Leaf cell wall properties and stomatal density influence oxygen isotope enrichment of leaf water.

Measurements of oxygen isotope enrichment of leaf water above source water (Δ18 OLW ) can improve our understanding of the interaction between leaf anatomy and physiology on leaf water transport. Models have been developed to predict Δ18 OLW such as the string-of-lakes model, which describes the mixing of leaf water pools, and the Péclet effect model, which incorporates transpiration rate and the mixing length between unenriched xylem and enriched mesophyll water in the mesophyll (Lm ) or veins (Lv ). Here we compare measurements and models of Δ18 OLW on two cell wall composition mutants grown under two light intensities and relative humidities to evaluate cell wall properties on leaf water transport. In maize (Zea mays), the compromised ultrastructure of the suberin lamellae in the bundle sheath of the ALIPHATIC SUBERIN FERULOYL TRANSFERASE mutant (Zmasft) reduced barriers to apoplastic water movement, resulting in higher E and, potentially, Lv and, consequently, lower Δ18 OLW . The difference in Δ18 OLW in cellulose synthase-like F6 (CslF6) mutants and wild-type of rice (Oryza sativa) grown under two light intensities co-varied with stomatal density. These results show that cell wall composition and stomatal density influence Δ18 OLW and that stable isotopes can facilitate the development of a physiologically and anatomically explicit water transport model.

Leaf water oxygen isotope measurement by direct equilibration.

The oxygen isotope composition of leaf water imparts a signal to a range of molecules in the atmosphere and biosphere, but has been notoriously difficult to measure in studies requiring a large number of samples as a consequence of the labour-intensive extraction step. We tested a method of direct equilibration of water in fresh leaf samples with CO2 , and subsequent oxygen isotope analysis on an optical spectrometer. The oxygen isotope composition of leaf water measured by the direct equilibration technique was strongly linearly related to that of cryogenically extracted leaf water in paired samples for a wide range of species with differing anatomy, with an R(2) of 0.95. The somewhat more enriched values produced by the direct equilibration method may reflect lack of full equilibration with unenriched water in the vascular bundles, but the strong relationship across a wide range of species suggests that this difference can be adequately corrected for using a simple linear relationship.

The impact of an inverse climate-isotope relationship in soil water on the oxygen-isotope composition of Larix gmelinii in Siberia.

Stable oxygen isotope ratios (δ(18) O) in trees from high latitude ecosystems are valuable sources of information for recent and past environmental changes, but the interpretation is hampered by the complex hydrology of forests growing under permafrost conditions, where only a shallow layer of soil thaws in summer. We investigated larch trees (Larix gmelinii) at two sites with contrasting soil conditions in Siberia and determined δ(18) O of water from different soil depths, roots, twigs, and needles as well as δ(18) O of soluble carbohydrates regularly over two growing seasons. A comparison of results from the 2 yrs revealed an unexpected 'inverse' climate-isotope relationship, as dry and warm summer conditions resulted in lower soil and root δ(18) O values. This was due to a stronger uptake of isotopically depleted water pools originating from melted permafrost or previous winter snow. We developed a conceptual framework that considers the dependence of soil water profiles on climatic conditions for explaining δ(18) O in needle water, needle soluble carbohydrates and stem cellulose. The negative feedback of drought conditions on the source isotope value could explain decreasing tree-ring δ(18) O trends in a warming climate and is likely relevant in many ecosystems, where a soil isotope gradient with depth is observed.

Measurements of transpiration isotopologues and leaf water to assess enrichment models in cotton.

