics Surveys to extract information on plant metrics were done in May, August 2019 and October 2020. For each individual plant, at each sampling event, we recorded the number of leaves per tree. We used the number of leaves as a proxy for plant biomass increase in response to soil fertilization (Throop & Lerdau, 2004 ), which serves as a more direct indicator of food availability for the herbivores studied here. When the plant was introduced in the experiment, we collected information on their height (stem length, cm). Plant height was used in the analyses of number of leaves to control for variations in size due to differences in age and planting times. Herbivores and predators Herbivory metrics considered in this study were: leaf herbivory by external large leaf-feeding invertebrate herbivores (e.g., ants, caterpillars, beetles and grasshoppers, no mammal herbivores were observed in the study areas) and density of phytophagous mites. In each sampling event, leaf herbivory was measured as the percentage of plant leaves showing signs of leaf tissue consumption by herbivores. If a leaf or leaflet was completely missing, this information was not counted as herbivory. Then, for each individual plant that had at least 12 leaves, we collected three leaves. If an individual had fewer than 12 leaves, the collection was adjusted so that we would never collect more than 25% of the total number of leaves of every individual. To avoid choosing which leaves to collect, we decided a priori to collect the 4th, 5th and 6th leaves, counting from the apex. For species with composed leaves with large leaflets (i.e., I. vera ), leaflets were collected. Leaves were stored in vials with ethanol 70%. Then, under laboratorial conditions, all the collected leaves were washed by shaking vigorously the recipient for 30 seconds to release all mites. Samples were then analyzed under stereoscopic microscope and all mites found were mounted on slides with Hoyer's medium (Moraes & Flechtmann, 2008 ). The mites were then quantified and identified to the lowest possible taxonomic level (Moraes and Flechtmann, 2008 ) and separated into morphogroups under phase contrast microscopy. Slides with mites were deposited in the collection of Acari of the Laboratory of Taxonomy, Ecology and Interaction of Arachnids (TEIA) at the Federal University of Goiás (UFG). The mite community was classified into three groups based on the predominant feeding habits of each group: 1) phytophagous, 2) predators and 3) undetermined/unknown (see Table S3). In the latter, we placed mites that were not identified to species level (e.g., immature or damaged individuals) and belong to families harboring species with different guilds. Groups that do not have species that feed on plants (Oribatida and Winterschimidtiida) like mycophagous species (which made up only a very small proportion of the mites collected) were excluded from analysis. Data analysis To test the effects of different fertilization treatments on number of leaves, leaf herbivory, phytophagous mite density, and predatory mite occurrence (presence/absence) for each plant species we used generalized linear mixed models (GLMMs). We used N and P levels as fixed categorical variables, also considering the interaction between the two nutrients. As some plant individuals were included after November 2018 (to compensate for high mortality, see above) the number of fertilization events that each plant received varied. Therefore, ‘number of fertilization events’ to which the plant was submitted was included as a covariate. To control for differences in plant size at the start of the experiment, in the model of plant number of leaves we included initial height (i.e., height in the first fertilization event to which the plant was submitted) as covariate. When analyzing herbivory (leaf herbivory (%) and phytophagous mite density) and predation, to disentangle between effects of resource abundance from resource quality, we included ‘number of leaves’ as a covariate in the models. A significant effect of N or P over and above the effect of ‘number of leaves’ is then an indication of an effect driven by changes in quality of plant resources. As we had multiple measurements per plant individual, to account for the spatial and temporal structure of the data, we included “sampling date” and “plant identity nested within study site” as random effects. This structure controls for repeated measurements on the same individual over time, as well as potential site-level variation. For each metric, we also attempted to run a single GLMM for all species, with plant species and its interaction with P and N as terms in the model. Yet, our statistical power was not sufficient for such large model. For number of leaves, we assumed a negative binomial error structure (and log-link function). To analyze leaf herbivory (%) and abundance of phytophagous mites we used Tweedie error structure (and log-link function). For phytophagous mites analyses we considered only the individuals classified as phytophagous, but as a sensitivity test, we re-run the analyses also considering the mites with undetermined diet (e.g., juveniles, mites without full taxonomic ID from families with mixed feeding strategies). A binomial model was used to analyze the probability of occurrence of predatory mites. For each plant species, a posteriori analyses were performed to compare results under different combinations of P and N, using the \"glht\" function version 1.4–25 from the ‘multcomp’ package (Hothorn et al., 2008 ). All analyses were performed using “glmmTMB” v.1.1.4 (Brooks et al., 2017 ), in the R program version 4.2.1 (R Core Team, 2023 ). Results Effects of fertilization on plant vegetative metrics Overall, N had a positive effect on the number of leaves, mean values being higher at the intermediate and/or highest N dosages for most species (Fig. 1 ). Yet, some species were more affected than others (significant for S. terebinthifolia , C. aeugenioides , P. guineense and E. involucrata , Fig. 1 ; Table S4), and no difference was detected between intermediate and highest levels of N (Fig. 1 ). No statistically significant influence of P levels on the effect of N was detected for most species (i.e., interaction term had no significant effect), but increases on P levels significantly increased the number of leaves of S. terebinthifolia and P. guineense (Fig. 1 b, e; Table S4). Effects of fertilization on herbivores and higher trophic levels Effects of fertilization treatments differed between herbivore types. For large foliar herbivores (mostly ants, caterpillars and beetles), the S. lycocarpum showed the highest proportion of leaves with signs of herbivory (mean ± sd: 38 ± 36%), the addition of N led to a reduction in herbivory in intermediate N level, while the presence of P increased herbivory in highest N level (Fig. 2 c). I. vera also had high levels of herbivory (31 ± 25% of the leaves affected), but no clear effect of fertilization treatment was detected (Fig. 2 a; Table S4). The other species showed less than 15% of leaves with signs of herbivory [ C. eugenioides (13 ± 21%), P. guianeense (9 ± 8%), E. involucrata (8 ± 17%), S. terebinthifolia (6 ± 10%)]. Of these, only P. guianeense showed effects of fertilization; no effects were detected in the other species. Yet, contrary to what was detected for S. lycocarpum , N addition reduced herbivory, this effect being significant when N was added in high dosages and combined with P (Fig. 2 e). As for mites, although their density was generally low (Table S6), effects of fertilizers were more frequently detected than in large invertebrate herbivores (Fig. 3 , Table S4). I. vera was the species with the highest density of phytophagous mites (79% of all mites detected were found in this species), followed by S. terebinthifolia (12%), and the three species of Myrtaceae showing the lowest densities. In total, we collected 11 976 mites belonging to 35 morphogroups in 16 families (Table S3). The phytophagous mites were the most abundant group, making up 86% of mites collected. Most (73%) mites belonged to the Tenuipalpidae family, followed by the Tetranychidae family (8%), both families of phytophagous mites, and by the predatory mite family Phytoseiidae (4%). The effect of N input on the density of phytophagous mites was dependent on the levels of P, positively for S. terebinthifolia and C. aeugenioides (Fig. 3 b; d, Table S5). For P. guineense , the high levels of N increased mite density only when P was not added, with the effect turning negative when P was introduced (Fig. 3 e, Table S5). For I. vera , the addition of P increased mite density, but its interaction with N reduced this effect (Fig. 3 a, Table S5). For S. lycocarpum N addition in the presence of P was also negative to phytophagous mites density (Fig. S2 e, Table S5). Most predatory mites were detected in I. vera (42% of the total number of predatory mites), S. lycocarpum (26%), S. terebinthifolia (19%), C. eugenioides (9%), being less frequent in E. involucrata (2%) and P. guineense (1%). As for the proportion of mites that were predatory (in relation to the total number of mites) E. involucrata (41% of their total number of mites), S. lycocarpum (39%) and C. eugenioides (27%) were the plant species with higher values, predatory mites being less frequent in P. guineense (12%), S. terebinthifolia (12%) and I. vera (4%). Due to the low number of mites in P. guineense , and E. involucrata these two species were not used to analyze the effect of fertilization on predatory mites. Among the remaining four plant species, I. vera , with the presence of P, the addition of intermediate N level also resulted in an increase in the occurrence of predatory mites (Fig. 4 a; Table S5). Discussion Our results show that the effects of fertilization on Cerrado tree species and their interactions with herbivores depend greatly on the identity of the plant species considered. Below we discuss possible mechanisms that regulate the interspecific variability of the effects detected here and the potential consequences for the conservation and restoration of Cerrado biodiversity. Effects of