The Responses of Stomatal Density to CO2 Partial Pressure

Experiments on a range of species of tree, shrub and herb haveshown that stomatal density and stomatal index increase as thepartial pressure of CO₂ decreases over the range from the currentlevel of 34 Pa to 22.5 Pa. Stomatal density responds to thereduced partial pressure of CO₂ in a simulation of high altitude(3000 m), when the CO₂ mole fraction is unchanged. When the partial pressure of CO₂ is increased from 35 to 70Pa stomatal density decreases slightly, with a response to unitchange in CO₂ which is about 10% of that below 34 Pa. Measurements of gas exchange on leaves which had developed indifferent CO₂ partial pressures, but at low saturation vapourpressure deficits in the range of 0.7 to 0.9 kPa, indicatedlower photosynthetic rates but higher stomatal conductancesat reduced CO₂ partial pressures. Experiments on populations of Nardus stricta originating fromaltitudes of 366 m and 810 m in Scotland, indicated geneticdifferences in the responses of stomatal density to CO₂ in pressuressimulating altitudes of sea level and 2 000 m. Plants from thehigher altitude showed greater declines in stomatal densitywhen the CO₂ partial pressure was increased. Key words: Stomata, CO₂, gas exchange, altitude, atmospheric pressure     

Effect of CO2 Concentration During Growth on a CO2 Concentrating Mechanism in White Clover as Predicted from Differential 14CO2/12CO2Uptake

Lehnherr, B. M?chler, F. and N?sberger, J. 1985. Effect of CO₂concentration during growth on a CO₂ concentrating mechanismin white clover as predicted from differential ¹⁴CO₂/¹²CO² uptake.-J. exp. Bot. 36: 1835-1841. White clover was grown at 20 and100 Pa p(CO₂). The CO₂ response of net photosynthesis and differentialuptake of ¹⁴CO₂ and ¹²CO₂ by leaves were measured at varioustemperatures and at various O₂ and CO₂ partial pressures andcompared with predictions from ribulose bisphosphate carboxylase/oxygenasekinetics. Discrepancies between the observed gas exchange characteristicsfor the leaves and those predicted from the enzyme kineticswere interpreted as being due to a CO₂ concentrating mechanism.Plants grown at 20 Pa p(CO₂) showed a higher affinity for CO₂than plants grown at 100 Pa p(CO₂) when measured at 10 ?C. Nodifference in affinity was found at 30 ?C. The postulated CO₂concentrating effect was greater in plants grown at low CO₂than in plants grown at high CO₂ concentration and occurredonly at low temperature and low CO₂ partial pressure. It issuggested that plants grown at the lower CO₂ partial pressurehave a higher affinity for CO₂ due to a more efficient CO₂ concentratingsystem than plants grown at the higher CO₂ partial pressure. Key words: Photosynthesis, CO₂, concentration, RuBP carboxylase/oxygenase     

Energetic Costs of Tissue Construction in Yellow-poplar and White Oak Trees Exposed to Long-term CO2Enrichment

Two methods were used to estimate construction costs for leaves,stems, branches and woody roots of yellow-poplar (LiriodendrontulipiferaL.) trees grown at ambient (35 Pa) and elevated (65Pa) CO₂for 2.7 years and trees of white oak (Quercus albaL.)grown at these same CO₂partial pressures for 4 years. Samplecombustion in a bomb calorimeter combined with measurementsof ash and nitrogen content provided the primary method of estimatingtissue construction costs (W_G; g glucose g^-1dry mass). Thesevalues were compared with a second, simpler method in whichcost estimates were derived from tissue ash, carbon and nitrogencontent (V_G). Estimates of W_Gwere lower for leaves, branchesand roots of yellow-poplar and for leaves of white oak grownat elevated compared with ambient CO₂partial pressures. TheseCO₂-induced differences in W_Granged from 3.7% in yellow-poplarroots to 2.1% in white oak leaves. Only in the case of yellow-poplarleaves, however, were differences in V_Gobserved between CO₂treatments.Leaf V_Gwas 1.46 g glucose g^-1dry mass in ambient-grown treescompared with 1.41 g glucose g^-1dry mass for CO₂-enriched trees.Although paired-estimates of W_Gand V_Gclustered about a 1:1 linefor leaves and branches, estimates of V_Gwere consistently lowerthan W_Gfor stems and roots. Construction costs per unit leafarea were 95 g glucose m^-2for yellow-poplar trees grown at ambientCO₂and 106 g glucose m^-2for trees grown at elevated CO₂partialpressures. No differences in area-based construction costs wereobserved for white oak. Whole-plant energy content was 1220g glucose per tree in ambient-grown white oak compared with2840 g glucose per tree for those grown at elevated CO₂partialpressures. These differences were driven largely by CO₂-inducedchanges in total biomass. We conclude that while constructioncosts were lower at elevated CO₂partial pressures, the magnitudeof this response argues against an increased efficiency of carbonuse in the growth processes of trees exposed to CO₂enrichment. Bomb calorimeter; construction costs; elevated CO₂; energy allocation; global change; growth respiration; heat of combustion; respiration; Liriodendron tulipifera; Quercus alba     

