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Leaf respiration in the dark (R-dark) is often measured at a single time during the day, with hot-acclimation lowering R-dark at a common measuring temperature. However, it is unclear whether the diel cycle influences the extent of thermal acclimation of R-dark, or how temperature and time of day interact to influence respiratory metabolites. To examine these issues, we grew rice under 25 degrees C : 20 degrees C, 30 degrees C : 25 degrees C and 40 degrees C : 35 degrees C day : night cycles, measuring R-dark and changes in metabolites at five time points spanning a single 24-h period. R-dark differed among the treatments and with time of day. However, there was no significant interaction between time and growth temperature, indicating that the diel cycle does not alter thermal acclimation of R-dark. Amino acids were highly responsive to the diel cycle and growth temperature, and many were negatively correlated with carbohydrates and with organic acids of the tricarboxylic acid (TCA) cycle. Organic TCA intermediates were significantly altered by the diel cycle irrespective of growth temperature, which we attributed to light-dependent regulatory control of TCA enzyme activities. Collectively, our study shows that environmental disruption of the balance between respiratory substrate supply and demand is corrected for by shifts in TCA-dependent metabolites.

The disruption of photorespiration in GDC knockdown rice plants alters leaf photorespiratory 13CO2 fractionation and carbon isotope exchange., The influence of reduced glycine decarboxylase complex (GDC) activity on leaf atmosphere CO2 and 13CO2 exchange was tested in transgenic Oryza sativa with the GDC H-subunit knocked down in leaf mesophyll cells. Leaf measurements on transgenic gdch knockdown and wild-type plants were carried out in the light under photorespiratory and low photorespiratory conditions (i.e. 18.4 kPa and 1.84 kPa atmospheric O2 partial pressure, respectively), and in the dark. Under approximately current ambient O2 partial pressure (18.4 kPa pO2), the gdch knockdown plants showed an expected photorespiratory-deficient phenotype, with lower leaf net CO2 assimilation rates (A) than the wild-type. Additionally, under these conditions, the gdch knockdown plants had greater leaf net discrimination against 13CO2 (Δo) than the wild-type. This difference in Δo was in part due to lower 13C photorespiratory fractionation (f) ascribed to alternative decarboxylation of photorespiratory intermediates. Furthermore, the leaf dark respiration rate (Rd) was enhanced and the 13CO2 composition of respired CO2 (δ13CRd) showed a tendency to be more depleted in the gdch knockdown plants. These changes in Rd and δ13CRd were due to the amount and carbon isotopic composition of substrates available for dark respiration. These results demonstrate that impairment of the photorespiratory pathway affects leaf 13CO2 exchange, particularly the 13C decarboxylation fractionation associated with photorespiration.

The engineering process of C4 photosynthesis into C3 plants requires an increased activity of phosphoenolpyruvate carboxylase (PEPC) in the cytosol of leaf mesophyll cells. The literature varies on the physiological effect of transgenic maize (Zea mays) PEPC (ZmPEPC) leaf expression in Oryza sativa (rice). Therefore, to address this issue, leaf-atmosphere CO2 and 13CO2 exchanges were measured, both in the light (at atmospheric O2 partial pressure of 1.84 kPa and at different CO2 levels) and in the dark, in transgenic rice expressing ZmPEPC and wild-type (WT) plants. The in vitro PEPC activity was 25 times higher in the PEPC overexpressing (PEPC-OE) plants (~20% of maize) compared to the negligible activity in WT. In the PEPC-OE plants, the estimated fraction of carboxylation by PEPC (β) was ~6% and leaf net biochemical discrimination against 13CO2[Formula: see text] was ~ 2‰ lower than in WT. However, there were no differences in leaf net CO2 assimilation rates (A) between genotypes, while the leaf dark respiration rates (Rd) over three hours after light-dark transition were enhanced (~ 30%) and with a higher 13C composition [Formula: see text] in the PEPC-OE plants compared to WT. These data indicate that ZmPEPC in the PEPC-OE rice plants contributes to leaf carbon metabolism in both the light and in the dark. However, there are some factors, potentially posttranslational regulation and PEP availability, which reduce ZmPEPC activity in vivo.

