12CO2 emission from different metabolic pathways measured in illuminated and darkened C3 and C4 leaves at low,atmospheric and elevated CO2 concentration

The detection of 12CO2 emission from leaves in air containing 13CO2 allows simple and fast determination of the CO2 emitted by different sources, which are separated on the basis of their labelling velocity. This technique was exploited to investigate the controversial effect of CO2 concentration on mitochondrial respiration. The 12CO2 emission was measured in illuminated and darkened leaves of one C4 plant and three C3 plants maintained at low (30-50 ppm), atmospheric (350-400 ppm) and elevated (700-800 ppm) CO2 concentration. In C3 leaves, the 12CO2 emission in the light (Rd) was low at ambient CO2 and was further quenched in elevated CO2, when it was often only 20-30% of the 12CO2 emission in the dark, interpreted as the mitochondrial respiration in the dark (Rn). Rn was also reduced in elevated CO2. At low CO2, Rd was often 70-80% of Rn, and a burst of 12CO2 was observed on darkening leaves of Mentha sativa and Phragmites australis after exposure for 4 min to 13CO2 in the light. The burst was partially removed at low oxygen and was never observed in C4 leaves, suggesting that it may be caused by incomplete labelling of the photorespiratory pool at low CO2. This pool may be low in sclerophyllous leaves, as in Quercus ilex where no burst was observed. Rd was inversely associated with photosynthesis, suggesting that the Rd/Rn ratio reflects the refixation of respiratory CO2 by photosynthesizing leaves rather than the inhibition of mitochondrial respiration in the light, and that CO2 produced by mitochondrial respiration in the light is mostly emitted at low CO2, and mostly refixed at elevated CO2. In the leaves of the C4 species Zea mays, the 12CO2 emission in the light also remained low at low CO2, suggesting efficient CO2 refixation associated with sustained photosynthesis in non-photorespiratory conditions. However, Rn was inhibited in CO2-free air, and the velocity of 12CO2 emission after darkening was inversely associated with the CO2 concentration. The emission may be modulated by the presence of post-illumination CO2 uptake deriving from temporary imbalance between C3 and C4 metabolism. These experiments suggest that this uptake lasts longer at low CO2 and that the imbalance is persistent once it has been generated by exposure to low CO2.     

A new measurement technique reveals rapid post-illumination changes in the carbon isotope composition of leaf-respired CO2

We describe an open leaf gas exchange system coupled to a tunable diode laser (TDL) spectroscopy system enabling measurement of the leaf respiratory CO(2) flux and its associated carbon isotope composition (delta(13)C(Rl)) every 3 min. The precision of delta(13)C(Rl) measurement is comparable to that of traditional mass spectrometry techniques. delta(13)C(Rl) from castor bean (Ricinus communis L.) leaves tended to be positively related to the ratio of CO(2) produced to O(2) consumed [respiratory quotient (RQ)] after 24-48 h of prolonged darkness, in support of existing models. Further, the apparent fractionation between respiratory substrates and respired CO(2) within 1-8 h after the start of the dark period was similar to previous observations. In subsequent experiments, R. communis plants were grown under variable water availability to provide a range in delta(13)C of recently fixed carbohydrate. In leaves exposed to high light levels prior to the start of the dark period, CO(2) respired by leaves was up to 11 per thousand more enriched than phloem sap sugars within the first 10-15 min after plants had been moved from the light into the dark. The (13)C enrichment in respired CO(2) then decreased rapidly to within 3-7 per thousand of phloem sap after 30-60 min in the dark. This strong enrichment was not observed if light levels were low prior to the start of the dark period. Measurements of RQ confirmed that carbohydrates were the likely respiratory substrate for plants (RQ > 0.8) within the first 60 min after illumination. The strong (13)C enrichment that followed a high light-to-dark transition coincided with high respiration rates, suggesting that so-called light-enhanced dark respiration (LEDR) is fed by (13)C-enriched metabolites.     

