Effects and Interactions of Ozone and the Anti-ozonant EDU at Different Stages of Radish {Raphanus sativus L.) Development

The use of the anti-ozonant ethylenediurea (EDU) to assess croplosses due to atmospheric ozone (O₃), is controversial becauseof incomplete knowledge on the optimal dosage, the extent ofprotection provided and potential effects on plant performanceof EDU per se. In a greenhouse experiment, radish (Raphanussativus L.) cv. ‘Cherry Belle’ plants were treatedwith EDU as a soil drench (150 mg1¹, 60 ml plant¹) at threestages of development: early, maximum and late hypocotyl thickening.Two days later the plants were exposed to O₃, mimicking an ambientpattern (70 nl 1¹ O₃, 7 h d¹, 5 d, and up to 120 nl 1¹ O₃ on2 d), or remained in filtered air. Assessment of leaf injury showed complete protection from ozone-inducedleaf damage at all stages of growth due to EDU. EDU did causeslight leaf margin necrosis and hyponastic leaf deformationirrespective of the atmospheric treatment, but biomass and relativegrowth rate (RGR) of the shoot and leaf number duration werenot altered and leaf area duration was increased when comparedto control plants. EDU did not affect dry weight accumulationnor RGR of the hypocotyl. Ozone-induced reductions in growthof below-ground organs (hypocotyl and root), which were mostprominent in the intermediate and early stage of growth, werelargely suppressed by the EDU-treatment. Decreases of hypocotylbiomass due to O₃ were transient and significant increases inRGR of the hypocotyl during the post-exposure period indicateda substantial capability of compensation during the period ofrecovery. Key words: Radish (Raphanus sativus), ozone, anti-ozonant, ethylenediurea (EDU), development, phytotoxicity     

Purification and Characterization of Thiol Proteinase as a Nitrate Reductase-Inactivating Factor from Leaves of Hordeum distichum L.

A thiol proteinase was purified 6,400-fold from leaves of Hordeumdistichum L. by a sequence of ammonium sulfate fractionation,gel filtration, ion exchange chromatography, hydrophobic chromatographyand chromatofocusing. This enzyme also had nitrate reductase(NR)-inactivating activity, which was associated with proteolyticactivity in almost constant proportions during the purificationprocedures. Its molecular weight was estimated as 74,000 bygel filtration, and it had an isoelectric point of 4.05 andan apparent K_m of 0.83 mg ml^–1 for azocasein. The respectiveoptimum pH for proteolytic and NR-inactivating activities were6.0 and 7.0. On electrophoresis, the proteinase gave a majorband that coincided with both activities; it also produced afaint band associated with no activity. Our purified thiol proteinase inactivated FMNH₂-NR and MVH-NRas well as NADH-NR, but it had only a slight effect on NADHcytochrome c reductase activity. This enzyme also inactivatedglutamine synthetase. (Received September 16, 1983; Accepted January 26, 1984)     

The Composition of Phloem Exudate and Xylem Sap from Broccoli (Brassica oleracea var. italica) Supplied with NH4+, NO""3 or NH4NO3

Shelp, B. J. 1987. The composition of phloem exudate and xylemsap from broccoli (Brassica oleracea var. italica) suppliedwith NH⁺₄, NO₃ or NH₄NO₃.—J. exp. Bot. 38: 1619–1636. The detailed composition of xylem sap and exudate from stemincisions of attached inflorescences of broccoli (Brassica oleraceavar. italica) was compared in plants supplied with NH⁺₄, NO₃or NH₄NO₃. A phloem origin for the exudate was suggested fromthe high levels of sugars (71–133 mg cm^-3), amino acids(8·1-26·7 mg cm³) and K. (2·3–3·8mg cm³), the low levels of NO₃ and Ca, the high C: N (w/w) ratios(8·3–33), and the alkaline pH (7·2–7·3).In contrast, the xylem sap was mildly acidic (pH 5·6–6·0),and possessed lower levels of all organic and inorganic solutesbut NO₃ and Ca, and lower ratios of K: Ca, Mg: Ca and C: N (0·6–4·4). Glutamine was the predominant o-phthalaldehyde-reactive aminocompound in both transport fluids with the next most abundantamino acids dependent on sap type and N-form. Together witharginine, -aminobutyric acid, which was found only in the xylemstream, was enhanced by NH⁺₄compared to NO₃ -nutrition suggestingthat glutamate metabolism was stimulated in the roots. Underlimiting N the amino acid concentrations in the transport fluidswere greater with NH⁺₄ than with NO₃. NO₃ reduction occurredin both the root and shoot with the latter site predominatingover the entire N range (0-300 mol m³). Even though the compositionof nitrogenous solutes in the xylem was dependent on cultivarand N source, the composition of the phloem streams supplyingthe developing inflorescence was relatively unaffected. The data on the element composition of organs and phloem sapare interpreted to suggest that, in spite of the restrictedmobility of some elements such as B and Mn, a significant proportionof their total supply to developing sinks is carried in thephloem stream. Key words: Transport fluid composition, plant nutrition, phloem mobility.     

