A BENEFICIAL GAS TRANSPORT SYSTEM IN NYMPHOIDES PELTATA

The aquatic vascular plant Nymphoides peltata (Gmel.) O. Kunze which inhabits anaerobic environments depends highly on the availability of oxygen for its submerged organs buried in the sediment of the lake. In tracer gas studies, carried out with shaded plants, it is shown that ethane is taken up by one of the youngest leaves in a whorl and transported down the petiole to the axis, returning to the surface via the older leaves. In sunlight, this gas diffusion through the plant is replaced by an effect which enhances the gas movement up to 1,200% due to the increased difference between leaf temperature and the surrounding air (ΔT = 1.7 K). The temperature difference is accompanied by a pressurization 50 Pascal above ambient inside the aerenchyma of the young leaves. These findings confirm that a pressurized flow-through system is established by N. peltata, whereby the oxygen supply to the rhizomes is improved. The temperature difference derived from irradiation energy initiates a circulating air stream, which transports air from the young leaves through the plant. Enhanced transport of a tracer gas as well as oxygen can be demonstrated by warming an excised young leaf with red-filtered light or warm water. A similar increase in gas transport is not detected through older leaves. The light energy needed to create a temperature difference can be substituted with warm water. Evidence is thus given, which shows that the increased oxygen emission from the petiole of young leaves is independent of photosynthesis. This gas transport is a result of thermo-osmosis under slip-flow conditions (Knudsen diffusion), limiting this effect to a temperature gradient between the surrounding air and the lacunar air of young leaves.     

A two-way gas transport system in Nelumbo nucifera

Abstract The aquatic vascular plant Nelumbo nucifera Gaertn. is able to improve its oxygen supply to the submerged and buried organs by a thermo-osmotic gas transport. Investigations with tracer gas and oxygen measurements have shown that thermo-osmotic gas transport exists in N. nucifera when there is a temperature difference between the lacunar air of the leaves and the surrounding atmosphere. The gas transport was increased by up to 935% when a temperature difference of 2.9 ± 1.0 K was detected. Lacunar pressure of up to 166 ± 44 Pa was measured in both young and old leaves. In contrast to the flow-through ventilation system recently described for Nuphar lutea and Nymphoides peltata, a two-way flow in separate air canals in the petioles of both young and old Nelumbo leaves may carry oxygen-rich air down to the rhizome and excess air back to the atmosphere. Anatomical investigations have shown that, in Nelumbo, the two largest air canals of the petiole end directly under the mesh system of the centre plate. These large air canals are proposed to be predominant in the upward flow of air in sunlight. The other air canals of the petiole veer into the leaf blade well below the centre plate. The gas flow system through fresh leaves may carry as much as 10.3 ± 4.5 cm³ air per minute to the buried rhizome.     

Localization of Thermo-Osmotically Active Partitions in Young Leaves of Nuphar lutea

The submerged roots and rhizomes of the aquatic vascular macrophyteNuphar lutea (L.) Sm. are aerated, at least in part, by pressurizedventilation. Depending on temperature differences of up to 5K between the inside of young, just-emerged leaves and the surroundingair, pressure differences of 79 to 100 Pa higher than atmosphericare detectable inside the lacunuous spongy parenchyma of theleaf blades. The pressurization is a consequence of structuralfeatures of leaf tissues separating the air filled spaces ofthe spongy parenchyma from the atmosphere. These tissues areacting as thermo-osmotic partitions. Whereas the dimensionsof the stomatal openings (about 5·6 x 2·4 µm)and of the intercellular spaces of the palisade parenchyma (diametersabout 15 µm) are too large, those of the monolayers ofcells separating the palisade and the spongy parenchyma (diameters:0·7–1·2 µm) are small enough to impedefree gaseous diffusion. This inner non-homogeneous partitioninggives rise to the so-called Knudsen diffusion, a physical phenomenonleading to pressurization of the warmer air inside the spongyparenchyma. The rising pressure difference is strong enoughto establish an air flow through the aerenchyma of the wholeplant and out of the most porous older leaves in which a temperatureinduced pressurization is never detectable. These thermo-osmoticallyactive leaves enhance the influx of air to the rhizome and thediffusion path for oxygen to the roots is shortened to the distancebetween rhizome and root tips. Therefore, pressurized ventilationin Nuphar is seen to be of considerable ecological importancefor plant life in anaerobic environments. Key words: Aeration, leaf anatomy, thermo-osmosis of gases, Nuphar lutea     