The engineering process of C4 photosynthesis into C3 plants requires an increased activity of phosphoenolpyruvate carboxylase (PEPC) in the cytosol of leaf mesophyll cells. The literature varies on the physiological effect of transgenic maize (Zea mays) PEPC (ZmPEPC) leaf expression in Oryza sativa (rice). Therefore, to address this issue, leaf–atmosphere CO2 and 13CO2 exchanges were measured, both in the light (at atmospheric O2 partial pressure of 1.84 kPa and at different CO2 levels) and in the dark, in transgenic rice expressing ZmPEPC and wild-type (WT) plants. The in vitro PEPC activity was 25 times higher in the PEPC overexpressing (PEPC-OE) plants (~20% of maize) compared to the negligible activity in WT. In the PEPC-OE plants, the estimated fraction of carboxylation by PEPC (β) was ~6% and leaf net biochemical discrimination against 13CO2\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\left( {\Delta_{\text{bio}} } \right)$$\end{document}Δbio was ~ 2‰ lower than in WT. However, there were no differences in leaf net CO2 assimilation rates (A) between genotypes, while the leaf dark respiration rates (Rd) over three hours after light–dark transition were enhanced (~ 30%) and with a higher 13C composition \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\left( {\delta^{ 1 3} {\text{C}}_{\text{Rd}} } \right)$$\end{document}δ13CRd in the PEPC-OE plants compared to WT. These data indicate that ZmPEPC in the PEPC-OE rice plants contributes to leaf carbon metabolism in both the light and in the dark. However, there are some factors, potentially posttranslational regulation and PEP availability, which reduce ZmPEPC activity in vivo. Electronic supplementary material The online version of this article (10.1007/s11120-019-00655-4) contains supplementary material, which is available to authorized users.

In the present study we investigated variations in leaf respiration in darkness (RD) and light (RL), and associated traits in response to season, and along a gradient of soil moisture, in Mediterranean woodland dominated by holm oak (Quercus ilex L.) in central and north-eastern Spain respectively. On seven occasions during the year in the central Spain site, and along the soil moisture gradient in north-eastern Spain, we measured rates of leaf RD, RL (using the Kok method), light-saturated photosynthesis (A) and related light response characteristics, leaf mass per unit area (MA) and leaf nitrogen (N) content. At the central Spain site, significant seasonal changes in soil water content and ambient temperature (T) were associated with changes in MA, foliar N, A and stomatal conductance. RD measured at the prevailing daily T and in instantaneous R-T responses, displayed signs of partial acclimation and was not significantly affected by time of year. RL was always less than, and strongly related to, RD, and RL/RD did not vary significantly or systematically with seasonal changes in T or soil water content. Averaged over the year, RL/RD was 0.66±0.05s.e. (n≤14) at the central Spain site. At the north-eastern Spain site, the soil moisture gradient was characterised by increasing MA and RD, and reduced foliar N, A, and stomatal conductance as soil water availability decreased. Light inhibition of R occurred across all sites (mean RL/RD≤0.69±0.01s.e. (n≤18)), resulting in ratios of RL/A being lower than for RD/A. Importantly, the degree of light inhibition was largely insensitive to changes in soil water content. Our findings provide evidence for a relatively constrained degree of light inhibition of R (RL/RD ∼ 0.7, or inhibition of ∼30%) across gradients of water availability, although the combined impacts of seasonal changes in both T and soil water content increase the range of values expressed. The findings thus have implications in terms of the assumptions made by predictive models that seek to account for light inhibition of R, and for our understanding of how environmental gradients impact on leaf trait relationships in Mediterranean plant communities.

