Improved method for measuring the apparent CO2 photocompensation point resolves the impact of multiple internal conductances to CO2 to net gas exchange

There is a growing interest in accurate and comparable measurements of the CO₂ photocompensation point (Γ*), a vital parameter to model leaf photosynthesis. The Γ* is measured as the common intersection of several CO₂ response curves, but this method may incorrectly estimate Γ* by using linear fits to extrapolate curvilinear responses and single conductances to convert intercellular photocompensation points (C_i*) to chloroplastic Γ*. To determine the magnitude and minimize the impact of these artefacts on Γ* determinations, we used a combination of meta‐analysis, modelling and original measurements to develop a framework to accurately determine C_i*. Our modelling indicated that the impact of using linear fits could be minimized based on the measurement CO₂ range. We also propose a novel method of analysing common intersection measurements using slope–intercept regression. Our modelling indicated that slope–intercept regression is a robust analytical tool that can help determine if a measurement is biased because of multiple internal conductances to CO₂. Application of slope–intercept regression to Nicotiana tabacum and Glycine max revealed that multiple conductances likely have little impact to C_i* measurements in these species. These findings present a robust and easy to apply protocol to help resolve key questions concerning CO₂ conductance through leaves.     

Zinc mediated effects on leaf CO2 diffusion conductances and net photosynthesis in Phaseolus vulgaris L.

Supra-optimal levels of zinc in primary leaves of Phaseolus vulgaris increased the CO₂ compensation point and inhibited net photosynthesis. Leaf morphology was modified: mesophyll intercellular area, stomatal slit length and interstomatal distance were reduced, but stomatal density increased. Internal and stomatal conductances to CO₂ diffusion decreased. These changes are discussed in relation to the observed effects on leaf gas exchange and to the previously reported inhibition of different photosynthetic and photorespiratory enzymes.     

Photorespiration in the context of Rubisco biochemistry,CO2 diffusion and metabolism

Photorespiratory metabolism is essential for plants to maintain functional photosynthesis in an oxygen‐containing environment. Because the oxygenation reaction of Rubisco is followed by the loss of previously fixed carbon, photorespiration is often considered a wasteful process and considerable efforts are aimed at minimizing the negative impact of photorespiration on the plant’s carbon uptake. However, the photorespiratory pathway has also many positive aspects, as it is well integrated within other metabolic processes, such as nitrogen assimilation and C₁ metabolism, and it is important for maintaining the redox balance of the plant. The overall effect of photorespiratory carbon loss on the net CO₂ fixation of the plant is also strongly influenced by the physiology of the leaf related to CO₂ diffusion. This review outlines the distinction between Rubisco oxygenation and photorespiratory CO₂ release as a basis to evaluate the costs and benefits of photorespiration.     

A dynamic model for photosynthesis by an aquatic plant, Egeria densa

Abstract This paper describes a dynamic model for photosynthesis by an aquatic plant, Egeria densa. The model takes into account an HCO^?₃ pump, high diffusion resistances and PEP carboxylase, and develops a set of differential equations to form the time-dependent solutions for photosynthesis. The predicted changes in pH, [CO₂]_aq and total inorganic carbon are compared with experimental data and the model is found to describe the data. The model is then used to examine the effect of O₂ on photosynthesis under these conditions, and shows that the increase in internal CO₂ concentration due to the recycling of photorespiratory CO₂ directly stimulates gross CO₂ fixation and can more than compensate for the O₂ inhibition of gross photosynthesis. The importance of the HCO^?₃ pump in O₂ inhibition is also examined. The CO₂ compensation point (where inorganic carbon influx and efflux are equal) is examined and the importance of the HCO^?₃ pump and PEP carboxylase in reducing the compensation concentration is discussed. The model was developed in order to study the photosynthesis of an aquatic weed, which will be reported in a later paper.     

