C3 plants enhance rates of photosynthesis by reassimilating photorespired and respired CO2

Photosynthetic carbon gain in plants using the C₃ photosynthetic pathway is substantially inhibited by photorespiration in warm environments, particularly in atmospheres with low CO₂ concentrations. Unlike C₄ plants, C₃ plants are thought to lack any mechanism to compensate for the loss of photosynthetic productivity caused by photorespiration. Here, for the first time, we demonstrate that the C₃ plants rice and wheat employ a specific mechanism to trap and reassimilate photorespired CO₂. A continuous layer of chloroplasts covering the portion of the mesophyll cell periphery that is exposed to the intercellular air space creates a diffusion barrier for CO₂ exiting the cell. This facilitates the capture and reassimilation of photorespired CO₂ in the chloroplast stroma. In both species, 24–38% of photorespired and respired CO₂ were reassimilated within the cell, thereby boosting photosynthesis by 8–11% at ambient atmospheric CO₂ concentration and 17–33% at a CO₂ concentration of 200 µmol mol^?1. Widespread use of this mechanism in tropical and subtropical C₃ plants could explain why the diversity of the world's C₃ flora, and dominance of terrestrial net primary productivity, was maintained during the Pleistocene, when atmospheric CO₂ concentrations fell below 200 µmol mol^?1.     

Estimation of Photorespiration of Green Plants and of their Mesophyll Resistance to CO2 Uptake

An estimate of photorespiration is obtained from the relationshipbetween the net exchange of CO₂ of the leaf and the internalCO₂ concentration, i.e. within the mesophyll intercellular spaces.The latter is obtained by calculation, taking into account thecombined epidermal and boundary-layer resistances between thebulk atmosphere and the mesophyll intercellular spaces. Thelinear part of this relationship (at low CO₂ concentrations)is extrapolated to zero internal concentration, at which noneof the intercellular photorespired CO₂ is available for reassimilation.The calculated output of CO₂ under such conditions providesan estimate of photorespiration, but, by failing to take intoaccount intracellular reassimilation of photorespired CO₂ underestimatesactual photorespiration. As the slope of this linear relationshiprepresents the mesophyll (intracellular) resistance to CO₂ uptake,this procedure was used to recalculate published data on effectsof light intensity and of oxygen concentration on net photosynthesis.The analysis showed that increased oxygen concentration anddecreased light intensity reduced photosynthesis largely byincreasing mesophyll resistance to CO₂ uptake. It is suggestedthat the CO₂ compensation point () is a function of both photorespiration(L) and mesophyll resistance (r_m): = L. r_m.     

Photosynthetic and photorespiratory characteristics of flaveria species

The genus Flaveria shows evidence of evolution in the mechanism of photosynthesis as its 21 species include C₃, C₃-C₄, C₄-like, and C₄ plants. In this study, several physiological and biochemical parameters of photosynthesis and photorespiration were measured in 18 Flaveria species representing all the photosynthetic types. The 10 species classified as C₃-C₄ intermediates showed an inverse continuum in level of photorespiration and development of the C₄ syndrome. This ranges from F. sonorensis with relatively high apparent photorespiration and lacking C₄ photosynthesis to F. Among the intermediates, the photosynthetic CO₂ compensation points at 30°C and 1150 micromoles quanta per square meter per second varied from 9 to 29 microbars. The values for the three C₄-like species varied from 3 to 6 microbars, similar to those measured for the C₄ species. The activities of the photorespiratory enzymes glycolate oxidase, hydroxypyruvate reductase, and serine hydroxymethyltransferase decreased progressively from C₃ to C₃-C₄ to C₄-like and C₄ species. On the other hand, most intermediates had higher levels of phosphenolpyruvate carboxylase and NADP-malic enzyme than C₃ species, but generally lower activities compared to C₄-like and C₄ species. The levels of these C₄ enzymes are correlated with the degree of C₄ photosynthesis, based on the initial products of photosynthesis. Another indication of development of the C₄ syndrome in C₃-C₄ Flaveria species was their intermediate chlorophyll a/b ratios. The chlorophyll a/b ratios of the various Flaveria species are highly correlated with the degree of C₄ photosynthesis suggesting that the photochemical machinery is progressively altered during evolution in order to meet the specific energy requirements for operating the C₄ pathway. In the progression from C₃ to C₄ species in Flaveria, the CO₂ compensation point decreased more rapidly than did the decrease in O₂ inhibition of photosynthesis or the increase in the degree of C₄ photosynthesis. These results suggest that the reduction in photorespiration during evolution occurred initially by refixation of photorespired CO₂ and prior to substantive reduction in O₂ inhibition and development of the C₄ syndrome. However, further reduction in O₂ inhibition in some intermediates and C₄-like species is considered primarily due to the development of the C₄ syndrome. Thus, the evolution of C₃-C₄ intermediate photosynthesis likely occurred in response to environmental conditions which limit the intercellular CO₂ concentration first via refixation of photorespired CO₂, followed by development of the C₄ syndrome.     

