The Occurrence of Nitrate Reduction in the Leaves of Woody Plants

Nitrate reductase activities greater than 02 µmol h^–1g^–1 f. wt, measured by an in vivo assay, occurred in 41per cent of a large sample (555 species) of woody plants. Ifseveral taxonomic groups (Gymnosperms, Ericaceae and Proteaceae)with consistently low activities were discounted activitiesgreater than 02 µmol h^–1 g^–1 f. wt occurredin 73 per cent of the species. This compares with 93 per centin herbaceous species, suggesting that leaf nitrate reductionis of common occurrence in woody plants. In a small sample ofspecies leaf nitrate reductase activity correlated with nitrateconcentration in the xylem sap. Low activities occurred consistentlyin the Gymnosperms, Ericaceae and Proteaceae. Feeding cut shootsof representatives of these groups with nitrate caused inductionof leaf nitrate reductase activity in the Gymnosperms and Proteaceae,but only limited induction in the Ericaceae. The Ericaceae,with the exception of two species, had low activities and lownitrate reductase inducibility. Root assimilation may predominatein the Gymnosperms and Proteaceae. It is suggested that nitratereduction generally occurs in the leaves of trees from a varietyof plant communities and that this may be related to the lowerenergy cost of leaf, as opposed to root, nitrate assimilation. Nitrate reductase, trees and shrubs, leaves, nitrate assimilation, nitrate translocation, nitrate reductase induction, energy cost, plant ecology     

Some effects of nitrate abundance and starvation on metabolism and accumulation of nitrogen in barley (Hordeum vulgare L. cv Sonja)

Nitrate and nitrite reductases were both induced by adding three concentrations of nitrate to the nutrient supply of nitrate-starved barley seedlings. Enzyme induction was not proportional to the amount of nitrate introduced. Glutamine synthetase also increased above a high endogenous activity but the increase did not differ significantly between any of the three nitrate treatments. Nitrate accumulated rapidly in leaves of plants given 4.0 mM or 0.5 mM nitrate but not with 0.1 mM nitrate. In all treatments, amino acids in leaves increased for 2 d, chiefly attributable to glutamine, then declined. Transferring plants from the three nitrate treatments to nitrate-free nutrient produced an immediate decline in nitrate reductase but nitrite reductase continued to increase for 2 d, before declining. Glutamine-synthetase activity was not affected by withdrawal of nitrate, nor did nitrate withdrawal retard plant growth during the 9-d period of the experiment. The disparity between accumulated nitrate and nitrate-reducing capacity and the rapid decrease in leaf nitrate when nutrient nitrate supply was removed, indicated the presence of a nitrate-storage pool that could be called upon to maintain amino-acid production in times of nitrogen starvation.Abbreviations GS glutamine synthetase - NR nitrate reductase - NiR nitrite reductase     

Determination of Nitrate Reductase Activity in Barley Leaves and Roots

The inactivation of nitrate reductase in the leaves and rootsof barley (Hordeum vulgare L. cv. Mazurka) during and afterextracting was investigated. At 0 °C in the absence of casein,25 per cent of ‘total’. i.e. maximal in vitro, nitratereductase activity was lost during the 2 min extraction process,followed by a slower loss of activity while the extract wasstored in ice. Activity was maintained by adding a minimum of1 per cent casein to the extraction medium containing 0·1M phosphate (pH 7·5), 1 mM EDTA and 1 mM dithiothreitol.Nitrate reductase was stable for several hours in these extracts,but declined in a first order manner in the absence of dithiothreitol.Casein also prevented the initial loss while making root extracts,but had less effect during storage. Using casein and thiols, nitrate reductase activity in light,(as product of maximal in vitro rates and wt g^–1) in leaveswas 98 per cent of the total activity in 31-day-old plants grownwith full nutrient in water culture and 60-day-old field-grownplants receiving no fertilizer. Field-grown plants, however,exhibited only 17 per cent of the activity of culture-grownplants. Nitrate reductase in leaves of barley plants grown in waterculture had a diurnal rhythm. During the first 3 h of the lightperiod, activity increased to 1·3 x the ‘dark’value. This was followed by a temporary decrease and then byanother increase to a maximum of 1·7 x the ‘dark’value, occurring about 8 h after illumination. Activity thendecreased during the rest of the light period and in darkness. Hordeum vulgare L., barley, nitrate reductase     

Effects of Nitrate Withdrawal and Resupply on the Assimilation of Nitrate in Leaves of Tomato

