Participation of Inorganic Orthophosphate in Regulation of the Ribulose-1,5-bisphosphate Carboxylase Activity in Response to Changes in the Photosynthetic Source-Sink Balance

Sink-limited conditions, defined as treatment with continuousillumination, cause a reduction in the rate of photosyntheticfixation of CO₂ in single-rooted leaves of soybean (Glycinemax. Merr. cv. Turunoko). We suggested previously that thisreduction is due to a deactivation of ribulose-1,5-bisphosphatecarboxylase (RuBPcase, E.C. 4.1.1.39 [EC] ) that is caused by a decreasein the level of P_i in the leaves [Sawada et al. (1989) PlantCell Physiol. 30: 691, Sawada et al. (1990) Plant Cell Physiol.31: 697]. In the present study, the mechanism of regulationof RuBPcase activity by P_i was examined. The activity of RuBPcasein the sink-limited leaves, exposed for 6 or 7 d to continuousillumination to alter the source/sink balance, was enhancedwith increasing concentrations of P_i, in a CO₂-free preincubationmedium in the presence of 5 mM MgCl₂ The maximum value [6.3µmole CO₂ (mg Chl)^–1 min^–1] was obtained atapproximately 5 mM P_i after a 5 min incubation, being 3 timesof the activity without the preincubation. The activity of acrude preparation of RuBPcase that had been deactivated by removalof CO₂ and Mg²⁺ ions by the gel filtration was 5.2–9.3nmole CO₂ (mg protein)^–1 min^–1 and was also enhancedby P_i plus Mg²⁺ ions. The maximum value [147–151 nmoleCO₂ (mg protein)^–1 min^–1] was attained at 5 mM P_iafter a 5 min incubation. The cycle of activation and inactivationof deactivated crude RuBPcase was perfectly reversible by additionof P_i to the enzyme and removal of P_i from the enzyme. Levelsof free P_i and of esterified phosphate in the sink-limited leaveswere 69% and 31% of the total phosphate, respectively. By contrast,in the control leaves, these values were 87% and 13%, respectively.These results support our previously stated hypothesis and indicatean important role for free P_i in the regulation of RuBPcaseactivity, in particular in sink-limited plants. (Received February 21, 1992; Accepted July 23, 1992)     

Stimulatory Effect of Chloride Ions on NADP Malic Enzyme from Leaves of two C4 Species of Cyperus

NADP malic enzyme (EC 1.1.1.40 [EC] ) from leaves of two C₄ speciesof Cyperus (C. rotundus and C. brevifolius var leiolepis) exihibiteda low level of activity in an assay mixture that contained lowconcentrations of Cl^–. This low level of activity wasmarkedly enhanced by increases in the concentration of NaClup to 200 mM. Since the activity of NADP malic enzyme was inhibitedby Na₂SO₄ and stimulated by relatively high concentration ofTris-HCl (50–100 mM, pH 7–8), the activation ofthe enzyme by NaCl appears to be due to Cl^–. Variationsin the concentration of Mg²⁺ affected the K_A (the concentrationof activator giving half-maximal activation) for Cl^–,which decreased from 500 mM to 80 mM with increasing concentrationsof Mg²⁺ from 0.5 mM to 7 mM. The K_m for Mg²⁺ was decreased from7.7 mM to 1.3 mM with increases in the concentration of NaClfrom zero to 200 mM, although the increase of V_max was not remarkable.NADP malic enzyme from Cyperus, being similar to that from otherC₄ species, was able to utilize Mn²⁺. The K_m for Mn²⁺ was 5mM, a value similar to that for Mg²⁺. The addition of 91 mMNaCl markedly decreased the K_m for Mn²⁺ to 20 +M. NADP malicenzyme from Setaria glauca, which contains rather less Cl^–than other C₄ species, was inactivated by concentrations ofNaCl above 20 mM, although slight activation of the enzyme wasobserved at low concentrations of NaCl at pH7.6. (Received February 20, 1989; Accepted June 12, 1989)     

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     

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)     

SPHAERIUM CORNEUM (L.) AND PISIDIUM SPP. PFEIFFER -- THE ECOLOGY OF FRESHWATER BIVALVE MOLLUSCS IN RELATION TO WATER CHEMISTRY

