Metabolism of <Superscript>14</Superscript>C-glycine as a substrate for photorespiration of the leaf at different developmental stages of ephemeroides

The metabolism of ¹⁴C-glycine (a substrate for photorespiration) was studied in the light and in darkness under natural CO₂ concentration (0.03%) in the leaves of ephemeroides Scilla sibirica Haw. and Ficaria verna Huds. at different developmental stages. Using one and the same sample, potential photosynthesis (at 1% CO₂), true photosynthesis (at 0.03% CO₂), and leaf respiratory capacity were measured by the radiometric and manometric methods, respectively. All measurements were performed at 15°C, an average temperature during ephemer growth. It was found that, in the white zone of the Scilla leaf, the rate of CO₂ evolution resulting from metabolization of exogenous ¹⁴C-glycine was similar in the light and in darkness. In the green zone of the Scilla leaf and in the green leaf of Ficaria, both ¹⁴C-glycine absorption and ¹⁴CO₂ evolution were lower in the light as compared with darkness, which is explained by CO₂ reassimilation. In all treatments of both plant species, a specific inhibitor of glycine decarboxylase complex (GDC), aminoacetonitrile (5 mM) suppressed CO₂ evolution by 20–40%. It was concluded that in ephemeroides mitochondrial GDC, responsible for CO₂ evolution in photorespiration, is formed at the earliest stage of leaf development. This indicates that photorespiration can occur simultaneously with the development of the leaf photosynthetic activity. On the basis of the assumption that carbon losses in the form of CO₂ evolved during photorespiration comprise 25% of true photosynthesis, it was calculated that, in ephemer leaves, the highest rates of photorespiration and photosynthesis were attained during flowering when the leaf area was the largest and the rate of dark respiration was reduced by 1.5–2.0 times. The highest rates of dark respiration were observed in the beginning of growth. In senescing leaves by the end of the plant vegetation, potential photosynthesis and true photosynthesis were reduced, whereas dark respiration remained essentially unchanged. It is concluded that the high rates of potential and true photosynthesis are characteristic of ephemeroides when they complete their short developmental program in early spring (at 15°C); theoretically, photorespiration also occurs at a high rate during this period, when this process provides for a defense against the threat of photoinhibition at low temperature and high insolation.     

Carbon balance in a monospecific stand of an annual herb <Emphasis Type="Italic">Chenopodium album</Emphasis> at an elevated CO<Subscript>2</Subscript> concentration

Elevated CO₂ enhances carbon uptake of a plant stand, but the magnitude of the increase varies among growth stages. We studied the relative contribution of structural and physiological factors to the CO₂ effect on the carbon balance during stand development. Stands of an annual herb Chenopodium album were established in open-top chambers at ambient and elevated CO₂ concentrations (370 and 700 μmol mol⁻¹). Plant biomass growth, canopy structural traits (leaf area, leaf nitrogen distribution, and light gradient in the canopy), and physiological characteristics (leaf photosynthesis and respiration of organs) were studied through the growing season. CO₂ exchange of the stand was estimated with a canopy photosynthesis model. Rates of light-saturated photosynthesis and dark respiration of leaves as related with nitrogen content per unit leaf area and time-dependent reduction in specific respiration rates of stems and roots were incorporated into the model. Daily canopy carbon balance, calculated as an integration of leaf photosynthesis minus stem and root respiration, well explained biomass growth determined by harvests (r ² = 0.98). The increase of canopy photosynthesis with elevated CO₂ was 80% at an early stage and decreased to 55% at flowering. Sensitivity analyses suggested that an alteration in leaf photosynthetic traits enhanced canopy photosynthesis by 40–60% throughout the experiment period, whereas altered canopy structure contributed to the increase at the early stage only. Thus, both physiological and structural factors are involved in the increase of carbon balance and growth rate of C. album stands at elevated CO₂. However, their contributions were not constant, but changed with stand development.     

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.

