Response of Wheat Seedlings to Low O2 Concentrations in Nutrient Solution: I. GROWTH, O2 UPTAKE AND SYNTHESIS OF FERMENTATIVE END-PRODUCTS BY ROOT SEGMENTS

Root growth of 7-d-old wheat (Triticum aestivum cv. Gamenya)seedlings was impaired at dissolved O₂ concentrations of 0.01and 0.055 mol m^–3 O₂, while growth at 0.115 mol m^–3O₂ was the same as that in continuously aerated controls (0.26mol m^–3 O2). Oxygen uptake by apical (0–2 mm), expanding (2–4mm) and expanded (10–12 mm) tissues of the roots decreasedbelow 0.16, 0.09 and 0.05 mol m^–3 O₂, respectively. Thishierarchy is consistent with the metabolic rates of these tissues.There was a small (c. 9%) inhibition of O₂ uptake and some netsynthesis of ethanol and alanine in root apices at 0.115 molm^–3 O₂. Significant amounts of anaerobic end-productsaccumulated at 0.055 mol m^–3 O₂ and even more so at 0.01mol m^–3 O₂, indicating that oxidative phosphorylationwas strongly inhibited. Net alanine synthesis increased in fully expanded (10–16mm) tissues exposed to <0.003–0.01 mol m^–3 O₂,and this increase was accompanied either by a proportionallysmaller increase in the concentration of other free amino acidsor by a net decrease in free amino acid levels excluding alanine.This suggests that alanine was synthesized as an end-productof anaerobic catabolism and did not accumulate simply becauseof decreased net protein synthesis. Comparing the carbon flow to CO₂, ethanol, lactate and alaninein roots at 0.01 mol m^–3 O₂ with carbon loss as CO₂ inaerated roots suggests that carbon flow to products of metabolismwas not greatly enhanced due to O₂ deficiency. This infers,but does not prove that, in wheat, generation of energy duringperiods of O₂ deficiency is not enhanced due to a Pasteur effect. Key words: Anaerobic, fermentation, oxygen, wheat     

The Relationship Between Growth and Oxygen Uptake in Hypoxic Rice Seedlings

Atwell, B. J. and Green way, H. 1987. The relationship betweengrowth and oxygen uptake in hypoxic rice seedlings.—J.exp. Bot. 38: 454–465. Rice seedlings (Oryza saliva L.) were grown in the dark forup to 4 d in solutions containing various concentrations ofO₂. Compared with seedlings grown at 0·250 mol O₂ m^–3,the dry weight of the growing seedling was 14% lower at 0·110mol O₂ m^–3 and 60% lower at 0 mol O₂ m^–3. Decreasesin fresh weight were similar but not identical to decreasesin dry weight, possibly because leaf growth was suppressed evenabove 0·110 mol O₂ m^–3. Oxygen deficiency inhibitedroot growth more severely than coleoptile growth. Coleoptiles from seedlings grown in aerated solution were exposedto an atmosphere of pure N₂ for 30 min. Anoxia caused a declinein ATP content and energy charge, suggestive of decreased oxidativephosphorylation. It is not clear whether the decline in oxidativephosphorylation was solely responsible for impaired growth inhypoxia. In seedlings growing at O₂ concentrations less than 0·110mol O₂ m^–3, significant amounts of ethanol were synthesized.The rate of O₂ uptake decreased markedly below 0·06 molO₂ m^–3; this was presumably near the external O₂ concentrationat which oxidative phosphorylation became limited by the supplyof O₂. The stage of development of the seedlings appeared toinfluence O₂ uptake, possibly through changes in conductanceof the tissue to O₂. Uncouplers were used to confirm that thecritical O₂ concentration was dependent on O₂ diffusion ratherthan enzyme kinetics. Impaired growth above 0·110 molO₂ m^–3 may have been due to a decreased activity of oxygenasesof relatively low affinity for O₂, which in turn altered cellmetabolism. Key words: Growth, oxygen uptake, rice seedlings, hypoxia     

Effects of Low Concentration of Ozone, Singly, and in Combination with Sulphur Dioxide on Net Photosynthesis Rates of Vicia faba L.

