Bahar, Nur HA, et al.Phosphorus deficiency alters scaling relationships between leaf gas exchange and associated traits in a wide range of contrasting Eucalyptus species”. Functional Plant Biology 45.8 (2018): , 45, 8, 813-826. Web. Publisher's VersionAbstract
Phosphorus (P) limitation is known to have substantial impacts on leaf metabolism. However, uncertainty remains around whether P deficiency alters scaling functions linking leaf metabolism to associated traits. We investigated the effect of P deficiency on leaf gas exchange and related leaf traits in 17 contrasting Eucalyptus species that exhibit inherent differences in leaf traits. Saplings were grown under controlled-environment conditions in a glasshouse, where they were subjected to minus and plus P treatments for 15 weeks. P deficiency decreased P concentrations and increased leaf mass per area (LMA) of newly-developed leaves. Rates of photosynthesis (A) and respiration (R) were also reduced in P-deficient plants compared with P-fertilised plants. By contrast, P deficiency had little effect on the temperature sensitivity of R. Irrespective of P treatment, on a log-log basis Aand R scaled positively with increasing leaf nitrogen concentration [N] and negatively with increasing LMA. Although P deficiency had limited impact on A-R-LMA relationships, rates of CO2 exchange per unit N were consistently lower in P-deficient plants. Our results highlight the importance of P supply for leaf carbon metabolism and show how P deficiencies (i.e. when excluding confounding genotypic and environmental effects) can have a direct effect on commonly used leaf trait scaling relationships.
Chaney, Nathaniel W., et al.Harnessing big data to rethink land heterogeneity in Earth system models”. Hydrology and Earth System Sciences 22 (2018): , 22, 3311-3330. Web. Publisher's VersionAbstract
The continual growth in the availability, detail, and wealth of environmental data provides an invaluable asset to improve the characterization of land heterogeneity in Earth system models – a persistent challenge in macroscale models. However, due to the nature of these data (volume and complexity) and computational constraints, these data are underused for global applications. As a proof of concept, this study explores how to effectively and efficiently harness these data in Earth system models over a 1/4° ( ∼  25 km) grid cell in the western foothills of the Sierra Nevada in central California. First, a novel hierarchical multivariate clustering approach (HMC) is introduced that summarizes the high-dimensional environmental data space into hydrologically interconnected representative clusters (i.e., tiles). These tiles and their associated properties are then used to parameterize the sub-grid heterogeneity of the Geophysical Fluid Dynamics Laboratory (GFDL) LM4-HB land model. To assess how this clustering approach impacts the simulated water, energy, and carbon cycles, model experiments are run using a series of different tile configurations assembled using HMC. The results over the test domain show that (1) the observed similarity over the landscape makes it possible to converge on the macroscale response of the fully distributed model with around 300 sub-grid land model tiles; (2) assembling the sub-grid tile configuration from available environmental data can have a large impact on the macroscale states and fluxes of the water, energy, and carbon cycles; for example, the defined subsurface connections between the tiles lead to a dampening of macroscale extremes; (3) connecting the fine-scale grid to the model tiles via HMC enables circumvention of the classic scale discrepancies between the macroscale and field-scale estimates; this has potentially significant implications for the evaluation and application of Earth system models.
Gerlein-Safdi, Cynthia, et al.The impact of dew-induced transpiration suppression on the water and isotope balance of Colocasia esculenta leaves. ”. Oecologia 187.4 (2018): , 187, 4, 1041-1051. Web. Publisher's VersionAbstract
Foliar uptake of water from the surface of leaves is common when rainfall is scarce and non-meteoric water such as dew or fog is more abundant. However, many species in more mesic environments have hydrophobic leaves that do not allow the plant to uptake water. Unlike foliar uptake, all species can benefit from dew- or fog-induced transpiration suppression, but despite its ubiquity, transpiration suppression has so far never been quantified. Here, we investigate the effect of dew-induced transpiration suppression on the water balance and the isotope composition of leaves via a series of experiments. Characteristically, hydrophobic leaves of a tropical plant, Colocasia esculenta, are misted with isotopically enriched water to reproduce dew deposition. This species does not uptake water from the surface of its leaves. We measure leaf water isotopes and water potential and find that misted leaves exhibit a higher water potential and a more depleted water isotope composition than dry leaves, suggesting a ∼ 30% decrease in transpiration rate compared to control leaves. We propose three possible mechanisms governing the interaction of water droplets with leaf energy balance: increase in albedo from the presence of dew droplets, decrease in leaf temperature from the evaporation of dew, and local decrease in vapor pressure deficit. Comparing previous studies on foliar uptake to our results, we conclude that transpiration suppression has an effect of similar amplitude, yet opposite sign to foliar uptake on leaf water isotopes.
Gauthier, Paul P. G., et al.Measurement of Gross Photosynthesis, Respiration in the Light and Mesophyll Conductance Using H218O Labeling”. Plant Physiology 177 (2018): , 177, 62-74. Web. Publisher's VersionAbstract