     

Variation in photorespiration. The effect of genetic differences in photorespiration on net photosynthesis in tobacco

The hypothesis that net photosynthesis is diminished in many plant species because of a high rate of CO₂ evolution in the light has been tested further. High rates of CO₂ output in CO₂-free air in comparison with dark respiration were found in Chlamydomonas reinhardi, wheat leaves, tomato leaves, and to a lesser extent in Chlorella pyrenoidosa by means of the ¹⁴C-photorespiration assay. In tobacco leaves high photorespiration was characteristic of a standard variety, Havana Seed, and a possibly still higher rate was found in a yellow heterozygous mutant, JWB Mutant. However, the dark homozygous sibling of the latter, JWB Wild, had a low photorespiration for the tobacco species. The relative rates of photorespiration were in the same sequence when measured by the ¹⁴CO₂ released in normal air from leaf disks supplied with glycolate-1-¹⁴C in the light.

As would be predicted by the hypothesis, the maximal net rate of photosynthesis at 300 ppm of CO₂ in the air in JWB Wild leaves was greater (24%) than in Havana Seed, while JWB Mutant had less CO₂ uptake than the standard variety (21%). At 550 ppm of CO₂ the differences in net photosynthesis were not as great between the 2 siblings as at 200 ppm. The relative leaf expansion rates of seedlings of the 3 tobacco varieties in a greenhouse had the same relationship as their rates of CO₂ assimilation.

Thus within the tobacco species, as in a comparison between tobacco and maize, low photorespiratory CO₂ evolution was correlated with higher photosynthetic efficiency. Therefore it seems that increased CO₂ uptake should be achieved by genetic interference with the process of photorespiration.

     

Seasonal Changes in the Photosynthetic Rate in Apple Trees : A Comparison between Fruiting and Nonfruiting Trees

Seasonal changes in photosynthesis of apple trees (Malus domestica Borkh.) were monitored to examine the effect of source-sink interactions on photosynthesis and photorespiration. Elevated photosynthetic rates were observed during two periods of the growing season and correlated with the fruiting process. The first period of increased photosynthetic rates was during the bloom period, when spur leaves on flowering shoots exhibited up to 25% higher photosynthetic rates than vegetative spur leaves on a leaf area basis. CO₂ assimilation rates were also higher in fruiting trees than nonfruiting trees during the period of rapid fruit growth from July to September. Photorespiration, dark respiration, leaf resistance, and transpiration exhibited no seasonal changes which correlated to the presence or absence of fruit. These data represent the first comprehensive examination of the effects of flowering/fruit formation on photosynthesis and photorespiration in perennial plants.     

Atmospheric CO2 mole fraction affects stand‐scale carbon use efficiency of sunflower by stimulating respiration in light

Plant carbon‐use‐efficiency (CUE), a key parameter in carbon cycle and plant growth models, quantifies the fraction of fixed carbon that is converted into net primary production rather than respired. CUE has not been directly measured, partly because of the difficulty of measuring respiration in light. Here, we explore if CUE is affected by atmospheric CO₂. Sunflower stands were grown at low (200 μmol mol^?1) or high CO₂ (1000 μmol mol^?1) in controlled environment mesocosms. CUE of stands was measured by dynamic stand‐scale ¹³C labelling and partitioning of photosynthesis and respiration. At the same plant age, growth at high CO₂ (compared with low CO₂) led to 91% higher rates of apparent photosynthesis, 97% higher respiration in the dark, yet 143% higher respiration in light. Thus, CUE was significantly lower at high (0.65) than at low CO₂ (0.71). Compartmental analysis of isotopic tracer kinetics demonstrated a greater commitment of carbon reserves in stand‐scale respiratory metabolism at high CO₂. Two main processes contributed to the reduction of CUE at high CO₂: a reduced inhibition of leaf respiration by light and a diminished leaf mass ratio. This work highlights the relevance of measuring respiration in light and assessment of the CUE response to environment conditions.     