Net photosynthesis rates (P_N) of Viciafaba plants were measuredin chambers containing either clean air or air containing between50 and 300 parts 10^–9 ozone (O₃) with or without 40 parts10^{– 9} SO₂. Exposure to O₃ concentrations greater than 50 parts 10^–9 for periods of 4 h resulted in reductions in PN with greaterreductions at higher concentrations. After exposure ceased,recovery of pre-exposure PN depended on O₃ concentration. Exposureto less than 90. 100 parts 10^{– 9} was followed by completerecovery after 20 h whereas 200-300 parts 10^{– 9} resultedin visible injury and irreversible depression of PN. The additionof 40 parts 10^{– 9} SO₂ to O₃ significantly decreased Psbut the effect of SO₂ declined with increasing O₃. The additionof SO₂ had no significant affect on recovery patterns. It is postulated that Vicia faba plants are able to toleratethe absorption of O, up to a threshold above which progressivedecreases in PN occur due to effects on photosynthetic processes.The slow and sometimes incompletely reversible effects of O₃on PN indicate cellular differences in the injury mechanismand repair capacity compared with the action of SO₂ alone. Lowconcentrations of O₃ with SO2 result in additive injury to PNsuggesting independent action of the pollutants, but the reductionin SO₂ action with increasing O₃ indicates a limit for potentialphysiological injury More the onset of visible injury.     

Nitrate transport in intact wheat roots:II. Long-term effects of NO3- concentration in the nutrient solution on NO3- unidirectional fluxes and distribution within the tissues5

We have examined the long-term effects of NO₃^– concentrationson NO₃^– (¹⁵NO₃^–) fluxes and cellular pool sizesin roots of intact 30-d-old wheat (Triticum aestivum cv. Courtot)grown hydroponically. Compartmental analysis was performed understeady-state conditions at five different levels of NO₃^–concentration (from 0.1 up to 5 mol m^–3 taking into accountmetabolism and secretion into the xylem (Devienne et al., 1994).Nitrate and reduced nitrogen levels in the tissues were largelyindependent of external NO₃^– concentration although below1.5 mol m^–3 NO₃^–; concentration limited plant growth.In the chamber, marked diurnal variations in net uptake occurredand, in the light, higher NO₃^– concentrations yieldedhigher NO₃^– uptake rates. After transfer of the plantsto the laboratory, the increase in net uptake linked to elevationof NO₃^–; concentrations was even larger (from 0.1 to 8.8µmolh^–1 g^–1 FW) as a result of a marked increase (x10–11) in the unidirectional influx at the plasmalemmawhile NO₃^– efflux was less enhanced (x 4–5). Underthese conditions, influx into the vacuole was also higher (x2–4) while efflux from the vacuole was little affected(x 1–3). NO₃^– concentrations within the cell compartmentswere estimated under the clas sical assumptions. The vacuolarconcentration was a little modified by NO₃^– availabilitywhereas that in the cytosol increased from about 10 mol m^–3to about 20 mol m^–3 indicating that (1) the absolute valuefor the cytosol was high and (2) it displayed only a small increasedespite very large changes in NO₃^– fluxes. NO₃^–distribution within the cells did not seem to involve an activeaccumulation of NO₃^– in the vacuole. Key words: Wheat, ion transport, nitrate, ¹⁵N, compartmentation     

Studies on the Movement of Water Through Apple Trees

Resistances to the flow of water through young potted appletrees were estimated by measuring the transpiration rate oftrees with and without root systems. Root system resistanceswere obtained by difference. Whole-plant resistances were ofthe order 10 x 10¹³ Pa s m^–3 and there was some evidencethat root resistances (R_r) varied with transpiration rate; theratio R_r:R_x (where R_x is resistance to water flow in the stemsystem) altered from 2:1 at relatively high transpiration ratesto 1:1 at lower rates. The trunk of a 9-year-old orchard tree (trunk diameter {smalltilde}7 cm, height {small tilde}2.5 m) was cut under water andestimates of the flow resistances in this tree were obtained.These were much lower than the resistances to flow in the pottedtrees. Capacitance (defined as the change in stored water content perunit change in plant water potential) values were calculatedfor the small trees and the large tree from measurements ofweight and water potential changes after the trees were removedfrom water. They were very similar on a weight basis (approx.2.0 x 10^–8 kg kg^–1 Pa^–1). Leaf capacitancevalues ({small tilde}1 x 10^–8 kg Pa^–1 m^–2)were also obtained. Stomatal conductances decreased with water potential and increasedwith short-wave radiation, but the relationships were not definitive.Estimates of boundary layer conductance in a greenhouse (verylow wind speeds) were of the same order ({small tilde}5 mm s^–1)as values obtained previously.     

