Lucas A. Cernusak

Heavy Water Fractionation during Transpiration

A small proportion of water molecules contain the heavier isotopes of hydrogen and oxygen. There is a tendency for these heavier molecules of water to accumulate in leaves during transpiration. This has several interesting repercussions, including effects on the isotopic composition of organic

Why are non-photosynthetic tissues generally 13C enriched compared with leaves in C3 plants? Review and synthesis of current hypotheses

Non-photosynthetic, or heterotrophic, tissues in C3 plants tend to be enriched in 13C compared with the leaves that supply them with photosynthate. This isotopic pattern has been observed for woody stems, roots, seeds and fruits, emerging leaves, and parasitic plants incapable of net CO2 fixation. Unlike in C3 plants, roots of herbaceous C4 plants are generally not 13C-enriched compared with leaves. We review six hypotheses aimed at explaining this isotopic pattern in C3 plants: (1) variation in biochemical composition of heterotrophic tissues compared with leaves; (2) seasonal separation of growth of leaves and heterotrophic tissues, with corresponding variation in photosynthetic discrimination against 13C; (3) differential use of day v. night sucrose between leaves and sink tissues, with day sucrose being relatively 13C-depleted and night sucrose 13C-enriched; (4) isotopic fractionation during dark respiration; (5) carbon fixation by PEP carboxylase; and (6) developmental variation in photosynthetic discrimination against 13C during leaf expansion. Although hypotheses (1) and (2) may contribute to the general pattern, they cannot explain all observations. Some evidence exists in support of hypotheses (3) through to (6), although for hypothesis (6) it is largely circumstantial. Hypothesis (3) provides a promising avenue for future research. Direct tests of these hypotheses should be carried out to provide insight into the mechanisms causing within-plant variation in carbon isotope composition.

Environmental and physiological determinants of carbon isotope discrimination in terrestrial plants

Stable carbon isotope ratios (δ(13) C) of terrestrial plants are employed across a diverse range of applications in environmental and plant sciences; however, the kind of information that is desired from the δ(13) C signal often differs. At the extremes, it ranges between purely environmental and purely biological. Here, we review environmental drivers of variation in carbon isotope discrimination (Δ) in terrestrial plants, and the biological processes that can either damp or amplify the response. For C3 plants, where Δ is primarily controlled by the ratio of intercellular to ambient CO2 concentrations (ci /ca ), coordination between stomatal conductance and photosynthesis and leaf area adjustment tends to constrain the potential environmentally driven range of Δ. For C4 plants, variation in bundle-sheath leakiness to CO2 can either damp or amplify the effects of ci /ca on Δ. For plants with crassulacean acid metabolism (CAM), Δ varies over a relatively large range as a function of the proportion of daytime to night-time CO2 fixation. This range can be substantially broadened by environmental effects on Δ when carbon uptake takes place primarily during the day. The effective use of Δ across its full range of applications will require a holistic view of the interplay between environmental control and physiological modulation of the environmental signal.

Water Relations Link Carbon and Oxygen Isotope Discrimination to Phloem Sap Sugar Concentration in Eucalyptus globulus

A strong correlation was previously observed between carbon isotope discrimination (Δ13C) of phloem sap sugars and phloem sap sugar concentration in the phloem-bleeding tree Eucalyptus globulus Labill. (J. Pate, E. Shedley, D. Arthur, M. Adams [1998] Oecologia 117: 312–322). We hypothesized that correspondence between these two parameters results from covarying responses to plant water potential. We expected Δ13C to decrease with decreasing plant water potential and phloem sap sugar concentration to increase, thereby maintaining turgor within sieve tubes. The hypothesis was tested with analyses of E. globulus trees growing on opposite ends of a rainfall gradient in southwestern Australia. The Δ13C of phloem sap sugars was closely related to phloem sap sugar concentration (r = −0.90,P < 0.0001, n = 40). As predicted, daytime shoot water potential was positively related to Δ13C (r = 0.70, P< 0.0001, n = 40) and negatively related to phloem sap sugar concentration (r = −0.86,P < 0.0001, n = 40). Additional measurements showed a strong correspondence between predawn shoot water potential and phloem sap sugar concentration measured at midday (r = −0.87, P < 0.0001, n = 30). The Δ13C of phloem sap sugars collected from the stem agreed well with that predicted from instantaneous measurements of the ratio of intercellular to ambient carbon dioxide concentrations on subtending donor leaves. In accordance, instantaneous ratio of intercellular to ambient carbon dioxide concentrations correlated negatively with phloem sap sugar concentration (r = −0.91, P < 0.0001, n = 27). Oxygen isotope enrichment (Δ18O) in phloem sap sugars also varied with phloem sap sugar concentration (r = 0.91,P < 0.0001, n = 39), consistent with predictions from a theoretical model of Δ18O. We conclude that drought induces correlated variation in the concentration of phloem sap sugars and their isotopic composition in E. globulus.