fertilization on plant vegetative metrics While we detected an overall positive trend of N input in leaf count, the variations in response intensity among plant species indicate that, under a scenario of soil nutrient enrichment in Cerrado, certain native plant species may have a competitive edge, and may increase their dominance, potentially driving others to local extinction. It is expected that negative impacts of nutrient addition would be greater for nitrophobous plants and positive impacts more accentuated for nitrophilous plants (David et al., 2019 ). For the species used in this study such classifications do not yet exist, but based on our results at least the two species of Myrtaceae for which positive impacts were more accentuated ( C. euginioides and P. guineense ) are likely to be nitrophilous. Other studies have shown that forest tree species of the Myrtaceae family exhibit a strong positive response to N with increasing survival (Nussbaumer et al., 2016 ) and biomass (Wooliver et al., 2016 ). Indeed, although the two Myrtaceae used in our study naturally in biomes known for having very nutrient-poor soils (i.e. Cerrado and Caatinga, see Table S2), they have very wide distributions within Brazil occurring in regions with naturally richer soils (e.g. Atlantic Forest, Table S2) and, for P. guineense , occurring also as exotic species in other continents (e.g. Asia and Africa, Senanayake et al. 2018 ). The results here presented may have implications for choices related to restoration programs. Plant responses to fertilization are closely related to their specific differences in nutritional requirement, such as co-limitation by different nutrients and differences in nutrient uptake and economy strategies, resulting in different intensities or even directions of responses (Kozovits et al., 2007 ; Wooliver et al., 2016 ). Yet, it is possible that such a beneficial effect only occurs when plants are protected from any competition effects with other species, as they were in our experimental setup. Further studies involving combinations of plants cooccurring and competing for soil resources would be needed to verify if such beneficial effect occurs in nature. Nevertheless, As P. guineense is a fast-growing species and C. euginioides is a slow-growing species (Table S2), in natural conditions where plants compete for space and resources, we can assume that the first has a higher risk of increasing dominance over other native species under a scenario of soil nutrient enrichment. For the slow-growing Myrtaceae species which appear later in the ecological succession ( C. eugenioide s and E. involucrata ) generally have a more conservative resource use strategy, being less able to respond to changes in nitrogen availability (Aidar et al., 2003 ; Báez and Homeier, 2017). Therefore, P. guineense is a species that may be better to avoid in restoration programs where soil is eutrophicated (e.g. due to nutrient leaching from farms). The lack of a significant increase in the number of leaves in I. vera , despite increased soil nutrient levels, may reflect its ability to regulate nitrogen uptake through symbiotic N₂ fixation with rhizobia, maintaining nitrogen balance even in nutrient-poor soils (see Table S2). Previous research suggests that increased N fertilization can lead to a reduction in biological N fixation (e.g., Weber et al., 2007 and Xia et al., 2017 ), which may explain the limited vegetative response observed in this species under higher N availability. Contrary to what we expected, we found no significant effect of P on the plant number of leaves of most plant species. Only S. terebinthifolia and P. guineense showed a significant response to increases in N and P in a non-additive manner. Other studies did not find effects of P and its interaction with N, which may be linked to factors ranging from low availability for use in the soil (Bucci et al., 2006 ) to the presence of P conservation mechanisms for some plant species adapted to dystrophic environments (Kozovits et al., 2007 ; Abrahão et al., 2018). Yet, it is important to note that we found an effect of P on other trophic levels. Thus, P was likely allocated to functions other than biomass production, potentially enhancing leaf nutritional quality by increasing tissue P concentration without affecting leaf quantity. Indeed, nutrient enrichment can alter several tree characteristics in addition to aerial biomass, such as root biomass, concentrations of photosynthetic pigments, tissue nitrogen, concentrations of proteins carbohydrates and secondary metabolites (Li et al., 2016 ) which in turn can affect interactions with higher trophic levels. Effects of fertilization on herbivores and higher trophic levels Given the great variability of effects of fertilization on plant growth, variability on herbivore responses to fertilization was expected. While previous studies on tropical regions show that, increasing soil N availability (alone or in combination with P) has negative impacts on the abundance of terrestrial tropical invertebrates (Nessel et al., 2021 ; 2022), other studies in temperate ecosystems demonstrated that the effects of soil fertilization on herbivores can be positive, negative, or neutral (Kyto et al., 1996 ; Butler et al., 2012 ; La Pierre and Smith, 2016 ; Nessel et al., 2021 ;2022), depending on the host plant species and herbivorous group considered, as observed in our study. The species I. vera showed the highest density of mites and a high leaf herbivory regardless of the nutrient treatment, while the other species showed intermediate to high resistance to herbivory. This reflected in differences between levels of herbivory in response to soil nutrient enrichment for each plant studied, making it difficult to detect general response patterns. As expected, the effects of nutrient supply were more pronounced for phytophagous mites than for plant growth. In general, we found a positive effect for plants. Other studies also point out that the effects can be magnified by trophic cascades (e.g., Carvalheiro et al., 2010 ). The fact that the effects of nutrients were more accentuated for mites than for herbivory resulting from large invertebrate could be related to different levels of diet specialization. Sucking herbivores, such as phytophagous mites, showed a much stronger response to fertilizers than chewing insects, such as large external leaf herbivores (Butler et al., 2012 ). Sucking herbivores tend to be more specialized feeding on a single species or phylogenetically close plant species, while chewing herbivores can be more generalist (Ali and Agrawal, 2012 ), and magnification of impacts through trophic chains or webs is mainly expected for species with more specialized diets (Carvalheiro et al., 2010 ). Differences in functional and behavioral traits (e.g., eating habit, mobility) could also influence the responses of herbivores to plant changes mediated by fertilizers (Kyto et al., 1996 ; Butler et al., 2012 ; Nessel et al., 2021 ). As in other studies (Butler et al., 2012 ; Nessel et al., 2021 ; 2022), here it was difficult to detect a consistent effect of phosphorus and its interaction with N on herbivores and predators. Although the literature describes an overall negative effect (Butler et al., 2012 ; Nessel et al., 2021 ; 2022), some studies indicate positive effects of increasing P availability on herbivores (Butler et al., 2012 ). However, the effect of increasing P, especially in isolation, still needs to be further investigated. Concluding remarks Environmental eutrophication is one of the major human-driven changes affecting biodiversity (Bobbink et al., 2010 ; Steffen et al., 2015 ). Due to its intensive use for agriculture, Cerrado is highly exposed to such effect (Bustamante et al., 2012 ). Our results clearly show that altered soil conditions will favor some plant species more than others and will have consequences for higher trophic levels. These findings highlight the urge for long term monitoring schemes to evaluate how ongoing soil nutrient enrichment is changing the dynamics between plant trees that naturally occur in Cerrado and their ecological interactions. These findings also have implications for restoration programs in Cerrado. Soil properties in areas that are subjected to restoration actions typically have highly degraded soils (Prescott et al., 2021 ), potentially being poorer in nutrients than normal soils or potentially being enriched in certain nutrients that are widely applied in crop fields (nitrogen and phosphorus). Future studies with a larger number of species that explore which plant traits explain their response to nutrient level changes are essential to identify how different plant species will perform when used in a restoration plan. Overall, the results of this work contribute to a better understanding of how different plant species, valuable for restoration, respond when planted in soils with different nutrient conditions. Declarations Authors contributions CFT and LGC conceived the project and designed methodology with help of CMCS and RDD. CFT collected and analyzed the data with help of LGC; CFT wrote the first draft; RDD, VLF, JAR and SRC identified the mite community. CFT wrote de first draft of the manuscript with help of LGC and SC. All authors revised the manuscript. Acknowledgements We thank F Venturoli, GM de Oliveira, AR Nascimento, CASB Braga, MCS Fioravanti, GA Ferreira for the spaces provided for setting up the experiment. We thank MBE Bernardes, PM Consorte and members of the Community Ecology laboratory at UFG for their help in setting up the experiment, and in collecting and sorting the data. Funding CFT was funded by CAPES (88882.347112/2019-01; 88887.800299/2022-00). LGC was funded by CNPq (307625/2021-4). Conflict of Interest Statement We declare there is no conflict of interest. Ethics approval Not applicable Consent to participate Not applicable Consent for publication Not applicable Availability of data and material The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request. Code availability Not applicable References Abrahão, A., Costa, P. de B., Lambers, H., Andrade, S. A. L., Sawaya, A. C. H. F., Ryan, M. H., & Oliveira, R. S. (2019). Soil types select for plants with matching nutrient-acquisition and -use traits in hyperdiverse and severely nutrient-impoverished campos rupestres and cerrado in Central Brazil. 