Bacterial Community Diversity in the Brazilian Atlantic Forest Soils

The aim of this study was to characterize the bacterial community diversity of the Brazilian Atlantic forest soil by means of both cultivation and 16S rRNA clone libraries. A collection of 86 representative isolates, obtained from six samples of Atlantic forest soils from the National Park of Serra dos Órgãos (PARNASO), belonged to the genera Arthrobacter, Bacillus, Burkholderia, Leifsonia, Paenibacillus, Pseudomonas, Ralstonia, Serratia, and Streptomyces according to the 16S rRNA sequences. Representative isolates from the different genera degraded cellulose and lignin. The culture-independent analysis based on 894 partial 16S rRNA gene sequences revealed that the most frequently retrieved groups belonged to the phyla Acidobacteria (29–54%), Proteobacteria (16–38%), and Verrucomicrobia (0.6–14%). The majority of the sequences (82.6%) were unidentified singletons and doubletons, indicating a high diversity of rare unique sequences. Chao1 estimator disclosed a high number of phyla (41–152) and species (263–446). This is the first survey on the Atlantic Forest soils using a combination of cultivation and culture-independent approaches. We conclude that the Brazilian Atlantic Forest soil represents a vast source of novel bacteria.     

Diversity of Tiphiidae (Insecta: Hymenoptera) in the fragmented Brazilian semi-deciduous Atlantic Forest

The Atlantic Forest is one of the most important areas of biodiversity in the world, but it has been largely replaced with agropastoral areas and at the present only 12.5 % of the original cover remains. Despite the ecological importance of insects, few studies have been used in conservation approaches for the Atlantic Forest, mainly due to a great taxonomic impediment. A group quite ecologically important but deeply neglected includes parasitoid wasps that control a great number of invertebrates, like tiphiid wasps that are parasitoids of underground coleopteran larvae. The present study aimed to estimate Tiphiidae species richness and diversity in 15 patches of a highly fragmented Atlantic Forest region, using factors that drive the diversity pool from a metacommunity, such as immigration and speciation probabilities. The parameters were estimated using the Neutral Biodiversity Theory, which is based on the total ecological equivalence of species at the same trophic level. Diversity values were molded to the area size, the immigration probabilities, and/or the speciation probability. Eight genera and 460 individuals of Thynninae, Myzininae and Tiphiinae were collected. Variation in species richness, estimated by both rarefaction and first-order jackknife methods, was explained by patch size and by immigration and speciation probabilities. These variables also explained the variation in Shannon diversity and species evenness. Variations in species richness and diversity of Tiphiidae are strongly associated with neutral processes, but they are also influenced by forest fragmentation and intensive agricultural activities.     

Amphibian Chytrid Fungus Broadly Distributed in the Brazilian Atlantic Rain Forest

To investigate the occurrence of the chytrid fungus Batrachochytrium dendrobatidis in Brazil, we conducted histological screenings of 96 preserved specimens of anurans collected at 10 sites in the Atlantic rain forest. Data show this fungus to be widely distributed. Infected specimens included Colostethus olfersioides (Dendrobatidae), Bokermannohyla gouveai and Hypsiboas freicanecae (Hylidae), as well as Thoropa miliaris and Crossodactylus caramaschii (Leptodactylidae), extending the area of B. dendrobatidis occurrence in Brazil approximately 1,600 km N, 200 km S, and 270 km E. The altitudinal range of the chytrid is broad, spanning from less than 100 m (Estação Ecológica Juréia-Itatins, Reserva Biológica do Tinguá) to about 2,400 m (Parque Nacional do Itatiaia). An infection record dating to 1981 roughly coincides with the time of the first observations of amphibian declines in the country. Widespread occurrence of B. dendrobatidis in the Atlantic Forest adds to the challenge of conserving an already endangered biome given the potential risk of further local biodiversity loss. Further research is needed to understand how environmental and genetic factors relate to chytridiomycosis in leading to or preventing local die-offs. Protected sites at mid and high elevations may be particularly threatened, while lowland populations may be functioning as reservoirs. Conservation efforts should also involve monitoring studies and habitat protection.     