Influence of certain environmental factors on photosynthesis and photorespiration in Simmondsia chinensis

The response of net photosynthesis and apparent light respiration to changes in [O₂], light intensity, and drought stress was determined by analysis of net photosynthetic CO₂ response curves. Low [O₂] treatment resulted in a large reduction in the rate of photorespiratory CO₂ evolution. Lightintensity levels influenced the maximum net photosynthetic rate at saturating [CO₂]. These results indicate that [CO₂], [O₂] and light intensity affect the levels of substrates involved in the enzymatic reactions of photosynthesis and photorespiration. Intracellular resistance to CO₂ uptake decreased in low [O₂] and increased at low leaf water potentials. This response reflects changes in the efficiency with which photosynthetic and photorespiratory substrates are formed and utilized. Water stress had no effect on the CO₂ compensation point or the [CO₂] at which net photosynthesis began to saturate at high light intensity. The relationship between these data and recently published in-vitro kinetic measurements with ribulose-diphosphate carboxylase is discussed.Abbreviations C _w intracellular CO₂ concentration - F _gross gross photosynthesis - F _net net photosynthesis - I light intensity - R _L light respiration rate - r _c carboxylation resistance - r ₈ leaf gas-phase resistance - r _i intracellular resistance; to CO₂ uptake - r _t resistance to CO₂ flux between the intercellular spaces and the carboxylation sites - T _L leaf temperature - _t leaf water potential - CO₂ compensation point     

Photosynthesis, Leaf Anatomy, and Morphology of Progeny from Hybrids between C(3) and C(3)/C(4)Panicum Species

Species in the Laxa group of Panicum have C₃ or C₃/C₄ photosynthesis based on leaf anatomical and CO₂ exchange characteristics. Hybrids were previously made between C₃/C₄ and C₃ species in this group (RH Brown et al. 1985 Plant Physiol 77: 653-658). In this paper, CO₂ exchange, morphological, and leaf anatomical characteristics of F₂ or F₅ progeny from colchicine-induced amphiploids of C₃/C₄ × C₃ hybrids (Panicum milioides Nees ex Trin. [C₃/C₄] × Panicum laxum Mez [C₃] and Panicum spathellosum Doell [C₃/C₄] × Panicum boliviense Hack. [C₃]) were studied.

There were no differences found in morphology or physiology between the amphiploids and the F₁ hybrids from which they were produced. In the segregating progeny, CO₂ compensation concentration and photorespiration values typical of C₃, but not of C₃/C₄ plants, were recovered. Progeny were found from both crosses which possessed O₂ inhibition of apparent photosynthesis typical of the parents, and in the case of the P. milioides × P. laxum cross, leaf anatomy and overall plant morphology typical of the parents were observed in some progeny. The progeny were found to possess recombinations of various traits associated with reduced photorespiration, so that no correlation existed among O₂ inhibition of apparent photosynthesis, CO₂ compensation concentration, and leaf anatomical traits. One plant was especially noteworthy in possessing leaf anatomy typical of C₃/C₄ plants, but with CO₂ exchange characteristics of C₃ plants.