Responses of Plant Dark Respiration to Doubled CO2 Concentration

Ten species of plants were grown at ambient (350μmol CO2·mol-1 air) and doubled (700 μmol CO2·mol-1 air) CO2 concentrations at ambient temperature and illumination in order to examine changes of dark respiration of whole seedlings or detached leaves. Effects of CO2 on dark respiration were determined by brief exposure ( ≤ 5 min) to corresponding CO2 concentration and temperatures ( 15,20,25,30 and 35 ℃ ) with infrared CO2 analyzer. The reductions in dark respiration on a weight base for leaves of East-Liaoning oak (Quercus liaotungensis Koidz. ) at 15,20 and 25 ℃ and of soybean ( Glycine max L. ) at 20,25,30 and 35 ℃ and for whole seedlings of three- tcoloured amaranth (Amaranthus tricolor L. ) at 15 and 20 ℃ and cucumber ( Cucumis sativus L. ) at 15 cE measured at elevated concentration relative to the ambient CO2 concentration were observed. No significant difference in respiration responded was observed to elevated or ambient CO2 concentrations at 15 ℃ in maize (Zea mays L. ) seedlings and alfalfa (Medicago sativa L. ) leaves, at 35 ℃ in East-Liaoning oak leaves and at 20,25 and 30 ℃ in three-coloured amaranth seedlings. However CO2 efflux in leaves of weeping willow (Salix babylonica L. ), simon poplar (Populus simonii Carr. ) and eucommia (Eucommia ulmoides Oliv. ) at 15,20,25,30 and 35 ℃, alfalfa at 20,25,30 and 35 ℃, East-Liaoning oak at 30 ℃, maize at 15 ℃, seedlings of common buckwheat (Fagotrytum esculentum Moench) at 15,20,25,30 and 35 ℃, cucumber and maize at 20,25,30 and 35 ℃ and three-coloured amaranth at 35 ℃ showed an increase at elevated in contrast to ambient CO2 concentration. In general, at lower temperatures (i. e. 15, 20 ℃ ) there was no significant difference between elevated and ambient CO2 concentration for dark respiration, while at higher temperatures (i. e. 30,35 ℃ ) elevated CO2 concentration positively stimulate clark respiretion. It has not yet been described that double CO2 concentration could enhance plant dark respiration at 30 and 35 ℃. Impacts of the characteristics in dark respiration on the future changes of vegetation and its mechanism were discussed.     

Carbon isotopes in terrestrial ecosystem pools and CO2 fluxes

Stable carbon isotopes are used extensively to examine physiological, ecological, and biogeochemical processes related to ecosystem, regional, and global carbon cycles and provide information at a variety of temporal and spatial scales. Much is known about the processes that regulate the carbon isotopic composition (delta(13)C) of leaf, plant, and ecosystem carbon pools and of photosynthetic and respiratory carbon dioxide (CO(2)) fluxes. In this review, systematic patterns and mechanisms underlying variation in delta(13)C of plant and ecosystem carbon pools and fluxes are described. We examine the hypothesis that the delta(13)C of leaf biomass can be used as a reference point for other carbon pools and fluxes, which differ from the leaf in delta(13)C in a systematic fashion. Plant organs are typically enriched in (13)C relative to leaves, and most ecosystem pools and respiratory fluxes are enriched relative to sun leaves of dominant plants, with the notable exception of root respiration. Analysis of the chemical and isotopic composition of leaves and leaf respiration suggests that growth respiration has the potential to contribute substantially to the observed offset between the delta(13)C values of ecosystem respiration and the bulk leaf. We discuss the implications of systematic variations in delta(13)C of ecosystem pools and CO(2) fluxes for studies of carbon cycling within ecosystems, as well as for studies that use the delta(13)C of atmospheric CO(2) to diagnose changes in the terrestrial biosphere over annual to millennial time scales.     