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Soil Science and Plant Nutrition, 63(5), 470–482 . https://doi.org/10.1080/00380768.2017.1370960 Supplementary Files SupportingInformationOecologia.docx Cite Share Download PDF Status: Published Journal Publication published 07 Feb, 2026 Read the published version in Oecologia → Version 1 posted Reviewers agreed at journal 07 Aug, 2025 Reviewers invited by journal 07 Aug, 2025 Editor assigned by journal 25 Jun, 2025 First submitted to journal 24 Jun, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-6968991\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":497018203,\"identity\":\"a6292f70-0aff-416d-b95d-826a412404a2\",\"order_by\":0,\"name\":\"Carla Faleiro Tinoco\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA7UlEQVRIiWNgGAWjYHACxgMMNgw8YOYHIGZjJ0LPAYY0A7AWxhkgLcxEagEzmMEaCWnhbz/74MCPhD8y5uzNDx/b/Nomz8fMwPjhYw5uLRJn0g0O9iQY8Fj2HDM2zu27bdjGzMAsOXMbbi0GDGkMB3h/GPAY3Mhhk87tuc0I1MLGzItPC/8zhoN/EqBaLHtu2xPWIpHGcJgHpoXhx+1EglokbjxjOCyTYMxjcOaYsWFvw+3kNmbGZrx+4e9PY3z4JkHO3uB488MHP/7ctp3f3nzww0c8WlABYxuYbCBWPQj8IUXxKBgFo2AUjBQAAMltTQ4WQqFkAAAAAElFTkSuQmCC\",\"orcid\":\"https://orcid.org/0000-0002-3200-5856\",\"institution\":\"UFG: Universidade Federal de Goias\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Carla\",\"middleName\":\"Faleiro\",\"lastName\":\"Tinoco\",\"suffix\":\"\"},{\"id\":497018204,\"identity\":\"6ddd771f-6d4b-43d3-bedc-d34121571643\",\"order_by\":1,\"name\":\"Sílvia Castro\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"University of Coimbra: Universidade de Coimbra\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Sílvia\",\"middleName\":\"\",\"lastName\":\"Castro\",\"suffix\":\"\"},{\"id\":497018205,\"identity\":\"4878fedb-7da3-4ced-8f0e-ca1a120af200\",\"order_by\":2,\"name\":\"Rodrigo Damasco Daud\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Federal University of Goias: Universidade Federal de Goias\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Rodrigo\",\"middleName\":\"Damasco\",\"lastName\":\"Daud\",\"suffix\":\"\"},{\"id\":497018206,\"identity\":\"feb8469f-6b2a-4543-bc61-2c4be6d4914f\",\"order_by\":3,\"name\":\"Vanessa Leonel Falchi\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Federal University of Goias: Universidade Federal de Goias\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Vanessa\",\"middleName\":\"Leonel\",\"lastName\":\"Falchi\",\"suffix\":\"\"},{\"id\":497018207,\"identity\":\"51fb9f43-1fdf-4fba-a785-ac5a75809f4f\",\"order_by\":4,\"name\":\"Júlia Almeida Reis\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Federal University of Goias: Universidade Federal de Goias\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Júlia\",\"middleName\":\"Almeida\",\"lastName\":\"Reis\",\"suffix\":\"\"},{\"id\":497018208,\"identity\":\"fa30c438-d694-42ac-b6e8-7131aca5ed57\",\"order_by\":5,\"name\":\"Stefany Ribeiro Constantino\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Federal University of Goias: Universidade Federal de Goias\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Stefany\",\"middleName\":\"Ribeiro\",\"lastName\":\"Constantino\",\"suffix\":\"\"},{\"id\":497018209,\"identity\":\"b1390636-7c99-409d-9129-c3ef2f7d83f4\",\"order_by\":6,\"name\":\"Carlos de Melo e Silva Neto\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Instituto Federal de Educação Ciência e Tecnologia Goiano: Instituto Federal Goiano\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Carlos\",\"middleName\":\"de Melo e Silva\",\"lastName\":\"Neto\",\"suffix\":\"\"},{\"id\":497018210,\"identity\":\"7a7fbb25-0546-4330-b5a4-fe282e2eec6c\",\"order_by\":7,\"name\":\"Luísa Gigante Carvalheiro\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Federal University of Goias: Universidade Federal de Goias\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Luísa\",\"middleName\":\"Gigante\",\"lastName\":\"Carvalheiro\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2025-06-24 21:51:24\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-6968991/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-6968991/v1\",\"draftVersion\":[],\"editorialEvents\":[{\"content\":\"https://doi.org/10.1007/s00442-026-05863-z\",\"type\":\"published\",\"date\":\"2026-02-07T15:57:17+00:00\"}],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":88910381,\"identity\":\"91754c21-b25a-4004-bdf2-860ea87770e9\",\"added_by\":\"auto\",\"created_at\":\"2025-08-12 15:09:44\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":219300,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eEffect of the different fertilization treatments on the number of leaves for each plant species. Estimates of mean values (back transformed values) and associated 95% confidence intervals are presented. N0= without addition of N, N1=60 kg N / ha, N2=130 kg N / ha, P0= without addition P, P1= 40 kg P / ha. Whenever the probability of N or P having a significant effect was higher than 90% (P-value \\u0026lt;0.1, Table S4), post hoc tests were done to compare values obtained under different nutrient combinations (values that have a probability of being different higher than 90%, i.e. P\\u0026lt;0.1, are indicated with distinct letters). Details on statistical analyses are presented in Table S4 and S5.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-6968991/v1/27a931cb8b8b8e0d202a0216.png\"},{\"id\":88910383,\"identity\":\"94dd65e5-9bd9-4c0d-84fb-362c4dc633d7\",\"added_by\":\"auto\",\"created_at\":\"2025-08-12 15:09:44\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":229620,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eEffect of the different fertilization treatments in the percentage of leaves with herbivory marks, for each plant species. Estimates of mean values (back transformed values) and associated 95% confidence intervals are presented. N0= without addition of N, N1=60 kg N / ha, N2=130 kg N / ha, P0= without addition P, P1= 40 kg P / ha. Whenever the probability of N or P having a significant effect was higher than 90% (P-value \\u0026lt;0.1, Table S4), post hoc tests were done to compare values obtained under different nutrient combinations (values that have a probability of being different higher than 90%, i.e. P\\u0026lt;0.1, are indicated with distinct letters). Details on statistical analyses are presented in Table S4 and S5.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-6968991/v1/f43066d5b29e00f4bffcc4d9.png\"},{\"id\":88910384,\"identity\":\"714ff280-208d-40c2-bdc9-2c068b408fb7\",\"added_by\":\"auto\",\"created_at\":\"2025-08-12 15:09:44\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":200768,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eEffect of the fertilization treatments on phytophagous mite density (individuals per leaf), assuming mites with indeterminate feeding habits are also phytophagous, detected on each plant species. Estimates of mean values (back transformed) and associated 95% confidence intervals are presented. N0= without addition of N, N1=60 kg N / ha, N2=130 kg N / ha, P0= without addition P, P1= 40 kg P / ha. N0= without addition of N, N1=60 kg N / ha, N2=130 kg N / ha, P0= without addition P, P1= 40 kg P / ha. Whenever the probability of N or P having a significant effect was higher than 90% (P-value \\u0026lt;0.1, Table S4), post hoc tests were done to compare values obtained under different nutrient combinations (values that have a probability of being different higher than 90%, i.e. P\\u0026lt;0.1, are indicated with distinct letters). Details on statistical analyses are presented in Table S4 and S5.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-6968991/v1/c5e141995b453081af117df1.png\"},{\"id\":88910392,\"identity\":\"0b2b12a9-e394-432b-89a2-5f476ca6c817\",\"added_by\":\"auto\",\"created_at\":\"2025-08-12 15:09:44\",\"extension\":\"png\",\"order_by\":4,\"title\":\"Figure 4\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":200660,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eEffect of the different fertilization treatments in predatory mite probability of occurrence for each plant species. Estimates of the mean values (back transformed) and associated 95% confidence intervals are presented. N0= without addition of N, N1=60 kg N / ha, N2=130 kg N / ha, P0= without addition P, P1= 40 kg P / ha. The species \\u003cem\\u003eP. guineense\\u003c/em\\u003e and \\u003cem\\u003eE. involucrate\\u003c/em\\u003e were not included in the analyses due to the low number of predatory mites detected. Whenever the probability of N or P having a significant effect was higher than 90% (P-value \\u0026lt;0.1, Table S4), post hoc tests were done to compare values obtained under different nutrient combinations (values that have a probability of being different higher than 90%, i.e. P\\u0026lt;0.1, are indicated with distinct letters). Details on statistical analyses are presented in Table S4 and S5.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"floatimage4.