Three new cecidogenous species of Palaeomystella Fletcher (Lepidoptera,Momphidae) from the Brazilian Atlantic Rain Forest

Three new cecidogenous species of Palaeomystella Fletcher (Lepidoptera, Momphidae) from the Brazilian Atlantic Rain Forest are described. Larvae of P. fernandesi Moreira & Becker, sp. n., P. rosaemariae Moreira & Becker, sp. n. and P. tavaresi Becker & Moreira, sp. n. induce galls, respectively, on Tibouchina sellowiana (Cham.) Cogn., T. asperior (Cham.) Cogn. and T. fissinervia (Schrank & Mart. ex DC.) Cogn. (Melastomataceae). Adults, immature stages and galls are illustrated, and data on life history and a preliminary analysis of mitochondrial DNA sequences, including related species, are also provided.     

A new arborescent Euphorbia (Euphorbiaceae) from northern Somalia

Euphorbia spissa, sp. nov., a tree known from a single population on a rocky limestone slope in northern Somalia, is described and illustrated. The species is compared with the widespread E. tirucalli, as well as with E. cameronii and E. bariensis (endemics in Somalia), E. arbuscula (endemic on Socotra), and E. uzmuk (endemic in northern Yemen).     

Long-term stability of the hawkmoth fauna (Lepidoptera,Sphingidae) in a protected area of Brazilian Atlantic Rain Forest

Hawkmoths are an important component of tropical ecosystems, with significant roles as herbivores and pollinators. These moths can be used as indicators in biodiversity assessments because they can be easily sampled and identified. However, hawkmoths have seldom been surveyed over the long term for this purpose, especially in the Neotropical region. Considering that long-term datasets are of indisputable importance for understanding and monitoring temporal changes in biodiversity, this study assessed long-term changes in the hawkmoth fauna in a protected Atlantic Rain Forest area over a period of 64 years. We used historical and recent empirical datasets to ask whether faunal-diversity patterns and species composition have changed over time. We used individual- and sample-based rarefaction and extrapolation curves based on Hill number (diversity order of q = 0) to compare species richness, and the probability version of the abundance-based Chao-Jaccard index to assess beta diversity over time. To assess changes in faunal composition, we conducted a nonmetric multidimensional scaling analysis and performed an analysis of similarities to test whether the community composition has changed. Our results clearly showed long-term stability of the hawkmoth community over the 64 years, despite the growing human-induced landscape changes that occurred in the region surrounding the study area during the last 6 decades. This study emphasizes the importance of large remnants of Atlantic Forest for long-term maintenance of both functional diversity and ecosystem functioning.     

Edge-Related Loss of Tree Phylogenetic Diversity in the Severely Fragmented Brazilian Atlantic Forest

Deforestation and forest fragmentation are known major causes of nonrandom extinction, but there is no information about their impact on the phylogenetic diversity of the remaining species assemblages. Using a large vegetation dataset from an old hyper-fragmented landscape in the Brazilian Atlantic rainforest we assess whether the local extirpation of tree species and functional impoverishment of tree assemblages reduce the phylogenetic diversity of the remaining tree assemblages. We detected a significant loss of tree phylogenetic diversity in forest edges, but not in core areas of small (<80 ha) forest fragments. This was attributed to a reduction of 11% in the average phylogenetic distance between any two randomly chosen individuals from forest edges; an increase of 17% in the average phylogenetic distance to closest non-conspecific relative for each individual in forest edges; and to the potential manifestation of late edge effects in the core areas of small forest remnants. We found no evidence supporting fragmentation-induced phylogenetic clustering or evenness. This could be explained by the low phylogenetic conservatism of key life-history traits corresponding to vulnerable species. Edge effects must be reduced to effectively protect tree phylogenetic diversity in the severely fragmented Brazilian Atlantic forest.     

Cave lithology determining the structure of the invertebrate communities in the Brazilian Atlantic Rain Forest

In Brazil, only limestone caves and a few caves in sandstone, iron ore and granite rocks had their invertebrate communities evaluated. Being such, the present study aimed to promote a comparative analysis of the structure of the invertebrate communities in caves associated to carbonatic, magmatic, siliciclastic and ferruginous rocks of the Brazilian Atlantic forest. Significant differences in the relative richness, abundance and diversity were observed between lithologies. The average relative richness was higher in the ferruginous caves (0.53 spp). The total number of troglomorphic species was significantly different among caves and the highest average richness occurred at ferruginous caves (5.79 spp/cave). Siliciclastic, carbonatic and magmatic caves presented a higher quantitative similarity of the fauna. Ferruginous caves revealed communities with a fauna composition different from the other lithologies. The total richness of invertebrates correlated significantly and positively with the linear development in the siliciclastic caves (R_s = 0.67, P < 0.05), carbonatic (R_s = 0.71, P < 0.05) and ferruginous (R_s = 0.74, P < 0.05). The rock type in which the cave is inserted can determine differences in the richness of invertebrate troglophyles and troglobites. Therefore, on creating value attributes, the size of the caves should always come related to their lithology by the fact that same sized caves associated to different lithologies, possess communities with quite diverse structures.     

