Margaret M. Barbour

Stable oxygen isotope composition of plant tissue: a review

With the development of rapid measurement techniques, stable oxygen isotope analysis of plant tissue is poised to become an important tool in plant physiological, ecological, paleoclimatic and forensic studies. Recent advances in mechanistic understanding have led to the improvement of process-based models that accurately predict variability in the oxygen isotope composition of plant organic material (δ18Op). δ18Op has been shown to reflect the isotope composition of soil water, evaporative enrichment in transpiring leaves, and isotopic exchange between oxygen atoms in organic molecules and local water in the cells in which organic molecules are formed. This review presents current theoretical models describing the influences on δ18Op, using recently published experimental work to outline strengths and weaknesses in the models. The potential and realised applications of the technique are described.

Mesophyll diffusion conductance to CO2: An unappreciated central player in photosynthesis

Mesophyll diffusion conductance to CO(2) is a key photosynthetic trait that has been studied intensively in the past years. The intention of the present review is to update knowledge of g(m), and highlight the important unknown and controversial aspects that require future work. The photosynthetic limitation imposed by mesophyll conductance is large, and under certain conditions can be the most significant photosynthetic limitation. New evidence shows that anatomical traits, such as cell wall thickness and chloroplast distribution are amongst the stronger determinants of mesophyll conductance, although rapid variations in response to environmental changes might be regulated by other factors such as aquaporin conductance. Gaps in knowledge that should be research priorities for the near future include: how different is mesophyll conductance among phylogenetically distant groups and how has it evolved? Can mesophyll conductance be uncoupled from regulation of the water path? What are the main drivers of mesophyll conductance? The need for mechanistic and phenomenological models of mesophyll conductance and its incorporation in process-based photosynthesis models is also highlighted.

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.

Expressing leaf water and cellulose oxygen isotope ratios as enrichment above source water reveals evidence of a Péclet effect.

There is an increasing ecological interest in understanding the gradients in H218O enrichment in leaf water (i.e. a Péclet effect), because an appreciation of the significance of the Péclet effect is important for improving our understanding of the mechanistic processes affecting the 18O composition of leaf water and plant organic material. In data sets where both source water and leaf water 18O data are available, we can evaluate the potential contribution of a Péclet effect. As an example, we recalculate data published earlier by Roden and Ehleringer (1999, Oecologia 121:467–477) as enrichments in leaf water (ΔL) and cellulose (Δcell) above source water. Based on these recalculations, we present support for the relevance of a Péclet effect in leaves. Further, we demonstrate that the subtle variations in ΔL and Δcell caused by a Péclet effect may be masked in experimental systems in which variation in the source water oxygen isotope ratio is considerable.

Sensitivity of plants to changing atmospheric CO2 concentration : from the geological past to the next century

The rate of CO(2) assimilation by plants is directly influenced by the concentration of CO(2) in the atmosphere, c(a). As an environmental variable, c(a) also has a unique global and historic significance. Although relatively stable and uniform in the short term, global c(a) has varied substantially on the timescale of thousands to millions of years, and currently is increasing at seemingly an unprecedented rate. This may exert profound impacts on both climate and plant function. Here we utilise extensive datasets and models to develop an integrated, multi-scale assessment of the impact of changing c(a) on plant carbon dioxide uptake and water use. We find that, overall, the sensitivity of plants to rising or falling c(a) is qualitatively similar across all scales considered. It is characterised by an adaptive feedback response that tends to maintain 1 - c(i)/c(a), the relative gradient for CO(2) diffusion into the leaf, relatively constant. This is achieved through predictable adjustments to stomatal anatomy and chloroplast biochemistry. Importantly, the long-term response to changing c(a) can be described by simple equations rooted in the formulation of more commonly studied short-term responses.

Oxygen isotope ratio of leaf and grain material correlates with stomatal conductance and grain yield in irrigated wheat

Theory (Craig and Gordon 1965; Dongmann et al. 1974; Sternberg et al. 1986; Farquhar and Lloyd 1993) suggests that the oxygen isotope ratio (d 18 O) of plant material should reflect the evaporative conditions under which the material was formed, so that differences in stomatal conductance should show up in plant d 18 O. To test this theory we measured the oxygen isotope ratio of organic matter from flag leaves at anthesis and grain at harvest from eight cultivars of spring wheat (Triticum aestivum L.) grown under irrigation in each of three seasons in Mexico. The cultivars ranged widely in stomatal conductance and in average grain yield, with which conductance was positively correlated. Supporting theory, the oxygen isotope ratio of flag leaves (d 18 Ol) was found to correlate negatively with stomatal conductance for two of the three seasons. The significant correlations are consistent with high conductance cultivars having lower leaf temperatures and kinetic fractionation factors, and higher vapour pressure fractionation factors and Peclet numbers, all of which combine to result in less enriched d 18 Ol. Yield (grain weight per square meter) was also found to be significantly negatively correlated with d 18Ol in two of the three seasons. d 18 Ol was as good a predictor of yield as stomatal conductance, and significantly better than carbon isotope discrimination. Correlations between grain d 18 O (d 18 Og) and physiological parameters were less clear. Significant negative correlations between d 18 Og and stomatal conductance, leaf temperature and yield were found only during the first season. By measuring the oxygen isotope ratio of cellulose extracted from leaf samples, the difference in fractionation factors (ecp) for cellulose and whole leaf tissue was assessed. ecp was found to be variable, and more negative when d 18 Oc and d 18 Ol were lower. Cultivar means for d 13 C and d 18 O of whole leaf material were found to be significantly positively related, and the factors required to produce such a relationship are discussed.