Populations of Sphaerium corneum (L.) and Pisidium spp. weresampled monthly for 13 months, at 11 sites with a wide rangeof water chemistry in N.W. England. Both genera showed almostyear-round reproduction, but the periods of maximum productionshowed between-population variation. S. corneum adults containedspat at different stages of development and spat size was relatedto number in the brood. Multiple regression analysis with 16water physicochemical variables showed the following to be highlysignificant factors (P<0.01) in the distribution of the molluses(factors in order of importance):- S. corneum: HCO₃, K^$, Cl^–, Mg^2$, temperature, Ca^2$, month; Pisidium spp: PO₄^3–, Mg^2$, mud, oxygen, temperature, month. (Received 25 January 1978;     

Evidence for Essential Maintenance Respiration of Leaves of Xanthium Strumarium at High Temperature

Mesophyll resistance to photosynthetic carboxylation (r'_m) wasused as a criterion for leaf integrity. It was measured, at25 °C, in the light, before and after periods of high temperature(3 h at 38 °C) in the dark. During the high temperatureperiods, respiration (R_D) of attached leaves of Xanthium strumariumwas suppressed from 27%-36% by either low [O₂] (1.04% or 0.21%v.v.) or high [CO₂] (840 µl 1^–1) in the ambientair. Neither treatment affected rates of R_D or photo-respirationduring the second period at 25 °C. There was no significant increase of r'_m when R_D was not suppressedduring the high temperature treatment. When R_D was suppressedat high temperatures, r'_m increased from about 3s cm^–1before, to about 26 s cm^–1 after the high temperaturetreatment. The increase depended upon the degree of suppression. It is concluded that increased R_D at high temperature in Xanthiumleaves is partly the result of an increase of energy demandingmaintenance. The subsequent rate of carbon dioxide fixationis reduced when this increase of maintenance-induced respirationis inhibited.     

Influence of Elevated CO2 and Temperature on the Photosynthesis and Respiration of White Clover Dependent on N2 Fixation

Single clonal plants of white clover (Trifolium repens L) grownfrom explants in a Perlite rooting medium, and dependent fornitrogen on N₂ fixation in root nodules, were grown for severalweeks in controlled environments which provided two regimesof CO₂, and temperature 23/18 °C day/night temperaturesat 680 µmol mol^–1 CO₂, (C680), and 20/15 °Cday/night temperatures at 340 µmol mol^–1 CO₂ (C340)After 3–4 weeks of growth, when the plants were acclimatedto the environmental regimes, leaf and whole-plant photosynthesisand respiration were measured using conventional infra-red gasanalysis techniques Elevated CO₂ and temperature increased ratesof photosynthesis of young, fully expanded leaves at the growthirradiance by 17–29%, despite decreased stomatal conductancesand transpiration rates Water use efficiency (mol CO₂ mol H₂O^–1)was also significantly increased Plants acclimated to elevatedCO₂, and temperature exhibited rates of leaf photosynthesisvery similar to those of C340 leaves ‘instantaneously’exposed to the C680 regime However, leaves developed in theC680 regime photosynthesised less rapidly than C340 leaves whenboth were exposed to a normal CO₂, and temperature environmentIn measurements where irradiance was varied, the enhancementof photosynthesis in elevated CO₂ at 23 °C increased graduallyfrom approx 10 % at 100 µmol m^–1 s^–1 to >27 % at 1170 µmol m^–2 s^–1 In parallel, wateruse efficiency increased by 20–40 % at 315 µmolm^–2 s^–1 In parallel, water use efficiency increasedby 20–40 % at 315 µmol m^–2 s^–1 In parallel,water use efficiency increased by 20–40 % at 315 µmolm^–2 s^–1 In parallel, water use efficiency increasedby 20–40 % at 315 µmol m^–2 s^–1 to approx100 % at the highest irradiance Elevated CO₂, and temperatureincreased whole-plant photosynthesis by > 40 %, when expressedin terms of shoot surface area or shoot weight No effects ofelevated CO₂ and temperature on rate of tissue respiration,either during growth or measurement, were established for singleleaves or for whole plants Dependence on N₂, fixation in rootnodules appeared to have no detrimental effect on photosyntheticperformance in elevated CO₂, and temperature Trifolium repens, white clover, photosynthesis, respiration, elevated CO₂, elevated temperature, water use efficiency, N₂ fixation     