Trypanocidal activity of extracts from Brazilian Atlantic Rain Forest plant species.

The trypanocidal activity of crude hydro alcoholic extracts and several fractions of 13 plants from Brazilian Atlantic Rain Forest were tested in vitro against epimastigote and trypomastigote forms of Trypanosoma cruzi, the etiological agent of Chagas disease. Crude ethanol extracts with promising in vitro activity (DL50 between 5-10 microg/ml) against epimastigotes were fractionated by solvent partition and further tested against bloodstream form of the parasite. Activity against bloodstream parasites was observed in both dichloromethane and hexane fractions of Polygala sabulosa and P. paniculata.     

First records of Synoeca septentrionalis Richards, 1978 (Hymenoptera, Vespidae, Epiponini) in the Brazilian Atlantic Rain Forest

Nests of Synoeca septentrionalis were collected in two Brazilian Atlantic Rain Forest localities (Itabuna and Santa Terezinha, in the state of Bahia and Alfredo Chaves in the state of Espírito Santo). Synoeca septentrionalis was previously recorded only from Central America and northwestern South America. This findingextends its geographical distribution to Northeast and Southeast regions of Brazil, and represents the first record for Synoeca septentrionalis in the Brazilian Atlantic Rain forest, raising to three the number of Synoeca species known from Bahia State.     

Influence of Soil Characteristics on the Diversity of Bacteria in the Southern Brazilian Atlantic Forest

The Brazilian Atlantic Forest is one of the 25 biodiversity hot spots in the world. Although the diversity of its fauna and flora has been studied fairly well, little is known of its microbial communities. In this work, we analyzed the Atlantic Forest ecosystem to determine its bacterial biodiversity, using 16S rRNA gene sequencing, and correlated changes in deduced taxonomic profiles with the physicochemical characteristics of the soil. DNAs were purified from soil samples, and the 16S rRNA gene was amplified to construct libraries. Comparison of 754 independent 16S rRNA gene sequences from 10 soil samples collected along a transect in an altitude gradient showed the prevalence of Acidobacteria (63%), followed by Proteobacteria (25.2%), Gemmatimonadetes (1.6%), Actinobacteria (1.2%), Bacteroidetes (1%), Chloroflexi (0.66%), Nitrospira (0.4%), Planctomycetes (0.4%), Firmicutes (0.26%), and OP10 (0.13%). Forty-eight sequences (6.5%) represented unidentified bacteria. The Shannon diversity indices of the samples varied from 4.12 to 3.57, indicating that the soils have a high level of diversity. Statistical analysis showed that the bacterial diversity is influenced by factors such as altitude, Ca²⁺/Mg²⁺ ratio, and Al³⁺ and phosphorus content, which also affected the diversity within the same lineage. In the samples analyzed, pH had no significant impact on diversity.The Brazilian Atlantic Forest is one of the 25 biodiversity hot spots in the world. Altogether, these hot spots contain more than 60% of the total terrestrial species of the planet (17). The Atlantic Forest is a dense ombrophilous forest with several variations, including coastal (3 to 50 m), submontane (50 to 500 m), montane (500 to 1,200 m), and high montane (1,200 to 1,400 m) forests, creating a vegetation gradient ranging from shrubs to well-developed montane forest (4). The Serra do Mar is a mountainous system that shelters the main remainder of the Atlantic Forest following the Brazilian east coast, from north to south along the coastal line, and it is divided into diverse sections of high and low blocks, which have regional denominations.The most important law-protected conservation area of the Brazilian Atlantic Forest is located in the Serra do Mar of the southern state of Paraná. This conservation area (∼5,000 km²) shelters 72% of the fauna and flora species that occur in Paraná and was declared a Biosphere Reserve by UNESCO in 1992. Much is known about the diversity of its fauna and flora, but little is known of its microbial diversity, particularly the soil microbial diversity and the soil characteristics that influence it.The soil microbial diversity is vast, and it is estimated that >99% of species remain unidentified (1, 28). Acidobacteria and Proteobacteria are the most abundant groups in soil (15). However, the Proteobacteria lineage is more diverse and stable than the Acidobacteria lineage, suggesting that the latter group is more susceptible to variation in soil properties and to disturbing factors (33). Seasonal, physical, and physicochemical factors can be relevant to the structure and diversity of microbial communities. For example, seasonal changes in vegetation and temperature led to replacement of dominant groups in a wheat field (25) and in grassland soils (16). The particle size also has an influence on the bacterial diversity of soils. The clay fraction has a more diverse bacterial community than do silt or sand fractions (23). Finally, analyses of communities from North and South American soils showed that pH plays a major role in bacterial diversity, with less diverse communities associated with a lower pH (9).Human activity can also change the microbial diversity of soils, both qualitatively and quantitatively. Analyses of microbial communities on coral atolls in the central Pacific Ocean under different degrees of human impact showed that the least-impacted atoll had autotrophs and heterotrophs equally distributed in the community, whereas the most-impacted atoll had a dominance of heterotrophs and about 10 times more microbial cells and virus-like particles in the water column, including a large percentage of potential pathogens (7). A comparison between bacterial communities in forest and pasture soil showed that there is a less diverse and more restricted community in pasture soils. The vegetation shift from forest to pasture resulted in changes to G+C% contents of soil bacterial DNA and amplified rRNA gene restriction analysis (ARDRA) profiles (18). Similar changes occurred with communities of soils submitted to agroindustrial treatments and pollutants (3, 30).In this work, we used a culture-independent approach based on 16S rRNA gene sequences to survey the bacterial community of the Atlantic Forest soils and determined the physicochemical factors affecting its bacterial biodiversity.     