Effect of local irradiance on CO(2) transfer conductance of mesophyll in walnut

The acclimation responses of walnut leaf photosynthesis to the irradiance microclimate were investigated by characterizing the photosynthetic properties of the leaves sampled on young trees (Juglans nigraxregia) grown in simulated sun and shade environments, and within a mature walnut tree crown (Juglans regia) in the field. In the young trees, the CO(2) compensation point in the absence of mitochondrial respiration (Gamma*), which probes the CO(2) versus O(2) specificity of Rubisco, was not significantly different in sun and shade leaves. The maximal net assimilation rates and stomatal and mesophyll conductances to CO(2) transfer were markedly lower in shade than in sun leaves. Dark respiration rates were also lower in shade leaves. However, the percentage inhibition of respiration by light during photosynthesis was similar in both sun and shade leaves. The extent of the changes in photosynthetic capacity and mesophyll conductance between sun and shade leaves under simulated conditions was similar to that observed between sun and shade leaves collected within the mature tree crown. Moreover, mesophyll conductance was strongly correlated with maximal net assimilation and the relationships were not significantly different between the two experiments, despite marked differences in leaf anatomy. These results suggest that photosynthetic capacity is a valuable parameter for modelling within-canopies variations of mesophyll conductance due to leaf acclimation to light.     

Changes in apoplastic pH and membrane potential in leaves in relation to stomatal responses to CO2, malate, abscisic acid or interruption of water supply

Low CO2 concentrations open CO2-sensitive stomata whereas elevated CO2 levels close them. This CO2 response is maintained in the dark. To elucidate mechanisms underlying the dark CO2 response we introduced pH- and potential-sensitive dyes into the apoplast of leaves. After mounting excised leaves in a gas-exchange chamber, changes in extracellular proton concentration and transmembrane potential differences as well as transpiration and respiration were simultaneously monitored. Upon an increase in CO2 concentration transient changes in apoplastic pH (occasionally brief acidification, but always followed by alkalinization) and in membrane potential (brief hyperpolarization followed by depolarization) accompanied stomatal closure. Alkalinization and depolarization were also observed when leaves were challenged with abscisic acid or when water flow was interrupted. During stomatal opening in response to CO2-free air the apoplastic pH increased while the membrane potential initially depolarized before it transiently hyperpolarized. To examine whether changes in apoplastic malate concentrations represent a closing signal for stomata, malate was fed into the transpiration stream. Although malate caused apoplastic alkalinization and membrane depolarization reminiscent of the effects observed with CO2 and abscisic acid, this dicarboxylate closed the stomata only partially and less effectively than CO2. Apoplastic alkalinization was also observed and stomata closed partially when KCl was fed to the leaves. Respiration increased on feeding of malate or KCl, or while abscisic acid closed the stomate. From these results we conclude that CO2 signals modulate the activity of plasma-membrane ion channels and of plasmalemma H+-ATPases during changes in stomatal aperture. Responses to potassium malate and KCl are not restricted to guard cells and neighbouring cells.     

Leaf O(2) uptake in the dark is independent of coincident CO(2) partial pressure

Elevated CO(2), in the dark, is sometimes reported to inhibit leaf respiration, with respiration usually measured as CO(2) efflux. Oxygen uptake may be a better gauge of respiration because non-respiratory processes can affect dark CO(2) efflux in elevated CO(2). Two methods of quantifying O(2) uptake indicated that leaf respiration was unaffected by coincident CO(2) level in the dark.     

大气CO2浓度倍增对植物暗呼吸的影响

以长期生长于350和700μmolCO_2·mol~(-1)空气的开顶式培养室的杜仲(Eucommia ulmoides Oliv.)、紫花苜蓿(Medicago sativa L.)、玉米(Zea mays L.)等10种植物的离体成熟叶片或整株为材料,研究不同测定温度(15～35℃)下,CO_2浓度倍增对植物暗呼吸的影响。结果表明:在较低温度(15℃、20℃)下,CO_2浓度倍增对植物暗呼吸没有显著效应,在较高温度(30℃、35℃)下多数被测植物的暗呼吸显著增强。讨论了实验所得结果在未来全球气候变化中的可能的意义。     