     

Carbon dioxide responsiveness mitigates rice yield loss under high night temperature

Increasing night-time temperatures are a major threat to sustaining global rice (Oryza sativa L.) production. A simultaneous increase in [CO₂] will lead to an inevitable interaction between elevated [CO₂] (e[CO₂]) and high night temperature (HNT) under current and future climates. Here, we conducted field experiments to identify [CO₂] responsiveness from a diverse indica panel comprising 194 genotypes under different planting geometries in 2016. Twenty-three different genotypes were tested under different planting geometries and e[CO₂] using a free-air [CO₂] enrichment facility in 2017. The most promising genotypes and positive and negative controls were tested under HNT and e[CO₂] + HNT in 2018. [CO₂] responsiveness, measured as a composite response index on different yield components, grain yield, and photosynthesis, revealed a strong relationship (R² = 0.71) between low planting density and e[CO₂]. The most promising genotypes revealed significantly lower (P < 0.001) impact of HNT in high [CO₂] responsive (HCR) genotypes compared to the least [CO₂] responsive genotype. [CO₂] responsiveness was the major driver determining grain yield and related components in HCR genotypes with a negligible yield loss under HNT. A systematic investigation highlighted that active selection and breeding for [CO₂] responsiveness can lead to maintained carbon balance and compensate for HNT-induced yield losses in rice and potentially other C₃ crops under current and future warmer climates.

Active selection for carbon dioxide responsiveness in rice and other C₃ crops can mitigate yield loss induced by high night temperature.     

Interactive Effects of Elevated CO2 Concentration and Irrigation on Photosynthetic Parameters and Yield of Maize in Northeast China

Maize is one of the major cultivated crops of China, having a central role in ensuring the food security of the country. There has been a significant increase in studies of maize under interactive effects of elevated CO₂ concentration ([CO₂]) and other factors, yet the interactive effects of elevated [CO₂] and increasing precipitation on maize has remained unclear. In this study, a manipulative experiment in Jinzhou, Liaoning province, Northeast China was performed so as to obtain reliable results concerning the later effects. The Open Top Chambers (OTCs) experiment was designed to control contrasting [CO₂] i.e., 390, 450 and 550 µmol·mol⁻¹, and the experiment with 15% increasing precipitation levels was also set based on the average monthly precipitation of 5–9 month from 1981 to 2010 and controlled by irrigation. Thus, six treatments, i.e. C₅₅₀W_+15%, C₅₅₀W₀, C₄₅₀W_+15%, C₄₅₀W₀, C₃₉₀W_+15% and C₃₉₀W₀ were included in this study. The results showed that the irrigation under elevated [CO₂] levels increased the leaf net photosynthetic rate (P _n) and intercellular CO₂ concentration (C _i) of maize. Similarly, the stomatal conductance (G _s) and transpiration rate (T _r) decreased with elevated [CO₂], but irrigation have a positive effect on increased of them at each [CO₂] level, resulting in the water use efficiency (WUE) higher in natural precipitation treatment than irrigation treatment at elevated [CO₂] levels. Irradiance-response parameters, e.g., maximum net photosynthetic rate (P _nmax) and light saturation points (LSP) were increased under elevated [CO₂] and irrigation, and dark respiration (R _d) was increased as well. The growth characteristics, e.g., plant height, leaf area and aboveground biomass were enhanced, resulting in an improved of yield and ear characteristics except axle diameter. The study concluded by reporting that, future elevated [CO₂] may favor to maize when coupled with increasing amount of precipitation in Northeast China.     

A Guide to Establishing Water Potential of Aqueous Two-Phase Solutions (Polyethylene Glycol plus Dextran) by Amendment with Mannitol

A practical guide to calculating the mannitol (MAN) amendment required to achieve the desired water potential (Ψ) of polyethylene glycol/dextran (PEG/DEX) aqueous two-phase systems for protoplast purification is presented. The empirically generated equation Ψ = 305[PEG′]²[MAN] + 0.74[PEG′][MAN]T − 103[PEG′][MAN] + 5.6[PEG′]²T − 623[PEG′]² − 0.25[PEG′]T + 12.7[PEG′] − 0.078[MAN]T − 22.75[MAN]accurately predicts experimental Ψ (in bars). [PEG′] indicates the presence of DEX where [DEX] = [PEG]/(0.6−0.4[PEG]). The equation is applicable for these ranges: [PEG′] from 0.047 to 0.13 gram per gram H₂O; [MAN] from 0 to 0.7 molal; T from 4.5 to 40°C. Actual Ψ should differ from derived Ψ by no more than 8% for the least negative values to 4% for the most negative values. The Ψ for solutions of MAN, of PEG, and of DEX were also determined. Equations to fit data for each were generated. Analyses indicated a significant synergistic effect on Ψ when MAN is added to PEG/DEX and, at certain concentrations, between PEG and DEX.     