In situ CO2 gas-exchange in fruits of a tropical tree,Durio zibethinus Murray

We examined the in situ CO₂ gas-exchange of fruits of a tropical tree, Durio zibethinus Murray, growing in an experimental field station of the Universiti Pertanian Malaysia. Day and night dark respiration rates were exponentially related to air temperature. The temperature dependent dark respiration rate showed a clockwise loop as time progressed from morning to night, and the rate was higher in the daytime than at night. The gross photosynthetic rate was estimated by summing the rates of daytime dark respiration and net photosynthesis. Photosynthetic CO₂ refixation, which is defined as the ratio of gross photosynthetic rate to dark respiration rate in the daytime, ranged between 15 and 45%. The photosynthetic CO₂ refixation increased rapidly as the temperature increased in the lower range of air temperature T _c (T _c <28.5 °C), while it decreased gradually as the temperature increased in the higher range (T _c 28.5 °C). Light dependence of photosynthetic CO₂ refixation was approximated by a hyperbolic formula, where light saturation was achieved at 100 mol m^–2 s^–1 and the asymptotic CO₂ refixation was determined to be 37.4%. The estimated gross photosynthesis and dark respiration per day were 1.15 and 4.90 g CO₂ fruit^–1, respectively. Thus the CO₂ refixation reduced the respiration loss per day by 23%. The effect of fruit size on night respiration rate satisfied a power function, where the exponent was larger than unity.     

Involvement of respiratory processes in the transient knockout of net CO2 uptake in Mimosa pudica upon heat stimulation

Leaf photosynthesis of the sensitive plant Mimosa pudica displays a transient knockout in response to electrical signals induced by heat stimulation. This study aims at clarifying the underlying mechanisms, in particular, the involvement of respiration. To this end, leaf gas exchange and light reactions of photosynthesis were assessed under atmospheric conditions largely eliminating photorespiration by either elevated atmospheric CO₂ or lowered O₂ concentration (i.e. 2000 μmol mol^?1 or 1%, respectively). In addition, leaf gas exchange was studied in the absence of light. Under darkness, heat stimulation caused a transient increase of respiratory CO₂ release simultaneously with stomatal opening, hence reflecting direct involvement of respiratory stimulation in the drop of the net CO₂ uptake rate. However, persistence of the transient decline in net CO₂ uptake rate under illumination and elevated CO₂ or 1% O₂ makes it unlikely that photorespiration is the metabolic origin of the respiratory CO₂ release. In conclusion, the transient knockout of net CO₂ uptake is at least partially attributed to an increased CO₂ release through mitochondrial respiration as stimulated by electrical signals. Putative CO₂ limitation of Rubisco due to decreased activity of carbonic anhydrase was ruled out as the photosynthesis effect was not prevented by elevated CO₂.     

Investigation on photorespiration with a sensitive C-assay

A leaf disk assay for photorespiration has been developed based on the rate of release of recently fixed ¹⁴CO₂ in light in a rapid stream of CO₂-free air at 30° to 35°. In tobacco leaves (Havana Seed) photorespiration with this assay is 3 to 5 times greater than the ¹⁴CO₂ output in the dark. In maize, photorespiration is only 2% of that in tobacco.

The importance of open leaf stomata, rapid flow rates of CO₂-free air, elevated temperatures, and oxygen in the atmosphere in order to obtain release into the air of a larger portion of the ¹⁴CO₂ evolved within the tissue in the light was established in tobacco. Photorespiration, but not dark respiration, was inhibited by α-hydroxy-2-pyridinemethanesulfonic acid, an inhibitor of glycolate oxidase, and by 3-(4-chlorophenyl)-1,1-dimethylurea (CMU), an inhibitor of photosynthetic electron transport, under conditions which did not affect the stomata. These experiments show that the substrates of photorespiration and dark respiration differ and also provide additional support for the role of glycolate as a major substrate of photorespiration. It was also shown that at 35° the quantity of ¹⁴CO₂ released in the assay may represent only 33% of the gross ¹⁴CO₂ evolved in the light, the remainder being recycled within the tissue.