Ternary effects on the gas exchange of isotopologues of carbon dioxide

The ternary effects of transpiration rate on the rate of assimilation of carbon dioxide through stomata, and on the calculation of the intercellular concentration of carbon dioxide, are now included in standard gas exchange studies. However, the equations for carbon isotope discrimination and for the exchange of oxygen isotopologues of carbon dioxide ignore ternary effects. Here we introduce equations to take them into account. The ternary effect is greatest when the leaf-to-air vapour mole fraction difference is greatest, and its impact is greatest on parameters derived by difference, such as the mesophyll resistance to CO(2) assimilation, r(m) . We show that the mesophyll resistance to CO(2) assimilation has been underestimated in the past. The impact is also large when there is a large difference in isotopic composition between the CO(2) inside the leaf and that in the air. We show that this partially reconciles estimates of the oxygen isotopic composition of CO(2) in the chloroplast and mitochondria in the light and in the dark, with values close to equilibrium with the estimated oxygen isotopic composition of water at the sites of evaporation within the leaf.

Measurement and Interpretation of the Oxygen Isotope Composition of Carbon Dioxide Respired by Leaves in the Dark

We measured the oxygen isotope composition (δ18O) of CO2 respired by Ricinus communis leaves in the dark. Experiments were conducted at low CO2 partial pressure and at normal atmospheric CO2 partial pressure. Across both experiments, the δ18O of dark-respired CO2 (δR) ranged from 44‰ to 324‰ (Vienna Standard Mean Ocean Water scale). This seemingly implausible range of values reflects the large flux of CO2 that diffuses into leaves, equilibrates with leaf water via the catalytic activity of carbonic anhydrase, then diffuses out of the leaf, leaving the net CO2 efflux rate unaltered. The impact of this process on δR is modulated by the δ18O difference between CO2 inside the leaf and in the air, and by variation in the CO2 partial pressure inside the leaf relative to that in the air. We developed theoretical equations to calculate δ18O of CO2 in leaf chloroplasts (δc), the assumed location of carbonic anhydrase activity, during dark respiration. Their application led to sensible estimates of δc, suggesting that the theory adequately accounted for the labeling of CO2 by leaf water in excess of that expected from the net CO2 efflux. The δc values were strongly correlated with δ18O of water at the evaporative sites within leaves. We estimated that approximately 80% of CO2 in chloroplasts had completely exchanged oxygen atoms with chloroplast water during dark respiration, whereas approximately 100% had exchanged during photosynthesis. Incorporation of the δ18O of leaf dark respiration into ecosystem and global scale models of C18OO dynamics could affect model outputs and their interpretation.

Oxygen isotope composition of phloem sap in relation to leaf water in Ricinus communis

We measured the oxygen isotope composition of both the water and dry matter components of phloem sap exported from photosynthesising Ricinus communis L. leaves. The 18O / 16O composition of exported dry matter matched almost exactly that expected for equilibrium with average lamina leaf water (leaf water exclusive of water associated with primary veins) with an isotope effect of αo=1.027, where αo=Ro / Rw , and Ro and Rw are 18O / 16O of organic molecules and water, respectively. Average lamina leaf water was enriched by 14-22‰ compared with source water under our experimental conditions, and depleted by 4-7‰, compared with evaporative site water. This showed that it is the average lamina leaf water 18O / 16O signal that is exported from photosynthesising leaves rather than a signal more closely related to that of evaporative site water or source water. Additionally, we found that water exported in phloem sap from photosynthesising leaves was enriched compared with source water; the mean phloem water enrichment observed for leaf petioles was 4.0 ± 1.5‰ (mean ± 1 s.d., n = 27). Phloem water collected from stem bases was also enriched compared with source water. However, the enrichment was approximately 0.8 times that observed for leaf petioles, suggesting some mixing between enriched phloem water and unenriched xylem water occurred during translocation. Results validated the assumption that organic molecules exported from photosynthesising leaves are enriched by 27‰ compared with average lamina leaf water. Furthermore, results suggest that the potential influence of enriched phloem water should be considered when interpreting the 18O / 16O signatures of plant organic material and plant cellulose.