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-6968991/v1/8630645da1d4b584c561cc0e.png\"},{\"id\":102234030,\"identity\":\"789abb18-88ef-405c-8ae3-3085cd00b763\",\"added_by\":\"auto\",\"created_at\":\"2026-02-09 16:04:32\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":1628383,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-6968991/v1/d108c652-f940-401b-9a8a-eca39ea987dd.pdf\"},{\"id\":88910389,\"identity\":\"da1bdc0a-361b-43f2-85d4-b398596e6844\",\"added_by\":\"auto\",\"created_at\":\"2025-08-12 15:09:44\",\"extension\":\"docx\",\"order_by\":4,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"supplement\",\"size\":1558853,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"SupportingInformationOecologia.docx\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-6968991/v1/ef57f926c35a0e1d1bdd9eaa.docx\"}],\"financialInterests\":\"\",\"formattedTitle\":\"Effects of soil nutrient enrichment on biomass, herbivores and their predators differ between tree species in the Brazilian Cerrado\",\"fulltext\":[{\"header\":\"Introduction\",\"content\":\"\\u003cp\\u003eBiogeochemical flows, especially those of nitrogen (N) and phosphorous (P), are greatly affected by anthropogenic activity, affecting ecosystems worldwide (Steffen et al., \\u003cspan citationid=\\\"CR41\\\" class=\\\"CitationRef\\\"\\u003e2015\\u003c/span\\u003e). While N is a limiting nutrient for plants in most environments, human-driven increases in its availability are leading to biodiversity losses and changes in ecosystem functioning (Bobbink et al., \\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e). Most studies that evaluate the impacts of ongoing soil nutrient enrichment on terrestrial communities tend to focus on community-level patterns of plants (Bobbink et al., \\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e; Stevens et al., \\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e). However, increased N availability can also lead to a variety of bottom-up effects, influencing trophic interactions through several mechanisms (Chen et al., \\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e). Such effects on plants and interacting partners likely depend on traits of the plant species that regulate how the plants make use of the existing nutrients (Barbosa et al., \\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e2014\\u003c/span\\u003e). Despite this, only a few studies investigate the interspecific variability of these effects (e.g. Barbosa et. al \\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e2014\\u003c/span\\u003e) and how such impacts propagate to higher trophic levels at species level (e.g., herbivores and their natural enemies; but see Chen et al., \\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e; P\\u0026ouml;yry et al., \\u003cspan citationid=\\\"CR34\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e; Stevens et al., \\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e; David et al., \\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e for community level analyses or reviews on propagation of impacts of nutrient enrichment). This knowledge gap is particularly pronounced in tropical regions and may limit our ability to predict the impacts of global change and plan appropriate management actions.\\u003c/p\\u003e\\u003cp\\u003eSavanna ecosystems, such as the Cerrado, have ancient geological formations with advanced soil weathering processes, limiting the development and productivity of its vegetation by the low availability of nutrients in the soil, mainly N and P (oligotrophic soils, Bustamante et al., \\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e2006\\u003c/span\\u003e; Haridasan, \\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e2008\\u003c/span\\u003e). Many species in these environments have specific and energetically costly strategies that increase access to soil nutrients (e.g., root structures like root dimorphism, resorption of nutrients before leaf senescence and symbioses with microorganisms; Haridasan, \\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e2008\\u003c/span\\u003e; Oliveira et al., \\u003cspan citationid=\\\"CR33\\\" class=\\\"CitationRef\\\"\\u003e2015\\u003c/span\\u003e). These strategies require a substantial investment and can, depending on plant traits (Barbosa et al., \\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e2014\\u003c/span\\u003e), result in less investment of plants in above-ground growth (Hoffmann \\u0026amp; Franco, \\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e2003\\u003c/span\\u003e; Lambers et al., \\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). Hence, in a scenario of increased nutrient availability, the negative impacts on plants above-ground growth may be more accentuated in these oligotrophic and highly biodiverse tropical ecosystems (e.g., Lambers et al., \\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). The change in plant nutritional quality, induced by increased N availability, may be more pronounced in soils co-limited by N and P and can affect consumers. Indeed, increased N availability tends to increase P limitation both in plants and their consumers (Vogels et al., \\u003cspan citationid=\\\"CR47\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e). Understanding the diversity of responses of plant trees in oligotrophic soils (e.g., Cerrado) to changes in nutrient soil levels is essential to better predict impacts of global environmental changes and to define adequate conservation and restoration practices.\\u003c/p\\u003e\\u003cp\\u003eEffects of increased N availability can vary between plant species depending on their nutritional requirements. In a scenario of increased nutrient availability, plant species adapted to N rich soils (nitrophilous) can better take advantage of such increase than species that are adapted to oligotrophic soils (nitrophobous) (Bobbink et al., \\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e; Stevens et al., \\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e). Such, N-driven changes in plant species composition (i.e., abundance, richness, evenness) will hence affect the availability of resources for primary consumers, such as abundance of leaves (Throop \\u0026amp; Lerdau, \\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e2004\\u003c/span\\u003e; Stevens et al., \\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e). Another important pathway through which soil nutrient enrichment can affect plant consumers is through changes in the nutritional quality of plant resources. Plant species naturally differ in their primary (e.g., protein and carbohydrate; Wilson et al., \\u003cspan citationid=\\\"CR49\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e) and secondary (e.g., alkaloids; Kessler \\u0026amp; Kalske, \\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e) compound content, and such variability is known to influence primary consumers performance and fitness (Throop \\u0026amp; Lerdau, \\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e2004\\u003c/span\\u003e; Nijssen et al., \\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e; Stevens et al., \\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e). The concentration of such compounds in leaves is affected by changes in soil N availability, especially when P is not a factor limiting N fixation (Throop \\u0026amp; Lerdau, \\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e2004\\u003c/span\\u003e, Vitousek et al., \\u003cspan citationid=\\\"CR46\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e), changing how herbivores perceive the quality of the plant as a food resource. Indeed, several amino acids necessary for herbivore growth and reproduction are solely obtained through diet (i.e., are essential amino acids), thus, an increase in N soil levels can change the quality and palatability of plants as a food resource (Throop \\u0026amp; Lerdau, \\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e2004\\u003c/span\\u003e; Li et al., \\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e; Stevens et al., \\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e). Furthermore, increasing N supply, can also significantly decrease concentrations of secondary C-based compounds (e.g., polyphenolics and phenolics), which act as herbivory defenses, reducing plant resistance to herbivory (Chen \\u0026amp; Ni, \\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e2011\\u003c/span\\u003e; Sun et al., \\u003cspan citationid=\\\"CR44\\\" class=\\\"CitationRef\\\"\\u003e2020\\u003c/span\\u003e). Consequently, such N-driven effects are likely to make the plant more susceptible to herbivory (Throop \\u0026amp; Lerdau, \\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e2004\\u003c/span\\u003e; Li et al., \\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e; Stevens et al., \\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003eThe effects of changes of soil N content in plants and herbivores can also propagate to higher trophic levels, e.g., changing the susceptibility of herbivorous insects to their natural enemies (i.e., predators, parasitoids, and pathogens; Throop \\u0026amp; Lerdau, \\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e2004\\u003c/span\\u003e) or changing the production of plant\\u0026rsquo;s volatile defense compounds that act as foraging signals for natural enemies (Chen \\u0026amp; Ni, \\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e2011\\u003c/span\\u003e). However, there is still little information on how changes in soil nutrient levels propagate from plants to herbivores and predators, particularly in tropical ecosystems.\\u003c/p\\u003e\\u003cp\\u003eIn this work, we used a controlled fertilization experiment to investigate the impacts of soil nutrient changes on plant biomass (number of leaves), herbivory, and predation in six tree species that naturally occur in Cerrado (but also occur in other biomes). Since the experimental plants were not exposed to competition with other plants, we expected that, moderated N increases would lead to a positive effect on number of leaves, these effects being more accentuated when P is not a limiting factor (expectation 1). We also expected herbivores to prefer plants grown in soils with moderate levels of N added than plants grown in low levels of N, these effects cascading to herbivore natural enemies (expectation 2). On the other hand, above certain thresholds of N and P levels, the plant may suffer a nutritional imbalance or start to invest more in toxic compounds (Throop \\u0026amp; Lerdau, \\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e2004\\u003c/span\\u003e; Chen et al., \\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e), reducing the herbivores and their natural enemies (expectation 3).