It was concluded that maize does not evolve appreciable quantities of CO₂ in the light and that this largely accounts for the greater efficiency of net photosynthesis exhibited by maize. Hence low rates of photorespiration may be expected to be correlated with a high rate of CO₂ uptake at the normal concentrations of CO₂ found in air and at higher light intensities.

     

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.     

Effect of carbon dioxide and temperature on photosynthetic CO2 uptake and photorespiratory CO2 evolution in sunflower leaves

Using an open gas-exchange system, apparent photosynthesis, true photosynthesis (TPS), photorespiration (PR) and dark respiration of sunflower (Helianthus annuus L.) leaves were determined at three temperatures and between 50 and 400 l/l external CO₂. The ratio of PR/TPS and the solubility ratio of O₂/CO₂ in the intercellular spaces both decreased with increasing CO₂. The rate of PR was not affected by the CO₂ concentration in the leaves and was independent of the solubility ratio of oxygen and CO₂ in the leaf cell. At photosynthesis-limiting concentrations of CO₂, the ratio of PR/TPS significantly increased from 18 to 30°C and the rate of PR increased from 4.3 mg CO₂ dm^-2 h^-1 at 18°C to 8.6 mg CO₂ dm^-2 h^-1 at 30°C. The specific activity of photorespired CO₂ was CO₂-dependent but temperature-independent, and the carbon traversing the glycolate pathway appeared to be derived both from recently fixed assimilate and from older reserve materials. It is concluded that PR as a percentage of TPS is affected by the concentrations of O₂ and CO₂ around the photosynthesizing cells, but the rate of PR may also be controlled by other factors.Abbreviations APS apparent photosynthesis (net CO₂ uptake) - PR photorespiration (CO₂ evolution in light) - RuBP ribulose-1,5-bisphosphate - TPS true photosynthesis (true CO₂ uptake)     

Effects of irradiance and temperature on photosynthesis in C3, C4 and C3/C4 Panicum species

Species in the Laxa and Grandia groups of the genus Panicum are adapted to low, wet areas of tropical and subtropical America. Panicum milioides is a species with C₃ photosynthesis and low apparent photorespiration and has been classified as a C₃/C₄ intermediate. Other species in the Laxa group are C₃ with normal photorespiration. Panicum prionitis is a C₄ species in the Grandia group. Since P. milioides has some leaf characteristics intermediate to C₃ and C₄ species, its photosynthetic response to irradiance and temperature was compared to the closely related C₃ species, P. laxum and P. boliviense and to P. prionitis. The response of apparent photosynthesis to irradiance and temperature was similar to that of P. laxum and P. boliviense, with saturation at a photosynthetic photo flux density of about 1 mmol m^-2 s^-1 at 30°C and temperature optimum near 30°C. In contrast, P. prionitis showed no light saturation up to 2 mmol m^-2 s^-1 and an optimum temperature near 40°C. P. milioides exhibited low CO₂ loss into CO₂-free air in the light and this loss was nearly insensitive to temperature. Loss of CO₂ in the light in the C₃ species, P. laxum and P. boliviense, was several-fold higher than in P. milioides and increased 2- to 5-fold with increases in temperature from 10 to 40°C. The level of dark respiration and its response to temperature were similar in all four Panicum species examined. It is concluded that the low apparent photorespiration in P. milioides does not influence its response of apparent photosynthesis to irradiance and temperature in comparison to closely related C₃ Panicum species.Abbreviations AP apparent photosynthesis - I CO₂ compensation point - gl leaf conductance; gm, mesophyll conductance - PPFD photosynthetic photon flux density - PR apparent photorespiration rate - RuBPC sibulose bisphosphate carboxylase     