Reactions of birch leaves to changes in light during early ontogeny: comparison between in vivo and in vitro techniques to measure carbon uptake

Trends in several photosynthetic parameters and their responseto changed growth light were followed for 15 d in leaves ofyoung birch saplings using a rapid-response gas exchange measuringequipment. These in vivo measurements were compared to biochemicalassays that were made from the same leaves after the gas exchangestudies. The measurements were made on leaves that were selectedprior to the study and were at that time of similar age. Forthe first 7 d the photosynthetic parameters were followed fromthe growth conditions of moderate light (200 µmol m^–2s^–1; referred to as controls later in the text). On day7 some of the saplings were transferred to grow either underhigh (450 µmol m^–2 s^–1; referred to as highlight plants) or low (75 µmol m^–2 s^–1; referredto as low light plants) light and the capability of the preselectedleaves for acclimation was followed for 6 d. For comparison,at the end of the experiment the measurements were made on bothcontrols and on young leaves that had developed under high andlow light. Generally the in vivo measured rate of CO₂ uptake (gross photosynthesis)both at 310 ppm CO₂ and 2000 ppm CO₂ corresponded very wellto the biochemically determined CO₂ fixation capacity in vitroafter rapid extraction (measured as the initial and total activityof Rubisco, respectively). However, if the flux of CO₂ intothe chloroplasts was limited by the closure of the stomata,as was the case of the high light plants, then the in vitromeasured Rubisco activity was greater than the in vivo measuredCO₂ uptake. V_max, calculated from the mesophyll conductanceat 1% O₂, exceeded the initial activity of Rubisco (assayedat saturating RuBP and CO₂) constantly by 60%. The catalyticactivity of Rubisco in birch leaves was overall very low, evenwhen calculated from the total activity of Rubisco (K_cat 0.63–1.18 s^–1), when compared to herbaceous C₃ species. Signs of light acclimation were not observed in most of thephotosynthetic parameters and in chloroplast structure whenmature birch leaves were subjected to changes in growth lightfor 6 d. However, the change of the growth light either to highor low light caused day-to-day fluctuations in most of the measuredphotosynthetic parameters and in the case of the high lightplants signs of photoinhibition and photodestruction were alsoobserved (decrease in the amount of chlorophyll and increasein chlorophyll a/b ratio). As a result of these fluctuationsthese plants achieved a new and lower steady-state conditionbetween the light and dark reactions, as judged from the molarratio of RuBP to Rubisco binding site. Key words: Acclimation, photosynthesis, light, Rubisco, birch     

Estimating the Excess Investment in Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase in Leaves of Spring Wheat Grown under Elevated CO2

Wheat (Triticum aestivum L.) was grown under CO₂ partial pressures of 36 and 70 Pa with two N-application regimes. Responses of photosynthesis to varying CO₂ partial pressure were fitted to estimate the maximal carboxylation rate and the nonphotorespiratory respiration rate in flag and preceding leaves. The maximal carboxylation rate was proportional to ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) content, and the light-saturated photosynthetic rate at 70 Pa CO₂ was proportional to the thylakoid ATP-synthase content. Potential photosynthetic rates at 70 Pa CO₂ were calculated and compared with the observed values to estimate excess investment in Rubisco. The excess was greater in leaves grown with high N application than in those grown with low N application and declined as the leaves senesced. The fraction of Rubisco that was estimated to be in excess was strongly dependent on leaf N content, increasing from approximately 5% in leaves with 1 g N m⁻² to approximately 40% in leaves with 2 g N m⁻². Growth at elevated CO₂ usually decreased the excess somewhat but only as a consequence of a general reduction in leaf N, since relationships between the amount of components and N content were unaffected by CO₂. We conclude that there is scope for improving the N-use efficiency of C₃ crop species under elevated CO₂ conditions.     

