Effect of Oxygen on the Contribution of Respiration to the CO(2) Compensation Point in Wheat and Bean Leaves

The CO₂ compensation point at 21% O₂ (Γ₂₁) and at 2% O₂ (Γ₂), and the rate of dark CO₂ efflux at 21% O₂ (R_n) were measured in adult wheat (Triticum aestivum L, cv Gabo) leaves at the end of the night and after a period of photosynthesis of 5 h at 800 μbar CO₂. The values of Γ₂₁ and R_n significantly increased after the light period, due to the stimulation of respiration by carbohydrates. In contrast, Γ₂ did not increase after the same period of photosynthesis, suggesting that the respiratory component of Γ₂ was not stimulated by carbohydrates. In a different experiment, Γ₂₁, Γ₂, and R_n were studied during the growth period of bean (Phaseolus vulgaris L, cv Hawkesbury Wonder) leaves. The values of Γ₂₁ and R_n were high in young leaves, and decreased rapidly in parallel during maturation. However, Γ₂ presented relatively low values in growing bean leaves, and a model predicted that the observed values of Γ₂ should have been considerably higher if their respiratory component was considered to be as large as that of Γ₂₁. The results suggest that the rate of respiration in the light contributing to the CO₂ compensation point in wheat and bean leaves is smaller at low O₂ levels than at ambient levels.     

Effect of Oxygen on Photosynthesis, Photorespiration and Respiration in Detached Leaves. II. Corn and other Monocotyledons

The effect of O₂ on the CO₂ exchange of detached leaves of corn (Zea mays), wheat (Triticum vulgare), oats (Avena sativa), barley (Hordeum vulgare), timothy (Phleum pratense) and cat-tail (Typha angustifolia) was measured with a Clark oxygen electrode and infrared carbon dioxide analysers in both open and closed systems.

Corn leaves did not produce CO₂ in the light at any O₂ concentration, as was shown by the zero CO₂ compensation point and the absence of a CO₂ burst in the first minute of darkness. The rate of photosynthesis was inhibited by O₂ and the inhibition was not completely reversible. On the other hand, the steady rate of respiration after a few minutes in the dark was not affected by O₂.

These results were interpreted as indicating the absence of any measurable respiration during photosynthesis. Twelve different varieties of corn studied all responded to O₂ in the same way.

The other 5 monocotyledons studied did produce CO₂ in the light. Moreover, the CO₂ compensation point increased linearly with O₂ indicating a stimulation of photorespiration.

The implications of the lack of photorespiration in studies of primary productivity are discussed.

     

Effect of oxygen on photosynthesis, photorespiration and respiration in detached leaves. I. Soybean

The effect of O₂ on the CO₂ exchange of detached soybean leaves was measured with a Clark oxygen electrode and infrared carbon dioxide analysers in both open and closed systems.

The rate of apparent photosynthesis was inhibited by O₂ while the steady rate of respiration after a few minutes in the dark was not affected. Part of the inhibition of apparent photosynthesis was shown to be a result of increased photorespiration. This stimulation of photorespiration by O₂ was manifested by an increase in the CO₂ compensation point.

The differential effects of O₂ on dark respiration (no effect) and photorespiration (stimulation) indicated that these were 2 different processes.

Moreover the extrapolation of the CO₂ compensation point to zero at zero O₂ indicated that dark respiration was suppressed in the light at least at zero O₂ concentration.

     

Response of Respiration of Tobacco Leaves in Light and Darkness and the CO(2) Compensation Concentration to Prior Illumination and Oxygen

The course of respiration in control leaves of tobacco (Nicotiana tabacum L.) that were illuminated 4 to 5 hours and then darkened 0.25 to 10 hours and in tobacco leaves starved of carbohydrate by 14 hours or more of darkness was measured as CO₂ efflux in light and darkness into CO₂-free atmospheres containing 0.04, 2.23, 21, 40, and 100% O₂.     