Nitrate Transport into Chara corallina Cells using 36ClO-3 as an Analogue for Nitrate: I. INTERACTION BETWEEN 36ClO-3 AND NO-3 AND CHARACTERIZATION OF 36CIO-3/NO-3 INFLUX

The use of chlorate as an analogue for NO^–₃ during nitrateuptake into Chara corallina cells has been investigated. NO^–₃inhibits ³⁶C1O^–₃ influx into Chara over the concentrationrange 0–1000 mmol m^–3. Lineweaver-Burke plots ofthe data are characteristic of competitive inhibition by NO^–3in the low concentration range (0–300 mmol m^–3 ClO^–₃)and apparent K_INO3 is 140 mmol m^–3 which is of a similarorder of magnitude as apparent K_mCIO3- 180 mmol m^–3. Athigher substrate concentrations the inhibition by NO^–₃was not characteristic of competitive or uncompetitive inhibition. ³⁶C1O^–₃/NO^–₃ influx was dependent on K⁺ and Ca²⁺in the external medium and inhibited by FCCP. NO^–₃ pretreatmentor N starvation increased subsequent ³⁶C1O^–₃/NO^–₃influx into Chara. A comparison between rates of net NO^–₃uptake and ³⁶C1O^–₃/NO^–₃ influx supported the previoushypothesis that NO^–₃ efflux is an important componentin the determination of overall uptake rates. Key words: Nitrate, Chara, ³⁶CIO^–₃     

Modification of the CO2 Responses of Maize Stomata by Abscisic Acid and by Naturally-Occurring and Synthetic Cytokinins

Fragments of maize leaves were incubated at controlled temperatureand irradiance either on distilled water or on one of threeconcentrations of cytokinin (10^–1, 10^–2 and 10^–3mol m^–3). The effects of zeatin or kinetin on stomatalaperture were determined by stripping abaxial epidermis fromthe fragments after incubation and immediately measuring stomatalapertures under the microscope. At each cytokinin concentrationleaf pieces were incubated at 5 or 350 µmol mol^–1CO₂ with or without ABA (10^–1 mol m^–3). At 5.0 µmolmol^–1 CO₂ increasing the concentrations of zeatin hada negligible effect upon stomatal aperture. When air containing350 umol mol^–1 CO₂ was bubbled through the incubationsolutions, apertures of stomata incubated on water were morethan halved. Increasing cytokinin concentrations reduced theeffect of CO₂ on stomata and incubation on 10^–1 mol m^–3zeatin completely removed any CO₂ response. The addition ofABA restored the effect of CO₂, even at the highest cytokininconcentration. Key words: Maize, CO₂ response, ABA, Cytokinins     

Response of Wheat Seedlings to Low O2 Concentrations in Nutrient Solution: II. K +/Na+ SELECTIVITY OF ROOT TISSUES9

This paper reports the effects of low O₂ concentration (0–01,0–055, and 0.115mol m^–3) in nutrient solutions onK⁺/Na⁺ selectivity of growing and mature root tissues of 6-to 8-d-old, intact, wheat (Triticum aestivum cv. Gamenya) seedlings. Increases in anaerobic catabolism and decreases in O₂ uptake,K⁺ uptake and K⁺/Na⁺ selectivity were all more pronounced and/oroccurred at higher external O₂ concentrations in the apex (0–2mm) than in the expanding tissues (2–4 mm); these growingtissues were, in turn, more affected than the expanded tissuesof the roots (4–12 mm). Selectivity for K⁺ over Na⁺ in roots and shoots was particularlysensitive to O₂ deficiency. For example, in apical tissues (0–2mm) K ⁺ /Na⁺ selectivity was already reduced at 0.115 mol m^–3O₂, yet at this O₂ concentration there was no effect on eithergrowth or (K⁺/Na⁺) uptake. Upon transfer from 0.01 to 0.26 mol m^–3 O₂, a detailedstudy of the 12 mm root tips showed that 70% of these tips regainedhigh (K⁺ + Na⁺) concentrations and K⁺/^Na+ ratios. In contrast,there was no recovery in the remaining 30% of the 12 mm roottips. Net K⁺ transport to the shoots during the period afterre-aeration was negative for the population as a whole. Theseverity of these effects supports the view that the root tipsand the stele were more susceptible to O2 deficiency than wasthe cortex of the fully-developed root tissues. Key words: Hypoxia, K⁺/Na⁺ selectivity, expanded and expanding tissues     