Gas Exchange and Carbohydrate and Nitrogen Concentrations in Leaves of Pascopyrum smithii (C3) and Bouteloua gracilis (C4) at Different Carbon Dioxide Concentrations and Temperatures

Pascopyrum smithii (C₃) andBouteloua gracilis (C₄) are importantforage grasses native to the Colorado shortgrass steppe. Thisstudy investigated photosynthetic responses of these grassesto long-term CO₂enrichment and temperature in relation to leafnonstructural carbohydrate (TNC) and [N]. Glasshouse-grown seedlingswere transferred to growth chambers and grown for 49 d at twoCO₂concentrations (380 and 750 µmol mol^-1) at 20 and 35°C, and two additional temperatures (25 and 30 °C) at750 µmol mol^-1CO₂. Leaf CO₂exchange rate (CER) was measuredat a plant's respective growth temperature and at two CO₂concentrationsof approx. 380 and 700 µmol mol^-1. Long-term CO₂enrichmentstimulated CER in both species, although the response was greaterin the C₃,P. smithii . Doubling the [CO₂] from 380 to 750 µmolmol^-1stimulated CER ofP. smithii slightly more in plants grownand measured at 30 °C compared to plants grown at 20, 25or 35 °C. CO₂-enriched plants sometimes exhibited lowerCER when compared to ambient-grown controls measured at thesame [CO₂], indicating photosynthetic acclimation to CO₂growthregime. InP. smithii , such reductions in CER were associatedwith increases in TNC and specific leaf mass, reductions inleaf [N] and, in one instance, a reduction in leaf conductancecompared to controls. InB. gracilis , photosynthetic acclimationwas observed more often, but significant changes in leaf metabolitelevels from growth at different [CO₂] were generally less evident.Temperatures considered optimal for growth (C₃: 20 °C; C₄:35 °C) sometimes led to CO₂-induced accumulations of TNCin both species, with starch accumulating in the leaves of bothspecies, and fructans accumulating only inP. smithii. Photosynthesisof both species is likely to be enhanced in future CO₂-enrichedand warmer environments, although responses will sometimes beattenuated by acclimation. Acclimation; blue grama (Bouteloua gracilis (H.B.K.) Lag ex Steud.); leaf nitrogen concentration; nonstructural carbohydrates; photosynthesis; western wheatgrass (Pascopyrum smithii (Rydb.) Love)     

Ethylene Exchange in Lycopersicon esculentum Mill. Leaves During Short- and Long-Term Exposures to CO2

The effects of long-term and transient exposure to elevatedCO₂ concentrations on photosynthetic gas exchange and ethylenerelease by tomato leaves were investigated. The net CO₂ assimilationrate was enhanced when leaf tissue grown at ambient (35 Pa CO₂)levels was assayed at 100 Pa CO₂. Leaf tissue grown at high(130 Pa) CO₂ exhibited a lower net CO₂ assimilation rate athigh CO₂ levels than leaf tissue grown at ambient (35 Pa) CO₂.This decrease in CO₂ exchange rate in response to growth athigh CO₂ is typical of C₃ species. Rates of endogenous and 1-aminocyclopropane-l-carboxylicacid (ACC)-stimulated ethylene release from leaf tissue wereenhanced by exposure to elevated CO₂ levels whether the leaftissue had been grown at ambient or enriched CO₂ levels. Thedata demonstrate that CO₂ enhanced C₂H₄ release from leaf tissuein response to both short-term perturbations in CO₂ concentrationand long-term growth and development under high CO₂. Prolongedgrowth at elevated CO₂ concentrations induced a higher endogenousrate of C₂H₄ release relative to that of leaf tissue grown atlower CO₂ levels. Leaf tissue from all leaf positions of plantsgrown at high CO₂ consistently evolved more C₂H₄ than correspondingtissue from ambient-grown plants when assayed under standardizedconditions. Endogenous (ACC) tissue contents and rates of ACC-stimulatedethylene release were also higher at all leaf positions in CO₂-enrichedtissue. Thus the higher rates appeared to be due to both higherendogenous precursor (ACC) levels in the tissue and greaterACC to C₂H₄ conversion capacity. Growth at elevated CO₂ levelsresulted in a persistent increase in the rate of endogenousC₂H₄ release in leaf tissue. The capacity for increased ethylenerelease in response to CO2 did not decline after prolonged growthat high CO₂. Key words: CO₂enrichment, ethylene, leaves, tomato     