Correlations between oxygen isotope ratios of wood constituents of Quercus and Pinus samples from around the world

The oxygen isotope compositions of three wood constituents (the solvent-extractable portion, lignin and α-cellulose) were measured for samples collected from Quercus and Pinus trees around the world. Among Pinus samples all wood constituents were positively related to modelled δ18O of source water at the site, while among Quercus samples whole wood, lignin and α-cellulose showed positive relationships. The data support the hypothesis that many oxygen atoms in lignin exchanged with unenriched stem water during synthesis, rather than retaining the full isotopic signal from the molecular oxygen added during hydroxylation of the aromatic ring. The data also suggest that extraction of α-cellulose from wood samples is unnecessary for isotope studies looking at correlations with site parameters.

Diffusional conductances to CO2 as a target for increasing photosynthesis and photosynthetic water-use efficiency

A key objective for sustainable agriculture and forestry is to breed plants with both high carbon gain and water-use efficiency (WUE). At the level of leaf physiology, this implies increasing net photosynthesis (AN) relative to stomatal conductance (gs). Here, we review evidence for CO2 diffusional constraints on photosynthesis and WUE. Analyzing past observations for an extensive pool of crop and wild plant species that vary widely in mesophyll conductance to CO2 (gm), gs, and foliage AN, it was shown that both gs and gm limit AN, although the relative importance of each of the two conductances depends on species and conditions. Based on Fick’s law of diffusion, intrinsic WUE (the ratio AN/gs) should correlate on the ratio gm/gs, and not gm itself. Such a correlation is indeed often observed in the data. However, since besides diffusion AN also depends on photosynthetic capacity (i.e., Vc,max), this relationship is not always sustained. It was shown that only in a very few cases, genotype selection has resulted in simultaneous increases of both AN and WUE. In fact, such a response has never been observed in genetically modified plants specifically engineered for either reduced gs or enhanced gm. Although increasing gm alone would result in increasing photosynthesis, and potentially increasing WUE, in practice, higher WUE seems to be only achieved when there are no parallel changes in gs. We conclude that for simultaneous improvement of AN and WUE, genetic manipulation of gm should avoid parallel changes in gs, and we suggest that the appropriate trait for selection for enhanced WUE is increased gm/gs.

Variability in mesophyll conductance between barley genotypes, and effects on transpiration efficiency and carbon isotope discrimination

Leaf internal, or mesophyll, conductance to CO(2) (g(m)) is a significant and variable limitation of photosynthesis that also affects leaf transpiration efficiency (TE). Genotypic variation in g(m) and the effect of g(m) on TE were assessed in six barley genotypes (four Hordeum vulgare and two H. bulbosum). Significant variation in g(m) was found between genotypes, and was correlated with photosynthetic rate. The genotype with the highest g(m) also had the highest TE and the lowest carbon isotope discrimination as recorded in leaf tissue (Delta(p)). These results suggest g(m) has unexplored potential to provide TE improvement within crop breeding programmes.

New constraints on atmospheric CO2 concentration for the Phanerozoic

Earths atmospheric CO2 concentration (ca) for the Phanerozoic Eon is estimated from proxies and geochemical carbon cycle models. Most estimates come with large, sometimes unbounded uncertainty. Here, we calculate tightly constrained estimates of ca using a universal equation for leaf gas exchange, with key variables obtained directly from the carbon isotope composition and stomatal anatomy of fossil leaves. Our new estimates, validated against ice cores and direct measurements of ca, are less than 1000 ppm for most of the Phanerozoic, from the Devonian to the present, coincident with the appearance and global proliferation of forests. Uncertainties, obtained from Monte Carlo simulations, are typically less than for ca estimates from other approaches. These results provide critical new empirical support for the emerging view that large (~2000-3000 ppm), long-term swings in ca do not characterize the post-Devonian and that Earths long-term climate sensitivity to ca is greater than originally thought. Key Points A novel CO2 proxy calculates past atmospheric CO2 with improved certainty CO2 is unlikely to have exceeded ~1000 ppm for extended periods post Devonian Earths long-term climate sensitivity to CO2 is greater than originally thought.