Effects of temperature and CO2 concentration on photosynthetic CO2 fixation by Chlorella

The rates of photosynthetic ¹⁴CO₂ fixation by Chlorella vulgarisllh, grown under high CO₂, were determined between 4 to 37°Cwith air containing from 300 to 13,000 ppm ¹⁴CO₂. When the CO₂level was increased, both the rate of photosynthesis and theoptimum temperature for maximum photosynthesis increased. Themaximum photosynthetic rate was reached at 12°C with 300ppm ^l4CO₂. Among the photosynthetic products fromed at 300 ppm ¹⁴CO₂, glycolatedecreased greatly when the temperature was raised from 20 to30°C. At 3,000 ppm ¹⁴CO₂ an insignificant amount of glycolatewas formed at all temperatures, whereas ¹⁴C-incorporation intothe insoluble fraction, sucrose, and the lipid fraction wassignificantly higher than at 300 ppm ¹⁴CO₂. The ¹⁴C in sucrosewas greatly increased and the radioactivity in the insolublefraction decreased when the temperature was raised from 28 to36°C. (Received April 8, 1980; )     

Growth and Partitioning in Pascopyrum smithii (C3) and Bouteloua gracilis(C4) as Influenced by Carbon Dioxide and Temperature

This study investigated how CO₂and temperature affect dry weight(d.wt) accumulation, total nonstructural carbohydrate (TNC)concentration, and partitioning of C and N among organs of twoimportant grasses of the shortgrass steppe,Pascopyrum smithiiRydb. (C₃) andBouteloua gracilis(H.B.K.) Lag. ex Steud. (C₄).Treatment combinations comprised two temperatures (20 and 35°C)at two concentrations of CO₂(380 and 750 µmol mol^-1),and two additional temperatures of 25 and 30°C at 750 µmolmol^-1CO₂. Plants were maintained under favourable nutrient andsoil moisture and harvested following 21, 35, and 49d of treatment.CO₂-induced growth enhancements were greatest at temperaturesconsidered favourable for growth of these grasses. Comparedto growth at 380 µmol mol^-1CO₂, final d.wt of CO₂-enrichedP.smithiiincreased 84% at 20°C, but only 4% at 35°C. Finald.wt ofB. graciliswas unaffected by CO₂at 20°C, but wasenhanced by 28% at 35°C. Root:shoot ratios remained relativelyconstant across CO₂levels, but increased inP. smithiiwith reductionin temperature. These partitioning results were adequately explainedby the theory of balanced root and shoot activity. Favourablegrowth temperatures led to CO₂-induced accumulations of TNCin leaves of both species, and in stems ofP. smithii, whichgenerally reflected responses of above-ground d.wt partitioningto CO₂. However, CO₂-induced decreases in plant tissue N concentrationswere more evident forP. smithii. Roots of CO₂-enrichedP. smithiihadgreater total N content at 20°C, an allocation of N below-groundthat may be an especially important adaptation for C₃plants.Tissue N contents ofB. graciliswere unaffected by CO₂. Resultssuggest CO₂enrichment may lead to reduced N requirements forgrowth in C₃plants and lower shoot N concentration, especiallyat favourable growth temperatures. Acclimation to CO₂; blue grama; Bouteloua gracilis ; carbohydrate; climate change; global change; grass; growth; growth temperature optima; nitrogen; N uptake; Pascopyrum smithii; western wheatgrass     

Preliminary characterization of 1-aminocyclopropane-1-carboxylate oxidase properties from embryonic axes of chick-pea (Cicer arietinum L.) seeds