Prior illumination and the respiration of maize leaves in the dark

The course of respiration of attached maize (Zea mays L.) leaves was measured by infrared gas analysis of CO₂ efflux in the dark following illumination in atmospheres of 300 microliters of CO₂ per liter of air, CO₂-free air, and CO₂-free N₂ containing 400 microliters of O₂ per liter. CO₂ efflux from control leaves started 3 to 4 minutes after darkening, increased to a maximum after about 20 minutes, and returned to a steady minimum after 2 to 3 hours. Respiration was quantitatively related to prior illumination, independent of net CO₂ fixation in the light, and depressed by N₂. Light, but not air, was required to produce a substrate for respiration in the subsequent dark period; air was required for oxidation of the substrate to CO₂. The stimulation of respiration by prior illumination in maize leaves differs in its slower onset and greater duration from the postillumination burst of photorespiration.     

A Comparison of Dark Respiration between C(3) and C(4) Plants

Lower respiratory costs were hypothesized as providing an additional benefit in C₄ plants compared to C₃ plants due to less investment in proteins in C₄ leaves. Therefore, photosynthesis and dark respiration of mature leaves were compared between a number of C₄ and C₃ species. Although photosynthetic rates were generally greater in C₄ when compared to C₃ species, no differences were found in dark respiration rates of individual leaves at either the beginning or after 16 h of the dark period. The effects of nitrogen on photosynthesis and respiration of individual leaves and whole plants were also investigated in two species that occupy similar habitats, Amaranthus retroflexus (C₄) and Chenopodium album (C₃). For mature leaves of both species, there was no relationship between leaf nitrogen and leaf respiration, with leaves of both species exhibiting a similar rate of decline after 16 h of darkness. In contrast, leaf photosynthesis increased with increasing leaf nitrogen in both species, with the C₄ species displaying a greater photosynthetic response to leaf nitrogen. For whole plants of both species grown at different nitrogen levels, there was a clear linear relationship between net CO₂ uptake and CO₂ efflux in the dark. The dependence of nightly CO₂ efflux on CO₂ uptake was similar for both species, although the response of CO₂ uptake to leaf nitrogen was much steeper in the C₄ species, Amaranthus retroflexus. Rates of growth and maintenance respiration by whole plants of both species were similar, with both species displaying higher rates at higher leaf nitrogen. There were no significant differences in leaf or whole plant maintenance respiration between species at any temperature between 18 and 42°C. The data suggest no obvious differences in respiratory costs in C₄ and C₃ plants.     

Effect of Temperature, CO(2) Concentration, and Light Intensity on Oxygen Inhibition of Photosynthesis in Wheat Leaves

The effect of 21% O₂ and 3% O₂ on the CO₂ exchange of detached wheat leaves was measured in a closed system with an infrared carbon dioxide analyzer. Temperature was varied between 2° and 43°, CO₂ concentration between 0.000% and 0.050% and light intensity between 40 ft-c and 1000 ft-c. In most conditions, the apparent rate of photosynthesis was inhibited in 21% O₂ compared to 3% O₂. The degree of inhibition increased with increasing temperature and decreasing CO₂ concentration. Light intensity did not alter the effect of O₂ except at light intensities or CO₂ concentrations near the compensation point. At high CO₂ concentrations and low temperature, O₂ inhibition of apparent photosynthesis was absent. At 3% O₂, wheat resembled tropical grasses in possessing a high rate of photosynthesis, a temperature optimum for photosynthesis above 30°, and a CO₂ compensation point of less than 0.0005% CO₂. The effect of O₂ on apparent photosynthesis could be ascribed to a combination of stimulation of CO₂ production during photosynthesis, and inhibition of photosynthesis itself.     

Effects of CO(2) Enrichment and Carbohydrate Content on the Dark Respiration of Soybeans

During the period of most active leaf expansion, the foliar dark respiration rate of soybeans (Glycine max cv Williams), grown for 2 weeks in 1000 microliters CO₂ per liter air, was 1.45 milligrams CO₂ evolved per hour leaf density thickness, and this was twice the rate displayed by leaves of control plants (350 microliters CO₂ per liter air). There was a higher foliar nonstructural carbohydrate level (e.g. sucrose and starch) in the CO₂ enriched compared with CO₂ normal plants. For example, leaves of enriched plants displayed levels of nonstructural carbohydrate equivalent to 174 milligrams glucose per gram dry weight compared to the 84 milligrams glucose per gram dry weight found in control plant leaves. As the leaves of CO₂ enriched plants approached full expansion, both the foliar respiration rate and carbohydrate content of the CO₂ enriched leaves decreased until they were equivalent with those same parameters in the leaves of control plants. A strong positive correlation between respiration rate and carbohydrate content was seen in high CO₂ adapted plants, but not in the control plants.