In tropical mountains, trees are the dominant life form from sea level to above 4,000-m altitude under highly variable thermal conditions (range of mean annual temperatures: 28°C). How light-saturated net photosynthesis of tropical trees adapts to variation in temperature, atmospheric CO2 concentration, and further environmental factors, that change along elevation gradients, is not precisely known. With gas exchange measurements in mature trees, we determined light-saturated net photosynthesis at ambient temperature (T) and [CO2] (Asat) of 40 tree species from 21 families in tropical mountain forests at 1000-, 2000-, and 3000-m elevation in southern Ecuador. We tested the hypothesis that stand-level averages of Asat and leaf dark respiration (RD) per leaf area remain constant with elevation. Stand-level means of Asat were 8.8, 11.3, and 7.2 μmol CO2 m−2 s−1; those of RD 0.8, 0.6, and 0.7 μmol CO2 m−2 s−1 at 1000-, 2000-, and 3000-m elevation, respectively, with no significant altitudinal trend. We obtained coefficients of among-species variation in Asat and RD of 20–53% (n = 10–16 tree species per stand). Examining our data in the context of a pan-tropical Asat data base for mature tropical trees (c. 170 species from 18 sites at variable elevation) revealed that area-based Asat decreases in tropical mountains by, on average, 1.3 μmol CO2 m−2 s−1 per km altitude increase (or by 0.2 μmol CO2 m−2 s−1 per K temperature decrease). The Asat decrease occurred despite an increase in leaf mass per area with altitude. Local geological and soil fertility conditions and related foliar N and P concentrations considerably influenced the altitudinal Asat patterns. We conclude that elevation is an important influencing factor of the photosynthetic activity of tropical trees. Lowered Asat together with a reduced stand leaf area decrease canopy C gain with elevation in tropical mountains. peerReviewed

Quercus ilex L. seedlings were exposed in open-top chambers for one growing season to three levels of ozone (O3 ): charcoal filtered air, non-filtered air supplemented with +30% or +74% ambient air O3 . Key functional parameters related to photosynthetic performance and stomatal density were measured to evaluate the response mechanisms of Q. ilex to chronic O3 exposure, clarifying how ecophysiological traits are modulated during the season in an ozone-enriched environment. Dark respiration showed an early response to O3 exposure, increasing approximately 45% relative to charcoal-filtered air in both O3 enriched treatments. However, at the end of the growing season, maximum rate of assimilation (Amax ) and stomatal conductance (gs ) showed a decline (-13% and -36%, for Amax and gs , respectively) only in plants under higher O3 levels. Photosystem I functionality supported the capacity of Q. ilex to cope with oxidative stress by adjusting the energy flow partitioning inside the photosystems. The response to O3 was also characterised by increased stomatal density in both O3 enriched treatments relative to controls. Our results suggest that in order to improve the reliability of metrics for O3 risk assessment, the seasonal changes in the response of gs and photosynthetic machinery to O3 stress should be considered.

Determining climate change feedbacks from tropical rainforests requires an understanding of how carbon gain through photosynthesis and loss through respiration will be altered. One of the key changes that tropical rainforests may experience under future climate change scenarios is reduced soil moisture availability. In this study we examine if and how both leaf photosynthesis and leaf dark respiration acclimate following more than 12 years of experimental soil moisture deficit, via a through-fall exclusion experiment (TFE) in an eastern Amazonian rainforest. We find that experimentally drought-stressed trees and taxa maintain the same maximum leaf photosynthetic capacity as trees in corresponding control forest, independent of their susceptibility to drought-induced mortality. We hypothesise that photosynthetic capacity is maintained across all treatments and taxa to take advantage of short-lived periods of high moisture availability, when stomatal conductance (gs ) and photosynthesis can increase rapidly, potentially compensating for reduced assimilate supply at other times. Average leaf dark respiration (Rd ) was elevated in the TFE-treated forest trees relative to the control by 28.2±2.8% (mean ± one standard error). This mean Rd value was dominated by a 48.5±3.6% increase in the Rd of drought-sensitive taxa, and likely reflects the need for additional metabolic support required for stress-related repair, and hydraulic or osmotic maintenance processes. Following soil moisture deficit that is maintained for several years, our data suggest that changes in respiration drive greater shifts in the canopy carbon balance, than changes in photosynthetic capacity. This article is protected by copyright. All rights reserved. (Less)