Effects of a Range of O2 Concentrations on Porosity of Barley Roots and on their Sugar and Protein Concentrations

Barley was grown at a range of oxygen concentrations (0.5–9mg l^–1), in nutrient solutions. Growth of both shootsand seminal roots was restricted by O₂ concentrations lowerthan 2–3 mg l^–1) but nodal root growth was not. Root porosities were increased even at those O₂ concentrationswhich did not restrict growth, and were inversely proportionalto the protein levels of the roots. Sugar concentrations increasedappreciably only at those O₂ concentrations which also restrictedgrowth. Hordeum vulgare L., barley, root porosity, sugar, protein, oxygen concentration     

L-Leucine Transport in Isolated Protoplasts of Vinca Suspension Culture: Characterization of Uptake

The uptake of L-leucine into Vinca protoplasts was studied undervarious conditions. The uptake was highly pH-dependent, withthe optimal pH between 3.0 and 4.0. The uptake was also energydependent, since azide, 2,4-dinitrophenol (DNP), carbonyl cyanidem-chlorophenyl hydrazone (CCCP), and iodoacetate inhibited theuptake. Oligomycin, N,N'-dicycIohexyI carbodiimide (DCCD) andvanadate, but not ouabain, inhibited the uptake, suggestingthat ATPase for H⁺ electrogenic extrusion was necessary to theuptake of L-leucine. The uptake showed stereospecificity, butwas partially inhibited by other L-amino acids. A kinetic studyof the uptake showed that the uptake was multiphasic with threesaturable phases and one unsaturable phase which occurred atconcentrations of L-leucine over 1 mM. The K_m values of thethree affinity sites were 1.4 x 10^–3 M, 1.3 x 10^–4M, 4.3 x 10^–5 M; the maximum velocity values were 3.3x 10^–8, 4.5 x 10^–9, 1.8 x 10_–9 mol/10 min/4x 10⁶ cells. (Received April 18, 1981; Accepted August 25, 1981)     

Carbohydrate Metabolism of Rice Seedlings Grown in Oxygen Deficient Solution

Atwell, B. J. and Greenway, H. 1987. Carbohydrate metabolismof rice seedlings grown in oxygen deficient solution.—J.exp. Bot. 38: 466–478. Rice seedlings (Oryza sativa L.) were grown in the dark forup to 4 d in solutions containing various concentrations ofO₂. The rate of depletion of the endosperm was most rapid inaerated solution (0·25 mol O₂ m^–3), largely dueto the inhibition of growth of seedlings at very low O₂ concentrations.Earlier suggestions that there is a deficit of sugars for growthand energy generation in O₂ deficient coleoptiles were tested. Coleoptiles, shaking in aerated solution, respired about one-thirdof the endogenous sugars to CO₂ and incorporated the rest intostructural compounds. In contrast, the proportion of carbonwhich went to growth in anoxia was very low. Consistent withthese results, endogenous sugar levels were generally highestat low O₂ concentrations. Even so, coleoptiles grown and testedas low as 0·03 mol O₂ m^–3 showed appreciable metabolismof exogenous ¹⁴C-glucose to CO₂, soluble and insoluble compounds,suggesting that a minimal O₂ supply was sufficient to sustainsome growth. Furthermore, glucose feeding caused little or norise in O₂ uptake or tissue sugar levels. Similarly, the specificactivity of the evolved CO₂ was not markedly different in coleoptilesgrowing at 0·03 and 0·25 mol O₂ m^–3 Further evidence was obtained to show that endogenous substrateswere adequate for growth and respiration at both low and highO₂ concentrations. Exogenous glucose and malate did not stimulateO₂ uptake at any stage of growth in aerated coleoptiles. Therewas sufficient endogenous substrate to sustain a 35–45%rise in O₂ uptake induced by uncoupling and enrichment withO₂. Exogenous glucose did not stimulate growth of intact seedlingsat any O₂ concentration. Key words: Rice seedlings, carbohydrate metabolism, oxygen deficient solution     

Leaf Abscission and Stem Lesions (Intumescences) on Poplar Clones after SO2 and O3 Fumigation: A Link with Ethylene Release?