Two photosynthetic mechanisms mediating the low photorespiratory state in submersed aquatic angiosperms

The submersed angiosperms Myriophyllum spicatum L. and Hydrilla verticillata (L.f.) Royal exhibited different photosynthetic pulse-chase labeling patterns. In Hydrilla, over 50% of the ¹⁴C was initially in malate and aspartate, but the fate of the malate depended upon the photorespiratory state of the plant. In low photorespiration Hydrilla, malate label decreased rapidly during an unlabeled chase, whereas labeling of sucrose and starch increased. In contrast, for high photorespiration Hydrilla, malate labeling continued to increase during a 2-hour chase. Thus, malate formation occurs in both photorespiratory states, but reduced photorespiration results when this malate is utilized in the light. Unlike Hydrilla, in low photorespiration Myriophyllum, ¹⁴C incorporation was via the Calvin cycle, and less than 10% was in C₄ acids.

Ethoxyzolamide, a carbonic anhydrase inhibitor and a repressor of the low photorespiratory state, increased the label in glycolate, glycine, and serine of Myriophyllum. Isonicotinic acid hydrazide increased glycine labeling of low photorespiration Myriophyllum from 14 to 25%, and from 12 to 48% with high photorespiration plants. Similar trends were observed with Hydrilla. Increasing O₂ increased the per cent [¹⁴C]glycine and the O₂ inhibition of photosynthesis in Myriophyllum. In low photorespiration Myriophyllum, glycine labeling and O₂ inhibition of photosynthesis were independent of the CO₂ level, but in high photorespiration plants the O₂ inhibition was competitively decreased by CO₂. Thus, in low but not high photorespiration plants, glycine labeling and O₂ inhibition appeared to be uncoupled from the external [O₂]/[CO₂] ratio.

These data indicate that the low photorespiratory states of Hydrilla and Myriophyllum are mediated by different mechanisms, the former being C₄-like, while the latter resembles that of low CO₂-grown algae. Both may require carbonic anhydrase to enhance the use of inorganic carbon for reducing photorespiration.

     

Fixation of O(2) during Photorespiration: Kinetic and Steady-State Studies of the Photorespiratory Carbon Oxidation Cycle with Intact Leaves and Isolated Chloroplasts of C(3) Plants

Mass spectrometric techniques were used to trace the incorporation of [¹⁸O]oxygen into metabolites of the photorespiratory pathway. Glycolate, glycine, and serine extracted from leaves of the C₃ plants, Spinacia oleracea L., Atriplex hastata, and Helianthus annuus which had been exposed to [¹⁸O]oxygen at the CO₂ compensation point were heavily labeled with ¹⁸O. In each case one, and only one of the carboxyl oxygens was labeled. The abundance of ¹⁸O in this oxygen of glycolate reached 50 to 70% of that of the oxygen provided after only 5 to 10 seconds exposure to [¹⁸O]oxygen. Glycine and serine attained the same final enrichment after 40 and 180 seconds, respectively. This confirms that glycine and serine are synthesized from glycolate.

The labeling of photorespiratory intermediates in intact leaves reached a mean of 59% of that of the oxygen provided in the feedings. This indicates that at least 59% of the glycolate photorespired is synthesized with the fixation of molecular oxygen. This estimate is certainly conservative owing to the dilution of labeled oxygen at the site of glycolate synthesis by photosynthetic oxygen. We examined the yield of ¹⁸O in glycolate synthesized in vitro by isolated intact spinach chloroplasts in a system which permitted direct sampling of the isotopic composition of the oxygen at the site of synthesis. The isotopic enrichment of glycolate from such experiments was 90 to 95% of that of the oxygen present during the incubation.

The carboxyl oxygens of 3-phosphoglycerate also became labeled with ¹⁸O in 20- and 40-minute feedings with [¹⁸O]oxygen to intact leaves at the CO₂ compensation point. Control experiments indicated that this label was probably due to direct synthesis of 3-phosphoglycerate from glycolate during photorespiration. The mean enrichment of 3-phosphoglycerate was 14 ± 4% of that of glycine or serine, its precursors of the photorespiratory pathway, in 10 separate feeding experiments. It is argued that this constant dilution of label indicates a constant stoichiometric balance between photorespiratory and photosynthetic sources of 3-phosphoglycerate at the CO₂ compensation point.