A fundamental challenge in plant physiology is independently determining the rates of gross O2 production by photosynthesis and O2 consumption by respiration, photorespiration, and other processes. Previous studies on isolated chloroplasts or leaves have separately constrained net and gross O2 production (NOP and GOP, respectively) by labeling ambient O2 with 18O while leaf water was unlabeled. Here, we describe a method to accurately measure GOP and NOP of whole detached leaves in a cuvette as a routine gas-exchange measurement. The petiole is immersed in water enriched to a d18O of ~9,000‰, and leaf water is labeled through the transpiration stream. Photosynthesis transfers 18O from H2O to O2. GOP is calculated from the increase in d18O of O2 as air passes through the cuvette. NOP is determined from the increase in O2/N2. Both terms are measured by isotope ratio mass spectrometry. CO2 assimilation and other standard gas-exchange parameters also were measured. Reproducible measurements are made on a single leaf for more than 15 h. We used this method to measure the light response curve of NOP and GOP in French bean (Phaseolus vulgaris) at 21% and 2% O2. We then used these data to examine the O2/CO2 ratio of net photosynthesis, the light response curve of mesophyll conductance, and the apparent inhibition of respiration in the light (Kok effect) at both oxygen levels. The results are discussed in the context of evaluating the technique as a tool to study and understand leaf physiological traits.

Tcherkez, Guillaume G. B., et al.Leaf day respiration: low CO2 flux but high significance for metabolism and carbon balance ”. Transley Review (2017). Web. Publisher's VersionAbstract
It has been 75 yr since leaf respiratory metabolism in the light (day respiration) was identified as a low-flux metabolic pathway that accompanies photosynthesis. In principle, it provides carbon backbones for nitrogen assimilation and evolves CO2 and thus impacts on plant carbon and nitrogen balances. However, for a long time, uncertainties have remained as to whether techniques used to measure day respiratory efflux were valid and whether day respiration responded to environmental gaseous conditions. In the past few years, significant advances have been made using carbon isotopes, ‘omics’ analyses and surveys of respiration rates in mesocosms or ecosystems. There is substantial evidence that day respiration should be viewed as a highly dynamic metabolic pathway that interacts with photosynthesis and photorespiration and responds to atmospheric CO2 mole fraction. The view of leaf day respiration as a constant and/or negligible parameter of net carbon exchange is now outdated and it should now be regarded as a central actor of plant carbon-use efficiency.
Taylor, Benton N, et al.Growth and physiology of a dominant understory shrub, Hamamelis virginiana, following canopy disturbance in a temperate hardwood forest”. Canadian Journal of Forest Research 47 (2017): , 47, 193-202. Web. Publisher's VersionAbstract