Mitochondria, isolated simultaneously from the leaves of CO₂ enriched and control plants, showed no difference in NADH or malate-glutamate dependent O₂ uptake, and there were no observed differences in the specific activities of NAD⁺ linked isocitrate dehydrogenase and cytochrome c oxidase. Since the mitochondrial O₂ uptake and total enzyme activities were not greater in young enriched leaves, the increase in leaf respiration rate was not caused by metabolic adaptations in the leaf mitochondria as a response to long term CO₂ enrichment. It was concluded, that the higher respiration rate in the enriched plant's foliage was attributable, in part, to a higher carbohydrate status.

     

Effect of photosynthesis and carbohydrate status on respiratory rates and the involvement of the alternative pathway in leaf respiration

In spinach (Spinacia oleracea Hybrid 102 [New World seeds]) and wheat (Triticum aestivum L. cv Gabo) leaves, O₂ uptake rates in the dark were faster after the plants had been allowed to photosynthesize for a period of several hours. Alternative path activity also increased following a period of photosynthesis in these leaves. No such effects were observed with isolated mitochondria. In spinach and wheat leaves, the level of fructose plus glucose decreased during a period of darkness. In pea (Pisum sativum cv Alaska) leaves, the level of these sugars did not vary significantly during the day, and respiratory rates were also constant. In slices cut from wheat leaves harvested at the end of the night, addition of sugars increased the rate of respiration and engaged the previously latent alternative oxidase. In pea leaves, O₂ uptake in the first few minutes following illumination was faster than that observed before illumination, but declined during the next 15 to 20 minutes. Adding the alternative oxidase inhibitor salicylhydroxamic acid, or imposing high bicarbonate concentrations during the period of photosynthesis, prevented the rise in O₂ uptake rate during the immediate post illumination period.

We conclude that the level of respiratory substrate in leaves determines their rate of O₂ uptake, and the degree to which the alternative path contributes to that O₂ uptake.

     

Photorespiratory phenomena in maize: oxygen uptake, isotope discrimination, and carbon dioxide efflux

Concurrent O₂ evolution, O₂ uptake, and CO₂ uptake by illuminated maize (Zea mays) leaves were measured using ¹³CO₂ and ¹⁸O₂. Considerable O₂ uptake occurred during active photosynthesis. At CO₂ compensation, O₂ uptake increased. Associated with this increase was a decrease in O₂ release such that a stoichiometric exchange of O₂ occurred. The rate of O₂ exchange at CO₂ compensation was directly related to O₂ concentration in the atmosphere at least up to 8% (v/v).     

Evaluation of the light/dark C assay of photorespiration: tobacco leaf disk studies with glycidate and glyoxylate

Preincubation of illuminated tobacco (Nicotiana tabacum L.) leaf disks in glycidate (2,3-epoxypropionate) or glyoxylate inhibited photorespiration by about 40% as determined by the ratio of ¹⁴CO₂ evolved into CO₂-free air in light and in darkness. However, under identical preincubation conditions used for the light/dark ¹⁴C assays, the compounds failed to reduce photorespiration or stimulate net photosynthesis in tobacco leaf disks based on other CO₂ exchange parameters, including the CO₂ compensation concentration in 21% O₂, the inhibitory effect of 21% O₂ on net photosynthesis in 360 microliters per liter of CO₂ and the rate of net photosynthetic ¹⁴CO₂ uptake in air.

The effects of both glycidate and glyoxylate on the ¹⁴C assay are inconsistent with other measures of photorespiratory CO₂ exchange in tobacco leaf disks, and thus these data question the validity of the light to dark ratio of ¹⁴CO₂ efflux as an assay for relative rates of photorespiration (Zelitch 1968, Plant Physiol 43: 1829-1837). The results of this study specifically indicate that neither glycidate nor glyoxylate reduces photorespiration or stimulates net photosynthesis by tobacco leaf disks under physiological conditions of pO₂ and pCO₂, contrary to previous reports.