Oxygen uptake sufficient to account for about half of the rate of ¹⁸O fixation into glycine in the intact leaves was observed with intact spinach chloroplasts. Oxygen uptake and production by intact leaves at the CO₂ compensation point indicate about 1.9 oxygen exchanged per glycolate photorespired. The fixation of molecular oxygen into glycolate plus the peroxisomal oxidation of glycolate to glyoxylate and the mitochondrial conversion of glycine to serine can account for up to 1.75 oxygen taken up per glycolate.

These studies provide new evidence which supports the current formulation of the pathway of photorespiration and its relation to photosynthetic metabolism. The experiments described also suggest new approaches using stable isotope techniques to study the rate of photorespiration and the balance between photorespiration and photosynthesis in vivo.

     

Internal CO(2) Measured Directly in Leaves : Abscisic Acid and Low Leaf Water Potential Cause Opposing Effects

Observations of nonuniform photosynthesis across leaves cast doubt on internal CO₂ partial pressures (p_i) calculated on the assumption of uniformity and can lead to incorrect conclusions about the stomatal control of photosynthesis. The problem can be avoided by measuring p_i directly because the assumptions of uniformity are not necessary. We therefore developed a method that allowed p_i to be measured continuously in situ for days at a time under growth conditions and used it to investigate intact leaves of sunflower (Helianthus annuus L.), soybean (Glycine max L. Merr.), and bush bean (Phaseolus vulgaris L.) subjected to high or low leaf water potentials (ψ_w) or high concentrations of abscisic acid (ABA). The leaves maintained a relatively constant differential (Δp) between ambient CO₂ and measured p_i throughout the light period when water was supplied. When water was withheld, ψ_w decreased and the stomata began to close, but measured p_i increased until the leaf reached a ψ_w of −1.76 (bush bean), −2.12 (sunflower) or −3.10 (soybean) megapascals, at which point Δp = 0. The increasing p_i indicated that stomata did not inhibit CO₂ uptake and a Δp of zero indicated that CO₂ uptake became zero despite the high availability of CO₂ inside the leaf. In contrast, when sunflower leaves at high ψ_w were treated with ABA, p_i did not increase and instead decreased rapidly and steadily for up to 8 hours even as ψ_w increased, as expected if ABA treatment primarily affected stomatal conductance. The accumulating CO₂ at low ψ_w and contrasting response to ABA indicates that photosynthetic biochemistry limited photosynthesis at low ψ_w but not at high ABA.     

Nitrogen assimilation and transpiration: key processes conditioning responsiveness of wheat to elevated [CO2] and temperature

Although climate scenarios have predicted an increase in [CO₂] and temperature conditions, to date few experiments have focused on the interaction of [CO₂] and temperature effects in wheat development. Recent evidence suggests that photosynthetic acclimation is linked to the photorespiration and N assimilation inhibition of plants exposed to elevated CO₂. The main goal of this study was to analyze the effect of interacting [CO₂] and temperature on leaf photorespiration, C/N metabolism and N transport in wheat plants exposed to elevated [CO₂] and temperature conditions. For this purpose, wheat plants were exposed to elevated [CO₂] (400 vs 700 µmol mol^?1) and temperature (ambient vs ambient + 4°C) in CO₂ gradient greenhouses during the entire life cycle. Although at the agronomic level, elevated temperature had no effect on plant biomass, physiological analyses revealed that combined elevated [CO₂] and temperature negatively affected photosynthetic performance. The limited energy levels resulting from the reduced respiratory and photorespiration rates of such plants were apparently inadequate to sustain nitrate reductase activity. Inhibited N assimilation was associated with a strong reduction in amino acid content, conditioned leaf soluble protein content and constrained leaf N status. Therefore, the plant response to elevated [CO₂] and elevated temperature resulted in photosynthetic acclimation. The reduction in transpiration rates induced limitations in nutrient transport in leaves of plants exposed to elevated [CO₂] and temperature, led to mineral depletion and therefore contributed to the inhibition of photosynthetic activity.     