As global climatic changes increase plant susceptibility to large-scale disturbances such as drought and pathogens, understory responses to these disturbances will become increasingly important to long-term forest dynamics. To better understand understory responses to canopy disturbance, we measured changes in the growth and physiology of the dominant understory shrub, American witch-hazel (Hamamelis virginiana L.), in response to girdling of canopy oaks in a temperate hardwood forest of the northeastern United States. Changes in the growth and physiology of H. virginiana may be important to the regeneration of northeastern temperate forests, as this common shrub largely shapes the microenvironment for seedlings on the forest floor where it occurs. Canopy disturbance by girdling resulted in significant increases in light and soil nitrogen availability. In response to these environmental changes, basal-area growth of H. virginianaincreased by an average 334%. This growth increase corresponded to significant increases in foliar nitrogen, respiration, and leaf chlorophyll and carotenoid concentrations. These findings indicate improved environmental conditions and increased growth for this understory shrub following the loss of dominant canopy trees. This study suggests that following large-scale canopy disturbance, H. virginiana and shrubs like it may play an important role in competing for soil N and shading seedlings of regenerating canopy species.

Gerlein-Safdi, Cynthia, et al.Leaf water 18O and 2H maps show directional enrichment discrepancy in Colocasia esculenta”. Plant, Cell and Environment 40 (2017): , 40, 2095-2108. Web. Publisher's VersionAbstract

Spatial patterns of leaf water isotopes are challenging to predict because of the intricate link between vein and lamina water. Many models have attempted to predict these patterns, but to date most have focused on monocots with parallel veins. These provide a simple system to study, but do not represent the majority of plant species. Here, a new protocol is developed using a Picarro induction module coupled to a cavity ringdown spectrometer to obtain maps of the leaf water isotopes (18O and D). The technique is applied to Colocasia esculenta leaves. The results are compared to isotope ratio mass spectrometry. In C. esculenta, a large enrichment in the radial direction is observed, but not in the longitudinal direction. The string-of-lakes model fails to predict the observed patterns, while the Farquhar-Gan model is more successful, especially when enrichment is accounted for along the radial and longitudinal direction together. Our results show that reticulate veined leaves experience a larger enrichment along the axis of the secondary veins than along the midrib. We hypothesize that this is due to the higher minor/major vein ratio that leads to longer pathways between major veins and sites of evaporation. 

Tcherkez, Guillaume G. B., et al.Tracking the origins of the Kok effect, 73 years after its discovery”. New Phytolologist 214 (2017): , 214, 506-510. Web. Publisher's VersionAbstract

The 18th New Phytologist workshop was dedicated to possible causes of the Kok effect, the typical break in the light response curve of net photosynthesis. Available data obtained since its discovery in 1948 show that the effect is not purely caused by a down-regulation of respiration, contrary to the commonly accepted view. However, estimates of leaf respiratory rates obtained in various ecosystems with techniques including the Kok method appear to be widely consistent across different studies, suggesting that Kok-derived values can be used as a surrogate for actual leaf respiration rates in the light. 

Atkin, Owen K, et al.Global variability in leaf respiration in relation to climate, plant functional types and leaf traits”. New Phytologist 206.2 (2015): , 206, 2, 614-636. Web. Publisher's VersionAbstract
  • Leaf dark respiration (Rdark) is an important yet poorly quantified component of the global carbon cycle. Given this, we analyzed a new global database of Rdark and associated leaf traits.
  • Data for 899 species were compiled from 100 sites (from the Arctic to the tropics). Several woody and nonwoody plant functional types (PFTs) were represented. Mixed-effects models were used to disentangle sources of variation in Rdark.
  • Area-based Rdark at the prevailing average daily growth temperature (T) of each site increased only twofold from the Arctic to the tropics, despite a 20°C increase in growing T (8–28°C). By contrast, Rdark at a standard T (25°C, Rdark25) was threefold higher in the Arctic than in the tropics, and twofold higher at arid than at mesic sites. Species and PFTs at cold sites exhibited higher Rdark25 at a given photosynthetic capacity (Vcmax25) or leaf nitrogen concentration ([N]) than species at warmer sites. Rdark25 values at any given Vcmax25 or [N] were higher in herbs than in woody plants.
  • The results highlight variation in Rdark among species and across global gradients in T and aridity. In addition to their ecological significance, the results provide a framework for improving representation of Rdark in terrestrial biosphere models (TBMs) and associated land-surface components of Earth system models (ESMs).
Gauthier, Paul PG, et al.Drought increases heat tolerance of leaf respiration in Eucalyptus globulus saplings grown under both ambient and elevated atmospheric [CO2] and temperature”. Journal of experimental Botany (2014). Web. Publisher's VersionAbstract