The space‐time continuum: the effects of elevated CO2 and temperature on trees and the importance of scaling

To predict how forests will respond to rising temperatures and atmospheric CO₂ concentrations, we need to understand how trees respond to both of these environmental factors. In this review, we discuss the importance of scaling, moving from leaf‐level responses to those of the canopy, and from short‐term to long‐term responses of vegetation to climate change. While our knowledge of leaf‐level, instantaneous responses of photosynthesis, respiration, stomatal conductance, transpiration and water‐use efficiency to elevated CO₂ and temperature is quite good, our ability to scale these responses up to larger spatial and temporal scales is less developed. We highlight which physiological processes are least understood at various levels of study, and discuss how ignoring differences in the spatial or temporal scale of a physiological process impedes our ability to predict how forest carbon and water fluxes forests will be altered in the future. We also synthesize data from the literature to show that light respiration follows a generalized temperature response across studies, and that the light compensation point of photosynthesis is reduced by elevated growth CO₂. Lastly, we emphasize the need to move beyond single factorial experiments whenever possible, and to combine both CO₂ and temperature treatments in studies of tree performance.     

Calculation of CO2 gas phase diffusion in leaves and its relation to stomatal resistance

A new theory and experimental method was developed to measure the diffusion resistance to CO₂ in the gas phase of mesophyll leaf tissue. Excised leaves were placed in a chamber and their net evaporation and CO₂ assimilation rates measured at two different ambient pressures. These data were used to calculate CO₂ gas phase diffusion resistances. A variety of field grown leaves were tested and the effects of various experimental errors considered. Increasing the gas phase diffusion resistance decreased transpiration more than it decreased CO₂ assimilation. It was concluded that gas phase diffusion resistance associated with CO₂ assimilation may sometimes be 100 or 200 s·m^-1 greater than the resistance implied by transpiration rates. This may be due to longer path lengths for the CO₂ diffusion, constricted in places by the shape and arrangement of mesophyll cells.     

Ozone effects on wheat in relation to CO2: modelling short‐term and long‐term responses of leaf photosynthesis and leaf duration

A combined stomatal–photosynthesis model was extended to simulate the effects of ozone exposure on leaf photosynthesis and leaf duration in relation to CO₂. We assume that ozone has a short‐term and a long‐term effect on the Rubisco‐limited rate of photosynthesis, A_c. Elevated CO₂ counteracts ozone damage via stomatal closure. Ozone is detoxified at uptake rates below a threshold value above which A_c decreases linearly with the rate of ozone uptake. Reduction in A_c is transient and depends on leaf age. Leaf duration decreases depending on accumulated ozone uptake. This approach is introduced into the mechanistic crop simulation model AFRCWHEAT2. The derived model, AFRCWHEAT2‐O3, is used to test the capability of these assumptions to explain responses at the plant and crop level. Simulations of short‐term and long‐term responses of leaf photosynthesis, leaf duration and plant and crop growth to ozone exposure in response to CO₂ are analysed and compared with experimental data derived from the literature. The model successfully reproduced published responses of leaf photosynthesis, leaf duration, radiation use efficiency and final biomass of wheat to elevated ozone and CO₂. However, simulations were unsatisfactory for cumulative radiation interception which had some impact on the accuracy of predictions of final biomass. There were responses of leaf‐area index to CO₂ and ozone as a result of effects on tillering which were not accounted for in the present model. We suggest that some model assumptions need to be tested, or analysed further to improve the mechanistic understanding of the combined effects of changes in ozone and CO₂ concentrations on leaf photosynthesis and senescence. We conclude that research is particularly needed to improve the understanding of leaf‐area dynamics in response to ozone exposure and elevated CO₂.     