Nitrate reduction in leaves of tomato occurred at the same ratein plants grown in 8.0 mol m^–3 nitrate as in plants grownin 2.0 mol m^–3 nitrate, but at a much slower rate in plantsgrown in 0.1 mol m^–3 nitrate. However, the plants grownin 8.0 mol m^–3 nitrate had a larger leaf system than theplants grown in 2.0 mol m^–3 nitrate, and so the totalcapacity to assimilate nitrate was greater in the plants grownin the higher concentration. It was shown that plants grownin 8.0 mol m^–3 nitrate were better buffered against nitratewithdrawal than plants grown in 2.0 mol m^–3 nitrate asthe rate of nitrate reduction declined more slowly when plantswere transferred to 0.1 mol m^–3 nitrate from the higherconcentration than from the lower concentration. Furthermore,leaf expansion continued in the plants transferred from thehigher concentration, whereas it ceased abruptly in the plantstransferred from the lower concentration. It was concluded thatboth continuing expansion and continuing nitrate reduction wereaccompanied, and possibly caused by, a release of nitrate fromstorage pools in the lower part of the stem or the roots. Duringwithdrawal of nitrate the leaves were shown to maintain potentialactivity of the enzyme nitrate reductase although there wasno nitrate to be reduced. When nitrate was resupplied it couldbe reduced very quickly and reduction in the leaves was seento increase within 5 h of resupply. By 3 d after resupply furtherenzyme activity had been induced. Key words: Lycopersicon esculentum Mill, nitrate assimilation, nitrate reductase activity, nitrate withdrawal     

Nitrate Nutrition and Temperature Effects on Wheat: Enzyme Composition, Nitrate and Total Amino Acid Content of Leaves

Lawlor, D. W., Boyle, F. A., Kendall, A. C. and Keys, A. J.1987. Nitrate nutrition and temperature effects on wheat: Enzymecomposition, nitrate and total amino acid content of leaves.—J.exp. Bot. 38: 378–392. Wheat plants were grown in controlled environments in two temperatureregimes with two rates of nitrate fertilization. In some experimentstwo light intensities were combined with the nitrogen and temperaturetreatments. The composition of the third leaf was studied fromsoon after emergence until early senescence. The amounts ofchlorophyll, soluble protein, ribulose bisphosphate carboxylase-oxygenase(RuBPc-o) protein, nitrate, and total amino acids were measuredtogether with the activities of RuBPc-o, fructose- 1,6-bisphosphatase,glycolate oxidase, carbonic anhydrase, nitrate reductase, glutaminesynthetase and serine- and glutamate-glyoxylate aminotransferases.Additional nitrate supply increased the amounts, per unit leafarea, of chlorophyll, total soluble protein and RuBPc-o proteinand the activities of all the enzymes. The ratio of RuBP carboxylaseto RuBP oxygenase activity, when measured at constant CO²/O²ratio and temperature, was unaffected by growth conditions orleaf age. Leaves grown at the lower temperature, especiallywith more nitrate, contained much more soluble protein, nitratereductase, fructose bisphosphatase and free amino acids perunit area than the plants grown in the warmer conditions. However,young leaves grown in the warm contained more nitrate than thosegrown in the cool. Amounts of protein, amino acids and chlorophylland most enzyme activities reached maxima near full leaf expansionand decreased with age; additional nitrate slowed the decreaseand senescence was delayed. Nitrate content and nitrate reductaseactivities were highest in leaves before full expansion andthen fell rapidly after full expansion. Increased light intensityincreased the content of RuBPc-o protein at the higher rateof nitrate supply. Chloroplast components and, to a lesser extent,peroxisomal enzymes associated with photosynthetic nitrogenassimilation changed in proportion with different treatmentsbut nitrate reductase activity was not closely related to chloroplastenzymes. Control of tissue composition in relation to environmentalconditions is discussed. Key words: Nitrate nutrition, temperature, wheat, enzyme, amino acid, leaves, ribulose bisphosphate carboxylase oxygenase, nitrate reductase     

Comparative studies of diurnal variations of nitrate reductase activity in wheat, oat and barley