The two-pool and Péclet effect models represent two theories describing mechanistic controls underlying leaf water oxygen isotope composition at the whole-leaf level (δ(18) OL ). To test these models, we used a laser spectrometer coupled to a gas-exchange cuvette to make online measurements of δ(18) O of transpiration (δ(18) Otrans ) and transpiration rate (E) in 61 cotton (Gossypium hirsutum) leaves. δ(18) Otrans measurements permitted direct calculation of δ(18) O at the sites of evaporation (δ(18) Oe ) which, combined with values of δ(18) OL from the same leaves, allowed unbiased estimation of the proportional deviation of enrichment of δ(18) OL from that of δ(18) Oe (f) under both steady-state (SS) and non-steady-state (NSS) conditions. Among all leaves measured, f expressed relative to both δ(18) O of transpired water (ftrans ) and source water (fsw ) remained relatively constant with a mean ± SD of 0.11 ± 0.05 and 0.13 ± 0.05, respectively, regardless of variation in E spanning 0.8-9.1 mmol m(-2)  s(-1) . Neither ftrans nor fsw exhibited a significant difference between the SS and NSS leaves at the P < 0.05 level. Our results suggest that the simpler two-pool model is adequate for predicting cotton leaf water enrichment at the whole-leaf level. We discuss the implications of adopting a two-pool concept for isotopic applications in ecological studies.

Seasonal transfer of oxygen isotopes from precipitation and soil to the tree ring: source water versus needle water enrichment.

For accurate interpretation of oxygen isotopes in tree rings (δ(18) O), it is necessary to disentangle the mechanisms underlying the variations in the tree's internal water cycle and to understand the transfer of source versus leaf water δ(18) O to phloem sugars and stem wood. We studied the seasonal transfer of oxygen isotopes from precipitation and soil water through the xylem, needles and phloem to the tree rings of Larix decidua at two alpine sites in the Lötschental (Switzerland). Weekly resolved δ(18) O records of precipitation, soil water, xylem and needle water, phloem organic matter and tree rings were developed. Week-to-week variations in needle-water (18) O enrichment were strongly controlled by weather conditions during the growing season. These short-term variations were, however, not significantly fingerprinted in tree-ring δ(18) O. Instead, seasonal trends in tree-ring δ(18) O predominantly mirrored trends in the source water, including recent precipitation and soil water pools. Modelling results support these findings: seasonal tree-ring δ(18) O variations are captured best when the week-to-week variations of the leaf water signal are suppressed. Our results suggest that climate signals in tree-ring δ(18) O variations should be strongest at temperate sites with humid conditions and precipitation maxima during the growing season.

Reconstructing the δ(18) O of atmospheric water vapour via the CAM epiphyte Tillandsia usneoides: seasonal controls on δ(18) O in the field and large-scale reconstruction of δ(18) Oa.

Using both oxygen isotope ratios of leaf water (δ(18) OL ) and cellulose (δ(18) OC ) of Tillandsia usneoides in situ, this paper examined how short- and long-term responses to environmental variation and model parameterization affected the reconstruction of the atmospheric water vapour (δ(18) Oa ). During sample-intensive field campaigns, predictions of δ(18) OL matched observations well using a non-steady-state model, but the model required data-rich parameterization. Predictions from the more easily parameterized maximum enrichment model (δ(18) OL-M ) matched observed δ(18) OL and observed δ(18) Oa when leaf water turnover was less than 3.5 d. Using the δ(18) OL-M model and weekly samples of δ(18) OL across two growing seasons in Florida, USA, reconstructed δ(18) Oa was -12.6 ± 0.3‰. This is compared with δ(18) Oa of -12.4 ± 0.2‰ resolved from the growing-season-weighted δ(18) OC . Both of these values were similar to δ(18) Oa in equilibrium with precipitation, -12.9‰. δ(18) Oa was also reconstructed through a large-scale transect with δ(18) OL and the growing-season-integrated δ(18) OC across the southeastern United States. There was considerable large-scale variation, but there was regional, weather-induced coherence in δ(18) Oa when using δ(18) OL . The reconstruction of δ(18) Oa with δ(18) OC generally supported the assumption of δ(18) Oa being in equilibrium with precipitation δ(18) O (δ(18) Oppt ), but the pool of δ(18) Oppt with which δ(18) Oa was in equilibrium - growing season versus annual δ(18) Oppt - changed with latitude.