Anaerobic conditions strongly promote extension by stems of overwintering tubers of Potamogeton pectinatus L

Elongation by dark-grown shoots of Potamogeton pectinatus tuberswas enhanced by the absence of oxygen. This promoting effectwas located in the stem and was stronger under water than ina gas phase and also stronger in unsparged water compared tosparged water. Anaerobic shoots elongated under water by almost13 cm in 5 d. This was the outcome of longer cells of the steminternodes and of some cell division coupled with leaf extensionwhich continued in the absence of oxygen, but at a slower rate.Continued attachment to a starch-filled tuber was required forsignificant anaerobic elongation, which could be sustained forat least 14 d. Switching intact, growing tubers from aerobicto anaerobic conditions stimulated stem extension within 24h. Conversely, stem elongation was slowed when tubers were transferredfrom an anaerobic to an aerobic environment. A marked gravitropicresponse occurred in anaerobic conditions, which involved bothstem and leaf tissue, and indicated that active internal growth-regulatingmechanisms continued to operate without oxygen. Shoot extension by tubers was also stimulated by hypo-aerobicconditions (5–8 kPa O₂) compared with fully aerated solutions(20.8 kPa O₂). This acceleration was smaller than that obtainedby the complete removal of oxygen, but still involved stem ratherthan leaf growth. Unlike elongation by apical shoots of tubers,that by shoot tips of rhizomes taken from mature light-grownplants was strongly inhibited by lack of oxygen, and in somecases shoot tips died within 5 d. All shoots and leaves werehighly aerenchymatous and the gas-filled lacunae were connectedby side-pores. Key words: Water plants, environmental stress, Potamogeton pectinatus, elongation, anaerobiosis, gravitropism     

Characterization of a thermo-osmotic gas transport mechanism in Alnus glutinosa (L.) Gaertn.

Summary A gas transport system based upon the physico-chemical effect of thermo-osmosis of gases in described for the black alder, Alnus glutinosa (L.) Gaertn. Air is transported through the alder's stem to the roots, thus improving O₂ supply to respiring tissues of the root system. The gas transport system is investigated by means of a tracer gas technique (11% ethane in air, v/v). Gas transport depends on any source of radiant heat generating a temperature difference between the tree's stems and the atmosphere. The amount of gas transported in leafless trees is four times higher than the amount of gas reaching the roots by gas diffusion. Two-thirds of the gas is transported in the wood, only one-third in the bark. Intercellular spaces inside the porous lenticels of the bark are responsible for this kind of gas transport. Their diameters are estimated by the effusion rates of different tracer gases to be in the range of 1 m.     

Photosynthetic Electron Transport During Senescence of the Primary Leaves of Phaseolus vulgaris L.: II. THE ACTIVITY OF PHOTOSYSTEMS ONE AND TWO, AND A NOTE ON THE SITE OF REDUCTION OF FERRICYANIDE

The activity of photosystems one and two (PS I and PS II) wasmeasured in chloroplasts isolated from the primary leaves ofPhaseolus vulgaris. During foliar senescence, the rates of electrontransport through PS I and PS II declined by approximately 25%and 33% respectively. These losses of activity could not accountfor the decrease of 80% in the rate of coupled, non-cyclic electrontransport during senescence. It is therefore suggested thatan impairment of electron flow between the photosystems limitednon-cyclic electron transport in chloroplasts from older leaves.In this study the activity of PS II was measured using oxidizedp-phenylenediamine as the electron acceptor, and trifluralinas an inhibitor of electron transport between PS II and PS I.In chloroplasts from young leaves the reduction of ferricyanidewas a measure of non-cyclic electron transport, but in preparationsfrom older leaves ferricyanide received a large proportion ofelectrons from PS II.     