Differences in premature leaf abscission and in visible steminjury in genetic lines of poplar followed continuous fumigationswith air pollutant levels of SO₂ (90–100 nl l^–1)and O₃ (70–80 nl l^–1) either separately or together.The clones used were: Populus deltoides var. missiouriensisMarsh., P. nigra cv. ‘italicd’ L., and the hybridsP. nigra cv. ‘italica’ xP. deltoides (He-X/3) andP. nigra cv.‘italica’ x P. nigra cv. ‘Serres’(He-K/7). While most leaf abscission occurred within 20 d fromthe start of fumigation, stem lesions (intumescences), appearedonly after 72 d. Their anatomical characteristics include theformation of lysigenous aerenchyma in the lower parts of theintumescence, the sloughing of superficial cells from the injuredarea, and the development of crystalline formations on the surfaceof the lesions. P. deltoides exhibited the least morphologicalresponse to the gases. Ethylene released from fumigated leaves was determined at thesame gas concentration of SO₂ (100 nl l^–1), O₃ (75 nll^–1) and their mixture. Leaves of P. deltoides consistentlyshowed the lowest ethylene production after the gas treatments.P. ‘italica’ production was higher but was littlealtered by the treatments. The two hybrids He-X/3 and He-K/7showed the greatest increases in ethylene evolution with time.With He-K/7 exposed to the gas mixture the production of ethylenedecreased after the initial sharp rise during days 1–2,and reflected the considerable leaf damage observed after day3. The results suggest that sensitivity to air pollution, (as shownby leaf abscission and the formation of stem intumescences)can be correlated with the level of pollutant-induced ethyleneevolution from leaves. Initially high levels could induce abscission,whilst prolonged production could be responsible for intumescenceinitiation. The discussion proposes a series of events fromSO₂ and/or O₃ entry into the leaf and the physiological reasonsfor the clonal differences. Key words: Sulphur dioxide, ozone, ethylene, poplar, leaf abscission, stem lesions     

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     

Stomatal Responses to Applied ABA and CO2 in Epidermis Detached from Well-Watered and Water-Stressed Plants of Commelina communis L.

Epidermal strips from either well-watered or water-stressedplants of Commelina communis L. were subjected to a range ofABA concentrations (10^–6–10^–3 mol m^–3)in the presence (330 parts 10^–6 in air) or virtual absence(3 parts 10–6 in air) of CO₂. The stomatal response toCO₂ was greater in epidermis from water-stressed plants, althoughthere was a distinct CO₂ response in epidermis from well-wateredplants. Additions of ABA via the incubation medium had littleeffect on the relative CO₂ response. Stomata responded to ABAboth in the presence and virtual absence of CO₂, but the relativeresponse to ABA was greatest in the high CO₂ treatment. Whenwell-watered plants were sprayed with a 10^–1 mol m^–3ABA solution 1 d prior to use, the stomatal response of detachedepidermis to both CO₂ and ABA was very similar to that of epidermisdetached from water-stressed leaves. It is hypothesized thata prolonged exposure to ABA is necessary before there is anymodification of the CO₂ response of stomata.     

Measuring the Canopy Net Photosynthesis of Glasshouse Crops

A null balance method is described for measuring net photosynthesisof mature canopies of cucumber and other protected crops overperiods of 10 min in a single-span glasshouse (c. 9m x 18m inarea). Accuracy of control of the CO₂ concentration in the greenhouseatmosphere is within ±10 vpm of the normal ambient level(c. 350 vpm). The amounts of CO₂ used in canopy net photosynthesisare measured with linear mass flowmeters accurate to within±0.80g. The total errors incurred in measuring canopynet photosynthesis at an ambient CO₂ level are estimated tobe of the order of ± 1·2% in bright light (350W m^–2, PAR)and ±3·6% in dull light (100W m^–2, PAR). Measurements of the rates of net photosynthesis of a maturecanopy of a cucumber crop were made at near-ambient CO₂ concentrationsover a range (0–350 W m^–2) of natural light fluxdensities. A model of light absorption and photosynthesis applicableto row crops was used to obtain a net photosynthesis versuslight response curve for the cucumber crop. At a light fluxdensity of 350 W m^–2 the fitted value of canopy net photosynthesiswas 2.65 mg CO₂ m^–2s^–1 (equivalent to over 95 kgCO₂ ha^–1h^–1). The results are discussed in relationto the need for CO₂ supplements to avoid depletion in both ventilatedand unventilated glasshouses during late spring and summer. Key words: Glasshouse crops, cucumber, measurement, canopy photosynthesis, light, CO₂     