Light Dependency of Carboxylation Efficiency and Ribulose-1, 5-Bisphosphate Carboxylase Activation in Trifolium subterraneum L. Leaves

The effect of photosynthetic photon flux density (PPFD) on carboxylationefficiency, estimated as the initial slope (IS) of net CO₂ assimilationrate versus intercellular CO₂ partial pressure response curve,as well as on ribulose-1, 5-bisphosphate carboxylase (Rubisco)activation was measured in Trifolium subterraneum L. leavesunder field conditions. The relationship between IS and PPFDfits a logarithmic curve. Rubisco activation accounts for theIS increase only up to a PPFD of 550 µmol photons m^-2s^-1. Further IS increase, between 550 and 1000 µmol photonsm^-2 s^-1, could be related to a higher ribulose fcwphosphate(RuBP) availability. The slow, but sustained IS increase above1000 µmol photons m^-2 s^-1 could be explained by the mesophyllCO₂ diffusion barriers associated with the high chlorophylland protein content in field developed leaves. Key words: Photosynthesis, initial slope, ribulose-1, 5-bissphosphate carboxylase activation, light response, Trifolium subterraneum L     

The Damping and Reinitiation of the Circadian Rhythm of CO2 Output in Bryophyllum Leaves in Relation to their Malate Content

The circadian rhythm of CO₂ output in leaves of Bryophyllumfedtschenkoi damps out after 3–4 d in continuous darknessand a CO₂-free air stream at 15°C. The rhythm is reinitiatedafter a single exposure to white light of 2, 4, 6 or 8 h duration,damps out again after a further 3–4 d and can be reinitiatedfor a second time by a further exposure to light. During the exposure to light there is a burst of CO₂ outputconsistent with the decarboxylation of malate, and the rhythmbegins afterwards with an initial high rate of CO₂ fixation.Malate gradually accumulates in the leaves in continuous darknessto attain a maximum value (35 mol m^–3) at the time whenthe circadian rhythm disappears, and decreases to a low value(19 mol m^–3) after a 4 h exposure to light which reinitiatesrhythmicity. These results support the hypothesis that damping of the rhythmof CO₂ output in continuous darkness is due to the accumulationof malate in the leaf cells, eventually reaching such a levelthat its removal from the cytoplasm into the vacuole cannottake place, with the result that PEPc activity, upon which therhythm of CO₂ output depends, remains allosterically inhibited. Key words: CAM, circadian rhythm, Bryophyllum, CO₂-fixation, malate metabolism     

Ribulose 1, 5-bisphosphate Carboxylase/ Oxygenase Activities in Leaves of Greenhouse Roses

An enzyme assay was developed to measure the initial and Mg²⁺–CO₂activated forms of Ribulose 1,5-bisphosphate Carboxylase/Oxygenase(Rubisco) in rose leaves. The assay was verified by co-extractionof the leaflets with partially purified spinach Rubisco andthrough correlation with net photosynthetic rates of individualleaflets (r²=0.7324). Changes in activities were measured asa function of depth of leaves in the canopy for two cultivarsof greenhouse hybrid tea roses. Initial Rubisco activity declinedwith increasing canopy depth for both cultivars. The activatedform of the enzyme, however, remained constant with canopy depthfor cv. Red Success; but increased with canopy depth, then declinedafter mid-canopy in the cv. Royalty. Rubisco activities werealso measured in the cv. Red Success grown in CO₂ enriched environments(100 mm³ dm^–3) at three humidity levels. The activitieswere not significantly affected by humidity treatment. However,there was a trend for plants grown at lower humidity to havehigher activated activities. Key words: Humidity, Rubisco, Rosa ? hybrida, Royalty, Red Success     

Growth and Physiology of One-year old Poplar (Populus) Under Elevated Atmospheric CO2 Levels