Fine litterfall in the Brazilian Atlantic Forest

Litterfall is an essential component of tropical forest productivity, transferring nutrients from the vegetation back to soils. Here, we summarize the data from 105 estimates of fine litterfall production from 45 sites in the Atlantic Forest domain, including two types of forests, evergreen and seasonal, and two successional stages, secondary and old growth. The overall litterfall average was 8.0 ± 2.5 Mg/ha. Litterfall was significantly in higher seasonal forests than in evergreen forests and in old growth versus secondary forests. Leaves were the major component of litterfall, contributing 68 percent to the total. The second most important component was branches, contributing 22 percent, followed by reproductive organs (flowers and fruits), at 6 percent. Accurate measurements of tropical forest productivity are crucial for estimating their role in sequestering atmospheric carbon, and we suggest some ways to standardize litterfall sampling to obtain better estimates.     

Skipper Richness (Hesperiidae) Along Elevational Gradients in Brazilian Atlantic Forest

Hesperiidae are claimed to be a group of elusive butterflies that need major effort for sampling, thus being frequently omitted from tropical butterfly surveys. As no studies have associated species richness patterns of butterflies with environmental gradients of high altitudes in Brazil, we surveyed Hesperiidae ensembles in Serra do Mar along elevational transects (900–1,800 m above sea level) on three mountains. Transects were sampled 11–12 times on each mountain to evaluate how local species richness is influenced by mountain region, vegetation type, and elevational zones. Patterns were also analyzed for the subfamilies, and after disregarding species that exhibit hilltopping behavior. Species richness was evaluated by the observed richness, Jacknife2 estimator and Chao 1 estimator standardized by sample coverage. Overall, 155 species were collected, but extrapolation algorithms suggest a regional richness of about 220 species. Species richness was far higher in forest than in early successional vegetation or grassland. Richness decreased with elevation, and was higher on Anhangava mountain compared with the two others. Patterns were similar between observed and extrapolated Jacknife2 richness, but vegetation type and mountain richness became altered using sample coverage standardization. Hilltopping species were more easily detected than species that do not show this behavior; however, their inclusion did neither affect estimated richness nor modify the shape of the species accumulation curve. This is the first contribution to systematically study highland butterflies in southern Brazil where all records above 1,200 m are altitudinal extensions of the known geographical ranges of skipper species in the region.     