Characteristics of the Cl- Action Site in the O2 Evolving Reaction in PS II Particles: Electrostatic Interaction with Ions

The role of Cl^– in the reactivation of O₂ evolution inphotosystem II (PS II) particles derived from spinach chloroplastswas studied in the presence of various salts. Multivalent ion(especially anion) salts were found to strongly suppress thereactivation of O₂ evolution by Cl^– in the Cl^–-depletedPS II particles in a competitive manner. The effectiveness ofanions in the suppression of Cl^–-supported O₂ evolutionwas in the order of trivalent>divalent>monovalent ones.Multivalent anions similarly suppressed O₂ evolution in theuntreated PS II particles under low and moderate Cl^– concentrations.pH dependence of the Cl^–-affinity (K_m) value for Cl^–)was also studied. Within the pH range 5.5 to 8 the K_m valuebecame higher as the pH of the medium increased. These resultssuggest that the membrane surface in the vicinity of the Cl^–action site is net positively charged and attracts Cl^–electrostatically, and that the site is almost freely accessibleto various anions. The origin and role of the local net positivedomain and the role of peripheral proteins are discussed. (Received May 27, 1985; Accepted October 8, 1985)     

The Effect of External pH on the Apparent CO2 Affinity of Chlorella saccharophila

The carbon dioxide compensation point of the unicellular greenalga, Chloretla saccharophila, was determined in aqueous mediumby a gas chromatographic method. Compensation points decreasedmarkedly from 63 cm³ m^–3 at an external pH of 4.0 to 3.2cm³ m^–3 at pH 8.0 and were not affected by the O₂ concentrationof the medium. The calculated CO² concentration required tosupport the half-maximum photosynthetic rate of the algal cellsranged from 6.0 mmol m_–3 at an external pH of 60 to 1.5mmol m^–3 at pH 8.0 and these values were not affectedby O₂ concentration. The K_m(CO₂) of nbulose-l,5-bisphosphatecarboxylase isolated from cells grown either at pH 4.0 or pH8.0 was determined to be 64 mmol m^–3. These results indicatethat loss of CO₂ by photorespiration does not occur in C. saccharophilacells at acid pH and the disparity between the apparent affinityfor CO₂ of the intact cells and that of the carboxylase indicatesthe operation of a ‘CO₂ concentrating mechanism’in this alga at acid pH. Key words: Acidophilic alga, bicarbonate transport, Chlorella saccharophila, compensation point, CO₂ affinity, PH, RuBP carboxylase     

Further Characteristics of Salt-Dependent Bicarbonate Use by the Seagrass Zostera muelleri

Millhouse, J. and Strother, S. 1987. Further characteristicsof salt-dependent bicarbonate use by the seagrass Zostera muelleri.—J.exp. Bot. 38: 1055–1068. The contribution of HCO^–₃to photosynthetic O₂ evolutionin the seagrass Zostera muelleri Irmisch ex Aschers. increasedwith increasing salinity of the bathing seawater when the inorganiccarbon concentration was kept constant. K_1/2 (seawater salts)for HCO^–₃ -dependent photosynthesis was 66% of seawatersalinity. Both short- and long-term pretreatment at low salinitiesstimulated photosynthesis in full strength seawater. Twentyfour hours pre-incubation of seagrass plants in 3·0 molm^–3 NaHCO₃ resulted in increased photosynthesis at allsalinities, apparently due to stimulation of HCO^–₃ use(K_1/2 (seawater salts) = 26%). V_max (HCO^–₃) was not affectedby low salinity pretreatment. The kinetics of HCO^–₃ stimulationby the major seawater cations was investigated. Ca²⁺ was themost effective cation with the highest V_max (HCO^–₃) andwith K_1/2(Ca²⁺) = 14 mol m^–3. Mg²⁺ was also very effectiveat less than 50 mol m^–3 but higher concentrations wereinhibitory. This inhibition cannot be accounted for solely byprecipitation of MgCO₃. Na⁺ and K⁺ were both capable of stimulatingHCO^–₃ use. Stimulation was in two distinct parts. Up to500 mol m^–3, both citrate and chloride salts gave similarresults (K_1/2(Na⁺) 81 mol m^–3, V_max(HCO^–₃) 0·26µmol O₂ mg^–1 chl min^–1), but use of citratesalts above 500 mol m^–2 caused a second stimulation ofHCO^–₃ use (K_1/2(Na⁺) 830 mol m^–3, V_max(HCO^–₃)0·68 µmol O₂ mg^–1 chl min^–1). V_max(HCO^–₃)for the second-phase Na⁺ or K⁺ stimulation was of the same orderas for Ca²⁺-stimulated HCO^–₃ use. To further characterizesalt-dependent HCO^–₃ use, the sensitivity of photosynthesisto Tris and TES buffers was investigated. The effects of Trisappear to be due to the action of Tris⁺ causing stimulationof HCO^–₃ -dependent photosynthesis in the absence of salt,but inhibition of HCO^–₃ use in saline media. TES has noeffect on photosynthesis. External carbonic anhydrase, althoughimplicated in salt-dependent HCO^–₃ use in Z. muelleri,could not be detected in whole leaves. Key words: Zostera muelleri, HCO^–₃ use, salinity     