The present study explores the potential contribution of the energy requirements associated with nocturnal carbohydrate export to (1) the fraction of dark respira­ tion correlating with leaf nitrogen concentration and (2) the dark respiration of mature source leaves. To this end, we determined the nocturnal carbohydrate-export rates from leaves with an optimal nitrogen supply, and the correlation between the nitrogen concentration and the dark respiration of leaves. The specific energy costs of carbohydrate export from starch-storing source leaves were determined both experimentally and theor­ etically. The present estimate of the specific energy cost involved in carbohydrate export as obtained by linear regression (0. 70 mol C02 (mol sucrose) - 1 ), agrees well with both literature data obtained by different methods (0.4 7 to 1.26 mol C02 (mol sucrose) - 1 ) and the theoretically calculated range for starch-storing species (0.40 to 1.20 mol C02 (mol sucrose) - 1 ). The conversion of starch in the chloroplast to sucrose in the cytosol is a major energy-requiring process. Maximally 42 to '107'% of the slope of the relationship between respiration rate and organic nitrogen concentration of primary bean leaves, may be ascribed to the energy costs associated with nocturnal export of carbohyd­ rates. Total energy costs associated with export were derived from the product of the specific costs of carbo- hydrate export and the export rates, either measured on full-grown (primary) leaves of potato and bean or derived from the literature. These export costs account, on average, for 29% of the dark respiration rate in starch-storing species. We conclude that nocturnal carbohydrate export is a major energy-requiring pro­ cess in starch-storing species.

To study the effects of warming and declining species richness on the carbon balance of grassland communities, model ecosystems containing one, three or nine species were exposed to ambient and elevated (ambient +3°C) air temperature. In this paper, we analyze measured ecosystem CO2 fluxes to test whether ecosystem photosynthesis and respiration had acclimated to warming after 28 months of continuous heating, and whether the degree of acclimation depended on species richness. In order to test whether acclimation occurred, short term temperature response curves were established for all communities in both treatments. At similar temperatures, lower flux rates in the heated communities as compared to the unheated communities would indicate thermal acclimation. Because plant cover was significantly higher in the heated treatment, we normalized the data for plant cover. Subsequently, down-regulation of both photosynthesis and respiration was observed. Although CO2 fluxes were larger in communities with higher species richness, species richness did not affect the degree of acclimation to warming. These results imply that models need to take thermal acclimation into account to simulate photosynthesis and respiration in a warmer world., This research was funded by the Fund for Scientific Research – Flanders (Belgium) as project “effects of biodiversity loss and climate warming on carbon sequestration mechanisms in terrestrial ecosystems”, contract #G.0434.03N. H. J. De Boeck holds a grant from the Institute for the Promotion of Innovation by science and Technology in Flanders. P. Serrano-Ortiz benefits from a pre-doctoral grant from the Junta de Andalucía.

The engineering process of C4 photosynthesis into C3 plants requires an increased activity of phosphoenolpyruvate carboxylase (PEPC) in the cytosol of leaf mesophyll cells. The literature varies on the physiological effect of transgenic maize (Zea mays) PEPC (ZmPEPC) leaf expression in Oryza sativa (rice). Therefore, to address this issue, leaf-atmosphere CO2 and 13CO2 exchanges were measured, both in the light (at atmospheric O2 partial pressure of 1.84 kPa and at different CO2 levels) and in the dark, in transgenic rice expressing ZmPEPC and wild-type (WT) plants. The in vitro PEPC activity was 25 times higher in the PEPC overexpressing (PEPC-OE) plants (~20% of maize) compared to the negligible activity in WT. In the PEPC-OE plants, the estimated fraction of carboxylation by PEPC (β) was ~6% and leaf net biochemical discrimination against 13CO2[Formula: see text] was ~ 2‰ lower than in WT. However, there were no differences in leaf net CO2 assimilation rates (A) between genotypes, while the leaf dark respiration rates (Rd) over three hours after light-dark transition were enhanced (~ 30%) and with a higher 13C composition [Formula: see text] in the PEPC-OE plants compared to WT. These data indicate that ZmPEPC in the PEPC-OE rice plants contributes to leaf carbon metabolism in both the light and in the dark. However, there are some factors, potentially posttranslational regulation and PEP availability, which reduce ZmPEPC activity in vivo.