A Transient Burst of CO(2) from Geranium Leaves during Illumination at Various Light Intensities as a Measure of Photorespiration

A transient CO₂ burst is exhibited by irradiated leaves of the C₃ plant geranium (Pelargonium X hortorum, Bailey) after the irradiance is quickly lowered. The light CO₂ burst appears to be related to photorespiration because of its irradiance dependency and its sensitivity to other environmental components such as CO₂ and O₂ concentration. The term post-lower-irradiance CO₂ burst or PLIB is used to describe the phenomenon. The PLIB appears to be a quantitative measurement of photorespiration with intact geranium leaves. The PLIB has been observed with intact leaves of other C₃ plants but not with C₄ leaves. Therefore, it is proposed that, after maximizing intact leaf photosynthetic rates and leaf chamber gas measuring conditions, photorespiration can be measured with intact C₃ leaves such as geranium as a transient post-lower-irradiance CO₂ burst.     

Degree of C(4) Photosynthesis in C(4) and C(3)-C(4)Flaveria Species and Their Hybrids : II. Inhibition of Apparent Photosynthesis by a Phosphoenolpyruvate Carboxylase Inhibitor

Hybrids have been made between species of Flaveria exhibiting varying levels of C₄ photosynthesis. The degree of C₄ photosynthesis expressed in four interspecific hybrids (Flaveria trinervia [C₄] × F. linearis [C₃-C₄], F. brownii [C₄-like] × F. linearis, and two three-species hybrids from F. trinervia × [F. brownii × F. linearis]) was estimated by inhibiting phosphoenolpyruvate carboxylase in vivo with 3,3-dichloro-2-dihydroxyphosphinoylmethyl-2-propenoate (DCDP). The inhibitor was fed to detached leaves at a concentration of 4 mm, and apparent photosynthesis was measured at atmospheric levels of CO₂ and at 20 and 210 mL L⁻¹ of O₂. Photosynthesis at 210 mL L⁻¹ of O₂ was inhibited 32% by DCDP in F. linearis, by 60% in F. brownii, and by 87% in F. trinervia. Inhibition in the hybrids ranged from 38 to 52%. The inhibition of photosynthesis by 210 mL L⁻¹ of O₂ was increased when DCDP was used, except in the C₄ species, F. trinervia, in which photosynthesis was insensitive to O₂. Except for F. trinervia, control plants with less O₂ sensitivity (more C₄-like) exhibited a progressively greater change in O₂ inhibition of photosynthesis when treated with DCDP. This increased O₂ inhibition probably resulted from decreased CO₂ concentrations in bundle sheath cells due to inhibition of phosphoenolpyruvate carboxylase. The inhibition of photosynthesis by DCDP is concluded to underestimate the degree of C₄ photosynthesis in the interspecific hybrids because increased direct assimilation of atmospheric CO₂ by ribulose bisphosphate carboxylase may compensate for inhibition of phosphoenolpyruvate carboxylase.     

Photorespiration-deficient Mutants of Arabidopsis thaliana Lacking Mitochondrial Serine Transhydroxymethylase Activity

Three allelic mutants of Arabidopsis thaliana which lack mitochondrial serine transhydroxymethylase activity due to a recessive nuclear mutation have been characterized. The mutants were shown to be deficient both in glycine decarboxylation and in the conversion of glycine to serine. Glycine accumulated as an end product of photosynthesis in the mutants, largely at the expense of serine, starch, and sucrose formation. The mutants photorespired CO₂ at low rates in the light, but this evolution of photorespiratory CO₂ was abolished by provision of exogenous NH₃. Exogenous NH₃ was required by the mutants for continued synthesis of glycine under photorespiratory conditions. These and related results with wild-type Arabidopsis suggested that glycine decarboxylation is the sole site of photorespiratory CO₂ release in wild-type plants but that depletion of the amino donors required for glyoxylate amination may lead to CO₂ release from direct decarboxylation of glyoxylate. Photosynthetic CO₂ fixation was inhibited in the mutants under atmospheric conditions which promote photorespiration but could be partially restored by exogenous NH₃. The magnitude of the NH₃ stimulation of photosynthesis indicated that the increase was due to the suppression of glyoxylate decarboxylation. The normal growth of the mutants under nonphotorespiratory atmospheric conditions indicates that mitochondrial serine transhydroxymethylase is not required in C₃ plants for any function unrelated to photorespiration.     