Electron transport is the functional limitation of photosynthesis in field-grown Pima cotton plants at high temperature

Restrictions to photosynthesis can limit plant growth at high temperature in a variety of ways. In addition to increasing photorespiration, moderately high temperatures (35–42 °C) can cause direct injury to the photosynthetic apparatus. Both carbon metabolism and thylakoid reactions have been suggested as the primary site of injury at these temperatures. In the present study this issue was addressed by first characterizing leaf temperature dynamics in Pima cotton (Gossypium barbadense) grown under irrigation in the US desert south‐west. It was found that cotton leaves repeatedly reached temperatures above 40 °C and could fluctuate as much as 8 or 10 °C in a matter of seconds. Laboratory studies revealed a maximum photosynthetic rate at 30–33 °C that declined by 22% at 45 °C. The majority of the inhibition persisted upon return to 30 °C. The mechanism of this limitation was assessed by measuring the response of photosynthesis to CO₂ in the laboratory. The first time a cotton leaf (grown at 30 °C) was exposed to 45 °C, photosynthetic electron transport was stimulated (at high CO₂) because of an increased flux through the photorespiratory pathway. However, upon cooling back to 30 °C, photosynthetic electron transport was inhibited and fell substantially below the level measured before the heat treatment. In the field, the response of assimilation (A) to various internal levels of CO₂ (C_i) revealed that photosynthesis was limited by ribulose‐1,5‐bisphosphate (RuBP) regeneration at normal levels of CO₂ (presumably because of limitations in thylakoid reactions needed to support RuBP regeneration). There was no evidence of a ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) limitation at air levels of CO₂ and at no point on any of 30 A–C_i curves measured on leaves at temperatures from 28 to 39 °C was RuBP regeneration capacity measured to be in substantial excess of the capacity of Rubisco to use RuBP. It is therefore concluded that photosynthesis in field‐grown Pima cotton leaves is functionally limited by photosynthetic electron transport and RuBP regeneration capacity, not Rubisco activity.     

Changes in photosynthetic capacity,carboxylation efficiency,and CO2 compensation point associated with midday stomatal closure and midday depression of net CO2 exchange of leaves of Quercus suber

The carbon-dioxide response of photosynthesis of leaves of Quercus suber, a sclerophyllous species of the European Mediterranean region, was studied as a function of time of day at the end of the summer dry season in the natural habitat. To examine the response experimentally, a standard time course for temperature and humidity, which resembled natural conditions, was imposed on the leaves, and the CO₂ pressure external to the leaves on subsequent days was varied. The particular temperature and humidity conditions chosen were those which elicited a strong stomatal closure at midday and the simultaneous depression of net CO₂ uptake. Midday depression of CO₂ uptake is the result of i) a decrease in CO₂-saturated photosynthetic capacity after light saturation is reached in the early morning, ii) a decrease in the initial slope of the CO₂ response curve (carboxylation efficiency), and iii) a substantial increase in the CO₂ compensation point caused by an increase in leaf temperature and a decrease in humidity. As a consequence of the changes in photosynthesis, the internal leaf CO₂ pressure remained essentially constant despite stomatal closure. The effects on capacity, slope, and compensation point were reversed by lowering the temperature and increasing the humidity in the afternoon. Constant internal CO₂ may aid in minimizing photoinhibition during stomatal closure at midday. The results are discussed in terms of possible temperature, humidity, and hormonal effects on photosynthesis.Abbreviations and symbols CE carboxylation efficiency - NP net photosynthesis rate - PAR photosynthetically active radiation - P_i leaf internal CO₂ partial pressure - W water vapor mole fraction difference between leaf and air - T CO₂ compensation pressure Dedicated to Professor Dr. Hubert Ziegler on the occasion of his 60th birthday     

Measurement of the photorespiratory activity of the submerged aquatic plant Myriophyllum spicatum L.

Abstract The CO₂ compensation concentrations (points) of leaves of the submerged vascular aquatic plant Myriophyllum spicatum L. were determined in a closed aqueous system at pH 7.0 by a gas chromatographic technique and over the range 10–30deg;C were found to range from 36 to 46 cm³m^?3 in medium equilibrated with 21% O₂ (0.03 kgm^?3), and 25 to 35 cm³m^?3 in medium equilibrated with 2% O₂ (0.03 kgm^?3). The rates of true (TPS) and apparent (APS) photosynthesis of leaves were measured in medium equilibrated with 21% O₂ and buffered at pH 7.0, at subsaturating concentrations (12.8–18.8 mmol m^?3) of dissolved inorganic carbor. (DIC) containing H¹⁴CO₃, by determining the initial rates of uptake by the leaves of DIC and ¹⁴C-activity from the medium. The rate of photorespiration, the difference between TPS and APS, was 7.0–13.3% of TPS over the range of 10–25°C and rose to 29% of TPS at 35°C. The magnitude of the compensation point of this plant is therefore similar to, but is much less O₂-sensitive than, those of C₃ plants, and the photorespiratory rate, at DIC concentrations near the CO₂ compensation point, is very low compared to that of C₃ plants.     