Anatomy of the gas canal system of Nelumbo nucifera

Nelumbo nucifera (Gaertn.) grows by extending a creeping rhizome through anaerobic sediments. Nodes form at intervals along the rhizome, each producing a single leaf, and gas canals channel air from the leaves throughout the petioles and rhizomes. The gas flow pathway was mapped by casting the canals in growing shoots with silicone and by blowing air through complexes of rhizomes and petioles. Air from a leaf flows to a rhizome through one of two petiolar canal pairs, joining with the lowermost of three canal pairs in the rhizome through a chamber in the node. The lowermost canal pair links these nodal chambers along the length of a rhizome, allowing air from a node to flow both forward, toward a growing shoot, and backward, toward preceding leaves. These linked chambers also connect with the middle pair of canals on their proximal side, enabling flow to proceed backward along the rhizome to an adjacent node. A chamber in the next node then diverts the flow into the upper canal pair. This pair leads to a third node and chamber from which the air vents to the atmosphere through the second petiolar canal pair. Thus, pressurised air from one leaf must flow backward through two nodes before it returns to the atmosphere. Forward flow also ventilates a shoot's growing tip, with air from the lowermost canal pair entering a chamber in the developing node which, as described above, connects with the middle canal. This allows the air to reverse direction at the tip and enter the vent flow pathway.     

15N Studies on the In Vivo Assay of Nitrate Reductase in Leaves: Occurrence of Underestimation of the Activity Due to Dark Assimilation of Nitrate and Nitrite

The reduction of nitrate and nitrite in leaf disks from sevendi- and two monocotyledonous species under in vivo nitrate reductaseassay conditions was studied using 15N-labeled substrates. Significantreduction of both nitrate and nitrite into ammonia and aminoacids was detected under aerobic conditions (in an atmosphereof air): in some cases, the amount of nitrate-N reduced to ammoniaand amino acids was more than that remaining as nitrite. Anaerobicincubation (under an atmosphere of N₂ gas) enhanced the accumulationof nitrite, but the subsequent reduction to the basic nitrogencompounds was 40 to 180% of the aerobic rates. The present examinationindicates that in vivo assays of nitrate reductase under aerobicconditions may give greatly underestimated results due to nitritereduction and that exclusion of oxygen from the in vivo assaymixture is desirable in terms of the quantity of nitrite formed.n-Propanal treatment increased nitrite accumulation under aerobicbut not under anaerobic conditions, and depressed the incorporationof nitrate-N into basic fractions under both conditions. Therefore,addition of n-propanol may be desirable for assays under aerobicconditions. No significant difference in the reduction of nitratesupplied as sodium and potassium salts was observed on the nitriteformation and on the incorporation of nitrate-N into basic fractions. ¹⁵N experiments on dark assimilation of nitrate, nitrite andammonia into amino acids in wheat leaves showed that these threenitrogen sources were assimilated through the same route andthat the glutamine synthetase/glutamate synthase pathway wasthe major route. With anaerobic treatment, the incorporationof nitrogen into alanine and serine remained at relatively high,but the incorporation into aspartate and asparagine was muchlower than in the cases of aerobic treatment. (Received July 11, 1981; Accepted October 3, 1981)     

Irrungen,Wirrungen? The Mehler reaction in relation to cyclic electron transport in C3 plants