The relationship of leaf photosynthetic traits – Vcmax and Jmax – to leaf nitrogen, leaf phosphorus, and specific leaf area: a meta‐analysis and modeling study

Great uncertainty exists in the global exchange of carbon between the atmosphere and the terrestrial biosphere. An important source of this uncertainty lies in the dependency of photosynthesis on the maximum rate of carboxylation (Vcmax) and the maximum rate of electron transport (Jmax). Understanding and making accurate prediction of C fluxes thus requires accurate characterization of these rates and their relationship with plant nutrient status over large geographic scales. Plant nutrient status is indicated by the traits: leaf nitrogen (N), leaf phosphorus (P), and specific leaf area (SLA). Correlations between Vcmax and Jmax and leaf nitrogen (N) are typically derived from local to global scales, while correlations with leaf phosphorus (P) and specific leaf area (SLA) have typically been derived at a local scale. Thus, there is no global-scale relationship between Vcmax and Jmax and P or SLA limiting the ability of global-scale carbon flux models do not account for P or SLA. We gathered published data from 24 studies to reveal global relationships of Vcmax and Jmax with leaf N, P, and SLA. Vcmax was strongly related to leaf N, and increasing leaf P substantially increased the sensitivity of Vcmax to leaf N. Jmax was strongly related to Vcmax, and neither leaf N, P, or SLA had a substantial impact on the relationship. Although more data are needed to expand the applicability of the relationship, we show leaf P is a globally important determinant of photosynthetic rates. In a model of photosynthesis, we showed that at high leaf N (3 gm−2), increasing leaf P from 0.05 to 0.22 gm−2 nearly doubled assimilation rates. Finally, we show that plants may employ a conservative strategy of Jmax to Vcmax coordination that restricts photoinhibition when carboxylation is limiting at the expense of maximizing photosynthetic rates when light is limiting.

Leaf nitrogen to phosphorus ratios of tropical trees: experimental assessment of physiological and environmental controls

We investigated the variation in leaf nitrogen to phosphorus ratios of tropical tree and liana seedlings as a function of the relative growth rate, whole-plant water-use efficiency, soil water content and fertilizer addition. First, seedlings of 13 tree and liana species were grown individually in 38-l pots prepared with a homogeneous soil mixture. Second, seedlings of three tree species were grown in 19-l pots at high or low soil water content, and with or without added fertilizer containing nitrogen, phosphorus and potassium. For plants grown under common soil conditions, leaf nitrogen to phosphorus ratios showed a unimodal, or hump-shaped, relationship with the relative growth rate. The leaf nitrogen to phosphorus ratio increased in response to low soil water content in three species, and increased in response to fertilizer addition in two of the three species. Across all species and treatments, the leaf nitrogen to phosphorus ratio was positively correlated with the water-use efficiency. The results suggest that the within-site variation among tropical tree species in the leaf nitrogen to phosphorus ratio may be caused by associations between this ratio and the relative growth rate. Modification of the soil environment changed the leaf nitrogen to phosphorus ratio, but underlying associations between this ratio and the relative growth rate were generally maintained. The observed correlation between the leaf nitrogen to phosphorus ratio and water-use efficiency has implications for linking nutrient stoichiometry with plant transpiration.

Tropical forest responses to increasing atmospheric CO2: current knowledge and opportunities for future research

Elevated atmospheric CO2 concentrations (ca) will undoubtedly affect the metabolism of tropical forests worldwide; however, critical aspects of how tropical forests will respond remain largely unknown. Here, we review the current state of knowledge about physiological and ecological responses, with the aim of providing a framework that can help to guide future experimental research. Modelling studies have indicated that elevated ca can potentially stimulate photosynthesis more in the tropics than at higher latitudes, because suppression of photorespiration by elevated ca increases with temperature. However, canopy leaves in tropical forests could also potentially reach a high temperature threshold under elevated ca that will moderate the rise in photosynthesis. Belowground responses, including fine root production, nutrient foraging and soil organic matter processing, will be especially important to the integrated ecosystem response to elevated ca. Water use efficiency will increase as ca rises, potentially impacting upon soil moisture status and nutrient availability. Recruitment may be differentially altered for some functional groups, potentially decreasing ecosystem carbon storage. Whole-forest CO2 enrichment experiments are urgently needed to test predictions of tropical forest functioning under elevated ca. Smaller scale experiments in the understorey and in gaps would also be informative, and could provide stepping stones towards stand-scale manipulations.