Degradation of Storage Photosynthates in Isolated Spinach Mesophyll Cells in CO2-Free System and Its Regulatory Factors

Isolated spinach (spinacia oleracea L.) mesophyll cells, after being preilluminated for 40 min for storage photosynthates loading (14C), were shifted into CO2-free mixture for investigation of storage photosynthates degradation and its regulatory factors. The loss of fixed carbon (14C) after 30 min incubation in CO2-free mixture in light was about 20%, which was much higher than that in dark (about 4.5%). The loss of fixed carbon was apparently stimulated by the increase of light intensity or O2 concentration and depressed by DCMU or NH4C1. Moreover, the loss of fixed carbon, especially the soluble fraction, was reduced by the addition of glycolate in light but not in dark. The obvious degradation of starch in spinach mesophyll cells in CO2-free system was monitored both in light and in dark. Unlike in dark, starch degradation in light was not associated with a detectable accumulation of the soluble products (PGA, Ribulose5-phosphate, sucrose etc.). Based on the above results, the different metabolic patterns for the products of starch degradation in light and in dark were discussed and the possible physiological roles were also proposed.     

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 the effects of carbon dioxide concentration and temperature on respiration, translocation and nitrate reduction in darkened soybean leaves

BACKGROUND AND AIMS: Respiration of autotrophs is an important component of their carbon balance as well as the global carbon dioxide budget. How autotrophic respiration may respond to increasing carbon dioxide concentrations, [CO(2)], in the atmosphere remains uncertain. The existence of short-term responses of respiration rates of plant leaves to [CO(2)] is controversial. Short-term responses of respiration to temperature are not disputed. This work compared responses of dark respiration and two processes dependent on the energy and reductant supplied by dark respiration, translocation and nitrate reduction, to changes in [CO(2)] and temperature. METHODS: Mature soybean leaves were exposed for a single 8-h dark period to one of five combinations of air temperature and [CO(2)], and rates of respiration, translocation and nitrate reduction were determined for each treatment. KEY RESULTS: Low temperature and elevated [CO(2)] reduced rates of respiration, translocation and nitrate reduction, while increased temperature and low [CO(2)] increased rates of all three processes. A given change in the rate of respiration was accompanied by the same change in the rate of translocation or nitrate reduction, regardless of whether the altered respiration was caused by a change in temperature or by a change in [CO(2)]. CONCLUSIONS: These results make it highly unlikely that the observed responses of respiration rate to [CO(2)] were artefacts due to errors in the measurement of carbon dioxide exchange rates in this case, and indicate that elevated [CO(2)] at night can affect translocation and nitrate reduction through its effect on respiration.     

Respiratory oxygen uptake is not decreased by an instantaneous elevation of [CO2], but is increased with long-term growth in the field at elevated [CO2

Averaged across many previous investigations, doubling the CO2 concentration ([CO2]) has frequently been reported to cause an instantaneous reduction of leaf dark respiration measured as CO2 efflux. No known mechanism accounts for this effect, and four recent studies have shown that the measurement of respiratory CO2 efflux is prone to experimental artifacts that could account for the reported response. Here, these artifacts are avoided by use of a high-resolution dual channel oxygen analyzer within an open gas exchange system to measure respiratory O2 uptake in normal air. Leaf O2 uptake was determined in response to instantaneous elevation of [CO2] in nine contrasting species and to long-term elevation in seven species from four field experiments. Over six hundred separate measurements of respiration failed to reveal any decrease in respiratory O2 uptake with an instantaneous increase in [CO2]. Respiration was found insensitive not only to doubling [CO2], but also to a 5-fold increase and to decrease to zero. Using a wide range of species and conditions, we confirm earlier reports that inhibition of respiration by instantaneous elevation of [CO2] is likely an experimental artifact. Instead of the expected decrease in respiration per unit leaf area in response to long-term growth in the field at elevated [CO2], there was a significant increase of 11% and 7% on an area and mass basis, respectively, averaged across all experiments. The findings suggest that leaf dark respiration will increase not decrease as atmospheric [CO2] rises.     