Short‐term effects of CO2 and O2 on citrate metabolism in illuminated leaves

Although there is now a considerable literature on the inhibition of leaf respiration (CO₂ evolution) by light, little is known about the effect of other environmental conditions on day respiratory metabolism. In particular, CO₂ and O₂ mole fractions are assumed to cause changes in the tricarboxylic acid pathway (TCAP) but the amplitude and even the direction of such changes are still a matter of debate. Here, we took advantage of isotopic techniques, new simple equations and instant freeze sampling to follow respiratory metabolism in illuminated cocklebur leaves (Xanthium strumarium L.) under different CO₂/O₂ conditions. Gas exchange coupled to online isotopic analysis showed that CO₂ evolved by leaves in the light came from ‘old’ carbon skeletons and there was a slight decrease in ¹³C natural abundance when [CO₂] increased. This suggested the involvement of enzymatic steps fractionating more strongly against ¹³C and thus increasingly limiting for the metabolic respiratory flux as [CO₂] increased. Isotopic labelling with ¹³C₂‐2,4‐citrate lead to ¹³C‐enriched Glu and 2‐oxoglutarate (2OG), clearly demonstrating poor metabolism of citrate by the TCAP. There was a clear relationship between the ribulose‐1,5‐bisphosphate oxygenation‐to‐carboxylation ratio (v_o/v_c) and the ¹³C commitment to 2OG, demonstrating that 2OG and Glu synthesis via the TCAP is positively influenced by photorespiration.     

Effects of temperature on the gas exchange of leaves in the light and dark

Summary Evolution of CO₂ into CO₂-free air was measured in the light and in the dark over a range of temperatures from 15 to 50°. Photosynthetic rates were measured in air and O₂-free air over the same range of temperatures. Respiration in the light had a different sensitivity to temperature compared with respiration in the dark. At the lower temperatures the rate of respiration in the light was higher than respiration in the dark, whereas at temperatures above 40° the reverse was observed. For any one species the maximum rates of photosynthesis and photorespiration occur at about the same temperature. The maximum rate for dark respiration generally is found at a temperature about 10° higher. Zea mays and Atriplex nummularia showed no enhancement of photosynthesis in O₂-free air nor any evolution of CO₂ in CO₂-free air at any of the temperatures.     

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     

Leaf development and phloem transport in Cucurbita pepo: Carbon economy

Summary Net photosynthesis, dark respiration and growth for leaf 5 of Cucurbita pepo L. plants grown under controlled conditions were measured and the data used for an assessment of the changes in carbon balance during growth of the leaf through expansion to maturity. The blade is first capable of net CO₂ fixation when ca. 8% expanded but the initial rapid growth during this period is sustained almost entirely through imported nutrients. When the growth rate starts to decline rapidly the net photosynthetic capacity of the blade begins to increase. This increase is accompanied by an expansion of the intercellular spaces and by decreasing dark respiration measured at night and in dark periods during the day. The blade becomes completely independent of phloem imported nutrients and begins to export excess photosynthate when the phase of rapid decrease in relative growth rate is almost complete at about 45% expansion. Maximum net photosynthesis of ca. 11 mg CO₂ h^-1 dm^-2 is achieved at 70% expansion. The first detectable synthesis of the transport sugars stachyose and raffinose in the blade coincides with the beginning of intralaminar phloem transport from the tip to the base of the leaf. The synthesis of sucrose, the other major transport sugar, is detectable at all stages of leaf development.     

Effects of daytime carbon dioxide concentration on dark respiration in rice

Rising atmospheric carbon dioxide concentration ([CO₂]) has generated considerable interest in the response of agricultural crops to [CO₂]. The objectives of this study were to determine the effects of a wide range of daytime [CO₂] on dark respiration of rice (Oryza sativa L. cv. IR-30). Rice plants were grown season-long in naturally sunlit plant growth chambers in subambient (160 and 250), ambient (330), or super-ambient (500, 660 and 900 μmol CO₂ mol^?1 air) [CO₂] treatments. Canopy dark respiration, expressed on a ground area basis (R_d) increased with increasing [CO₂] treatment from 160 to 500 μmol mol^?1 treatments and was very similar among the superambient treatments. The trends in R_d over time and in response to increasing daytime [CO₂] treatment were associated with and similar to trends previously described for photosynthesis. Specific respiration rate (R_dw) decreased with time during the growing season and was higher in the subambient than the ambient and superambient [CO₂] treatments. This greater R_dw in the subambient [CO₂] treatments was attributed to a higher specific maintenance respiration rate and was associated with higher plant tissue nitrogen concentration.     