Plants not only evolve but also reduce oxygen in photosynthesis. Considerable oxygen uptake occurs during photorespiration of C3 plants. Controversies exist on whether direct oxygen reduction in the Mehler reaction together with associated electron transport is also a major sink of electrons when leaves are exposed to sunlight. Here, preference is given to the view that it is not. Whereas photorespiration consumes ATP, the Mehler reaction does not. In isolated chloroplasts photosynthesizing in the presence of saturating bicarbonate, the Mehler reaction is suppressed. In the water – water cycle of leaves, which includes the Mehler reaction, water is oxidized and electrons flow through Photosystems II and I to oxygen producing water. The known properties of coupled electron transport suggest that the water – water cycle cannot act as an efficient electron sink. Rather, by contributing to thylakoid acidification it plays a role in the control of Photosystem II activity. Cyclic electron transport competes with the Mehler reaction for electrons. Both pathways can help to defray possible ATP deficiencies in the chloroplast stroma, but play a more important role by making intrathylakoid protein protonation possible. This is a necessary step for the dissipation of excess excitation energy as heat. Linear electron flow to oxygen relieves the inhibition of cyclic electron transport, which is observed under excessive reduction of intersystem electron carriers. In turn, cyclic electron transport replaces functions of the linear pathway in the control of Photosystem II when oxygen reduction is decreased at low temperatures or, experimentally, when the oxygen concentration of the gas phase is low. Thus, cyclic electron flow acts in flexible relationship with the water–water cycle to control Photosystem II activity. This revised version was published online in June 2006 with corrections to the Cover Date.     

Pressurized gas transport in two Japanese alder species in relation to their natural habitats

Oxygen uptake measurements have shown that pressurized gas transport, resulting from the physical effect of thermo-osmosis of gases, improves oxygen supply to the roots of the seedlings in two alder speciesAlnus japonica (Thunb.) Steud. andAlnus hirsuta (Spach) Rupr., which are both native in Japan. When gas transport conditions were established by irradiation of the tree stems the internal aeration was increased to a level nearly equal to the oxygen demand of the root system in leafless seedlings ofA. hirsuta, but was higher inA. japonica so that excess oxygen was excreted into the environment. An increase of superoxide dismutase (SOD) activity, which protects plants from toxic oxygen radicals and post-anoxic injury, has been observed in root tissues ofA. japonica when the seedlings were flooded for 3 days. The increase of SOD activity, in concert with high gas transport rates, may enable this tree species to grow in wet sites characterized by low oxygen partial pressure in the soil and by varying water tables. A less effective gas transport, flood-induced reduction of SOD activity in root tissues, and reduced height growth in waterlogged soil may be responsible for the fact thatA. hirsuta is unable to inhabit wettland sites.     

Preliminary Experiments on the Effect of Temperature on the Movement of 14C-labelled Assimilates through the Phloem of Willow

Experiments were performed on both young shoots of willow (3–5weeks old) and mature stems which had been growing for a periodof 2–3 years. ¹⁴CO₂ was supplied to the leaves, and themass transport of the labelled assimilates through a portionof the stem enclosed in a temperature-controlled jacket wasmeasured by determining the slope of the rise in the activityof huneydew collected from a colony of the aphid Tuberolachnussalignus (Gmelin). It was found that there was a marked difference in the behaviourof the young shoots and mature stems. In the former, a fallin temperature slowed the transport, whilst in the latter acomparable fall in temperature increased the transport. Possiblecauses for these effects have been suggested, and the resultsare discussed in relation to published work on temperature effects.     

Changes in the Root System of Wheat Seedlings Following Root Anaerobiosis: III. Oxygen Concentration in the Roots