\\u003c/p\\u003e\"},{\"header\":\"Materials and methods\",\"content\":\"\\u003cp\\u003eThis study was carried out in five study sites within the Campus (Samambaia) of Federal University of Goi\\u0026aacute;s (Table \\u003cspan refid=\\\"MOESM1\\\" class=\\\"InternalRef\\\"\\u003eS1\\u003c/span\\u003e), Goi\\u0026acirc;nia, Goi\\u0026aacute;s, Brazil. The study sites were at least 500 m apart. Due to the limited dispersal ability of mites (e.g., Jung, \\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e2005\\u003c/span\\u003e) and insect herbivores (Ricketts et al., \\u003cspan citationid=\\\"CR39\\\" class=\\\"CitationRef\\\"\\u003e2008\\u003c/span\\u003e; Zurbuchen et al., \\u003cspan citationid=\\\"CR51\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e) this distance ensured that plants in each site were exposed to independent communities of these guilds. The sites are within a region of Brazilian Cerrado, with climate classified as Aw (K\\u0026ouml;ppen), where rainfall ranges from 1600 to 1900 mm year\\u003csup\\u003e\\u0026minus;\\u0026thinsp;1\\u003c/sup\\u003e, with an average annual temperature between 20 \\u0026ordm;C and 22 \\u0026ordm;C (Alvares et al., \\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e2013\\u003c/span\\u003e). Besides sharing the same climatic regime, the areas exhibit a similar pattern of insolation, rainfall, and winds due to their proximity.\\u003c/p\\u003e\\u003cdiv id=\\\"Sec3\\\" class=\\\"Section2\\\"\\u003e\\u003ch2\\u003eFocal plant species\\u003c/h2\\u003e\\u003cp\\u003eThe study focused on six tree species that naturally occur in the Cerrado biome, being relatively common. These species also occur in other Brazilian biomes, having different spatial ranges within South America, some being more widespread than others (Table S2). The selected species were chosen considering the availability of individuals in local commercial nurseries and cover a variety of strategies of nutrient acquisition (Table S2). Four of the selected species are fast-growing: \\u003cem\\u003eSchinus terebinthifolia\\u003c/em\\u003e Raddi (Anacardiaceae) and \\u003cem\\u003eSolanum lycocarpum\\u003c/em\\u003e A.St.-Hil. (Solanaceae) being pioneer species and \\u003cem\\u003eInga vera subsp. affinis (DC.) T.D.Penn.\\u003c/em\\u003e and \\u003cem\\u003ePsidium guineense\\u003c/em\\u003e Sw. (Myrtaceae) being early secondary successional plants. \\u003cem\\u003eS. terebinthifola\\u003c/em\\u003e and \\u003cem\\u003eI. vera\\u003c/em\\u003e are also known to occur in biomes where nutrients are typically not a limitation (Pampas, Pantanal, Amazonia, and Atlantic Forest, Table S2). Two species are slow growing typical of later secondary successional stages, \\u003cem\\u003eCampomanesia eugenioides\\u003c/em\\u003e (Cambess.) D.Legrand ex Landrum (Myrtaceae), and \\u003cem\\u003eEugenia involucrata\\u003c/em\\u003e DC. (Myrtaceae), the last occurring in biomes where nutrients are typically not a limitation (Pampas and Atlantic Forest, Table S2).\\u003c/p\\u003e\\u003cp\\u003eAll \\u003cem\\u003eS. lycocarpum\\u003c/em\\u003e seedlings were obtained from the sowing of seeds collected from trees in natural areas. Four seeds were added to each pot and, before the addition of fertilizer, only the largest plant was left in each pot. For the other five plant species, saplings were directly acquired from local nurseries.\\u003c/p\\u003e\\u003cp\\u003eA total of 730 individuals were reared in pots throughout the entire experiment. In October 2018, 350 individuals (70 individuals of \\u003cem\\u003eS. terebinthifolia\\u003c/em\\u003e, \\u003cem\\u003eI. vera\\u003c/em\\u003e, \\u003cem\\u003eE. involucrate\\u003c/em\\u003e, \\u003cem\\u003eC. eugenioides\\u003c/em\\u003e and \\u003cem\\u003eS. lycocarpum\\u003c/em\\u003e) were transferred to 11 L pots. The individuals of \\u003cem\\u003eI. vera\\u003c/em\\u003e were planted in 18 L pots, due to its growing requirements. Due to high mortality, a further 180 individuals (45 saplings of \\u003cem\\u003eC. eugenioides\\u003c/em\\u003e, 40 of \\u003cem\\u003eE. involucrate\\u003c/em\\u003e, 25 of \\u003cem\\u003eI. vera\\u003c/em\\u003e e 40 of \\u003cem\\u003eS.terebinthifolia\\u003c/em\\u003e and 30 seeds of \\u003cem\\u003eS. lycocarpum\\u003c/em\\u003e) were acquired in February 2019 and 200 more individuals were added in November 2019 (50 saplings of \\u003cem\\u003eS.terebinthifolia\\u003c/em\\u003e, 30 of \\u003cem\\u003eI. vera\\u003c/em\\u003e and 60 seeds of \\u003cem\\u003eS. lycocarpum\\u003c/em\\u003e), including a new plant species, \\u003cem\\u003eP. guineense\\u003c/em\\u003e (60 saplings of \\u003cem\\u003eP. guineense\\u003c/em\\u003e). Due to lack of availability of seedlings of \\u003cem\\u003eC. eugenioides\\u003c/em\\u003e and \\u003cem\\u003eE. involucrata\\u003c/em\\u003e in the nursery, no extra saplings of these species were included in the experiment.\\u003c/p\\u003e\\u003cp\\u003eThe soil used in the experiment was red underground soil (oxisol) commercially purchased, which was impoverished in terms of organic matter (0.75%, i.e., 3 to 6 times less than normal levels in Cerrado having between 2 and 5% of organic matter; Resck et al., \\u003cspan citationid=\\\"CR38\\\" class=\\\"CitationRef\\\"\\u003e1991\\u003c/span\\u003e; Ruggiero et al., \\u003cspan citationid=\\\"CR40\\\" class=\\\"CitationRef\\\"\\u003e2002\\u003c/span\\u003e; Lopes \\u0026amp; Guilherme, 2016). Soil P levels are like those found in preserved natural soil from the Cerrado [ca. 1.83 mg/dm\\u003csup\\u003e3\\u003c/sup\\u003e, which corresponds to 3.66 kg/ha, normal levels below 2 mg/dm\\u003csup\\u003e3\\u003c/sup\\u003e (4 kg/ha), Lopes \\u0026amp; Guilherme, 2016].\\u003c/p\\u003e\\u003c/div\\u003e\\n\\u003ch3\\u003eExperimental set up\\u003c/h3\\u003e\\n\\u003cp\\u003eIn each of the five study sites (Table \\u003cspan refid=\\\"MOESM1\\\" class=\\\"InternalRef\\\"\\u003eS1\\u003c/span\\u003e) in a complete randomized block design arrangement (Figure \\u003cspan refid=\\\"MOESM1\\\" class=\\\"InternalRef\\\"\\u003eS1\\u003c/span\\u003e), with six treatments with different levels of nutrients (see below). Each treatment contained three pots from each of the plant species (the exception being \\u003cem\\u003eP. guineense\\u003c/em\\u003e that due to lack of available seedlings only had two pots). The position of each treatment in each study site and the order of the plants in each treatment was randomized. A drip irrigation system has been installed and regulated to provide approximately 750 ml of water per day for each sapling.\\u003c/p\\u003e\\u003cp\\u003eTreatments combined three different levels of N (N0\\u0026thinsp;=\\u0026thinsp;without addition of N, N1\\u0026thinsp;=\\u0026thinsp;60 kg.ha\\u003csup\\u003e\\u0026minus;\\u0026thinsp;1\\u003c/sup\\u003eper application, N2\\u0026thinsp;=\\u0026thinsp;130 kg.ha\\u003csup\\u003e\\u0026minus;\\u0026thinsp;1\\u003c/sup\\u003e per application) and two levels of P (P0\\u0026thinsp;=\\u0026thinsp;without addition of P, P1\\u0026thinsp;=\\u0026thinsp;40 kg.ha\\u003csup\\u003e\\u0026minus;\\u0026thinsp;1\\u003c/sup\\u003e per application). The application of N was done in the form of Urea [CO(NH\\u003csub\\u003e2\\u003c/sub\\u003e)\\u003csub\\u003e2\\u003c/sub\\u003e] and for P we used simple superphosphate [Ca(H\\u003csub\\u003e2\\u003c/sub\\u003ePO\\u003csub\\u003e4\\u003c/sub\\u003e)\\u003csub\\u003e2\\u003c/sub\\u003e]. The nutrient concentration used aims to replicate the practices recommended for fertilizer use agricultural crops farms. The application of 60 kg.ha\\u003csup\\u003e\\u0026minus;\\u0026thinsp;1\\u003c/sup\\u003eof N and 40 kg.ha\\u003csup\\u003e\\u0026minus;\\u0026thinsp;1\\u003c/sup\\u003eof P corresponds to a recommended level for some of the most common crop species in the study region (i.e., common bean, see Ramos et al., \\u003cspan citationid=\\\"CR37\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e), and 130 kg.ha\\u003csup\\u003e\\u0026minus;\\u0026thinsp;1\\u003c/sup\\u003e of N corresponds to an excessive level of N, but is frequently applied by farmers in Cerrado environments (Ramos et al., \\u003cspan citationid=\\\"CR37\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e). The amount of fertilizer (N and P) applied in each pot was calculated based on their concentration in the applied elemental form (45% of N in Urea and 21% of P in simple superphosphate for P). The nutrients were applied directly to the soil in each pot approximately every three months since November 2018. \\u003cp\\u003ePlant vegetative metrics\\u003c/b\\u003e\\u003c/p\\u003e\\u003cp\\u003eSurveys to extract information on plant metrics were done in May, August 2019 and October 2020. For each individual plant, at each sampling event, we recorded the number of leaves per tree. We used the number of leaves as a proxy for plant biomass increase in response to soil fertilization (Throop \\u0026amp; Lerdau, \\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e2004\\u003c/span\\u003e), which serves as a more direct indicator of food availability for the herbivores studied here.\\u003c/p\\u003e\\u003cp\\u003eWhen the plant was introduced in the experiment, we collected information on their height (stem length, cm). Plant height was used in the analyses of number of leaves to control for variations in size due to differences in age and planting times.\\u003c/p\\u003e\\n\\u003ch3\\u003eHerbivores and predators\\u003c/h3\\u003e\\n\\u003cp\\u003eHerbivory metrics considered in this study were: leaf herbivory by external large leaf-feeding invertebrate herbivores (e.g., ants, caterpillars, beetles and grasshoppers, no mammal herbivores were observed in the study areas) and density of phytophagous mites. In each sampling event, leaf herbivory was measured as the percentage of plant leaves showing signs of leaf tissue consumption by herbivores. If a leaf or leaflet was completely missing, this information was not counted as herbivory. Then, for each individual plant that had at least 12 leaves, we collected three leaves. If an individual had fewer than 12 leaves, the collection was adjusted so that we would never collect more than 25% of the total number of leaves of every individual.