For a deeper understanding of the germination of chick–pea(Cicer arietinum) seeds, which is dependent upon ethylene synthesis,a crude extract containing authentic ACC oxidase (ACCO) activitywas isolated in soluble form from the embryonic axes of seedsgerminated for 24 h. Under our optimal assay conditions (200mM HEPES at pH 7.0, 4µM FeS0₄, 6 mM Na–ascorbate,1 mM ACC, 20% 0₂, 3% CO₂ , and 10%glycerol) this enzyme was5–fold more active than under the conditions we used initiallyin the present work. The enzyme has the following K_m: 28 µMfor ACC (approximately 4–fold less than in vivo), 1.2%for O₂ (in the presence of an optimal CO₂ concentration of 3%),and 1% for CO₂ in the presence of O₂ (20%). The enzyme is inhibitedby phenanthroline (PNT) (specific chelating agent of ferrousion), and competitively inhibited (K₁, =0.5 mM) by 2–aminoisobutyricacid (AIB), and the enzymatic activity was not detectable inthe absence of CO₂. Under optimal assay conditions, the enzymehas two optimum temperatures (28 C and 35 C) and is inhibitedby divalent metal cations (Zn²⁺> CO²⁺>Ni²⁺>Cu²⁺>Mn²⁺>Mg²⁺) and by salicylic acid, propylgallate, carbonyl cyanidem–chlorophenyl hydrazone (CCCP), dinitrophenol (DNP),and Na–benzoate. The in vitro ACCO activity which we recoveredin soluble form is equivalent to approximately 80–85%of the apparent activity evaluated in vivo. Key words: ACC oxidase, Cicer arietinum, ethylene, germination, seeds     

Effect of Root Zone Temperature on the Mineral Composition of Xylem Sap and Plasma Membrane K+-Mg++-ATPase Activity of Grafted-Cucumber and -Figleaf Gourd Root Systems

Cucumber (Cucumis sativus L.) seedlings were grafted onto cucumber-(CG) or figleaf gourd- (FG, Cucurbita ficifolia Bouché)seedlings in order to determine the effect of solution temperature(12, 22, and 32°C) on the mineral composition of xylem sapand the plasma membrane K⁺-Mg⁺⁺-ATPase activities of the roots.Low solution temperature (12°C) lowered the concentrationof NO^–₃ and H₂PO^–₄ in xylem sap of CG plants butnot of FG plants. Concentrations of K⁺, Ca⁺⁺ and Mg⁺⁺ in xylemsap were less affected than anions by solution temperature.The plasma membrane of FG plants grown in 12°C solutiontemperature showed the highest K⁺- Mg⁺⁺-ATPase activity at allATP concentrations up to 3 mM and at low reaction temperatureup to 12°C, indicating resistance of figleaf gourd to lowroot temperature. (Received December 27, 1994; Accepted March 10, 1995)     

RNA Synthesis in Phaseolus Chloroplasts: I. RIBONUCLEIC ACID SYNTHESIS IN CHLOROPLAST PREPARATIONS FROM PHASEOLUS VULGARIS L. LEAVES AND SOLUBILIZATION OF THE RNA POLYMERASE

Chloroplast preparations from the young primary leaves of Phaseolusvulgaris L. cv. Canadian Wonder carry out the DNA-dependentincorporation of UTP into RNA at rates between 8 and 14 pmolUTP µg^–1 chlorophyll h^–1. It is estimatedthat 90% of the activity was localized in the chloroplasts.The incorporation proceeded for between 20 and 30 min at 35°C. The maximum rates of RNA synthesis were attained atpH 8.3, in the presence of 15 mM MgCl₂. Chloroplasts were alsoactive, to a lesser extent, with 1.5 mM MnCl₂. The simultaneouspresence of MnCl₂ and MgCl₂ resulted in inhibition of activity.Nuclear material prepared from young P. vulgaris leaves incorporatedUTP at a rate of about 12 pmol UTP µg^–1 DNA h^–1.On a chloroplast (Tritonsoluble) DNA basis chloroplast activitywas over 40-fold that of nuclei. Methods of solubilizing chloroplastRNA polymerase were explored. Yields of over 75% were achieved,but methods suitable for one species were not always successfulwhen applied to another. The highest yields of the P. vulgarisenzyme were obtained using EDTA and KCl. All methods resultedin solubilization of DNA. RNA synthesis by the soluble P. vulgarisenzyme proceeded for more than 40 min at 35 °C.     