The metabolic origins of non-photorespiratory CO2 release during photosynthesis: a metabolic flux analysis

Respiration in the light (R_L) releases CO₂ in photosynthesizing leaves and is a phenomenon that occurs independently from photorespiration. Since R_L lowers net carbon fixation, understanding R_L could help improve plant carbon-use efficiency and models of crop photosynthesis. Although R_L was identified more than 75 years ago, its biochemical mechanisms remain unclear. To identify reactions contributing to R_L, we mapped metabolic fluxes in photosynthesizing source leaves of the oilseed crop and model plant camelina (Camelina sativa). We performed a flux analysis using isotopic labeling patterns of central metabolites during ¹³CO₂ labeling time course, gas exchange, and carbohydrate production rate experiments. To quantify the contributions of multiple potential CO₂ sources with statistical and biological confidence, we increased the number of metabolites measured and reduced biological and technical heterogeneity by using single mature source leaves and quickly quenching metabolism by directly injecting liquid N₂; we then compared the goodness-of-fit between these data and data from models with alternative metabolic network structures and constraints. Our analysis predicted that R_L releases 5.2 μmol CO₂ g⁻¹ FW h⁻¹ of CO₂, which is relatively consistent with a value of 9.3 μmol CO₂ g⁻¹ FW h⁻¹ measured by CO₂ gas exchange. The results indicated that ≤10% of R_L results from TCA cycle reactions, which are widely considered to dominate R_L. Further analysis of the results indicated that oxidation of glucose-6-phosphate to pentose phosphate via 6-phosphogluconate (the G6P/OPP shunt) can account for >93% of CO₂ released by R_L.     

Effects of Elevated [CO2] and Low Soil Moisture on the Physiological Responses of Mountain Maple (Acer spicatum L.) Seedlings to Light

Global climate change is expected to affect how plants respond to their physical and biological environments. In this study, we examined the effects of elevated CO₂ ([CO₂]) and low soil moisture on the physiological responses of mountain maple (Acer spicatum L.) seedlings to light availability. The seedlings were grown at ambient (392 µmol mol⁻¹) and elevated (784 µmol mol⁻¹) [CO₂], low and high soil moisture (M) regimes, at high light (100%) and low light (30%) in the greenhouse for one growing season. We measured net photosynthesis (A), stomatal conductance (g _s), instantaneous water use efficiency (IWUE), maximum rate of carboxylation (V _cmax), rate of photosynthetic electron transport (J), triose phosphate utilization (TPU)), leaf respiration (R _d), light compensation point (LCP) and mid-day shoot water potential (Ψ_x). A and g _s did not show significant responses to light treatment in seedlings grown at low soil moisture treatment, but the high light significantly decreased the C _i/C _a in those seedlings. IWUE was significantly higher in the elevated compared with the ambient [CO₂], and the effect was greater at high than the low light treatment. LCP did not respond to the soil moisture treatments when seedlings were grown in high light under both [CO₂]. The low soil moisture significantly reduced Ψ_x but had no significant effect on the responses of other physiological traits to light or [CO₂]. These results suggest that as the atmospheric [CO₂] rises, the physiological performance of mountain maple seedlings in high light environments may be enhanced, particularly when soil moisture conditions are favourable.     

An examination of the advantages of C3-C4 intermediate photosynthesis in warm environments