Diurnal variations of in vitro and in vivo (intact tissue assay) nitrate reductase (EC 1.6.6.1) activity and stability were examined in leaves of wheat ( Triticum aestivum L. cv. Runar), oat ( Avcna saliva L. cv. Mustang) and barley ( Hordeum vulgure L. cv. Agneta and cv. Gunillu). Nitrate reductase activity was generally higher for wheat than for oat and barley. However, the diurnal variations of nitrate reductase activity and stability were principally the same for all species, e.g. the high activity during the photoperiod was associated with low stability. All species showed a rapid (30-60 min) increase in the in vitro and in vivo activity when the light was switched on. When light was switched off the in vitro activity decreased rapidly whereas decrease in in vivo activity was slower. These experiments support the hypothesis that an activation/ deactivation mechanism is involved in the regulation of diurnal variations in nitrate reductase activity. Red light enhanced nitrate reductase activity in etiolated wheat and barley leaves. In green leaves, however, the daily increase in nitrate reductase activity was not induced by a brief red light treatment. Indications of different regulation mechanisms for the diurnal variations of nitrate reductase activity among the cereals were not found.     

Low CO(2) Prevents Nitrate Reduction in Leaves

The correlation between CO₂ assimilation and nitrate reduction in detached spinach (Spinacia oleracea L.) leaves was examined by measuring light-dependent changes in leaf nitrate levels in response to mild water stress and to artificially imposed CO₂ deficiency. The level of extractable nitrate reductase (NR) activity was also measured. The results are: (a) In the light, detached turgid spinach leaves reduced nitrate stored in the vacuoles of mesophyll cells at rates between 3 and 10 micromoles per milligram of chlorophyll per hour. Nitrate fed through the petiole was reduced at similar rates as storage nitrate. Nitrate reduction was accompanied by malate accumulation. (b) Under mild water stress which caused stomatal closure, nitrate reduction was prevented. The inhibition of nitrate reduction observed in water stressed leaves was reversed by external CO₂ concentrations (10-15%) high enough to overcome stomatal resistance. (c) Nitrate reduction was also inhibited when turgid leaves were kept in CO₂-free air or at the CO₂-compensation point or in nitrogen. (d) When leaves were illuminated in CO₂-free air, activity of NR decreased rapidly. It increased again, when CO₂ was added back to the system. The half-time for a 50% change in activity was about 30 min. It thus appears that there is a rapid inactivation/activation mechanism of NR in leaves which couples nitrate reductase to net photosynthesis.     

Response of Indoor-Grown Cassava to Water Deficits and Recovery of Leaf Water Potential and Stomatal Activity after Water Stress

Wilting of the leaves occurred in acropetal succession at leafwater potentials between –0.9 and –1.1 MPa. Onlysevere water stress caused the discoloration and abscissionof the basal leaves. Leaf resistance was independent of leafwater potential above –0.5 MPa but increased as the potentialdropped below this value. When the stressed plants were rewatered,leaf water potentials recovered rapidly within the first h.Subsequently, the rate of recovery declined gradually. The maximumvalue of leaf water potential after rewatering was dependenton the severity of the water stress.     

Effect of Nitrogen on Nitrate Reductase Activity in the Nodules and Leaves of Summer Moong (Vigna radiata)

The relation of the in vivo nitrate reductase (NR) activityto growth period was studied in the nodules and the leaves ofthe summer moong (Vigna radiata). The maximum NR activity wasobserved 31 days after sowing (DAS) in the leaves and 28 DASin the case of the nodules. In a pot experiment, the effectof the various nitrogen concentrations, namely 0, 3, 6, 9 and12 mg kg^–1 was studied on NR activity at three growthstages. The maximum NR activity was observed at 6 mg kg^–1N during the pre-flowering stage (26 DAS). Though the noduleshave higher NR activity, its expression was limited by substrateavailability. The NR activity in the leaf could be used as anindex of NR activity in the nodules. Nitrate reductase, nitrogen, nitrate, moong, Vigna radiata     

Triadimenol Increases Nitrate Levels and Nitrate Reductase Activity in Canola Leaves