The oxygen status in roots of wheat seedlings (Triticum aestivum)was determined by a volumetric micro-absorption method. Plantsgrew in nutrient solution (aerated or nitrogen-flushed) or onflooded sand up to the 10th day. The roots were then exposedto aerated or hypoxic conditions for several hours before gaswas extracted by reducing the pressure within a concentratedsalt solution or by physical crushing. The oxygen content ofthe extracted gas bubbles was measured with pyrogallol. Comparativeexperiments with the helophytes Phalaris arundinacea and Carexacutiformis yielded similar oxygen concentrations to those alreadydescribed in literature. The concentrations of oxygen (13–16%)in young wheat roots were surprisingly high when exposed tonutrient solution flushed with nitrogen gas. Removal of the shoots decreased the oxygen concentration inthe roots, indicating some internal oxygen transport from shootsto roots. Detached, submerged roots of wheat still contained6% oxygen following 20 h of submergence in nitrogen-flushedsolution. A linear relationship was found between the oxygenconcentration in roots of Triticum aestivum, Zea mays and thetwo helophytes and the volume of extractable gas per volumeof root. This ratio corresponded to the extent of aerenchymaformation. Hence, a certain amount of oxygen may have been adsorbedonto the inner surfaces of the lacunae of the roots. However, the large amount of oxygen in the roots of intact wheatplants suggest that some parts of the root system are unlikelyto suffer from the oxygen shortage imposed by oxygen-deficientexternal conditions such as flooded soil. Triticum aestivum L. cv. Hatri, wheat, helophytes, roots, micro-absorption method, oxygen concentration, hypoxia, intercellular space     

Oxygen as an alternative electron acceptor in the photosynthetic electron transport chain of C3 plants

This study deals with effects of oxygen on the kinetics of P(700) photoinduced redox transitions and on induction transients of chlorophyll fluorescence in leaves of C(3) plants Hibiscus rosa-sinensis and Vicia faba. It is shown that the removal of oxygen from the leaf environment has a conspicuous effect on photosynthetic electron transport. Under anaerobic conditions, the concentration of oxidized P700 centers in continuous white light was substantially lower than under aerobic conditions. The deficiency of oxygen released non-photochemical quenching of chlorophyll fluorescence, thus indicating a decrease in the trans-thylakoid pH gradient (DeltapH). Quantitative analysis of experimental data within the framework of an original mathematical model has shown that the steady-state electron flux toward oxygen in Chinese hibiscus leaves makes up to approximately 40% of the total electron flow passing through photosystem 1 (PS1). The decrease in P700+ content under anaerobic conditions can be due to two causes: i) the retardation of electron outflow from PS1, and ii) the release of photosynthetic control (acceleration of electron flow from PS2 to P700+) owing to lower acidification of the intra-thylakoid space. At the same time, cyclic electron transport around PS1 was not stimulated in the oxygen-free medium, although such stimulation seemed likely in view of possible rearrangement of electron flows on the acceptor side of PS1. This conclusion stems from observations that the rates of P700+ reduction in DCMU-poisoned samples, both under aerobic and anaerobic conditions, were negligibly small compared to rates of electron flow from PS2 toward P700+ in untreated samples.     

Comparison of some Processes in Homogenates and Sections of Healthy Bean Leaves Resistant and Susceptible to the Bacterial Halo-blight Disease

In unifoliates of eight lines of bean (Phaseolus vulgaris),four with a pole and four with a bush habit, symptoms of resistanceand susceptibility, according to line, developed quickly whenplants were inoculated with Pseudomonas phaseolicola on theday when the leaves opened. When older leaves were inoculatedsymptoms developed more slowly, but the resistant/susceptibledifference was retained. Oxygen uptakes by homogenates of healthy unifoliates of thefour resistant lines were 30 to 75 per cent. higher than theuptakes by homogenates of those of the four susceptible lineson the day when the leaves opened. Subsequently the uptakesfluctuated somewhat, although those of resistant leaves werealways higher. Using leaf sections, rates of respiration and the extents ofdark CO₂ fixation were found to be the same in both resistantand susceptible unifoliates at all stages of development. Old healthy resistant leaves decarboxylated indoleacetic acidless rapidly than did old healthy susceptible leaves, but therewas no difference in young leaves. The oxygen uptake by homogenates is, therefore, the most promisingreaction to pursue in order that the difference between resistantand susceptible leaves might be understood.     