The effects of elevated atmospheric CO₂ concentrations on theecophysiological responses (gas exchange, chlorophyll a fluorescence,Rubisco activity, leaf area development) as well as on the growthand biomass production of two poplar clones (i.e. Populus trichocarpax P. deltoides clone Beaupré and P. x euramericana cloneRobusta) were examined under open top chamber conditions. Theelevated CO₂ treatment (ambient + 350 µmol mol^-1) stimulatedabove-ground biomass of clones Robusta and Beaupré afterthe first growing season by 55 and 38%, respectively. This increasedbiomass production under elevated CO₂ was associated with asignificant increase in plant height, the latter being the resultof enhanced internode elongation rather than an increased productionof leaves or internodes. Both an increased leaf area index (LAI)and a stimulated net photosynthesis per unit leaf contributedto a significantly higher stem biomass per unit leaf area, andthus to the increased above-ground biomass production underthe elevated CO₂ concentrations in both clones. The larger LAIwas caused by a larger individual leaf size and leaf growthrate; the number of leaves was not altered by the elevated CO₂treatment. The higher net leaf photosynthesis was the resultof an increase in the photochemical (maximal chlorophyll fluorescenceFm and photochemical efficiency Fv/Fm) as well as in the biochemical(increased Rubisco activity) process capacities. No significantdifferences were found in dark respiration rate, neither betweenclones nor between treatments, but specific leaf area significantlydecreased under elevated CO₂ conditions.Copyright 1995, 1999Academic Press Biomass, chlorophyll a fluorescence, elevated CO₂, growth, Populus, poplar, photosynthesis, respiration, Rubisco     

Photosynthetic acclimation to elevated CO2 in poplar grown in glasshouse cabinets or in open top chambers depends on duration of exposure

The effects of elevated CO₂ were studied on the photosyntheticgas exchange behaviour and leaf physiology of two contrastingpoplar (Populus) hybrids grown and treated in open top chambers(OTCs in Antwerp, Belgium) and in closed glasshouse cabinets(GHCs in Sussex, UK). The CO₂ concentrations used in the OTCswere ambient and ambient +350 µmol mol^–1 while inthe GHCs they were c. 360 µmol mol^–1 versus 719µmol mol^–1. Measurements of photosynthetic gas exchangewere made for euramerican and interamerican poplar hybrids incombination with measurements of dark respiration rate and Rubiscoactivity. Significant differences in the leaf anatomy and structure(leaf mass per area and chlorophyll content) were observed betweenthe leaves grown in the OTCs and those grown in the GHCs. ElevatedCO₂ stimulated net photosynthesis in the poplar hybrids after1 month in the GHCs and after 4 months in the OTCs, and therewas no evidence of downward acclimation (or down-regulation)of photosynthesis when the plants in the two treatments weremeasured in their growth CO₂ concentration. There was also noevidence of down-regulation of Rubisco activity and there wereeven examples of increases in Rubisco activity. Rubisco exerteda strong control over the light-saturated rate of photosynthesis,which was demonstrated by the close agreement between observednet photosynthetic rates and those that were predicted fromRubisco activities and Michaelis-Menten kinetics. After 17 monthsin elevated CO₂ in the OTCs there was a significant loss ofRubisco activity for one of the hybrid clones, i.e. Beaupr,but not for clone Robusta. The effect of the CO₂ measurementconcentration (i.e. the short-term treatment effect) on netphotosynthesis was always larger than the effect of the growthconcentration in both the OTCs or GHCs (i.e. the longterm growthCO₂ effect), with one exception. For the interamerican hybridBeaupr dark respiration rates in the OTCs were not significantlyaffected by the elevated CO₂ concentrations. The results suggestthat for rapidly growing tree species, such as poplars, thereis little evidence for downward acclimation of photosynthesiswhen plants are exposed to elevated CO₂ for up to 4 months;longer term exposure reveals loss of Rubisco activity. Key words: Elevated CO₂, Populus, Rubisco, photosynthesis, chlorophyll content     

Extreme Phosphate Deficiency Decreases the In Vivo CO2/O2 Specificity Factor of Ribulose 1,5-Bisphosphate Carboxylase-Oxygenase in Intact Leaves of Sunflower