Climate change is resulting in increasing atmospheric [CO2], rising growth temperature (T), and greater frequency/severity of drought, with each factor having the potential to alter the respiratory metabolism of leaves. Here, the effects of elevated atmospheric [CO2], sustained warming, and drought on leaf dark respiration (Rdark), and the short-term T response of Rdark were examined in Eucalyptus globulus. Comparisons were made using seedlings grown under different [CO2],

T, and drought treatments. Using high resolution T–response curves of Rdark measured over the 15–65 °C range, it was found that elevated [CO2], elevated growth T, and drought had little effect on rates of Rdark measured at T <35 °C and that there was no interactive effect of [CO2], growth T, and drought on T response of Rdark. However, drought increased Rdark at high leaf T typical of heatwave events (35–45 °C), and increased the measuring T at which maximal rates of Rdark occurred (Tmax) by 8 °C (from 52 °C in well-watered plants to 60 °C in drought-treated plants). Leaf starch and soluble sugars decreased under drought and elevated growth T, respectively, but no effect was found under elevated [CO2]. Elevated [CO2] increased the Q10 of Rdark (i.e. proportional rise in Rdark per 10 °C) over the 15–35 °C range, while drought increased Q10 values between 35 °C and 45 °C. Collectively, the study highlights the dynamic nature of the dependence of Rdark in plants experiencing future climate change scenarios, particularly with respect to drought and elevated [CO2]. 

Boex-Fontvieille, E, et al.A new anaplerotic respiratory pathway involving lysine biosynthesis in isocitrate dehydrogenase-deficient Arabidopsis mutants”. New Phytologist (2013). Web. Publisher's VersionAbstract

The cornerstone of carbon (C) and nitrogen (N) metabolic interactions – respiration – is presently not well understood in plant cells: the source of the key intermediate 2-oxoglutarate (2OG), to which reduced N is combined to yield glutamate and glutamine, remains somewhat unclear.
We took advantage of combined mutations of NAD- and NADP-dependent isocitrate dehydrogenase activity and investigated the associated metabolic effects in Arabidopsis leaves (the major site of N assimilation in this genus), using metabolomics and 13C-labelling techniques.
We show that a substantial reduction in leaf isocitrate dehydrogenase activity did not lead to changes in the respiration efflux rate but respiratory metabolism was reorchestrated: 2OG production was supplemented by a metabolic bypass involving both lysine synthesis and degradation.
Although the recycling of lysine has long been considered important in sustaining respiration, we show here that lysine neosynthesis itself participates in an alternative respiratory pathway. Lys metabolism thus contributes to explaining the metabolic flexibility of plant leaves and the effect (or the lack thereof) of respiratory mutations.

Gauthier, Paul PG, et al.Metabolic origin of d15N values in nitrogenous compounds from Brassica napus L. leaves”. Plant, Cell & Environment 36 (2013): , 36, 128-137. Web. Publisher's VersionAbstract

Nitrogen isotope composition (d15N) in plant organic matter is currently used as a natural tracer of nitrogen acquisition efficiency. However, the d15N value of whole leaf material does not properly reflect the way in which N is assimilated because isotope fractionations along metabolic reactions may cause substantial differences among leaf compounds. In other words, any change in metabolic com- position or allocation pattern may cause undesirable vari- ability in leaf d15N. Here, we investigated the d15N in different leaf fractions and individual metabolites from rapeseed (Brassica napus) leaves. We show that there were substantial differences in d15N between nitrogenous com- pounds (up to 30‰) and the content in (15N enriched) nitrate had a clear influence on leaf d15N. Using a simple steady-state model of day metabolism, we suggest that the d15N value in major amino acids was mostly explained by isotope fractionation associated with isotope effects on enzyme-catalysed reactions in primary nitrogen metabo- lism. d15N values were further influenced by light versus dark conditions and the probable occurrence of alternative biosynthetic pathways. We conclude that both biochemical pathways (that fractionate between isotopes) and nitrogen sources (used for amino acid production) should be consid- ered when interpreting the d15N value of leaf nitrogenous compounds.

