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Leaf water isotopic composition is imprinted in several biomarkers of interest and it is imperative that we understand the isotopic enrichment of leaf water. Here, we test the effect of stomatal density and leaf water content on the oxygen isotopic composition of leaf water in transgenic Arabidopsis plants expressing different stomatal densities, and several other species showing a range of stomatal density. We grew Arabidopsis plants hydroponically and collected other species in the field. Stomatal density and leaf water content were determined for each plant. We measured transpiration and extracted leaf water for isotopic determination. Using these measurements and the current leaf water isotope model, we calculated several of the parameters related to leaf water isotopic enrichment. High stomatal density promoted leaf water isotope enrichment. No conclusion, however, can be drawn regarding the effect of leaf water content on leaf water isotope enrichment. Factors such as transpiration might mask the effect of stomatal density on leaf water isotopic enrichment. We propose a method by which stomatal density can be incorporated in the current Peclet model of leaf water isotope enrichment. These findings have important applications in the use of plant-based metabolic proxies in paleoclimate studies.

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During plant-transpiration, water molecules having the lighter stable isotopes of oxygen and hydrogen evaporate and diffuse at a faster rate through the stomata than molecules having the heavier isotopes, which cause isotopic enrichment of leaf water. Although previous models have assumed that leaf water is well-mixed and isotopically uniform, non-uniform stomatal closure, promoting different enrichments between cells, and different pools of water within leaves, due to morpho-physiological traits, might lead to inaccuracies in isotopic models predicting leaf water enrichment. We evaluate the role of leaf morpho-physiological traits on leaf water isotopic enrichment in woody species occurring in a coastal vegetation of Brazil known as restinga. Hydrogen and oxygen stable isotope values of soil, plant stem and leaf water and leaf traits were measured in six species from restinga vegetation during a drought and a wet period. Leaf water isotopic enrichment relative to stem water was more homogeneous among species during the drought in contrast to the wet period suggesting convergent responses to deal to temporal heterogeneity in water availability. Average leaf water isotopic enrichment relative to stem water during the drought period was highly correlated with relative apoplastic water content. We discuss this observation in the context of current models of leaf water isotopic enrichment as a function of the Péclet effect. We suggest that future studies should include relative apoplastic water content in isotopic models.

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Leaf-cutting ants (Atta spp.) are known for their extensive defoliation in neo-tropical forests and savannahs. Debate about the costs and benefits of their activities has been largely dominated by their detrimental effects on agriculture and agroforestry. However, the large accumulation of nutrients and changes in soil properties near their nests might benefit plants growing near them. Here, we test whether trees use nutrients that accumulate in debris piles near, or refuse chambers within, leaf-cutting ant nests. At two tropical sites (a moist tropical forest site in Panama and a savannah site in Brazil), we fed leaves labelled with the stable isotope 15N to two species of leaf-cutting ants (Atta colombica and Atta laevigata) and traced the stable isotope label in plants surrounding the two nests. Thus, we show that plants in both sites access resources associated with Atta nests. In addition, leaf tissue of trees near the nests labelled with 15N had significantly higher calcium concentrations than those of distal, unlabelled conspecifics. It has been documented that calcium is a limiting macronutrient in tropical forests and savannahs. Atta may thus play an important ecological role through their long-distance transport, redistribution and concentration of critical macronutrients.

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As evidence mounts that sea levels are rising, it becomes increasingly important to understand the role of ocean water within terrestrial ecosystem dynamics. Coastal sand dunes are ecosystems that occur on the interface of land and sea. They are classic ecotones characterized by zonal distribution of vegetation in response to strong gradients of environmental factors from the ocean to the inland. Despite the proximity of the dune ecosystem to the ocean, it is generally assumed that all vegetation utilizes only freshwater and that water sources do not change across the ecotone. Evidence of ocean water uptake by vegetation would redefine the traditional interpretation of plant-water relations in the dune ecosystem and offer new ideas for assessing maritime influences on function and spatial distribution of plants across the dune. The purpose of this study was to identify sources of water (ocean, ground, and rain) taken up by vegetation using isotopic analysis of stem water and to evaluate water uptake patterns at the community level based on the distribution and assemblage of species. Three coastal dune systems located in southern Florida, USA, and the Bahamian bank/platform system were investigated. Plant distributions across the dune were zonal for 61-94% of the 18 most abundant species at each site. Species with their highest frequency on the fore dune (nearest the ocean) indicate ocean water uptake as evidenced by delta 18O values of stem water. In contrast, species most frequent in the back dune show no evidence of ocean water uptake. Analysis of species not grouped by frequency, but instead sampled along a transect from the ocean toward the inland, indicates that individuals from the vegetation assemblage closest to the ocean had a mixed water-harvesting strategy characterized by plants that may utilize ocean, ground-, and/or rainwater. In contrast, the inland vegetation relies mostly on rainwater. Our results show evidence supporting ocean water use by dune vegetation and demonstrate an exciting relationship between seawater and ecotonal shifts in plant function of a terrestrial ecosystem.

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The variability of δ13C values was measured in leaf, stem and root tissues of several tree species growing in closed natural and plantation forests in the Luquillo mountains of Puerto Rico. Results confirm a significant decrease of δ13C values from the tree canopy to the forest floor. The values measured in understory plants growing in gaps were not significantly different from the average for plants growing under the forest shade. Seedling leaf values tended to be more positive than those of saplings, probably reflecting the contribution of organic matter from the mother tree. Photosynthetic independence on the forest floor results in a reduction in °13C value. Stem and root tissue values of seedlings and saplings were less negative than those of the leaves of the same plants. It is suggested that this difference results from the slower change in isotopic composition experienced by the woody tissue, as the seedlings become photosynthetically independent in the forest floor.