Nitrate reductase activity in the first true leaves of canola(Brassica napus L.) seedlings grown in one-quarter strengthHoagland's solution from seeds pretreated with triadimenol (0.3or 30 g (a.i.) kg^–1 of seed) was higher than controlsduring the growth period of 15 to 25 d after planting. Triadimenolalso increased chlorophyll levels, the increase being more pronouncedat its lower concentration. The treatment also increased theweight and nitrate content of the leaves. When seedlings weregrown in nutrient solution containing 1 to 20 mM nitrate, theincrease in nitrate reductase activity by triadimenol was higherat lower rather than at higher nitrate concentrations. The nitratelevels and Kjeldahl nitrogen in the triadimenol-treated leaveswas higher than the controls at concentrations of added nitrateabove 2 mM. Addition of nitrate to plants grown in ammonium,increased nitrate reductase activity more in plants grown fromtriadimenol-treated seeds than controls. However, addition of10µM triadimenol for 24 h to ammonium-grown plants hadlittle effect on enzyme activity, both in the absence as wellas the presence of nitrate. This study demonstrates that triadimenolincreases nitrate reductase activity and nitrate accumulationin the leaves and at least part of the increased enzyme activityis independent of nitrate accumulation. Key words: Triazoles, nitrate content, nitrate reductase activity     

Nitrate Reduction in Solanum tuberosum L.: Development of Nitrate Reductase Activity in Field-grown Plants

Nitrate reductase activity (in vivo method, substrate non-limiting)in unshaded leaves from the top of the canopy has been determinedfor field-grown potato plants over the course of the growingseason. The pattern of change was almost identical for plantsreceiving no added fertilizer and those receiving 24 g N m^–2.Activity increased to a peak at about 90 days after plantingand declined thereafter. On a fresh weight basis activity wasalways higher in fertilized plants. Nitrate reductase activitywas positively and significantly correlated with leaf proteincontent in high N plants (r² = 0.71; P = 0.05), but poorly correlatedwith both the nitrate content of the leaf lamina and the nitrateconcentration in petiole sap. Up until 90 days after planting(mid-July) there appeared to be a positive relationship betweenincreased activity of nitrate reductase and solar radiation.However, results obtained over two seasons showed that the declinein activity after this time was not consistently linked witha fall in the level of solar radiation. Remobilization of reduced-Nand stored nitrate from leaves and stems accompanied this declinein nitrate reductase activity and in the latter part of theseason appeared to account for all of the N gained by growingtubers. In unfertilized plants nitrate-N accounted for 5 per cent orless of total plant N. Fertilized plants contained up to 25per cent nitrate-N. While nitrate availability limited growthin unfertilized plants, sub-optimal rates of nitrate assimilationin fertilized plants, particularly during the early stages ofpost-emergence growth, may contribute to inefficient use ofacquired nitrate. The carbohydrate status of leaf lamina and petiole sap weremodified by N supply. The soluble sugar and starch contentsof low N leaves were higher than in their high N counterparts.By contrast, the concentration of soluble sugars in petiolesap increased to a higher value in high N samples. Althoughsap sugar levels declined in both treatments towards the endof the season, N application delayed this decline for severalweeks. Solanum tuberosum, nitrate reductase, nitrate assimilation, senescence     

Synthesis and degradation of barley nitrate reductase

Nitrate and light are known to modulate barley (Hordeum vulgare L.) nitrate reductase activity. The objective of this investigation was to determine whether barley nitrate reductase is regulated by enzyme synthesis and degradation or by an activation-inactivation mechanism. Barley seedling nitrate reductase protein (cross-reacting material) was determined by rocket immunoelectrophoresis and a qualitative immunochemical technique (western blot) during the induction and decay of nitrate reductase activity. Nitrate reductase cross-reacting material was not detected in root or shoot extracts from seedlings grown without nitrate. Low levels of nitrate reductase activity and cross-reacting material were observed in leaf extracts from plants grown on nitrate in the dark. Upon nitrate induction or transfer of nitrate-grown etiolated plants to the light, increases in nitrate reductase activity were positively correlated with increases in immunological cross-reactivity. Root and shoot nitrate reductase activity and cross-reacting material decreased when nitrate-induced seedlings were transferred to a nitrate-free nutrient solution or from light to darkness. These results indicate that barley nitrate reductase levels are regulated by de novo synthesis and protein degradation.     

Nitrate Reductase Activity of Wheat Seedlings during Exposure to and Recovery from Water Stress and Salinity

Nitrate reductase activity was inhibited as a result of reduced soil moisture potentials or application of NaCI to nutrient solutions. The decrease in enzyme activity of wheat seedlings exposed to salinity, was found 24 hours after exposure to stress. The effect of stress on nitrate reductase was found in cell-free extracts as well as in riro in assays of intact leaf sections. A recovery in enzyme activity was found after irrigation or after removal of seedlings from salinity. While relative water content of the leaves was restored within 3 hours after removal of stress, full recovery of enzyme activity occurred only after 24 hours. Cycloheximide and chloramphenicol suppressed the activity of nitrate reductase in non-stressed seedlings, but had no effect on the activity of plants exposed to salinity. However, during removal of stress, cycloheximide prevented completely the recovery of nitrate reductase, while chloramphenicol did not interfere with the recovery of the inhibited enzyme activity. It is concluded that a fraction of nitrate reductase may be located in the cytoplasm and lost activity during stress, probably due to inhibited protein synthesis. Another fraction which may be associated with chloroplasts, was inhibited by stress due to conformational changes or partial denaturation.     