\\u003c/p\\u003e\\u003cp\\u003eTo avoid choosing which leaves to collect, we decided a priori to collect the 4th, 5th and 6th leaves, counting from the apex. For species with composed leaves with large leaflets (i.e., \\u003cem\\u003eI. vera\\u003c/em\\u003e), leaflets were collected. Leaves were stored in vials with ethanol 70%. Then, under laboratorial conditions, all the collected leaves were washed by shaking vigorously the recipient for 30 seconds to release all mites. Samples were then analyzed under stereoscopic microscope and all mites found were mounted on slides with Hoyer's medium (Moraes \\u0026amp; Flechtmann, \\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e2008\\u003c/span\\u003e). The mites were then quantified and identified to the lowest possible taxonomic level (Moraes and Flechtmann, \\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e2008\\u003c/span\\u003e) and separated into morphogroups under phase contrast microscopy. Slides with mites were deposited in the collection of Acari of the Laboratory of Taxonomy, Ecology and Interaction of Arachnids (TEIA) at the Federal University of Goi\\u0026aacute;s (UFG). The mite community was classified into three groups based on the predominant feeding habits of each group: 1) phytophagous, 2) predators and 3) undetermined/unknown (see Table S3). In the latter, we placed mites that were not identified to species level (e.g., immature or damaged individuals) and belong to families harboring species with different guilds. Groups that do not have species that feed on plants (Oribatida and Winterschimidtiida) like mycophagous species (which made up only a very small proportion of the mites collected) were excluded from analysis.\\u003c/p\\u003e\\u003cdiv id=\\\"Sec6\\\" class=\\\"Section2\\\"\\u003e\\u003ch2\\u003eData analysis\\u003c/h2\\u003e\\u003cp\\u003eTo test the effects of different fertilization treatments on number of leaves, leaf herbivory, phytophagous mite density, and predatory mite occurrence (presence/absence) for each plant species we used generalized linear mixed models (GLMMs). We used N and P levels as fixed categorical variables, also considering the interaction between the two nutrients. As some plant individuals were included after November 2018 (to compensate for high mortality, see above) the number of fertilization events that each plant received varied. Therefore, \\u0026lsquo;number of fertilization events\\u0026rsquo; to which the plant was submitted was included as a covariate. To control for differences in plant size at the start of the experiment, in the model of plant number of leaves we included initial height (i.e., height in the first fertilization event to which the plant was submitted) as covariate. When analyzing herbivory (leaf herbivory (%) and phytophagous mite density) and predation, to disentangle between effects of resource abundance from resource quality, we included \\u0026lsquo;number of leaves\\u0026rsquo; as a covariate in the models. A significant effect of N or P over and above the effect of \\u0026lsquo;number of leaves\\u0026rsquo; is then an indication of an effect driven by changes in quality of plant resources. As we had multiple measurements per plant individual, to account for the spatial and temporal structure of the data, we included \\u0026ldquo;sampling date\\u0026rdquo; and \\u0026ldquo;plant identity nested within study site\\u0026rdquo; as random effects. This structure controls for repeated measurements on the same individual over time, as well as potential site-level variation. For each metric, we also attempted to run a single GLMM for all species, with plant species and its interaction with P and N as terms in the model. Yet, our statistical power was not sufficient for such large model.\\u003c/p\\u003e\\u003cp\\u003eFor number of leaves, we assumed a negative binomial error structure (and log-link function). To analyze leaf herbivory (%) and abundance of phytophagous mites we used Tweedie error structure (and log-link function). For phytophagous mites analyses we considered only the individuals classified as phytophagous, but as a sensitivity test, we re-run the analyses also considering the mites with undetermined diet (e.g., juveniles, mites without full taxonomic ID from families with mixed feeding strategies). A binomial model was used to analyze the probability of occurrence of predatory mites. For each plant species, \\u003cem\\u003ea posteriori\\u003c/em\\u003e analyses were performed to compare results under different combinations of P and N, using the \\\"glht\\\" function version 1.4\\u0026ndash;25 from the \\u0026lsquo;multcomp\\u0026rsquo; package (Hothorn et al., \\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e2008\\u003c/span\\u003e). All analyses were performed using \\u0026ldquo;glmmTMB\\u0026rdquo; v.1.1.4 (Brooks et al., \\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e), in the R program version 4.2.1 (R Core Team, \\u003cspan citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e2023\\u003c/span\\u003e).\\u003c/p\\u003e\\u003c/div\\u003e\"},{\"header\":\"Results\",\"content\":\"\\u003cdiv id=\\\"Sec8\\\" class=\\\"Section2\\\"\\u003e\\u003ch2\\u003eEffects of fertilization on plant vegetative metrics\\u003c/h2\\u003e\\u003cp\\u003eOverall, N had a positive effect on the number of leaves, mean values being higher at the intermediate and/or highest N dosages for most species (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e). Yet, some species were more affected than others (significant for \\u003cem\\u003eS. terebinthifolia\\u003c/em\\u003e, \\u003cem\\u003eC. aeugenioides\\u003c/em\\u003e, \\u003cem\\u003eP. guineense\\u003c/em\\u003e and \\u003cem\\u003eE. involucrata\\u003c/em\\u003e, Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e; Table S4), and no difference was detected between intermediate and highest levels of N (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e). No statistically significant influence of P levels on the effect of N was detected for most species (i.e., interaction term had no significant effect), but increases on P levels significantly increased the number of leaves of \\u003cem\\u003eS. terebinthifolia\\u003c/em\\u003e and \\u003cem\\u003eP. guineense\\u003c/em\\u003e (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eb, e; Table S4).\\u003c/p\\u003e\\u003cp\\u003e\\u003c/p\\u003e\\u003c/div\\u003e\\n\\u003ch3\\u003eEffects of fertilization on herbivores and higher trophic levels\\u003c/h3\\u003e\\n\\u003cp\\u003eEffects of fertilization treatments differed between herbivore types. For large foliar herbivores (mostly ants, caterpillars and beetles), the \\u003cem\\u003eS. lycocarpum\\u003c/em\\u003e showed the highest proportion of leaves with signs of herbivory (mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;sd: 38\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;36%), the addition of N led to a reduction in herbivory in intermediate N level, while the presence of P increased herbivory in highest N level (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003ec). \\u003cem\\u003eI. vera\\u003c/em\\u003e also had high levels of herbivory (31\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;25% of the leaves affected), but no clear effect of fertilization treatment was detected (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003ea; Table S4). The other species showed less than 15% of leaves with signs of herbivory [\\u003cem\\u003eC. eugenioides\\u003c/em\\u003e (13\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;21%), \\u003cem\\u003eP. guianeense\\u003c/em\\u003e (9\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;8%), \\u003cem\\u003eE. involucrata\\u003c/em\\u003e (8\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;17%), \\u003cem\\u003eS. terebinthifolia\\u003c/em\\u003e (6\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;10%)]. Of these, only \\u003cem\\u003eP. guianeense\\u003c/em\\u003e showed effects of fertilization; no effects were detected in the other species. Yet, contrary to what was detected for \\u003cem\\u003eS. lycocarpum\\u003c/em\\u003e, N addition reduced herbivory, this effect being significant when N was added in high dosages and combined with P (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003ee).\\u003c/p\\u003e\\u003cp\\u003e\\u003c/p\\u003e\\u003cp\\u003eAs for mites, although their density was generally low (Table S6), effects of fertilizers were more frequently detected than in large invertebrate herbivores (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e, Table S4). \\u003cem\\u003eI. vera\\u003c/em\\u003e was the species with the highest density of phytophagous mites (79% of all mites detected were found in this species), followed by \\u003cem\\u003eS. terebinthifolia\\u003c/em\\u003e (12%), and the three species of Myrtaceae showing the lowest densities.\\u003c/p\\u003e\\u003cp\\u003e\\u003c/p\\u003e\\u003cp\\u003eIn total, we collected 11 976 mites belonging to 35 morphogroups in 16 families (Table S3). The phytophagous mites were the most abundant group, making up 86% of mites collected. Most (73%) mites belonged to the Tenuipalpidae family, followed by the Tetranychidae family (8%), both families of phytophagous mites, and by the predatory mite family Phytoseiidae (4%).\\u003c/p\\u003e\\u003cp\\u003eThe effect of N input on the density of phytophagous mites was dependent on the levels of P, positively for \\u003cem\\u003eS. terebinthifolia\\u003c/em\\u003e and \\u003cem\\u003eC. aeugenioides\\u003c/em\\u003e (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003eb; d, Table S5). For \\u003cem\\u003eP. guineense\\u003c/em\\u003e, the high levels of N increased mite density only when P was not added, with the effect turning negative when P was introduced (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003ee, Table S5). For \\u003cem\\u003eI. vera\\u003c/em\\u003e, the addition of P increased mite density, but its interaction with N reduced this effect (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003ea, Table S5). For \\u003cem\\u003eS. lycocarpum\\u003c/em\\u003e N addition in the presence of P was also negative to phytophagous mites density (Fig. S2 e, Table S5).