Measurements of 14C Incorporation by Illuminated Intact Leaves of Coffee Plants from Gas Mixtures Containing 14CO2

A study was made of the incorporation of ¹⁴C by intact leavesof Coffea arabica (cultivars Mundo Novo, Catuai, 1130–13,and H 6586–2) and Coffea canephora (cultivar Guarini)supplied with gas mixtures containing ¹⁴CO₂ under controlledconditions. Samples of the leaves were combusted and the ¹⁴Cin the CO₂ produced measured using a liquid scintillation counter.The results were used to estimate photosynthetic rates. Theeffects of changing the partial pressures of O₂ and CO₂ on thephotosynthetic rate were studied and estimates made of the CO₂compensation point and photorespiration. The data obtained show differences between the mean net photosyntheticrates of the C. arabica cultivars (6·14 mg CO₂ dm^–2h^–1) and the mean rate for the C. canephora cultivar (3·96mg CO₂ dm^–2 h^–1). The cultivar of the latter speciesphotorespired more rapidly than the cultivar Catuai of C. arabica.Rates of photosynthesis in coffee measured using the ¹⁴CO₂ methodwere similar to rates obtained by others using an infrared gasanalyser. The ¹⁴CO₂ method proved to be reliable for photosyntheticmeasurements and the apparatus is suitable for use in fieldconditions.     

The Effect of Temperature and Light on Gas Exchange and Acid Accumulation in the C3-CAM Plant Clusia minor L

Gas exchange and organic acid accumulation of the C₃-CAM intermediateClusia minor L. were investigated in response to various day/nighttemperatures and two light regimes (low and high PAR). For bothlight levels equal day/night temperatures between 20°C and30°C caused a typical C₃ gas exchange pattern with all CO₂uptake occurring during daylight hours. A day/ night temperatureof 15°C caused a negative CO₂ balance over a 24 h periodfor low-PAR-grown plants while high-PAR-grown plants showeda CAM gas exchange pattern with most CO₂ uptake taking placeduring the dark period. However, there was always a considerablenight-time accumulation of malic acid which increased when thenight-time temperature was lowered and had its maximum (54 mmolm^–2) at day/night temperature of 30/15°C. A significantamount of malic acid accumulation (23 mmol m^–2) in low-PAR-grownplants was observed only at 30/15°C. Recycling of respiratoryCO₂ in terms of malic acid accumulation reached between 2·0and 21·5 mmol m_–2 for high-PAR-grown plants whilethere was no significant recycling for low-PAR-grown plants.Both low and high-PAR-grown plants showed considerable night-timeaccumulation of citric acid. Indeed under several temperatureregimes low-PAR-grown plants showed day/night changes in citricacid levels whereas malic acid levels remained approximatelyconstant or slightly decreased. It is hypothesized that lowand high-PAR-grown plants have different requirements for citrate.In high-PAR-grown plants, the breakdown of citrate preventsphotoinhibition by increasing internal CO₂ levels, whereas inlow-PAR-grown plants the night-time accumulation of citric acidmay function as an energy and carbon saving mechanism. Key words: C. minor, C₃, CAM, citric acid, light intensity     

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     

Purification and Storage of Ribulose 1,5-bisphosphate Carboxylase from Rice Leaves

Ribulose 1,5-bisphosphate (RuBP) carboxylase was purified fromrice leaves. By using a buffer containing 12.5% (v/v) glycerolthroughout purification, the enzyme was protected from coldlability and was obtained at a high yield (5.5 mg/g fresh wt).The purified enzyme exhibited different rates of CO₂/Mg²⁺-activationby temperature pretreatment/storage. The purified enzyme was stable for at least one year in phosphatebuffer containing 12.5% (v/v) glycerol at 4°C or 50% (v/v)glycerol at –20°C. (Received March 1, 1983; Accepted June 27, 1983)     

Apparatus for the Simultaneous Measurement of Water Vapour and Carbon Dioxide Exchanges of Single Leaves

Equipment is described which delivers air with concentrationsof CO₂ and water vapour closely controlled in the ranges 0 to2500 ppm and 5 to 15 mb respectively, at flow rates of up to10 1 min^-1, to each four leaf chambers. The leaf temperatureis controlled to ±0.5 °C and, with a light intensityof 0.3 cal cm^-2 min^-1 visible radiation (0.4 to 0.7 µm)leaf temperature can be maintained at 17.5 °C.The apparatusused to measure the concentration differences between the watervapour and CO₂ entering and leaving the leaf chamber (used tocalculate transpiration, photosynthetic, and respiration rates)is described in detail.Results of tests, which show the necessityfor mounting a fan within the leaf chamber, are reported.Typicallight- and CO₂-response curves are given for kale leaves (Brassicaoleracca var. acephala) and an attempt is made to quantify theerrors in the measurement of photosynthesis and transpiration.     