Comparative Photosynthesis in Developing Leaves of Brachystegia spiciformis Benth

Photosynthesis and dark respiration were studied during andafter the spring flush in Brachystegia spiciformis Benth. Variousparameters were examined including anthocyanin and chlorophyllcontent, Hill reaction activity, and gas exchange. Total chlorophyllcontent steadily increased reaching a constant value in fullyexpanded (25-d-old) leaves, whereas anthocyanin concentrationincreased as the chlorophylls but rapidly declined after 23d. Hill reaction measurements confirmed that leaf material fromevery stage of the flush (bud burst to mature leaves) was photosyntheticallyfunctional. The activity was low in flushing leaves with highanthocyanin content and then dramatically increased as leafanthocyanin content declined. Oxygen exchange measurements showedflushing leaves to have lower photosynthetic rates but higherrespiratory activity than mature leaves (60% and 120%, respectively).Gas exchange and in vitro electron transport were also generallycorrelated; Hill reaction activity was 128% of gas exchangein mature leaves and 92% in flushing leaves. It is concludedthat although photosynthetic rates are lower and respirationrates are higher in flushing leaves than in mature, fully expandedleaves, flushing leaves are fully photosynthetically competentand apparently require no net input of carbon for growth anddevelopment. Key words: Brachystegia, Hill reaction, photosynthesis, Zimbabwe     

Gas transfer in floating-leaved plants

Pressurized gas transport with flow rates of 1.1 to 1.81 gas h^-1 plant^-1 have been detected in the floating-leaved aquatic macrophyte Euryale ferox on sunny days. The younger leaves gave the highest pressurization, but the gas flow was initiated mainly by the middle-aged leaves of the plants. The gas through-flow was shown to be highly beneficial for floating-leaved plants. It improved the oxygen level in the aerenchyma of the submerged organs of Nymphaea alba and Nymphoides peltata, and doubled the ATP level in the root tissues. On the other hand, the sulphur dioxide fumigation experiments on Nymphaea odorata indicated that the pressurized gas transport may adversely affect the floating-leaved plants, due to an increased contamination of the leaves leading to increased photo bleaching and impaired photosynthesis.Abbreviations ATP adenosine triphosphate - FW fresh weight - SO₂ sulphur dioxide     

Responses of Alnus glutinosa to anaerobic conditions--mechanisms and rate of oxygen flux into the roots

Upon exposure to waterlogged growing conditions two-year-old alder trees reduced total root mass. Roots were concentrated in the uppermost soil horizon, and only few coarse roots penetrated into deeper soil layers. Root porosity was only slightly affected and did not exceed 8 % in fine roots. Porosity of coarse roots was higher (27 %) but unaffected by growing conditions. The stem base area covered by lenticels increased strongly and so did the cross section diameter of the stem base. The latter showed a highly significant correlation with O (2) transport into the roots, measured by a Clark type oxygen electrode. Exposure of the lower 5 cm of the stem base, where lenticels were concentrated, to pure N (2) led to a cessation of O (2) transport, confirming that lenticels were the major site of air entry into the stem. In alder plants grown under waterlogged conditions, temperature had a pronounced effect on O (2) gas exchange of the root system. The temperature compensation point, i.e., the temperature where O (2) transport equals O (2) consumption by respiration, was 10.5 degrees C for the entire root system, when measured in a range of 0.15 - 0.20 mmol dissolved O (2) L (-1), which is typical for an open water surface equilibrated with air. O (2) net flow was inversely related to O (2) concentration in the rooting media, indicating that higher root and microbial respiration induced higher net fluxes of O (2) into the root system. With 0.04 mmol dissolved O (2) L (-1) nutrient solution, the temperature compensation point increased to 20 degrees C. Measurement of O (2) gradients in the rhizosphere of agar-embedded roots using O (2) microelectrodes showed a preference for O (2) release in the tip region of coarse roots. Increasing stem temperature over air temperature by 5 degrees C stimulated O (2) flux into the roots as suggested by the model of thermo-osmotic gas transport. However determination of stem and air temperature in a natural alder swamp in northern Germany revealed that within the experimental period of almost one year, temperature gradients required for thermo-osmotic gas transport were very seldom. From this it is concluded that under natural conditions in northern Germany, oxygen diffusion along the stem into the root system is driven by O (2) concentration gradients rather than by thermo-osmosis.     