Floristics and environmental factors determining the geographic distribution of epiphytic bromeliads in the Brazilian Atlantic Rain Forest

We investigated how epiphytic species and subfamilies of Bromeliaceae change along the extent of the Atlantic Rain Forest, to answer the questions: (i) How do the epiphytic genera and subfamilies of Bromeliaceae change along the domain? (ii) How similar are the different regions of the Atlantic Rain Forest in relation to the epiphytic species of bromeliads? (iii) Which environmental variables are the most important factors in determining species composition along the domain? We found 114 species of Bromelioideae and 73 of Tillandsioideae. The predominance of Bromelioideae was unexpected, because they are not wind-dispersed as would be expected for most epiphytes. The smaller number of species of Tillandsioideae, and the high frequency of species of Vriesea with limited geographic distributions indicated that epiphytes with rather limited geographic distributions predominate in this domain. Species similarity was divided into one block of south–southeastern localities, and a second block of northeastern–southeastern localities. These results suggest that the distribution of epiphytic bromeliad species resembles that of the phorophyte trees, more than a previous pattern suggested for all epiphytes in the domain. Latitude, temperature and altitude were important factors affecting the species composition along the domain. In general, our results differ from those of other studies in Latin America, and we suggest that historical and evolutionary events generated these differences.     

Canopy Openness Enhances Diversity of Ant–Plant Interactions in the Brazilian Amazon Rain Forest

In closed‐canopy tropical forest understory, light availability is a significant determinant of habitat diversity because canopy structure is highly variable in most tropical forests. Consequently, variation in canopy cover affects the composition and distribution of plant species via creating variable light environments. Nevertheless, little is known about how variation in canopy openness structures patterns of plant–animal interactions. Because of the great diversity and dominance of ants in tropical environments, we used ant–plant interactions as a focal network to evaluate how variation in canopy cover influences patterns of plant–insect interactions in the Brazilian Amazon rain forest. We observed that small increases in canopy openness are associated with increased diversity of ant–plant interactions in our study area, and this change is independent of plant or ant species richness. Additionally, we found smaller niche overlap for both ants and plants associated with greater canopy openness. We hypothesize that enhanced light availability increases the breadth of ant foraging sources because variation in light availability gives rise to plant resources of different quality and amounts. Moreover, greater light availability promotes vegetative growth in plants, creating ant foraging ‘bridges’ between plants. In sum, our results highlight the importance of environmental heterogeneity as a determinant of ant–plant interaction diversity in tropical environments.     

Distribution of Leguminosae tree species in different altitudinal levels along the Atlantic Rain Forest in the Brazilian coast

The Atlantic Forest is one of the most threatened tropical forests in the world. Leguminosae, by its great richness and dominance among arboreal stratum elements, is of major importance in the floristic composition and structure of this forest. We investigated the distribution of legume species on an altitudinal gradient to find out the altitudinal zones with higher richness of species; the altitudinal zones with greater floristic similarity; the possible presence of species that may be exclusive to certain vegetation types and the altitudinal amplitudes of those species, as well as the occurrence of species substitution along the altitudinal gradient. Therefore, thirty one studies conducted in different altitudinal levels between 5° S and 29° S were analyzed. A matrix with 142 tree species distributed in altitudinal zones (every 100 m) from sea level to 2100 m was built. The greatest species richness was observed in the Submontane Forest (50–500 m) with 92 species. The cluster analysis revealed a strong dissimilarity of the 1400–2100 m (Upper Montane) and 0–10 m zones (Restinga Forest). The Submontane and the Montane Forest share the highest number of species (38 ssp.). Forty species are unique to Submontane. Substitution of species was verified. Some species have their preferred habitat located at a specific altitudinal amplitude, as is the case of Inga laurina and I. subnuda (0–10 m), I. lanceifolia and Machaerium scleroxylon (800–1200 m). The Leguminosae, although well adapted to the first colonization and establishment of diverse environment, was poorly represented above 1500 m altitude.     