Nutrient Solution pH Control using Dipolar Buffers in Studies of Trifolium repens L. Nitrogen Nutrition

In studies of Trifolium repens nitrogen nutrition, the controlof nutrient solution pH using dipolar buffers, was evaluatedin tube culture under sterile conditions. Five buffers; MES,ADA, ACES, BES and MOPS with pK₂s (20 °C) of 6.15, 6.60,6.90, 7.15 and 7.20 respectively, at a concentration of 2.0mol m^–3, were provided to inoculated Trifolium repensgrowing in nutrient solution containing 7.13 mol m^–3 nitrogenas (NH₄)₂SO₄. Initial pH of each solution was adjusted to theappropriate buffer pK₂ Two buffers, ADA and ACES completelyinhibited plant growth. The remaining buffers had little effectin limiting pH change, although plant dry matter was higherand nodule numbers lower in the presence of these buffers. MESand MOPS were supplied to nutrient solutions with and without7.13 mol m^–3 (NH₄)₂SO₄, at concentrations ranging from0–12 mol m^–3. MES at 9 mol m^–3 and 12 molm^–3 reduced growth of plants reliant on the symbiosisfor providing nitrogen. The provision of MES to plants providedwith NH₄⁺ significantly increased plant yield and reduced nodulenumber at all concentrations. MOPS did not affect plant yieldor nodule number. The use of dipolar buffers in legume nitrogennutrition studies is considered in terms of buffering capacity,and the side effects on plant growth and symbiotic development. Key words: Ammonium, Dipolar buffer, Nitrogen nutrition, pH control, Symbiosis, Trifolium repens     

Effects of CO2 and Photosynthetic Photon Flux on Yield, Gas Exchange and Growth Rate of Lactuca Sativa L. 'Waldmann's Green'

Knight, S. L. and Mitchell, C. A. 1988. Effects of CO₂ and photosyntheticphoton flux on yield, gas exchange and growth rate of Lactucasativa L. ‘Waldmann’s Green'.—J. exp. Bot.39: 317–328. Enrichment of CO₂ to 46 mmol m^–3 (1 000 mm³ dm^–3)at a moderate photosynthetic photon flux (PPF) of 450 µmolm^–2 s^–1 stimulated fresh and dry weight gain oflettuce leaves 39% to 75% relative to plants at 16 mmol m^–3CO₂ (350 mm³ dm^–3). Relative growth rate (RGR) was stimulatedonly during the first several days of exponential growth. ElevatingCO₂ above 46 mmol m^–3 at moderate PPF had no further benefit.However, high PPF of 880–900 µmol m^–2 s^–1gave further, substantial increases in growth, RGR, net assimilationrate (NAR) and photosynthetic rate (Pn), but a decrease in leafarea ratio (LAR), at 46 or 69 mmol m^–3 (1000 or 1500 mm³dm^–3) CO², the differences being greater at the higherCO₂ level. Enrichment of CO₂ to a supraoptimal level of 92 mmolm^–3 (2000 mm³ dm^–3) at high PPF increased leaf areaand LAR, decreased specific leaf weight, NAR and Pn and hadno effect on leaf, stem and root dry weight or RGR relativeto plants grown at 69 mmol m^–3 CO₂ after 8 d of treatment.The results of the study indicate that leaf lettuce growth ismost responsive to a combination of high PPF and CO₂ enrichmentto 69 mmol m^–3 for several days at the onset of exponentialgrowth, after which optimizing resources might be conserved. Key words: Photosynthesis, relative growth rate, CO₂ enrichment