Sunflower plants were grown under controlled environmental conditionswith either 0 or 10 mol m^–3 phosphate (Pi). From steady-statemeasurements of gas exchange and chlorophyll fluorescence madeon intact leaves, the in vivo CO₂/O₂ specificity factor (invivo K_sp) of ribulose 1,5-Aisphosphate carboxylase-oxygenase(Rubisco) was determined following two methods based on modelsof C₃ photosynthesis by Brooks and Farquhar (1985) and Peterson(1989). The two methods gave in vivo K_sp values for controlsunflower leaves which were similar to published values forhigher plants. Extreme Pi deficiency decreased in vivo K_sp,in sunflower leaves compared to adequate Pi. This suggests thatPi deficiency affected photorespiration less than photosynthesis.The decrease in in vivo K_sp may be due to a real change in theenzyme kinetics favouring oxygenation more than carboxylationor due to an increase in the number of CO₂ molecules releasedper oxygenation; in which case the observed decrease in thein vivo K_sp determined on intact leaves will not agree numericallywith the true K_sp of Rubisco determined in vitro using purifiedenzyme from the same leaf. We discuss the implications of therelatively large photorespiration in Pi-deficient sunflowerleaves with respect to the increased dissipation of photosyntheticelectrons and photorespiratory recycling of Pi in thechloroplaststroma. Although our results on in vivo K_sp suggested a relativelylarger photorespiratory potential in Pi-deficient than controlsunflower leaves, photosynthesis was insensitive to O₂ in Pi-deficientleaves; the possible reasons for this phenomenon are discussed.Under extreme Pi deficiency, O₂ sensitivity of photosynthesisis not a reflection of the in vivo photorespiratory rates. Determinationof in vivo K_sp of Rubisco is a useful approach to study thephotorespiratory potential of intact leaves. Key words: Chlorophyll fluorescence, phosphate deficiency, photorespiration, photosynthesis, PSII quantum yield, Rubisco specificity factor     

Photosynthesis and Light Activation of Ribulose 1, 5-bisphosphate Carboxylase in the Presence of Starch

Owing to a typographical error three equations were omittedfrom page 1294. The correct paragraphs are set out below. The component K₁ corrected for the difference in temperaturebetween the enzyme assay and the leaf and was calculated accordingto the Arrhemus equation. where v10 and v18 are the reaction velocities of carboxylationat 10?C and 18?C, respectively and A is the activation energy(A = 90 kJ mol^–1, as determined for purified wheat RuBPCOby M?chler, Keys and Cornelius, 1980) The components K2 corrected for the difference in CO₂ partialpressure between enzyme assay and leaf and for competitive inhibitionof carboxylation by O₂ and was calculated according to the modifiedMichaelis Menten equation where v_c, is the carboxylation velocity under leaf conditions,V_c. is the maximum carboxylation velocity as determined in theenzyme assay, K_c, and K_o are the Michaelis constants for carboxylationand oxygenation, respectively (K_o = 159 Pa CO₂. K_o = 35.3 kPaO₂, as interpolated for 18?C from spinach data as determinedby Jordan and Ogren, 1984), O is oxygen partial pressure inair and C₁ is intercellular CO₂ partial pressure in leaves (C₁= 29.1 ? 0.8 Pa (? s c , n = 15)) The component K3 corrected for the decrease in CO₂ fixationin leaves due to photorespiration and was calculated accordingto equation 3 Equation 3 is denved from the equation for the substrate specificityof RuBPCO, S= v_c/v_oC (Laing, Ogren, and Hageman, 1974), andfrom the equation for the stoichiometry of photorespiratoryCO₂ release, F=v_c–1/2 v_o, where v_c, and v_c are reactionvelocities of carboxylation and oxygenation, O and C are partialpressures of 02 and intercellular CO₂, F is net photosynthesisand S is the substrate specificity of RuBPCO (S= 3061 Pa/Pa,as interpolated for 18?C from spinach data as determined byJordan and Ogren, 1984)     

Effects of phosphorus nutrition on the response of photosynthesis to CO2 and O2, activation of ribulose bisphosphate carboxylase and amounts of ribulose bisphosphate and 3-phosphoglycerate in spinach leaves