Radioactive Oxygen in the Study of Gas Transport down the Root of Vicia faba

The movement of labelled oxygen down the main root of Viciafaba seedlings was studied. The rise of radioactivity in theroots at 15 and 18 cm. below the cotyledons follows the samekinetics as the transport of radioactive gas down a thin polythenetube open at one end. It is therefore consistent with diffusionof oxygen through continuous gas spaces in the roots. Loss ofoxygen to the surrounding tissue is small in the upper partsof the roots. The volume of the gas spaces was estimated fromthe equilibrium radioactivity and compared with data obtainedby filling the spaces with paraffin oil.     

Polyphenoloxidase and the Respiration of Ivy Leaves

1. Polyphenoloxidase is present in ivy leaves but not in thoseof Aucuba or Euonymus.
2. Respiration of intact ivy leaves wasmarkedly stimulated bycatechol (R.Q. approximately=I), gallicacid, caffeic acid,and dihydroxyphenylalanine. The stimulationwas not relatedto injury as far as could be detected and wasnot followed byinhibition. The extra oxygen consumption representsa many timesrepeated oxidation of the amount of catechol supplied.
3. The effect of catechol on the respiration of Aucuba and Euonymusleaves was very small.
4. Cupferron and phenylthiourea, whichinhibit polyphenoloxidasein vitro, nevertheless increased respirationwhen administeredto leaves through the petiole. On the otherhand, when appliedby infiltration, cupferron did cause inhibition,but in Aucubaand Euonymus as much as in ivy.

     

Influence of Temperature and O2 Concentration on Photosynthesis and Light Activation of Ribulosebisphosphate Carboxylase Oxygenase in Intact Leaves of White Clover (Trifolium repens L.)

Detached leaves of white clover (Trifolium repens L.) were keptfor 1 h under various conditions of temperature, oxygen concentrationand light intensity. Rates of photosynthesis were measured whereappropriate and then ribulosebisphosphate carboxylase oxygenase(RuBPCO) was extracted rapidly and its initial activity measuredimmediately. The extracted activity increased with increased intensity ofillumination of the leaves. Where leaves were pretreated atlow light intensity, the lower the temperature of the leavesthe higher the extracted activity of RuBPCO. At high light intensitytemperature did not affect the activity of subsequently extractedRuBPCO but the light intensity which was necessary for maximumactivity increased with temperature. Activity of RuBPCO fromleaves pretreated in the dark was least when CO₂ was low andtemperature high. Leaves, pretreated at low temperatures andhigh light intensity in 20% O₂, yielded higher activity in extractsthan leaves pretreated under similar conditions but in 2% O₂.A relatively weak temperature response of photosynthesis atlow irradiances was associated with a decrease in extractableRuBPCO activity with increasing temperature. A strong temperaturedependence of the oxygen inhibition of photosynthesis was associatedwith lower extractable RuBPCO activity in leaves pretreatedat low oxygen concentration at low temperatures. With leavesfrom plants grown at low temperatures prior to treatment ofleaves, oxygen inhibition of photosynthesis was less temperaturedependent and activity of RuBPCO in extracts was not decreasedby low O₂ at low temperatures. Differences in the activationof RuBPCO appear to influence photosynthesis and account foran absence of oxygen inhibition of photosynthesis at low temperaturesin plants grown in warm conditions. Key words: Ribulosebisphosphate carboxylase oxygenase activation, Photosynthesis, Temperature, O₂ effect, White clover