Effects of desiccation and CO2 concentrations on emersed photosynthesis in Porphyra haitanensis (Bangiales,Rhodophyta), a species farmed in China

The effects on photosynthesis of CO₂ and desiccation in Porphyra haitanensis were investigated to establish the effects of increased atmospheric CO₂ on this alga during emersion at low tides. With enhanced desiccation, net photosynthesis, dark respiration, photosynthetic efficiency, apparent carboxylating efficiency and light saturation point decreased, while the light compensation point and CO₂ compensation point increased. Emersed net photosynthesis was not saturated by the present atmospheric CO₂ level (about 350?ml?m^?3), and doubling the CO₂ concentration (700?ml?m^?3) increased photosynthesis by between 31% and 89% at moderate levels of desiccation. The relative enhancement of emersed net photosynthesis at 700?ml?m^?3 CO₂ was greater at higher temperatures and higher levels of desiccation. The photosynthetic production of Porphyra haitanensis may benefit from increasing atmospheric CO₂ concentration during emersion.     

Coordination between leaf CO2 diffusion and Rubisco properties allows maximizing photosynthetic efficiency in Limonium species

High photosynthetic efficiency intrinsically demands tight coordination between traits related to CO₂ diffusion capacity and leaf biochemistry. Although this coordination constitutes the basis of existing mathematical models of leaf photosynthesis, it has been barely explored among closely related species, which could reveal rapid adaptation clues in the recent past. With this aim, we characterized the photosynthetic capacity of 12 species of Limonium, possessing contrasting Rubisco catalytic properties, grown under optimal (WW) and extreme drought conditions (WD). The availability of CO₂ at the site of carboxylation (C_c) determined the photosynthetic capacity of Limonium under WD, while both diffusional and biochemical components governed the photosynthetic performance under WW. The variation in the in vivo caboxylation efficiency correlated with both the concentration of active Rubisco sites and the in vitro‐based properties of Rubisco, such as the maximum carboxylase turnover rate (k_cat^c) and the Michaelis–Menten constant for CO₂ (K_c). Notably, the results confirmed the hypothesis of coordination between the CO₂ offer and demand functions of photosynthesis: those Limonium species with high total leaf conductance to CO₂ have evolved towards increased velocity (i.e. higher k_cat^c), at the penalty of lower affinity for CO₂ (i.e. lower specificity factor, S_c/o).     

Variables Affecting the CO(2) Compensation Point

Some factors influencing dark respiration, photorespiration, and photosynthesis were examined for their effect on the CO₂ compensation point (70 μl/l) of detached soybean (Glycine max) leaf discs. A higher compensation point in young leaves decreased to the constant value after leaf expansion and maturation, but increased again during senescence. The compensation point was 40 to 50% higher in plants grown in the summer than in the winter. The compensation point and dark respiration increased with temperatures above 17 C. Below 17 C dark respiration continued to decrease, but the compensation point did not decrease further. Increasing light intensities did not affect the compensation point.     