Phosphorus-deficient spinach plants were grown by transferring them to nutrient solutions without PO₄. Photosynthetic rates were measured at a range of intercellular CO₂ partial pressures from 50–500 bar and then the leaves were freeze-clamped in situ to measure ribulose bisphosphate carboxylase (Rubisco) activity and metabolite concentrations. Compared with control leaves, deficient leaves had significantly lower photosynthetic rates, percentage activation of Rubisco, and amounts of ribulose bisphosphate and 3-phosphoglycerate at all CO₂ partial pressures. After feeding 10 mM PO₄ to the petioles of detached deficient leaves, all these measurements increased within 2 hours. At atmospheric CO₂ partial pressure the photosynthetic rate was stimulated in 19 mbar O₂ compared with 200 mbar. At higher CO₂ partial pressures this stimulation was less but the percentage stimulation in deficient leaves was no different from controls in either CO₂ partial pressure. It was concluded that phosphorus deficiency affects both Rubisco activity and the capacity for ribulose bisphosphate regeneration, and possible causes are discussed.Abbreviations A CO₂ assimilation rate - C_i intercellular CO₂ partial pressure - PGA 3-phosphoglycerate - RuP₂ ribulose 1,5-bisphosphate - Rubisco RuP₂ carboxylase/oxygenase     

Carbon Exchange Rate of Intact Individual Soya Bean Pods. 2. Ontogeny of Temperature Sensitivity

Diurnal temperature fluctuations induced change in soya bean-pod[Glycine max (L.) Merr.] carbon exchange rate (CER, where positiveCER represents CO₂ evolution). CER appeared to depend linearlyon temperature. Linear regressions of CER on temperature interceptedthe temperature axis at 5°C (i.e. zero CER at 5°C).Slopes of these regressions (i.e. temperature sensitivity) changedover the season. The CER-temperature sensitivity coefficient,K, (calculated from observed values of CER. pod temperatureand temperature intercept) rose from less than 0·02 mgCO₂ h^–1 pod^–1 °C^–1 during early pod-flll,peaked at over 0·04 mg CO₂ h^–1 pod^–1 °C^–1at mid pod-fill, and then declined during late pod-fill andmaturation. Glycine max (L.) Merr., Soya bean, carbon exchange rate, temperature     

Photosynthesis by Flag Leaves of Wheat in Relation to Protein, Ribulose Bisphosphate Carboxylase Activity and Nitrogen Supply

The effects of nitrate supply on the composition (cell numbers,protein and chlorophyll contents) of flag leaves of winter wheatgrown with two amounts of N fertilizer and of spring wheat grownin the glasshouse under controlled nitrate supply are describedand related to photosynthesis. Nitrogen deficiency decreasedthe size of leaves, mainly by reducing cell number and, to asmaller extent, by decreasing cell volume. Protein content perunit leaf area, per cell and per unit cell volume was largerwith abundant N. Total soluble protein, ribulose bisphosphatecarboxylase-oxygenase (RuBPc-o) protein and chlorophyll changedin proportion irrespective of nitrogen supply and leaf age.Photosynthesis per unit area of flag leaf and carboxylationefficiency in both winter and spring wheat were proportionalto the amount of total soluble protein up to 7.0 g m^–2and to the amount of RuBPc-o protein up to 4.0 g m^–2.However, photosynthesis did not increase in proportion to theamount of total soluble or RuBPc-o protein above these amounts.In young leaves with a high protein content the measured ratesof photosynthesis were lower than expected from the amount andactivity of RuBPc-o. Carboxylation per unit of RuBPc-o protein,measured in vitro, was slightly greater in N-deficient leavesof winter wheat but not of spring wheat. RuBPc-o activity perunit of RuBPc-o protein was similar in winter and spring wheatleaves and remained approximately constant with age, but increasedin leaves showing advanced senescence. RuBPc-o protein fromN-deficient leaves migrated faster on polyacrylamide gels thanprotein from leaves with high N content. Regulation of the rateof photosynthesis in leaves and chloroplasts with a high proteincontent is discussed. The conductance of the cell to the fluxof CO₂ from intercellular spaces to RuBPc-o active sites iscalculated, from cell surface areas and CO₂ fluxes, to decreasethe CO₂ partial pressure at the active site by less than 0.8Pa at an internal CO₂ partial pressure of 34 Pa. Thus the decreasein partial pressure of CO₂ is insufficient to account for theinefficiency of RuBPc-o in vivo at high protein contents. Otherlimitations to the rate of photosynthesis are considered. Key words: Wheat, photosynthesis, nitrogen, ribulose, bisphosphate carboxylase     

Photoinhibition under Atmospheric O2, the Activation State of RuBP Carboxylase and the Content of Photosynthetic Intermediates in Soybean and Wheat