W., Rundel P, et al.Predicting Polylepis distribution: vulnerable and increasingly important Andean woodlands”. Revista peruana de biologia 19.2 (2012): , 19, 2, 205-212. Print. Zutta et al 2012.pdf
Gauthier, Paul PG, et al.In folio isotopic tracing demonstrates that nitrogen assimilation into glutamate is mostly independent from current CO2 assimilation in illuminated leaves of Brassica napus”. New Phytologist 185 (2010): , 185, 988-999. Web. Publisher's VersionAbstract

• Nitrogen assimilation in leaves requires primary NH2 acceptors that, in turn, originate from primary carbon metabolism. Respiratory metabolism is believed to provide such acceptors (such as 2-oxoglutarate), so that day respiration is com- monly seen as a cornerstone for nitrogen assimilation into glutamate in illuminated leaves. However, both glycolysis and day respiratory CO2 evolution are known to be inhibited by light, thereby compromising the input of recent photosynthetic car- bon for glutamate production.
• In this study, we carried out isotopic labelling experiments with 13CO2 and 15N- ammonium nitrate on detached leaves of rapeseed (Brassica napus), and per- formed 13C- and 15N-nuclear magnetic resonance analyses.
• Our results indicated that the production of 13C-glutamate and 13C-glutamine under a 13CO2 atmosphere was very weak, whereas 13C-glutamate and 13C-gluta- mine appeared in both the subsequent dark period and the next light period under a 12CO2 atmosphere. Consistently, the analysis of heteronuclear (13C–15N) interac- tions within molecules indicated that most 15N-glutamate and 15N-glutamine mol- ecules were not 13C labelled after 13C ⁄ 15N double labelling. That is, recent carbon atoms (i.e. 13C) were hardly incorporated into glutamate, but new glutamate mol- ecules were synthesized, as evidenced by 15N incorporation.
• We conclude that the remobilization of night-stored molecules plays a signifi- cant role in providing 2-oxoglutarate for glutamate synthesis in illuminated rape- seed leaves, and therefore the natural day : night cycle seems critical for nitrogen assimilation.

Tcherkez, G, et al.In folio respiratory fluxomics revealed by 13-C-isotopic labeling and H/D isotope effects highlight the non-cyclic nature of the tricarboxylic acid "cycle" in illuminated leaves”. Plant physiology 151 (2009): , 151, 620 - 630. Web. Publisher's VersionAbstract

While the possible importance of the tricarboxylic acid (TCA) cycle reactions for leaf photosynthesis operation has been recognized, many uncertainties remain on whether TCA cycle biochemistry is similar in the light compared with the dark. It is widely accepted that leaf day respiration and the metabolic commitment to TCA decarboxylation are down-regulated in illuminated leaves. However, the metabolic basis (i.e. the limiting steps involved in such a down-regulation) is not well known. Here, we investigated the in vivo metabolic fluxes of individual reactions of the TCA cycle by developing two isotopic methods, 13C tracing and fluxomics and the use of H/D isotope effects, with Xanthium strumarium leaves. We provide evidence that the TCA “cycle” does not work in the forward direction like a proper cycle but, rather, operates in both the reverse and forward directions to produce fumarate and glutamate, respectively. Such a functional division of the cycle plausibly reflects the compromise between two contrasted forces: (1) the feedback inhibition by NADH and ATP on TCA enzymes in the light, and (2) the need to provide pH-buffering organic acids and carbon skeletons for nitrate absorption and assimilation.