     

The Respiration of Mature Field Bean (Vicia faba L.) Leaves During Prolonged Darkness

Dark respiration in attached and detached mature leaves of thefield bean (Vicia faba L.) was studied whilst leaves experiencedup to 60 h of darkness. The results showed: (1) the initialrespiration rate to vary according to the irradiance duringthe previous photoperiod; (2) the dark respiration rate (perunit area) of attached leaves to be essentially constant duringa normal 12 h night although there was a rapid loss in leafd. wt during this time; (3) after 12 h, the respiration rateof attached leaves decayed to an asymptotic value at about 36h; (4) the respiration rate of leaves detached at the end ofthe photoperiod and maintained in the dark on deionised water,decayed only after 36 h of darkness; (5) there was no differencebetween the respiration rate of attached and detached leavesduring the normal 12 h night. It is concluded that the dark respiration of attached fieldbean leaves is intially related to the synthesis and translocationof sucrose in addition to maintenance. After about 36 h, whenthe rate of CO₂ efflux is more or less steady, the CO₂ effluxreflects the intensity of maintenance processes only. The maintenancerespiration rate (determined after 60 h in the dark) rangedfrom 062 to 151 mg CO₂ (g d. wt)^–1 h^–1 but wasrelatively unaffected by several applied treatments. Vicia faba L., field bean, respiration, maintenance, nitrate, non-structural carbohydrate, export     

Is Transfer of CO2 to C3-Plants Purely by Diffusion?

The dependence of the CO₂ compensation concentration on O₂ partial pressure and the dependence of differential uptake of ¹⁴CO₂ and ¹²CO₂ on CO₂ and O₂ partial pressures are analyzed in illuminated white clover (Trifolium repens L.) leaves. The data show a deviation of the photosynthetic gas exchange from ribulose bisphosphate carboxylase oxygenase kinetics at 10°C but not at 30°C. This deviation is due to an effect of CO₂ partial pressure on the ratio of photosynthesis to photorespiration which can be explained if active inorganic carbon transport is assumed.     

Positive effects of night warming on physiology of coniferous trees in late growing season: Leaf and root

Previous studies about the effects of experimental warming on tree species have focused primarily on response of morphology and physiology in leaf and biomass allocation in the growing season, and a few studies considered the importance of roots. Based on the available evidence, it is unclear whether photosynthesis rate is enhanced by night warming in late autumn an issue that deserves further investigation. Thus, we exposed two coniferous species, Picea asperata and Abies faxoniana, to night warming continued throughout the year to investigate morphological and physiological responses of roots and leaves in the autumn. The results showed that night warming caused significant increases in net influxes of NH₄⁺ and NO₃⁻ in P. asperata seedlings corresponding well with net H⁺ efflux and net influx of O₂. Meanwhile, night warming had a positive effect on foliar gas exchange such as net photosynthesis rate, apparent quantum efficiency, dark respiration rate and maximum quantum efficiency of PS II, and nitrate reductase activity of roots. Additionally, root morphology such as total roots length, surface area, specific root area and specific root length was also stimulated by night warming. In contrast, night warming decreased concentrations of non-structural carbohydrate in leaves and roots of both species in autumn. The present study demonstrates that night warming would enhance late autumn leaf photosynthetic rate, and increase N uptake capacity of roots.     