Forest regeneration affects dung beetle assemblages (Coleoptera: Scarabaeinae) in the southern Brazilian Atlantic Forest

The Brazilian Atlantic Forest is one of the most diverse environments, but it is also one of the most threatened areas in terms of loss of biodiversity and ecosystem services. Assessment of changes in the community structure during the recovery of forests can be performed using indicator organisms. Dung beetles perform several ecological functions and show high sensitivity to natural and anthropogenic environmental changes. This study aimed to investigate the effect of regeneration time of Atlantic Forest sites on structure of Scarabaeinae assemblages. We sampled dung beetles using ten baited pitfall traps per site, in six sites grouped into three classes of forest regeneration time (~30, ~60 and >80 years) in the southern Brazilian Atlantic Forest, during January 2015. A total of 520 individuals belonging to 16 species and nine genera of dung beetles were sampled. Rarefied species richness did not differ between sites with different regeneration times. Average species richness and abundance of Scarabaeinae was smaller in areas of shorter recovery time. True alpha diversity was higher in areas with intermediate recovery whereas Shannon diversity showed higher values in areas of shorter recovery. Approximately 29?% of the variation in abundance data of Scarabaeinae was explained by environmental variables, with one-third of this variation explained also by spatial predictors. External factors such as landscape management and farming practices in the surroundings must be taken into consideration in management plans and the management of natural areas for the recovery of biodiversity in the Atlantic Forest. These external factors can considerably affect the structure of communities and lead to scenarios of greater diversity in intermediate regeneration sites due to the heterogeneity of the landscape.     

Phylobetadiversity among Forest Types in the Brazilian Atlantic Forest Complex

Phylobetadiversity is defined as the phylogenetic resemblance between communities or biomes. Analyzing phylobetadiversity patterns among different vegetation physiognomies within a single biome is crucial to understand the historical affinities between them. Based on the widely accepted idea that different forest physiognomies within the Southern Brazilian Atlantic Forest constitute different facies of a single biome, we hypothesize that more recent phylogenetic nodes should drive phylobetadiversity gradients between the different forest types within the Atlantic Forest, as the phylogenetic divergence among those forest types is biogeographically recent. We compiled information from 206 checklists describing the occurrence of shrub/tree species across three different forest physiognomies within the Southern Brazilian Atlantic Forest (Dense, Mixed and Seasonal forests). We analyzed intra-site phylogenetic structure (phylogenetic diversity, net relatedness index and nearest taxon index) and phylobetadiversity between plots located at different forest types, using five different methods differing in sensitivity to either basal or terminal nodes (phylogenetic fuzzy weighting, COMDIST, COMDISTNT, UniFrac and Rao’s H). Mixed forests showed higher phylogenetic diversity and overdispersion than the other forest types. Furthermore, all forest types differed from each other in relation phylobetadiversity patterns, particularly when phylobetadiversity methods more sensitive to terminal nodes were employed. Mixed forests tended to show higher phylogenetic differentiation to Dense and Seasonal forests than these latter from each other. The higher phylogenetic diversity and phylobetadiversity levels found in Mixed forests when compared to the others likely result from the biogeographical origin of several taxa occurring in these forests. On one hand, Mixed forests shelter several temperate taxa, like the conifers Araucaria and Podocarpus. On the other hand, tropical groups, like Myrtaceae, are also very representative of this forest type. We point out to the need of more attention to Mixed forests as a conservation target within the Brazilian Atlantic Forest given their high phylogenetic uniqueness.     

Mechanisms of Araucaria (Atlantic) Forest Expansion into Southern Brazilian Grasslands

Recent studies have shown that tropical and subtropical forests expanded during the late Holocene, but rates and mechanisms of expansion are still unknown. Here, we investigate how a forest–grassland mosaic changed over the past 10,000 years at the southernmost limit of the Brazilian Atlantic forest. We used soil organic matter carbon isotopes (δ¹³C and ¹⁴C) to quantify and date changes in vegetation, examining soil properties and leaf traits of tree species (nutrient content, δ¹³C, δ¹⁵N, and specific leaf area—SLA) to describe potential mechanisms of expansion. Our results show that after several millennia of stability, forests have been expanding over grasslands through continuous, but very slow, border dynamics and patch formation (<100 m since ~4,000 YBP). This process of expansion coincided with past changes in climate, but biotic feedback mechanisms also appear to be important for the long-term persistence and expansion of forests. Soil fertility and microbial biomass match current rather than past vegetation distribution, increasing progressively across the gradient: grasslands < isolated trees < forest patches < forests. Foliar δ¹⁵N values of trees that are able to colonize the grassland are consistently lower across this vegetation gradient, suggesting an increasingly greater reliance on symbiotic nutrient uptake from grasslands to forests. No significant relationships were found between soil and leaf nutrients, but SLA explained variation in leaf N, P, and K (positive relationships) and in leaf δ¹³C (negative relationship). These findings suggest that a tradeoff between tree growth and water use efficiency is an important regulator of forest–grassland dynamics in the study region.