Affixing the O to Rubisco: discovering the source of photorespiratory glycolate and its regulation

The source of glycolate in photorespiration and its control, a particularly active and controversial research topic in the 1970s, was resolved in large part by several discoveries and observations described here. George Bowes discovered that the key carboxylation enzyme Rubisco (ribulosebisphosphate carboxylase/oxygenase) is competitively inhibited by O₂ and that O₂ substitutes for CO₂ in the initial `dark' reaction of photosynthesis to yield glycolate-P, the substrate for photorespiration. William Laing derived an equation from basic enzyme kinetics that describes the CO₂, O₂, and temperature dependence of photosynthesis, photorespiration, and the CO₂ compensation point in C₃ plants. Jerome Servaites established that photosynthesis cannot be increased by inhibiting the photorespiratory pathway prior to the release of photorespiratory CO₂, and Douglas Jordan discovered substantial natural variation in the Rubisco oxygenase/carboxylase ratio. A mutant Arabidopsis plant with defective glycolate-P phosphatase, isolated by Chris Somerville, definitively established the role of O₂ and Rubisco in providing photorespiratory glycolate. Selection techniques to isolate photorespiration-deficient plants were devised by Jack Widholm and by Somerville, but no plants with reduced photorespiration were found. Somerville's approach, directed mutagenesis of Arabidopsis plants, was subsequently successful in the isolation of numerous other classes of mutants and revolutionized the science of plant biology. This revised version was published online in August 2006 with corrections to the Cover Date.     

Very high CO2 partially restores photosynthesis in sunflower at low water potentials

We re-examined the question of whether the stomata limit photosynthesis in dehydrated sunflower (Helianthus annuus L.) plants having low leaf water potentials. A gas-exchange apparatus was modified to operate at external CO₂ partial pressures as high as 3000 Pa (3%), which were much higher than previously achieved. This allowed photosynthesis and stomatal behavior to be monitored simultaneously at very high CO₂ in the same leaf. The data were compared with those from leaves treated with abscisic acid (ABA) where effects on photosynthesis are entirely stomatal. Photosynthesis was inhibited at low water potential and was only slightly enhanced by increasing the external CO₂ partial pressure from 34 Pa (normal air) to 300 Pa. Photosynthesis in ABA-treated leaves was similarly inhibited but recovered fully at 300 Pa. In both cases, the stomata closed to the same extent as judged from the average conductance of the leaves. Because the ABA effect resulted from diffusion limitation for CO₂ caused by stomatal closure, the contrasting data show that most of the dehydration effect was nonstomatal at low water potentials. When CO₂ partial pressures were raised further to 3000 Pa, photosynthesis increased somewhat at low water potentials but not in ABA-treated leaves. This indicates that some nonstomatal component of photosynthesis responded differently in leaves at low water potential and leaves treated with ABA. Because this component was only partially restored by very high CO₂, it was likely to be metabolic and was an important source of photosynthetic inhibition.Abbreviations and Symbol ABA abscisic acid - Chl chlorophyll - p_a external partial pressure of CO₂ - P_i intercellular partial pressure of CO₂ - _w water potential This work was supported by grant DE-FG02-87ER13776 from the Department of Energy and a grant from E.I. DuPont de Nemours and Company.     

Three‐dimensional microscale modelling of CO2 transport and light propagation in tomato leaves enlightens photosynthesis

We present a combined three‐dimensional (3‐D) model of light propagation, CO₂ diffusion and photosynthesis in tomato (Solanum lycopersicum L.) leaves. The model incorporates a geometrical representation of the actual leaf microstructure that we obtained with synchrotron radiation X‐ray laminography, and was evaluated using measurements of gas exchange and leaf optical properties. The combination of the 3‐D microstructure of leaf tissue and chloroplast movement induced by changes in light intensity affects the simulated CO₂ transport within the leaf. The model predicts extensive reassimilation of CO₂ produced by respiration and photorespiration. Simulations also suggest that carbonic anhydrase could enhance photosynthesis at low CO₂ levels but had little impact on photosynthesis at high CO₂ levels. The model confirms that scaling of photosynthetic capacity with absorbed light would improve efficiency of CO₂ fixation in the leaf, especially at low light intensity.     