Response of respiration of soybean leaves grown at ambient and elevated carbon dioxide concentrations to day-to-day variation in light and temperature under field conditions

BACKGROUND AND AIMS: Respiration is an important component of plant carbon balance, but it remains uncertain how respiration will respond to increases in atmospheric carbon dioxide concentration, and there are few measurements of respiration for crop plants grown at elevated [CO(2)] under field conditions. The hypothesis that respiration of leaves of soybeans grown at elevated [CO(2)] is increased is tested; and the effects of photosynthesis and acclimation to temperature examined. METHODS: Net rates of carbon dioxide exchange were recorded every 10 min, 24 h per day for mature upper canopy leaves of soybeans grown in field plots at the current ambient [CO(2)] and at ambient plus 350 micromol mol(-1) [CO(2)] in open top chambers. Measurements were made on pairs of leaves from both [CO(2)] treatments on a total of 16 d during the middle of the growing seasons of two years. KEY RESULTS: Elevated [CO(2)] increased daytime net carbon dioxide fixation rates per unit of leaf area by an average of 48 %, but had no effect on night-time respiration expressed per unit of area, which averaged 53 mmol m(-2) d(-1) (1.4 micromol m(-2) s(-1)) for both the ambient and elevated [CO(2)] treatments. Leaf dry mass per unit of area was increased on average by 23 % by elevated [CO(2)], and respiration per unit of mass was significantly lower at elevated [CO(2)]. Respiration increased by a factor of 2.5 between 18 and 26 degrees C average night temperature, for both [CO(2)] treatments. CONCLUSIONS: These results do not support predictions that elevated [CO(2)] would increase respiration per unit of area by increasing photosynthesis or by increasing leaf mass per unit of area, nor the idea that acclimation of respiration to temperature would be rapid enough to make dark respiration insensitive to variation in temperature between nights.     

Rapid changes in δ¹³C of ecosystem-respired CO₂ after sunset are consistent with transient ¹³C enrichment of leaf respired CO₂

The CO? respired by darkened, light-adapted, leaves is enriched in 13C during the first minutes, and this effect may be related to rapid changes in leaf respiratory biochemistry upon darkening. We hypothesized that this effect would be evident at the ecosystem scale. High temporal resolution measurements of the carbon isotope composition of ecosystem respiration were made over 28 diel periods in an abandoned temperate pasture, and were compared with leaf-level measurements at differing levels of pre-illumination. At the leaf level, CO? respired by darkened leaves that had been preadapted to high light was strongly enriched in 13C, but such a 13C-enrichment rapidly declined over 60-100 min. The 13C-enrichment was less pronounced when leaves were preadapted to low light. These leaf-level responses were mirrored at the ecosystem scale; after sunset following clear, sunny days respired CO? was first 13C enriched, but the 13C-enrichment rapidly declined over 60-100 min. Further, this response was less pronounced following cloudy days. We conclude that the dynamics of leaf respiratory isotopic signal caused variations in ecosystem-scale 12CO?/13) CO? exchange. Such rapid isotope kinetics should be considered when applying 13C-based techniques to elucidate ecosystem carbon cycling.     

Respiratory carbon metabolism following illumination in intact French bean leaves using (13)C/(12)C isotope labeling

The origin of the carbon atoms in the CO(2) respired by French bean (Phaseolus vulgaris) leaves in the dark has been studied using (13)C/(12)C isotopes as tracers. The stable isotope labeling was achieved through a technical device that uses an open gas-exchange system coupled online to an elemental analyzer and linked to an isotope ratio mass spectrometer. The isotopic analysis of the CO(2) respired in the dark after a light period revealed that the CO(2) was labeled, but the labeling level decreased progressively as the dark period increased. The pattern of disappearance depended on the amount of carbon fixed during the labeling and indicated that there were several pools of respiratory metabolites with distinct turnover rates. We demonstrate that the carbon recently assimilated during photosynthesis accounts for less than 50% of the carbon in the CO(2) lost by dark respiration and that the proportion is not influenced by leaf starvation in darkness before the labeling. Therefore, most of the carbon released by dark respiration after illumination does not come from new photosynthates.     

