Abstract: In tropical forests, free-living Biological nitrogen (N) fixation (BNF) in soil and litter tends to decrease when substrate N concentrations increase, whereas increasing phosphorus (P) and molybdenum (Mo) soil and litter concentrations have been shown to stimulate free-living BNF rates. Yet, very few studies explored the effects of adding N, P, and Mo together in a single large-scale fertilization experiment, which would teach us which of these elements constrain or limit BNF activities. At two distinct forest sites in French Guiana, we performed a 3-year in situ nutrient addition study to explore the effects of N, P, and Mo additions on leaf litter and soil BNF. Additionally, we conducted a short-term laboratory study with the same nutrient addition treatments (+N, +N+P, +P, +Mo, and +P+Mo). We found that N additions alone suppressed litter free-living BNF in the field, but not in the short-term laboratory study, while litter free-living BNF remained unchanged in response to N+P additions. Additionally, we found that P and P+Mo additions stimulated BNF in leaf litter, both in the field and in the lab, while Mo alone yielded no changes. Soil BNF increased with P and P+Mo additions in only one of the field sites, while in the other site soil BNF increased with Mo and P+Mo additions. We concluded that increased substrate N concentrations suppress BNF. Moreover, both P and Mo have the potential to limit free-living BNF in these tropical forests, but the balance between P versus Mo limitation is determined by site-specific characteristics of nutrient supply and demand.

Abstract: Across the evolutionary history of insects, the shift from nitrogen-rich carnivore/omnivore diets to nitrogen-poor herbivorous diets was made possible through symbiosis with microbes. The herbivorous turtle ants Cephalotes possess a conserved gut microbiome which enriches the nutrient composition by recycling nitrogen-rich metabolic waste to increase the production of amino acids. This enrichment is assumed to benefit the host, but we do not know to what extent. To gain insights into nitrogen assimilation in the ant cuticle we use gut bacterial manipulation, 15N isotopic enrichment, isotope-ratio mass spectrometry, and 15N nuclear magnetic resonance spectroscopy to demonstrate that gut bacteria contribute to the formation of proteins, catecholamine cross-linkers, and chitin in the cuticle. This study identifies the cuticular components which are nitrogen-enriched by gut bacteria, highlighting the role of symbionts in insect evolution, and provides a framework for understanding the nitrogen flow from nutrients through bacteria into the insect cuticle.

Abstract: Trees can modify their wood structure in response to changes in mechanical, hydraulic and storage demands during their life-cycles. Thus, examining radial variations in wood traits is important to expand our knowledge of tree functioning and species ecological strategies. Yet, several aspects of radial changes in wood functional traits are still poorly understood, especially in angiosperm trees from tropical humid forests. Here, we examined radial shifts in wood traits in trunks of tropical forest species and explored their potential ecological implications. We first examined radial variations in wood specific gravity (WSG). Then, we asked what anatomical traits drove radial variations in WSG, and whether WSG, vessel fraction and specific hydraulic conductivity vary independently from each other along the radius gradients. We measured WSG and eight wood anatomical traits, at different radial positions along the trunks, in 19 tree species with contrasting shade-tolerance from a lowland tropical forest in eastern Amazonia. Most species had significant radials shifts in WSG. Positive radial gradients in WSG (i.e., increments from pith to bark) were common among shade-intolerant species and were explained by different combinations of fiber and parenchyma traits, while negative radial shifts in WSG (e.g., decreases towards the bark) were present in shade-tolerants, but were generally weakly related to anatomical traits. We also found that, in general, WSG was unrelated to vessel fraction and specific hydraulic conductivity in any radial position. This study illustrates the contrasting radial variations in wood functional traits that occur in tree species from a humid lowland tropical forest. In particular, our results provide valuable insights into the anatomical traits driving WSG variations during tree development. These insights are important to expand our knowledge on tree ecological strategies by providing evidence on how wood allocation varies as trees grow, which in turn can be useful in studying trait-demography associations, and in estimating tree above-ground biomass.

Abstract: Questions Changes in the functional composition of tree communities along resource availability gradients have received attention, but it is unclear whether understorey and canopy guilds respond similarly to different light, biomechanical, and hydraulic constraints. Location An anthropically undisturbed, old-growth wet evergreen dipterocarp forest plot located in Karnataka State, India. Methods We measured leaf and wood traits of 89 tree species representing 99% of all individuals in a 10-ha permanent plot with varying topographic and canopy conditions inferred from Light Detection And Ranging (LiDAR) data. We assigned tree species to guilds of canopy and understorey species and assessed the variation of the guild-weighted means of functional-trait values with canopy height and topography. Results The functional-trait space did not differ between canopy and understorey tree species. However, environmental filtering led to significantly different functional composition of canopy and understorey guild assemblages. Furthermore, they responded differently along environmental gradients related to water, nutrients, light, and wind exposure. For example, the canopy guild responded to wind exposure while the understorey guild did not. Conclusions The pools of understorey and canopy species are functionally similar. However, fine-scale environmental heterogeneity impacts differently on these two guilds, generating striking differences in functional composition between understorey and canopy guild assemblages. Accounting for vertical guilds improves our understanding of forest communities' assembly processes.

Abstract: For highly specialized insect herbivores, plant chemical defenses are often co-opted as cues for oviposition and sequestration. In such interactions, can plants evolve novel defenses, pushing herbivores to trade off benefits of specialization with costs of coping with toxins? We tested how variation in milkweed toxins (cardenolides) impacted monarch butterfly ( Danaus plexippus ) growth, sequestration, and oviposition when consuming tropical milkweed ( Asclepias curassavica ), one of two critical host plants worldwide. The most abundant leaf toxin, highly apolar and thiazolidine ring–containing voruscharin, accounted for 40% of leaf cardenolides, negatively predicted caterpillar growth, and was not sequestered. Using whole plants and purified voruscharin, we show that monarch caterpillars convert voruscharin to calotropin and calactin in vivo, imposing a burden on growth. As shown by in vitro experiments, this conversion is facilitated by temperature and alkaline pH. We next employed toxin-target site experiments with isolated cardenolides and the monarch’s neural Na + /K + -ATPase, revealing that voruscharin is highly inhibitory compared with several standards and sequestered cardenolides. The monarch’s typical >50-fold enhanced resistance to cardenolides compared with sensitive animals was absent for voruscharin, suggesting highly specific plant defense. Finally, oviposition was greatest on intermediate cardenolide plants, supporting the notion of a trade-off between benefits and costs of sequestration for this highly specialized herbivore. There is apparently ample opportunity for continued coevolution between monarchs and milkweeds, although the diffuse nature of the interaction, due to migration and interaction with multiple milkweeds, may limit the ability of monarchs to counteradapt.