The acclimation of photosynthesis and respiration to temperature in the C3–C4 intermediate Salsola divaricata: induction of high respiratory CO2 release under low temperature

Photosynthesis in C₃–C₄ intermediates reduces carbon loss by photorespiration through refixing photorespired CO₂ within bundle sheath cells. This is beneficial under warm temperatures where rates of photorespiration are high; however, it is unknown how photosynthesis in C₃–C₄ plants acclimates to growth under cold conditions. Therefore, the cold tolerance of the C₃–C₄ Salsola divaricata was tested to determine whether it reverts to C₃ photosynthesis when grown under low temperatures. Plants were grown under cold (15/10 °C), moderate (25/18 °C) or hot (35/25 °C) day/night temperatures and analysed to determine how photosynthesis, respiration and C₃–C₄ features acclimate to these growth conditions. The CO₂ compensation point and net rates of CO₂ assimilation in cold‐grown plants changed dramatically when measured in response to temperature. However, this was not due to the loss of C₃–C₄ intermediacy, but rather to a large increase in mitochondrial respiration supported primarily by the non‐phosphorylating alternative oxidative pathway (AOP) and, to a lesser degree, the cytochrome oxidative pathway (COP). The increase in respiration and AOP capacity in cold‐grown plants likely protects against reactive oxygen species (ROS) in mitochondria and photodamage in chloroplasts by consuming excess reductant via the alternative mitochondrial respiratory electron transport chain.     

Photosynthetic responses of soybean (Glycine max L.) to heat‐induced electrical signalling are predominantly governed by modifications of mesophyll conductance for CO2

In recent years, the effect of heat‐induced electrical signalling on plant photosynthetic activity has been demonstrated for many plant species. However, the underlying triggers of the resulting transient inhibition of photosynthesis still remain unknown. To further investigate on this phenomenon, we focused in our present study on soybean (Glycine max L.) on the direct effect of signal transmission in the leaf mesophyll on conductance for CO₂ diffusion in the mesophyll (g_m) and detected a drastic decline in g_m following the electrical signal, whereas the photosynthetic electron transport rate (ETR) was only marginally affected. In accordance with the drop in net photosynthesis (A_N), energy dispersive X‐ray analysis (EDXA) revealed a shift of K, Mg, O and P on leaf chloroplasts. Control experiments under elevated CO₂ conditions proved the transient reduction of A_N, ETR, the chloroplast CO₂ concentration (C_c) and g_m to be independent of the external CO₂ regime, whereas the effect of the electrical signal on stomatal conductance for CO₂ (g_s) turned out much less distinctive. We therefore conclude that the effect of electrical signalling on photosynthesis in soybean is triggered by its immediate effects on g_m.     

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     

Interactive effects of light,leaf temperature,CO2 and O2 on photosynthesis in soybean

A biochemical model of C ₃photosynthesis has been developed by G.D. Farquhar et al. (1980, Planta 149, 78–90) based on Michaelis-Menten kinetics of ribulose-1,5-bisphosphate (RuBP) carboxylase-oxygenase, with a potential RuBP limitation imposed via the Calvin cycle and rates of electron transport. The model presented here is slightly modified so that parameters may be estimated from whole-leaf gas-exchange measurements. Carbon-dioxide response curves of net photosynthesis obtained using soybean plants (Glycine max (L.) Merr.) at four partial pressures of oxygen and five leaf temperatures are presented, and a method for estimating the kinetic parameters of RuBP carboxylase-oxygenase, as manifested in vivo, is discussed. The kinetic parameters so obtained compare well with kinetic parameters obtained in vitro, and the model fits to the measured data give r ²values ranging from 0.87 to 0.98. In addition, equations developed by J.D. Tenhunen et al. (1976, Oecologia 26, 89–100, 101–109) to describe the light and temperature responses of measured CO₂-saturated photosynthetic rates are applied to data collected on soybean. Combining these equations with those describing the kinetics of RuBP carboxylase-oxygenase allows one to model successfully the interactive effects of incident irradiance, leaf temperature, CO₂ and O₂ on whole-leaf photosynthesis. This analytical model may become a useful tool for plant ecologists interested in comparing photosynthetic responses of different C₃ plants or of a single species grown in contrasting environments.Abbreviations PCO photorespiratory carbon oxidation - PCR photosynthetic carbon reduction - PPFD photosynthetic photon-flux density - RuBP ribulose bisphosphate