Some Properties of Ribulose Bisphosphate Carboxylase Extracted from Tomato Leaves

Ribulose bisphosphate carboxylase (EC 4.1.1.39 [EC] ) activity wasvery low in tomato leaf extracts unless prepared in the presenceof Mg²⁺, and polyclar AT. With young leaves, but not with fully-expanded leaves, the RuBP carboxylase activityextracted was increased by prolonged illumination of the leaves(2 h). The main effect of the light treatment was to increasethe specific activity of the enzyme but there was also a smallincrease in RuBP carboxylase protein. Tomato leaf RuBP carboxylasein extracts had specific activities in the range 0.2–0–6µmol CO₂ min^–1 mg^–-1 total protein extracted,or 0.5–1.2 µmol CO₂ min^–1 mg^–1 RuBPcarboxylase, and an apparent K_m (CO₂) at 20 ?C of 9.3 ? 1.2µM (using a of 6.407). Key words: Tomato leaf, RuBP carboxylase, Properties     

Glucose 6-phosphate dehydrogenase from sweet potato: Effect of various ions and their ionic strength on enzyme activity

Activity of glucose 6-phosphate dehydrogenase (D-glucose 6-phosphate:NADP oxidoreductase, EC 1.1.1.49 [EC] ) preparation from sweet potatoroot tissue was markedly altered in the presence of variousions. Cations or anions were effective in the following order:Na^$, K^$>Tris^$>NH₄^$>Mg^2$>Ca^2$, or Cl^–>NO₃^–,HPO₄^2–>SO₄^2–>HCO₃^–. Activity was inhibitedat high concentrations of Ca^2$, and HCO₃^–,. In an investigationon the dependence of the activity on pH, two activity peakswere clearly observed at low ionic strength. Ionic strength altered both the Km and Vmax for glucose 6-phosphate(G6P). A Lineweaver-Burk plot for the enzyme, with respect toG6P, showed a bimodal nature at low ionic strength; suggestingnegative cooperativity. Deviation from linearity of the plotwas less with an increase in the ionic strength. ¹ Present address: Institute of Applied Microbiology, Universityof Tokyo, Bunkyo-ku, Tokyo 113. (Received September 18, 1971; )     

Long Term Effects of High CO2 Concentration on Photosynthesis of Water Hyacinth (Eichhornia crassipes (Mart.) Solms)

The photosynthetic response to CO₂ concentration, light intensityand temperature was investigated in water hyacinth plants (Eichhorniacrassipes (Mart.) Solms) grown in summer at ambient CO₂ or at10000 µmol(CO₂) mol^–1 and in winter at 6000 µmol(CO₂)mol^–1 Plants grown and measured at ambient CO₂ had highphotosynthetic rate (35 µmo1(CO₂) m^–2 s^–1),high saturating photon flux density (1500–2000) µmolm^–2 s^–1 and low sensitivity to temperature in therange 20–40 °C. Maximum photosynthetic rate (63 µmol(CO₂)m^–2 s^–1) was reached at an internal CO₂ concentrationof 800 µmol mol^–1. Plants grown at high CO₂ in summerhad photosynthetic capacities at ambient CO₂ which were 15%less than for plants grown at ambient CO₂, but maximum photosyntheticrates were similar. Photosynthesis by plants grown at high CO₂and high light intensity had typical response curves to internalCO₂ concentration with saturation at high CO₂, but for plantsgrown under high CO₂ and low light and plants grown under lowCO₂ and high light intensity photosynthetic rates decreasedsharply at internal CO₂ concentrations above 1000 µmol^–1. Key words: Photosynthesis, CO₂, enrichment, Eichhornia crassipes