On the 13C/12C isotopic signal of day and night respiration at the mesocosm level

While there is currently intense effort to examine the ¹³C signal of CO₂ evolved in the dark, less is known on the isotope composition of day‐respired CO₂. This lack of knowledge stems from technical difficulties to measure the pure respiratory isotopic signal: day respiration is mixed up with photorespiration, and there is no obvious way to separate photosynthetic fractionation (pure c_i/c_a effect) from respiratory effect (production of CO₂ with a different δ¹³C value from that of net‐fixed CO₂) at the ecosystem level. Here, we took advantage of new simple equations, and applied them to sunflower canopies grown under low and high [CO₂]. We show that whole mesocosm‐respired CO₂ is slightly ¹³C depleted in the light at the mesocosm level (by 0.2–0.8‰), while it is slightly ¹³C enriched in darkness (by 1.5–3.2‰). The turnover of the respiratory carbon pool after labelling appears similar in the light and in the dark, and accordingly, a hierarchical clustering analysis shows a close correlation between the ¹³C abundance in day‐ and night‐evolved CO₂. We conclude that the carbon source for respiration is similar in the dark and in the light, but the metabolic pathways associated with CO₂ production may change, thereby explaining the different ¹²C/¹³C respiratory fractionations in the light and in the dark.     

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.     

Would transformation of C3 crop plants with foreign Rubisco increase productivity? A computational analysis extrapolating from kinetic properties to canopy photosynthesis

Genetic modification of Rubisco to increase the specificity for CO₂ relative to O₂ (τ) would decrease photorespiration and in principle should increase crop productivity. When the kinetic properties of Rubisco from different photosynthetic organisms are compared, it appears that forms with high τ have low maximum catalytic rates of carboxylation per active site (k^c_c). If it is assumed that an inverse relationship between k^c_c and τ exists, as implied from measurements, and that an increased concentration of Rubisco per unit leaf area is not possible, will increasing τ result in increased leaf and canopy photosynthesis? A steady‐state biochemical model for leaf photosynthesis was coupled to a canopy biophysical microclimate model and used to explore this question. C₃ photosynthetic CO₂ uptake rate (A) is either limited by the maximum rate of Rubisco activity (V_cmax) or by the rate of regeneration of ribulose‐1,5‐bisphosphate, in turn determined by the rate of whole chain electron transport (J). Thus, if J is limiting, an increase in τ will increase net CO₂ uptake because more products of the electron transport chain will be partitioned away from photorespiration into photosynthesis. The effect of an increase in τ on Rubisco‐limited photosynthesis depends on both k^c_c and the concentration of CO₂ ([CO₂]). Assuming a strict inverse relationship between k^c_c and τ, the simulations showed that a decrease, not an increase, in τ increases Rubisco‐limited photosynthesis at the current atmospheric [CO₂], but the increase is observed only in high light. In crop canopies, significant amounts of both light‐limited and light‐saturated photosynthesis contribute to total crop carbon gain. For canopies, the present average τ found in C₃ terrestrial plants is supra‐optimal for the present atmospheric [CO₂] of 370 µmol mol^?1, but would be optimal for a CO₂ concentration of around 200 µmol mol^?1, a value close to the average of the last 400 000 years. Replacing the average Rubisco of terrestrial C₃ plants with one having a lower and optimal τ would increase canopy carbon gain by 3%. Because there are significant deviations from the strict inverse relationship between k^c_c and τ, the canopy model was also used to compare the rates of canopy photosynthesis for several Rubiscos with well‐defined kinetic constants. These simulations suggest that very substantial increases (> 25%) in crop carbon gain could result if specific Rubiscos having either a higher τ or higher k^c_c were successfully expressed in C₃ plants.