Ward, D. A. and Drake, B. G. 1987. Photoinhibition under atmosphericO₂, the activation state of RuBP carboxylase and the contentof photosynthetic intermediates in soybean and wheat.—J.exp. Bot. 38: 1937–1948. Associations between photosynthesis, the activation state ofRuBP carboxylase and the contents of photosynthetic intermediateswere compared in soybean and wheat leaves before and after exposureto photoinhibitory treatments in the presence of atmosphericO₂. Exposing attached leaves to a supra-saturating irradiance(3 800 µmol quanta m^{– 2} s^–1) for 2 h in CO_2-freeair decreased carboxylation efficiency and the light-saturatedphotosynthetic rate in air by approximately 50%. Exposure tothe photoinhibitory treatment for periods in excess of 2 h didnot cause a further decrease of photosynthesis in soybean. Althoughphotosynthesis was reduced, the initial and total (fully-activated)activities of ribulose 1,5-bisphosphate carboxylase (RuBPCase)in leaf extracts were unaltered in each species by the photoinhibitorytreatment. This was true for leaves sampled under both air andat a rate-limiting intercellular CO₂ partial pressure (C_i) of75 µPa Pa^–1. The contents of ribulose l,5-bisphosphate(RuBP) and 3-phosphoglyceric acid (3-PGA) were reduced by thephotoinhibitory treatment in soybean leaves sampled in air andat a rate-limiting C_i, although the RuBP/3-PGA ratio was unaffected.The relative reduction of RuBP content in soybean leaves atrate-limiting C₁ was similar to the corresponding reductionof carboxylation efficiency. For wheat,the relative reductionof RuBP content at rate-limiting C_i (–19%) caused by thephotoinhibitory treatment was considerably less than the correspondingdecrease of carboxylation efficiency (–49%).The RuBP/3-PGAratio of wheat was also increased significantly by the photoinhibitorytreatment The significance of these observations to the regulationof CO₂-limited photosynthesis in leaves experiencing photoinhibitionunder atmospheric oxygen is discussed. Consideration is alsogiven to the previous contention that contemporary measurementsof initial activity in crude extracts may provide a spuriousindication of the amount of the enzyme-CO₂-Mg_{2 +} form of RuBPcarboxylase present in the leaf. Key words: Carboxylation efficiency, RuBP carboxylase, photoinhibition, RuBP, 3-PGA     

Physiological and Anatomical Effects of Growth Temperature on Phaseolus vulgaris L. Cultivars

Two Phaseolus vulgaris L. cultivars were grown at 20/15, 25/20,and 30/25 °C day/night temperatures in growth chambers witha 16 h thermoperiod corresponding to the photoperiod. When thefirst trifoliolate leaf was fully expanded rates of CO₂ exchange(CER) were measured at 27 °C and saturating light usinginfrared gas analysis. Stomatal (r_s) and mesophyll resistances,CO₂ compensation points, activities of the enzymes ribulosebisphosphate carboxylase (RuBPCase), glycolate oxidase (GAO),malate dehydrogenase (MDH), and fructose-1, 6 diphosphate (FDP),chlorophyll content, Hill activities, and leaf anatomy at boththe light and electron microscope level were also investigatedin these leaves. Rates of CO₂ exchange in the light, transpiration rate, andchlorophyll content increased with increasing growth temperaturewhile leaf thickness, specific leaf weight, RuBPCase activity,compensation point, and stomatal resistance decreased. Mesophyllresistance also decreased when calculated assuming zero chloroplastCO₂ concentration (rm, o), but not when calculated assuminga chloroplast CO₂ concentration equal to the CO₂ compensationconcentration (rm, g). Average leaf size was maximal in 25/20°C plants while dark respiration, MDH activity, stomataldensity, and starch were minimal. The activities of GAO andFDP and Hill activity were not affected by temperature pretreatment.     

Non-Stomatal Inhibition Associated with Inactivation of Rubisco in Dehydrated Coffee Leaves under Unshaded and Shaded Conditions

The effect of rapid dehydration due to withholding of irrigationon leaf photosynthesis in coffee (Coffea arabica L.) was studiedby comparing the CO₂-dependent rate of photosynthesis in intactleaves (A/Ci curve), the amounts of Chl, total soluble proteinand ribulose-l,5-bisphosphate carboxylase (Rubisco) in leaves,and the activities of photochemical reactions of isolated chloroplastsand of Rubisco under unshaded and shaded conditions. The CO₂-saturatedrate of photosynthesis and the carboxylation efficiency (theinitial slope of A/Ci curve) decreased with decreasing leafwater potential (