Abstract. The photosynthetic responses to temperature in C₃, C₃-C₄ intermediate, and C₄ species in the genus Flaveria were examined in an effort to identify whether the reduced photorespiration rates characteristic of C₃-C₄ intermediate photosynthesis result in adaptive advantages at warm leaf temperatures. Reduced photorespiration rates were reflected in lower CO₂ compensation points at all temperatures examined in the C₃-C₄ intermediate, Flaveria floridana, compared to the C₃ species, F. cronquistii. The C₃-C₄ intermediate, F. floridana, exhibited a C₃-like photosynthetic temperature dependence, except for relatively higher photosynthesis rates at warm leaf temperatures compared to the C₃ species, F. cronquistii. Using models of C₃ and C₃-C₄ intermediate photosynthesis, it was predicted that by recycling photorespired CO₂ in bundle-sheath cells, as occurs in many C₃-C₄ intermediates, photosynthesis rates at 35°C could be increased by 28%, compared to a C₃ plant. Without recycling photorespired CO₂, it was calculated that in order to improve photosynthesis rates at 35°C by this amount in C₃ plants, (1) intercellular CO₂ partial pressures would have to be increased from 25 to 31 Pa, resulting in a 57% decrease in water-use efficiency, or (2) the activity of RuBP carboxylase would have to be increased by 32%, resulting in a 22% decrease in nitrogen-use efficiency. In addition to the recycling of photorespired CO₂, leaves of F. floridana appear to effectively concentrate CO₂ at the active site of RuBP carboxylase, increasing the apparent carboxylation efficiency per unit of in vitro RuBP carboxylase activity. The CO₂-concentrating activity also appears to reduce the temperature sensitivity of the carboxylation efficiency in F. floridana compared to F. cronquistii. The carboxylation efficiency per unit of RuBP carboxylase activity decreased by only 38% in F. floridana, compared to 50% in F. cronquistii, as leaf temperature was raised from 25 to 35°C. The C₃-C₄ intermediate, F. ramosissima, exhibited a photosynthetic temperature temperature response curve that was more similar to the C₄ species, F. trinervia, than the C₃ species, F. cronquistii. The C₄-like pattern is probably related to the advanced nature of C₄-like biochemical traits in F. ramosissima The results demonstrate that reductions in photorespiration rates in C₃-C₄ intermediate plants create photosynthetic advantages at warm leaf temperatures that in C₃ plants could only be achieved through substantial costs to water-use efficiency and/or nitrogen-use efficiency.     

Quantitation of the O(2)-Dependent, CO(2)-Reversible Component of the Postillumination CO(2) Exchange Transient in Tobacco and Maize Leaves

The postillumination transient of CO₂ exchange and its relation to photorespiration has been examined in leaf discs from tobacco (Nicotiana tabacum) and maize (Zea mays). Studies of the transients observed by infrared gas analysis at 1, 21, and 43% O₂ in an open system were extended using the nonsteady state model described previously (Peterson and Ferrandino 1984 Plant Physiol 76: 976-978). Cumulative CO₂ exchange equivalents (i.e. nanomoles CO₂) versus time were derived from the analyzer responses of individual transients. In tobacco (C₃), subtraction of the time course of cumulative CO₂ exchange under photorespiratory conditions (21 or 43% O₂) from that obtained under nonphotorespiratory conditions (1% O₂) revealed the presence of an O₂-dependent and CO₂-reversible component within the first 60 seconds following darkening. This component was absent in maize (C₄) and at low external O₂:CO₂ ratios (i.e. <100) in tobacco. The size of the component in tobacco increased with net photosynthesis as irradiance was increased and was positively associated with inhibition of net photosynthesis by O₂. This relatively simple and rapid method of analysis of the transient is introduced to eliminate some uncertainties associated with estimation of photorespiration based on the maximal rate of postillumination CO₂ evolution. This method also provides a useful and complementary tool for detecting variation in photorespiration.     

Effects of genetic manipulation of the activity of photorespiration on the redox state of photosystem I and its robustness against excess light stress under CO<Subscript>2</Subscript>-limited conditions in rice

Under CO₂-limited conditions such as during stomatal closure, photorespiration is suggested to act as a sink for excess light energy and protect photosystem I (PSI) by oxidizing its reaction center chlorophyll P700. In this study, this issue was directly examined with rice (Oryza sativa L.) plants via genetic manipulation of the amount of Rubisco, which can be a limiting factor for photorespiration. At low [CO₂] of 5 Pa that mimicked stomatal closure condition, the activity of photorespiration in transgenic plants with decreased Rubisco content (RBCS-antisense plants) markedly decreased, whereas the activity in transgenic plants with overproduction of Rubisco (RBCS-sense plants) was similar to that in wild-type plants. Oxidation of P700 was enhanced at [CO₂] of 5 Pa in wild-type and RBCS-sense plants. PSI was not damaged by excess light stress induced by repetitive saturated pulse-light (rSP) in the presence of strong steady-state light. On the other hand, P700 was strongly reduced in RBCS-antisense plants at [CO₂] of 5 Pa. PSI was also damaged by rSP illumination. These results indicate that oxidation of P700 and the robustness of PSI against excess light stress are hampered by the decreased activity of photorespiration as a result of genetic manipulation of Rubisco content. It is also suggested that overproduction of Rubisco does not enhance photorespiration as well as CO₂ assimilation probably due to partial deactivation of Rubisco.