Nitrogen Assimilation in Barley (Hordeum vulgare L. cv. Mazurka) in Response to Nitrate and Ammonium Nutrition

Barley plants (Hordeum vulgare L. cv. Mazurka) were grown inaerated solution cultures with 2 mM or 8 mM inorganic nitrogensupplied as nitrate alone, ammonium alone or 1:1 nitrate+ammonium.Activities of the principal inorganic nitrogen assimilatoryenzymes and nitrogen transport were measured. Activities ofnitrate and nitrite reductases, glutamine synthetase and glutamatesynthase were greater in leaves than in roots but glutamatedehydrogenase was most active in roots. Only nitrate and nitritereductases changed notably (4–10 times) in response tothe different nitrogen treatments. Nitrate reductase appearedto be rate-limiting for nitrate assimilation to glutamate inroots and also in leaves, where its total in vitro activitywas closely related to nitrate flux in the xylem sap and wasslightly in excess of that needed to reduce the transportednitrate. Xylem nitrate concentration was 13 times greater thanthat in the nutrient solution. Ammonium nitrogen was assimilatedalmost completely in the roots and the small amount releasedinto the xylem sap was similar for the nitrate and the ammoniumtreatments. The presence of ammonium in the nutrient decreasedboth export of nitrate to the xylem and its accumulation inleaves and roots. Nitrate was stored in stem bases and was releasedto the xylem and thence to the leaves during nitrogen starvation.In these experiments, ammonium was assimilated principally inthe roots and nitrate in the leaves. Any advantage of this divisionof function may depend partly on total conversion of inorganicnitrogen to amino acids when nitrate and ammonium are givenin optimal concentrations. Hordeum vulgare L., barley, nitrate, ammonium, nitrate reductase, nitrite reductase, glutamine synthetase, glutamate synthase, glutamate dehydrogenase, nitrogen transport     

Drought Influences the Activity of Enzymes of the Chloroplast Hydrogen Peroxide Scavenging System

Smirnoff, N. and Colomb?, S. V. 1988. Drought influences theactivity of enzymes of the chloroplast hydrogen peroxide scavengingsystem.—J. exp. Bot. 39: 1097–1108. The effect of drought on the activity of ascorbate peroxidase(AP), glutathione reductase (GR) and monodehydroascorbate reductase(MDAR) in leaves of barley (Hordeum vulgare) and tef {Eragrostistef) was studied. These enzymes are components of the chloroplasthydrogen peroxide scavenging system. Severe leaf water deficit(<–30 M Pa) resulted in increased activity (leaf dryweight basis) of GR and MDAR in barley and of AP and MDAR intef. The specific activity of all the scavenging enzymes wasgreater in droughted plants. The activities (dry weight basis)of two ‘control’ enzymes, malate dehydrogenase (barley)and phosphoenolpyruvate carboxylase (tef), not directly involvedin the scavenging system, were not affected by drought. These data suggest that drought may cause an increase in thecapacity of the hydrogen peroxide scavenging system and thatit may, therefore, increase the rate of hydrogen peroxide formationin chloroplasts. Key words: Water stress, hydrogen peroxide, enzyme activity     

Characterization of the 'In Vivo' Nitrate Reductase Activity in the Roots and Leaves of Eichhornia crassipes

Studies of in vivo nitrate reductase activity (NRA) in Eichhorniacrassipes showed that adding 5% isopropanol gave highest NRAin all parts of the plant, but it was also necessary to flushthe assay media containing leaves with N₁ to obtain the maximalactivity. There were differences in the pH optima for NRA, theroot optimum being 6·5 leaf 7·5–8·0,and the petiole showing a major peak at 6·5 and a minorpeak at 7·.5–8·0. Possible significancesof these differences are discussed. Nitrate reductase activity, Eichhornia crassipes     

Response of wheat seedlings to short-term drought stress with particular respect to nitrate utilization