Measurement and interpretation of the oxygen isotope composition of carbon dioxide respired by leaves in the dark

We measured the oxygen isotope composition (delta(18)O) of CO(2) respired by Ricinus communis leaves in the dark. Experiments were conducted at low CO(2) partial pressure and at normal atmospheric CO(2) partial pressure. Across both experiments, the delta(18)O of dark-respired CO(2) (delta(R)) ranged from 44 per thousand to 324 per thousand (Vienna Standard Mean Ocean Water scale). This seemingly implausible range of values reflects the large flux of CO(2) that diffuses into leaves, equilibrates with leaf water via the catalytic activity of carbonic anhydrase, then diffuses out of the leaf, leaving the net CO(2) efflux rate unaltered. The impact of this process on delta(R) is modulated by the delta(18)O difference between CO(2) inside the leaf and in the air, and by variation in the CO(2) partial pressure inside the leaf relative to that in the air. We developed theoretical equations to calculate delta(18)O of CO(2) in leaf chloroplasts (delta(c)), the assumed location of carbonic anhydrase activity, during dark respiration. Their application led to sensible estimates of delta(c), suggesting that the theory adequately accounted for the labeling of CO(2) by leaf water in excess of that expected from the net CO(2) efflux. The delta(c) values were strongly correlated with delta(18)O of water at the evaporative sites within leaves. We estimated that approximately 80% of CO(2) in chloroplasts had completely exchanged oxygen atoms with chloroplast water during dark respiration, whereas approximately 100% had exchanged during photosynthesis. Incorporation of the delta(18)O of leaf dark respiration into ecosystem and global scale models of C(18)OO dynamics could affect model outputs and their interpretation.     

Metabolic origin of carbon isotope composition of leaf dark-respired CO2 in French bean

The carbon isotope composition (delta(13)C) of CO(2) produced in darkness by intact French bean (Phaseolus vulgaris) leaves was investigated for different leaf temperatures and during dark periods of increasing length. The delta(13)C of CO(2) linearly decreased when temperature increased, from -19 per thousand at 10 degrees C to -24 per thousand at 35 degrees C. It also progressively decreased from -21 per thousand to -30 per thousand when leaves were maintained in continuous darkness for several days. Under normal conditions (temperature not exceeding 30 degrees C and normal dark period), the evolved CO(2) was enriched in (13)C compared with carbohydrates, the most (13)C-enriched metabolites. However, at the end of a long dark period (carbohydrate starvation), CO(2) was depleted in (13)C even when compared with the composition of total organic matter. In the two types of experiment, the variations of delta(13)C were linearly related to those of the respiratory quotient. This strongly suggests that the variation of delta(13)C is the direct consequence of a substrate switch that may occur to feed respiration; carbohydrate oxidation producing (13)C-enriched CO(2) and beta-oxidation of fatty acids producing (13)C-depleted CO(2) when compared with total organic matter (-27.5 per thousand). These results are consistent with the assumption that the delta(13)C of dark respired CO(2) is determined by the relative contributions of the two major decarboxylation processes that occur in darkness: pyruvate dehydrogenase activity and the Krebs cycle.     

Atmospheric CO2 concentration may directly affect leaf respiration measurement in tobacco,but not respiration itself*