Changes in the activation state of rubisco in bean leaves in relation to recovery of photosynthetic activity after various treatments

Changes in the activation level of rubisco from bean leaves were studied by measuring carboxylation rates after either a prolonged period in the dark after photoinhibitory treatment. Plants kept for 11 h in darkness were subjected to three stages of strong irradiance: stage I, photosynthesis in air; stage II, exposure in1% O₂ and CO₂-free N₂; stage III, reintroduction of air. Decrease in the rate of CO₂ uptake at stage III are due to photoinhibition. From the end of stage I to stage III, initial carboxylase activity, measured in extracts, exhibited large variations whereas changes for total activity were much smaller. Initial activity was four times lower at stage II compared to stage I. It increased in several minutes to high values at stage III where rates of CO₂ uptake were attained more gradually. No large amount of ribulose 1–5 bisp phosphate (RuBP) appeared to be bound to inactive enzyme at stage II. When darkened leaves were illuminated (stage I) the increase of both initial and total activity was slow and parallel to disappeance of the inhibitor carboxyarabinitol-1-phosphate (CA1P). Conversely, increase of CO₂ uptake rate was more rapid.     

Light Stimulation of Ethylene Release from Leaves of Gomphrena globosa L

The effect of light and CO₂ on both the endogenous and 1-aminocyclopropane-1-carboxylic acid (ACC)-dependent ethylene evolution from metabolically active detached leaves and leaf discs of Gomphrena globosa L. is reported. Treatment with varying concentrations of ACC did not appear to inhibit photosynthesis, respiration, or stomatal behavior. In all treatments, more ethylene was released into a closed flask from ACC-treated tissue, but the pattern of ethylene release with respect to light/dark/CO₂ treatments was the same.

Leaf tissue in the light with a source of CO₂ sufficient to maintain photosynthesis always generates 3 to 4 times more ethylene than tissue in the dark. Conversely, the lowest rate of ethylene release occurs when leaf tissue is illuminated and photosynthetic activity depletes the CO₂ to the compensation point. Ethylene release in the dark is also stimulated by CO₂ either added to the flask as bicarbonate or generated by dark respiration. Ethylene release increases dramatically and in parallel with photosynthesis at increasing light intensities in this C₄ plant. Ethylene release appears dependent on CO₂ both in the light and in the dark. Therefore, it is suggested that the important factor regulating the evolution of ethylene gas from leaves of Gomphrena may be CO₂ metabolism rather than light per se.

     

Photosynthesis, Respiration and Post-illumination Fixation of CO2 by Corn Leaves as Influenced by Light and Oxygen Concentration

The effect of oxygen concentration on the rate of CO₂-uptake in continuous and intermittent light was studied as well as the CO₂-fixation during a short dark period after light was turned off. In addition the dark respiration and the CO₂-compensation point of attached and detached corn leaves were determined. Leaves of 4 to 22-day old plants were used as experimental material. A closed circuit system of an infrared carbon dioxide analyzer was employed to measure the rate of CO₂-exchange. It was found that in an atmosphere consisting of 100 % oxygen, there was about 50 per cent inhibition of the rate of CO₂-uptake in continuous and intermittent light compared to that in an atmosphere consisting of 21% oxygen. The same was true of the rate of CO₂-fixation in darkness during a short period after the light was turned off. Since the response to oxygen concentration of the CO₂-uptake in light and of the CO₂-fixation in darkness after the light was turned off were similar, it is concluded that the fixation of CO₂ in the short dark period represents an over- shoot of photosynthesis. The rate of dark respiration was little affected by the oxygen concentration in the ranges used in the experiments. The carbon dioxide compensation point which has been observed in leaves of 4 to 14-day old plants was not influenced by either oxygen concentration or light intensity. Since the changes in the rate of CO₂-uptake due to changes in the concentration of oxygen and light intensity had no effect on the CO₂-compensation point, it is concluded that a reabsorption of respiratory CO₂ by photosynthesis could not account for the low value of this point. These results are interpreted as a further corroboration of the statement that the leaves of corn lack the process of photorespiration and that dark respiration is inhibited in light. It was observed that the rate of the CO₂-uptake gradually increased in plants which were from 4 to 22-days old. The inhibitory effect of high concentration of oxygen on the rate of CO₂-uptake was relatively higher in old plants than in young ones.     