\\u003c/p\\u003e\\u003cp\\u003eMost predatory mites were detected in \\u003cem\\u003eI. vera\\u003c/em\\u003e (42% of the total number of predatory mites), \\u003cem\\u003eS. lycocarpum\\u003c/em\\u003e (26%), \\u003cem\\u003eS. terebinthifolia\\u003c/em\\u003e (19%), \\u003cem\\u003eC. eugenioides\\u003c/em\\u003e (9%), being less frequent in \\u003cem\\u003eE. involucrata\\u003c/em\\u003e (2%) and \\u003cem\\u003eP. guineense\\u003c/em\\u003e (1%). As for the proportion of mites that were predatory (in relation to the total number of mites) \\u003cem\\u003eE. involucrata\\u003c/em\\u003e (41% of their total number of mites), \\u003cem\\u003eS. lycocarpum\\u003c/em\\u003e (39%) and \\u003cem\\u003eC. eugenioides\\u003c/em\\u003e (27%) were the plant species with higher values, predatory mites being less frequent in \\u003cem\\u003eP. guineense\\u003c/em\\u003e (12%), \\u003cem\\u003eS. terebinthifolia\\u003c/em\\u003e (12%) and \\u003cem\\u003eI. vera\\u003c/em\\u003e (4%). Due to the low number of mites in \\u003cem\\u003eP. guineense\\u003c/em\\u003e, and \\u003cem\\u003eE. involucrata\\u003c/em\\u003e these two species were not used to analyze the effect of fertilization on predatory mites. Among the remaining four plant species, \\u003cem\\u003eI. vera\\u003c/em\\u003e, with the presence of P, the addition of intermediate N level also resulted in an increase in the occurrence of predatory mites (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003ea; Table S5).\\u003c/p\\u003e\\u003cp\\u003e\\u003c/p\\u003e\"},{\"header\":\"Discussion\",\"content\":\"\\u003cp\\u003eOur results show that the effects of fertilization on Cerrado tree species and their interactions with herbivores depend greatly on the identity of the plant species considered. Below we discuss possible mechanisms that regulate the interspecific variability of the effects detected here and the potential consequences for the conservation and restoration of Cerrado biodiversity.\\u003c/p\\u003e\\u003cdiv id=\\\"Sec11\\\" class=\\\"Section2\\\"\\u003e\\u003ch2\\u003eEffects of fertilization on plant vegetative metrics\\u003c/h2\\u003e\\u003cp\\u003eWhile we detected an overall positive trend of N input in leaf count, the variations in response intensity among plant species indicate that, under a scenario of soil nutrient enrichment in Cerrado, certain native plant species may have a competitive edge, and may increase their dominance, potentially driving others to local extinction. It is expected that negative impacts of nutrient addition would be greater for nitrophobous plants and positive impacts more accentuated for nitrophilous plants (David et al., \\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e2019\\u003c/span\\u003e). For the species used in this study such classifications do not yet exist, but based on our results at least the two species of Myrtaceae for which positive impacts were more accentuated (\\u003cem\\u003eC. euginioides\\u003c/em\\u003e and \\u003cem\\u003eP. guineense\\u003c/em\\u003e) are likely to be nitrophilous. Other studies have shown that forest tree species of the Myrtaceae family exhibit a strong positive response to N with increasing survival (Nussbaumer et al., \\u003cspan citationid=\\\"CR32\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e) and biomass (Wooliver et al., \\u003cspan citationid=\\\"CR50\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e). Indeed, although the two Myrtaceae used in our study naturally in biomes known for having very nutrient-poor soils (i.e. Cerrado and Caatinga, see Table S2), they have very wide distributions within Brazil occurring in regions with naturally richer soils (e.g. Atlantic Forest, Table S2) and, for \\u003cem\\u003eP. guineense\\u003c/em\\u003e, occurring also as exotic species in other continents (e.g. Asia and Africa, Senanayake et al. \\u003cspan citationid=\\\"CR42\\\" class=\\\"CitationRef\\\"\\u003e2018\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003eThe results here presented may have implications for choices related to restoration programs. Plant responses to fertilization are closely related to their specific differences in nutritional requirement, such as co-limitation by different nutrients and differences in nutrient uptake and economy strategies, resulting in different intensities or even directions of responses (Kozovits et al., \\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e2007\\u003c/span\\u003e; Wooliver et al., \\u003cspan citationid=\\\"CR50\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e). Yet, it is possible that such a beneficial effect only occurs when plants are protected from any competition effects with other species, as they were in our experimental setup. Further studies involving combinations of plants cooccurring and competing for soil resources would be needed to verify if such beneficial effect occurs in nature. Nevertheless, As \\u003cem\\u003eP. guineense\\u003c/em\\u003e is a fast-growing species and \\u003cem\\u003eC. euginioides\\u003c/em\\u003e is a slow-growing species (Table S2), in natural conditions where plants compete for space and resources, we can assume that the first has a higher risk of increasing dominance over other native species under a scenario of soil nutrient enrichment. For the slow-growing Myrtaceae species which appear later in the ecological succession (\\u003cem\\u003eC. eugenioide\\u003c/em\\u003es and \\u003cem\\u003eE. involucrata\\u003c/em\\u003e) generally have a more conservative resource use strategy, being less able to respond to changes in nitrogen availability (Aidar et al., \\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2003\\u003c/span\\u003e; Báez and Homeier, 2017). Therefore, \\u003cem\\u003eP. guineense\\u003c/em\\u003e is a species that may be better to avoid in restoration programs where soil is eutrophicated (e.g. due to nutrient leaching from farms).\\u003c/p\\u003e\\u003cp\\u003eThe lack of a significant increase in the number of leaves in \\u003cem\\u003eI. vera\\u003c/em\\u003e, despite increased soil nutrient levels, may reflect its ability to regulate nitrogen uptake through symbiotic N₂ fixation with rhizobia, maintaining nitrogen balance even in nutrient-poor soils (see Table S2). Previous research suggests that increased N fertilization can lead to a reduction in biological N fixation (e.g., Weber et al., \\u003cspan citationid=\\\"CR48\\\" class=\\\"CitationRef\\\"\\u003e2007\\u003c/span\\u003e and Xia et al., \\u003cspan citationid=\\\"CR52\\\" class=\\\"CitationRef\\\"\\u003e2017\\u003c/span\\u003e), which may explain the limited vegetative response observed in this species under higher N availability.\\u003c/p\\u003e\\u003cp\\u003eContrary to what we expected, we found no significant effect of P on the plant number of leaves of most plant species. Only \\u003cem\\u003eS. terebinthifolia\\u003c/em\\u003e and \\u003cem\\u003eP. guineense\\u003c/em\\u003e showed a significant response to increases in N and P in a non-additive manner. Other studies did not find effects of P and its interaction with N, which may be linked to factors ranging from low availability for use in the soil (Bucci et al., \\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e2006\\u003c/span\\u003e) to the presence of P conservation mechanisms for some plant species adapted to dystrophic environments (Kozovits et al., \\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e2007\\u003c/span\\u003e; Abrahão et al., 2018). Yet, it is important to note that we found an effect of P on other trophic levels. Thus, P was likely allocated to functions other than biomass production, potentially enhancing leaf nutritional quality by increasing tissue P concentration without affecting leaf quantity. Indeed, nutrient enrichment can alter several tree characteristics in addition to aerial biomass, such as root biomass, concentrations of photosynthetic pigments, tissue nitrogen, concentrations of proteins carbohydrates and secondary metabolites (Li et al., \\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e) which in turn can affect interactions with higher trophic levels.\\u003c/p\\u003e\\u003c/div\\u003e\\u003cdiv id=\\\"Sec12\\\" class=\\\"Section2\\\"\\u003e\\u003ch2\\u003eEffects of fertilization on herbivores and higher trophic levels\\u003c/h2\\u003e\\u003cp\\u003eGiven the great variability of effects of fertilization on plant growth, variability on herbivore responses to fertilization was expected. While previous studies on tropical regions show that, increasing soil N availability (alone or in combination with P) has negative impacts on the abundance of terrestrial tropical invertebrates (Nessel et al., \\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; 2022), other studies in temperate ecosystems demonstrated that the effects of soil fertilization on herbivores can be positive, negative, or neutral (Kyto et al., \\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e1996\\u003c/span\\u003e; Butler et al., \\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e; La Pierre and Smith, \\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e2016\\u003c/span\\u003e; Nessel et al., \\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e;2022), depending on the host plant species and herbivorous group considered, as observed in our study. The species \\u003cem\\u003eI. vera\\u003c/em\\u003e showed the highest density of mites and a high leaf herbivory regardless of the nutrient treatment, while the other species showed intermediate to high resistance to herbivory. This reflected in differences between levels of herbivory in response to soil nutrient enrichment for each plant studied, making it difficult to detect general response patterns.