When atmospheric CO₂ concentration increases, various consequences for plant metabolism have been suggested, such as changes in photosynthesis, photorespiration or respiration which can affect growth and carbon sequestration. In addition to long‐term (indirect) effects on respiration, short‐term (direct) effects of CO₂ concentration on the respiration of leaves, shoots and roots are described in the literature. In most cases, respiration is reported to be inhibited by increased CO₂ concentration, but the mechanism(s) are not yet understood. It has been shown previously that, when the respective technical problems and properties of a gas exchange system are fully considered, a short‐term increase in CO₂ (up to 4200 µmol mol^?1) had no effect on respiration of Phaseolus or Populus leaves (Jahnke, Plant, Cell and Environment 24, 1139–1151, 2001). However, in the present study, large (apparent) CO₂ effects were found with mature Nicotiana leaves whereas, in young leaves, the effect was absent. The experimental results clearly show that the observed direct CO₂ effect on dark CO₂ efflux in the mature tobacco leaves was caused by leakage of CO₂ inside the leaves (and the magnitude of the effect was dependent on the size of the leakage). Nicotiana leaves are, in contrast to Phaseolus and Populus leaves (which are heterobaric), characterized by a homobaric anatomy in which intercellular air spaces are not compartmented and provide a continuous system of open pores in the lateral (paradermal) direction of the leaves. Mesophyll porosity increases with leaf development, which explains the differences between young and mature tobacco leaves. When internal leakage was experimentally restricted, the CO₂ inhibition on CO₂ efflux was no longer observed. It is concluded that the measured direct CO₂ effect(s) on leaf CO₂ efflux in the dark are artefactual, and that a true direct CO₂ effect on leaf respiration does not exist.     

Respiratory carbon metabolism in the high mountain plant species Ranunculus glacialis

Very little is known about the primary carbon metabolism of the high mountain plant Ranunculus glacialis. It is a species with C3 photosynthesis, but with exceptionally high malate content in its leaves, the biological significance of which remains unclear. 13C/12C-isotope ratio mass spectrometry (IRMS) and 13C-nuclear magnetic resonance (NMR) labelling were used to study the carbon metabolism of R. glacialis, paying special attention to respiration. Although leaf dark respiration was high, the temperature response had a Q10 of 2, and the respiratory quotient (CO2 produced divided by O2 consumed) was nearly 1, indicating that the respiratory pool is comprised of carbohydrates. Malate, which may be a large carbon substrate, was not respired. However, when CO2 fixed by photosynthesis was labelled, little labelling of the CO2 subsequently respired in the dark was detected, indicating that: (i) most of the carbon recently assimilated during photosynthesis is not respired in the dark; and (ii) the carbon used for respiration originates from (unlabelled) reserves. This is the first demonstration of such a low metabolic coupling of assimilated and respired carbon in leaves. The biological significance of the uncoupling between assimilation and respiration is discussed.     

Carbon dioxide concentration at night affects translocation from soybean leaves

Studies have indicated that the concentration of carbon dioxide [CO2] during the dark period may influence plant dry matter accumulation. It is often suggested that these effects on growth result from effects of [CO2] on rates of respiration, but responses of respiration to [CO2] remain controversial, and connections between changes in respiration rate and altered growth rate have not always been clear. The present experiments tested whether translocation, a major consumer of energy from respiration in exporting leaves, was sensitive to [CO2]. Nineteen-day-old soybean plants grown initially at a constant [CO2] of 350 micromol mol(-1) were exposed to three consecutive nights with a [CO2] of 220-1400 micromol mol(-1), with a daytime [CO2] of 350 micromol mol(-1). Change in dry mass of the individual second, third and fourth trifoliate leaves over the 3-d period was determined, along with rates of respiration and photosynthesis of second leaves, measured by net CO2 exchange. Translocation was determined from mass balance for second leaves. Additional experiments were conducted where the [CO2] around individual leaves was controlled separately from that of the rest of the plant. Results indicated that low [CO2] at night increased both respiration and translocation and elevated [CO2] decreased both processes, to similar relative extents. The effect of [CO2] during the dark on the change in leaf mass over 3 d was largest in second leaves, where the change in mass was about 50% greater at 1400 micromol mol(-1) CO2 than at 220 micromol mol(-1) CO2. The response of translocation to [CO2] was localized in individual leaves. Results indicated that effects of [CO2] on net carbon dioxide exchange rate in the dark either caused or reflected a change in a physiologically important process which is known to depend on energy supplied by respiration. Thus, it is unlikely that the observed effects of [CO2] on respiration were artefacts of the measurement process in this case.