Abstract. The effect of short-term changes in the water potential (from 0 to – 2.5 MPa) by addition of PEG 4000 to the nutrient solution was investigated with respect to nitrate uptake and reduction in 3-week-old wheat plants ( Triticum aestivum , cv Fidel). Plants were harvested at the end of 12-h treatments in the dark. The water potential of the mature leaves was similar to that of the medium down to – 0.8 MPa and was maintained at this level even though the external water potential was much lower. The medium water potential of 0.8 was a threshold level below which elongation of the youngest leaf was inhibited. Increase of the PEG concentration in the medium brought about a decrease of evapotranspiration and enhancement of nitrate uptake. No difference in the rate of nitrate reduction was observed, although the in vitro nitrate reductase activity was lowered. Nitrate accumulation in the shoot was ascribed both to the stimulation of net uptake from the medium, and to the mobilization and translocation of nitrate from the root. It is suggested that increase in the storage pool of nitrate in shoots was related to the role of NO₃⁻ as an osmoticum.     

The Effects of Irradiance on Uptake and Assimilation of Nitrate by Young Barley Seedlings

The nitrogen economy of barley plants growing in a range ofirradiances from full shade (less than 0·5 W m^–2)to 119 W ^m–2 has been examined by analysing levels oftotal, organic and nitrate nitrogen, and by determining nitratereductase activity in leaf extracts. It has been confirmed thatroot growth is reduced in low irradiances which are also associatedwith a lower level of total nitrogen in the plant, and hencewith a lower uptake of nitrate. In all parts of the plant thelevel of organic nitrogen is higher in high light intensitybut nitrate-nitrogen as a proportion of the total is greatestin low irradiances. In the first leaf accumulation of free nitrateis substantially greater in low irradiances. The data indicate a higher level of nitrate assimilation inhigh irradiances and nitrate reductase activity in leaf extractsis higher in such conditions. When the first leaf is shadednitrate reductase activity falls to undetectable levels afterabout 4 days, but in the case of the second leaf, where thisis shaded, some reductase activity is always found, althoughthis is substantially less than that in unshaded conditions. It is concluded that in vitro rates of nitrate reduction mayover-estimate nitrate assimilation determined as increase inorganic nitrogen.     

Absence of nitrate reductase activity in San 9789 bleached leaves of barley seedlings (Hordeum vulgare cv. Midas)

Abstract San 9789 (norflurazone) blocks carotenoid synthesis which allows chlorophyll bleaching in the light, and has been used recently as a tool to study phytochrome responses without interference from photosynthetic pigments. By using this herbicide, we have found that nitrate reductase activity and light dependent nitrite reduction were lost simultaneously from achlorophyllous areas of barley leaves, with the green areas of the leaf tip still showing high activities. By contrast nitrate reductase is still present in the roots of herbicide treated plants. We suggest that intact chloroplasts are required for the presence of nitrate reductase in barley leaves.     

The rate of development of water deficits affects Saxifraga cernua leaf respiration

We allowed plant water deficits to develop at two different rates following the cessation of watering in order to investigate the effects of water stress on cytochrome pathway and alternative pathway respiration in the leaves of the arctic herb Saxifraga cernua. Plants were pretreated by growth in either a commercial organic (CO) mixture or a vermiculite-perlite (VP) mixture, which allowed the complete development of water deficits in 19 and 8 days, respectively. The rate of water potential reduction was approximately 0.11 MPa day⁻¹ in the leaves of CO plants, compared to a reduction of 0.21 MPa day⁻¹ in leaves of VP plants. Osmotic adjustment occurred to a greater extent in leaves of CO plants and corresponded with an increase in ethanol-soluble sugars. In leaves of CO plants, cytochrome pathway activity gradually declined from that of control rates until day 11, and then declined more rapidly. In contrast, cytochrome pathway activity significantly increased in response to water deficits in leaves of VP plants. In leaves of both CO and VP plants, alternative pathway activity declined as water stress progressed. Relatively severe water deficits reduced alternative pathway capacity in leaves of both CO and VP plants. We also investigated the effect of previous exposure to water deficits on leaf respiration. In plants that had previously experienced three cycles of water stress, the increase in cytochrome pathway activity during the fourth water stress cycle was small compared to the increase observed in leaves of plants experiencing water stress for the first time. These results suggest that cytochrome pathway activity is differentially sensitive to the rate of development of plant water deficits and that respiratory responses to acute water stress are not necessarily similar to the responses to chronic water stress.