\\u003c/p\\u003e\\u003cp\\u003eAs expected, the effects of nutrient supply were more pronounced for phytophagous mites than for plant growth. In general, we found a positive effect for plants. Other studies also point out that the effects can be magnified by trophic cascades (e.g., Carvalheiro et al., \\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003eThe fact that the effects of nutrients were more accentuated for mites than for herbivory resulting from large invertebrate could be related to different levels of diet specialization. Sucking herbivores, such as phytophagous mites, showed a much stronger response to fertilizers than chewing insects, such as large external leaf herbivores (Butler et al., \\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e). Sucking herbivores tend to be more specialized feeding on a single species or phylogenetically close plant species, while chewing herbivores can be more generalist (Ali and Agrawal, \\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e), and magnification of impacts through trophic chains or webs is mainly expected for species with more specialized diets (Carvalheiro et al., \\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e). Differences in functional and behavioral traits (e.g., eating habit, mobility) could also influence the responses of herbivores to plant changes mediated by fertilizers (Kyto et al., \\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e1996\\u003c/span\\u003e; Butler et al., \\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e; Nessel et al., \\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003eAs in other studies (Butler et al., \\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e; Nessel et al., \\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; 2022), here it was difficult to detect a consistent effect of phosphorus and its interaction with N on herbivores and predators. Although the literature describes an overall negative effect (Butler et al., \\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e; Nessel et al., \\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e; 2022), some studies indicate positive effects of increasing P availability on herbivores (Butler et al., \\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e). However, the effect of increasing P, especially in isolation, still needs to be further investigated.\\u003c/p\\u003e\\u003c/div\\u003e\"},{\"header\":\"Concluding remarks\",\"content\":\"\\u003cp\\u003eEnvironmental eutrophication is one of the major human-driven changes affecting biodiversity (Bobbink et al., \\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e2010\\u003c/span\\u003e; Steffen et al., \\u003cspan citationid=\\\"CR41\\\" class=\\\"CitationRef\\\"\\u003e2015\\u003c/span\\u003e). Due to its intensive use for agriculture, Cerrado is highly exposed to such effect (Bustamante et al., \\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e2012\\u003c/span\\u003e). Our results clearly show that altered soil conditions will favor some plant species more than others and will have consequences for higher trophic levels. These findings highlight the urge for long term monitoring schemes to evaluate how ongoing soil nutrient enrichment is changing the dynamics between plant trees that naturally occur in Cerrado and their ecological interactions. These findings also have implications for restoration programs in Cerrado. Soil properties in areas that are subjected to restoration actions typically have highly degraded soils (Prescott et al., \\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e2021\\u003c/span\\u003e), potentially being poorer in nutrients than normal soils or potentially being enriched in certain nutrients that are widely applied in crop fields (nitrogen and phosphorus). Future studies with a larger number of species that explore which plant traits explain their response to nutrient level changes are essential to identify how different plant species will perform when used in a restoration plan. Overall, the results of this work contribute to a better understanding of how different plant species, valuable for restoration, respond when planted in soils with different nutrient conditions.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eAuthors contributions\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eCFT and LGC\\u003csup\\u003e\\u0026nbsp;\\u003c/sup\\u003econceived the project and designed methodology with help of CMCS and RDD. CFT collected and analyzed the data with help of LGC; CFT wrote the first draft; RDD, VLF, JAR\\u003csup\\u003e\\u0026nbsp;\\u003c/sup\\u003eand SRC identified the mite community. CFT wrote de first draft of the manuscript with help of LGC and SC. All authors revised the manuscript.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAcknowledgements\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eWe thank F Venturoli, GM de Oliveira, AR Nascimento, CASB Braga, MCS Fioravanti, GA Ferreira for the spaces provided for setting up the experiment. We thank MBE Bernardes, PM Consorte and members of the Community Ecology laboratory at UFG for their help in setting up the experiment, and in collecting and sorting the data.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eFunding\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eCFT was funded by CAPES (88882.347112/2019-01; 88887.800299/2022-00). LGC was funded by CNPq (307625/2021-4).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConflict of Interest Statement\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eWe declare there is no conflict of interest.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eEthics approval\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConsent to participate\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConsent for publication\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAvailability of data and material\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eCode availability\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eNot applicable\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n\\u003cli\\u003eAbrah\\u0026atilde;o, A., Costa, P. de B., Lambers, H., Andrade, S. 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Effects of nitrogen concentrations on nodulation and nitrogenase activity in dual root systems of soybean plants. \\u003cem\\u003eSoil Science and Plant Nutrition, 63(5), 470\\u0026ndash;482\\u003c/em\\u003e. https://doi.org/10.1080/00380768.2017.1370960\\u003c/li\\u003e\\n\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":true,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"oecologia\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"oeco\",\"sideBox\":\"Learn more about [Oecologia](https://www.springer.com/journal/442)\",\"snPcode\":\"442\",\"submissionUrl\":\"https://submission.nature.com/new-submission/442/3\",\"title\":\"Oecologia\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false},\"keywords\":\"plant-herbivore interaction, phytophagous mites, fertilization effects, soil nutrient enrichment, trees and shrubs\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-6968991/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-6968991/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eHuman-induced changes in nitrogen (N) and phosphorus (P) global cycles, significantly impact plant growth and nutritional composition, thereby affecting ecosystem dynamics. However, research on the effects of increased nutrient availability often focuses on plant community-level effects, overlooking interspecific variability and neglecting impacts on higher trophic levels. Using a controlled fertilization experiment with six tree species that naturally occur in the Cerrado biome (Brazilian savannas) we showed that there is significant interspecific variation in how plants responded to nutrient changes, indicating competitive advantages for certain native species under a scenario of increased soil nutrient availability. Such effects propagated to higher trophic levels (herbivores and their predators), also varying between plant host species. The strength and direction of N input effect depended on P levels and the type of herbivores. Large invertebrate leaf-herbivores were less affected than phytophagous mites. Impacts on higher trophic levels (predatory mites) were less pronounced than on phytophagous mites. Overall, we show that ongoing soil nutrient enrichment has the potential to alter interspecific competition dynamics in plant communities with consequences for ecological interaction partners. These findings have important implications for conservation and ecosystem management, especially in areas highly exposed to soil nutrient enrichment due to farming and industrial activities.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Effects of soil nutrient enrichment on biomass, herbivores and their predators differ between tree species in the Brazilian Cerrado\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2025-08-12 15:09:39\",\"doi\":\"10.21203/rs.3.rs-6968991/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"reviewerAgreed\",\"content\":\"\",\"date\":\"2025-08-07T17:46:11+00:00\",\"index\":0,\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2025-08-07T08:19:11+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2025-06-25T04:05:45+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Oecologia\",\"date\":\"2025-06-24T17:51:07+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"oecologia\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"oeco\",\"sideBox\":\"Learn more about [Oecologia](https://www.springer.com/journal/442)\",\"snPcode\":\"442\",\"submissionUrl\":\"https://submission.nature.com/new-submission/442/3\",\"title\":\"Oecologia\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false}}],\"origin\":\"\",\"ownerIdentity\":\"913bd931-12ca-4c2b-afbb-94fd3bd6ba17\",\"owner\":[],\"postedDate\":\"August 12th, 2025\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"published-in-journal\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2026-02-09T16:01:27+00:00\",\"versionOfRecord\":{\"articleIdentity\":\"rs-6968991\",\"link\":\"https://doi.org/10.1007/s00442-026-05863-z\",\"journal\":{\"identity\":\"oecologia\",\"isVorOnly\":false,\"title\":\"Oecologia\"},\"publishedOn\":\"2026-02-07 15:57:17\",\"publishedOnDateReadable\":\"February 7th, 2026\"},\"versionCreatedAt\":\"2025-08-12 15:09:39\",\"video\":\"\",\"vorDoi\":\"10.1007/s00442-026-05863-z\",\"vorDoiUrl\":\"https://doi.org/10.1007/s00442-026-05863-z\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-6968991\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-6968991\",\"identity\":\"rs-6968991\",\"version\":[\"v1\"]},\"buildId\":\"XKTyCvWXoU3ODBz1xrDgd\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}