Effects of Glucose Feeding on Respiration and Photosynthesis in Photoautotrophic Dianthus caryophyllus Cells: Mass Spectrometric Determination of Gas Exchange

When glucose (20 millimolar) was added to photoautotrophic cell suspension cultures of Dianthus caryophyllus, there was during the first 10 hours an accumulation of carbohydrates and phosphorylated compounds. These biochemical changes were accompanied by a progressive decrease of net photosynthesis and a twofold increase of the dark respiratory rate. The rise of respiration was associated with a rise of fumarase and cytochrome c oxidase activities, two mitochondrial markers. Gas exchange of illuminated cells were performed with a mass spectrometry technique and clearly established that during the first hours of glucose feeding, the decrease of net photosynthesis was essentially due to an increase of respiration in light, whereas the photosynthetic processes (gross O₂ evolution and gross CO₂ fixation) were almost not affected. However, after 24 hours of experiment, O₂ evolution and CO₂ fixation started to decline in turn. While ribulose-1,5-bisphosphate carboxylase activity was little affected during the first 48 hours of the experiment, the maximal light-induced phosphoribulokinase activity dramatically decreased with time and represented after 48 hours only 30% of its initial activity. It is postulated that the decrease in phosphoribulokinase activity was at least partially responsible for the decrease of CO₂ fixation and the metabolic events involved in this regulation are discussed.     

Photosynthesis: action spectra for leaves in normal and low oxygen

The action spectrum of apparent photosynthesis for attached radish (Raphanus sativus L. var. Early Scarlet Globe) and corn (Zea mays L. var. Pride V.) leaves was measured at 300 μl/l CO₂ and both 21% and 2% O₂. The spectra were measured at light intensities where apparent photosynthesis was proportional to intensity. For radish, a high compensation point plant, oxygen had an inhibiting effect on photosynthesis at all wavelengths from 402 to 694 mμ. If a constant rate of photosynthesis at 21% O₂ for the different wavelengths was chosen, then the percent increase in net CO₂ fixation at 2% O₂ was constant. For corn, a low compensation point plant, no inhibitory effect of oxygen concentration from 2% to 21% O₂ was found over the visible spectrum. The CO₂ compensation point for light intensities greater than the light compensation point was found to be constant and independent of wavelength for both radish and corn leaves. For radish, the lowering of the oxygen concentration from 21% to 2% at these intensities was found to reduce the CO₂ compensation point by the same amount for the wavelengths studied.     

Patterns of Stomatal Behaviour, Transpiration, and CO2 Exchange in Pea Following Infection by Powdery Mildew (Erysiphe pisi)

A comparison was made of stomatal behaviour, and related phenomena,between leaves of garden pea (Pisum sativum cv. Feltham First)inoculated with powdery mildew fungus (Erysiphe pisi) and uninfectedleaves on healthy plants. Twenty four hours after inoculation,stomata opened more widely in the light in infected leaves thanin healthy leaves. Thereafter, stomatal opening was progressivelyreduced by infection and stomata failed to close completelyin the dark until, 7 d after inoculation, all movements ceasedand stomata remained partly open. Transpiration in the lightfollowed closely the pattem of stomatal opening and, after anearly increase compared with healthy controls, was progressivelyreduced by infection. Evidence is presented that transpirationfrom the fungus was less than the reduction in transpiraationfrom the leaf which was caused when development of the myceliumincreased the boundary layer resistance of the leaf. Seven daysafter inoculation, transpiration in the dark was greater frominfected leaves than from healthy leaves because of partly openstomata in the dark. Net photosynthesis in infected leaves was reduced within 24h of inoculation to a level below that found in healthy leavesand thereafter it declined progressively. The initial reductionwas due to a transient increase in photorespiration, for whenthe glycolate pathway was inhibited by a 2% O₂ concentrationthere was no difference between the (gross) photosynthetic ratesof healthy and infected leaves. Changes in photorespirationrate were confirmed from the interpretation of the CO₂ burston darkening. Reduced stomatal opening was a contributory causeof the reduction in net photosynthesis in the later stages ofinfection. Since the rate of gross photosynthesis, but not therate of photorespiration, of infected plants fell below thatof healthy plants, and infected plants had a higher rate ofrelease of CO₂ in the dark than healthy plants from the thirdday after inoculation onwards, infected plants consume an increasinglygreater proportion of their photosynthate in respiratory processesthan do healthy plants. The CO₂ compensation point of infectedplants increased at every time of sampling after inoculation.