Comparison of the Effects of Root Temperature on Nitrate and Ammonium Nutrition of Oilseed Rape (Brassica napusL.) in Flowing Solution Culture: II. CATION-ANION BALANCE

Macduff, J. H., Hopper, M. J., Wild, A. and Trim, F. E. 1987.Comparison of the effects of root temperature on nitrate andammonium nutrition of oilseed rape (Brassica napusL.) in flowingsolution culture. II. Cation-anion balance.—J. exp. Bot.38: 1589-1602. The effects of root temperature and form of N nutrition (NH4or NOJ) on the mineral composition of the plant, the balanceof inorganic cation-anion uptake and on the apparent net effluxof H ⁺/OH^–ions from the roots were studied with 49-d-oldoilseed rape (Brassica napusL. cv. Bien venu) in flowing solutionculture. Plants were pre-treated for 14 d at a root temperatureof 5 °C prior to constant root temperatures of 3, 7, 11or 17°C for 14 d, with a common shoot temperature of 20/15°Cday/night. Nitrogen was supplied as NH⁺₄4 or NO^–₃ at 10mmol m³. Values of Q₁₀ (7-17°C) for mean unit absorptionrates of all the major nutrient ions (K⁺ , Mg⁺⁺ , NH⁺₄, SO₄,H₂PO₄, NO₃), except Ca⁺⁺, were > 2.0 over the first 5 d oftreatment but thereafter were < 1.5; the apparent effectof temperature on uptake rates diminished with time. Under NH⁺₄nutrition, inorganic cation uptake (Mg^{+ +} + K⁺+Ca^{+ +} +NH⁺₄)exceeded inorganic anion uptake (SO4 ^–₄+ H₂PO₄) over 14d at all temperatures, with the proportion of cation uptakeas NH4 remaining constant (0.67-0-68) irrespective of root temperature.The net efflux of H ⁺ from the roots approximately balancedNH⁺₄ uptake (1:1) over 14 d at each temperature and also balancedthe difference between the total uptake of inorganic cationsand inorganic anions. Under NO₃ nutrition, the sum of the netefflux of OH and the change in the carboxylate contents of plantsover 14 d approximately balanced the sum of NO₃ and SO ^–₄reduced in the plant. The majority of the negative charge associatedwith the reduction of NO₃ and SO ^–₄ was apparently effluxedas OH, but this fraction was lower at low root temperatures.The results are discussed in terms of mechanisms that have beenproposed to regulate the internal pH of plants. Key words: Brassica napus, oilseed rape, root temperature, cation-anion balance, H⁺/OH efflux.     

Wheat Response to CO2 Enrichment: Growth and CO2 Exchanges at Two Plant Densities

Du Cloux, H. C, André, M., Daguenet, A. and Massinuno,J. 1987. Wheat response to CO₂ enrichment: Growth and CO₂ exchangesat two plant densities.—J. exp. Bot. 38: 1421–1431. The vegetative growth of wheat (Triticum aestivum L., var. Capitole)was followed for almost 40 d after germination in controlledconditions. Four different treatments were carried out by combiningtwo air concentrations of CO₂, either normal (330 mm³ dm ³)or doubled (660 mm³ dm ³) with two plant densities, either 200plants m ² or 40 plants m ². Throughout the experiment the CO₂gas exchanges of each canopy were measured 24 h d¹. These provideda continuous growth curve for each treatment, which were comparedwith dry weights. After a small stimulation at the start (first13 d), no further effect of CO₂ enrichment was observed on relativegrowth rate (RGR). However, RGR was stimulated throughout theexperiment when plotted as a function of biomass. The finalstimulation ol dry weight at 660 mm³ dm ³ CO₂ was a factor of1·45 at high density and 1·50 at low density,contrary to other studies, no diminution of this CO₂ effecton dry weight was observed over time. Nevertheless, at low density,a transient additional enhancement of biomass (up to 1·70)was obtained at a leaf area index (LAI) below 1. This effectwas attributed to a different build up of the gain of carbonin the case of an isolated plant or a closed canopy. In theformer, the stimulation of leaf area and the net assimilationrate are both involved; in the latter the enhancement becomesindependent of the effect on leaf area because the canopy photosynthesisper unit ground area as a function of LAI reaches a plateau. Key words: Triticum aestuum, L. var. Capitole, Vegetative growth, Canopy     

Plant Characteristics and Nutrient Composition and Mobility of Broccoli (Brassica oleracea var. italica) Supplied with NH4+, NO3"" or NH4NO3

Shelp, B. J. 1987. Plant characteristics and nutrient compositionand mobility of broccoli (Brassica oleracea var. italica) suppliedwith NH⁺₄, NO₃ or NH₄NO₃.—J. exp. Bot. 38: 1603–1618. The effects of varying NH⁺₄, NO₃ or NH₄NO₃ concentration onthe final plant characteristics, element composition, and accumulationof NO₃-N, NH⁺₄-N and organic-N were evaluated in broccoli (Brassicaoleracea var. italicacv. Futura and Premium Crop) plants culturedin vermiculite under greenhouse conditions supplemented withlight. NH⁺₄-grown plants were stunted and exhibited signs ofmarginal necrosis on the old leaves, accompanied by an accumulationof NH4. The tissue levels of N, P, Mn, Cu, Zn and B were generallyincreased by NH⁺₄ versus NO₃ nutrition whereas the reverse wastrue for Ca; Mg and K were only slightly affected, if at all.These results are attributed to: changes in element availabilityresulting from reduced rhizosphere pH due to NH⁺₄uptake ratherthan NO ₃uptake; competition of Ca uptake by NH⁺₄; and dilutionof N by increased vegetative growth with NO₃-nutrition. Theelement concentrations of N, P or K were similar in all tissueswhereas Ca, B and Mn were markedly less in the florets and youngleaves compared to mature leaves; this supports literature indicatingthat the former elements are phloem-mobile whereas the latterare not. Assuming that the nutrient supply for mature leavesis delivered principally via the xylem stream, the data suggestthat nutrients for developing leaves and florets are suppliedpredominantly in the phloem. If so, under our experimental conditions.Zn and Cu were also readily mobile in the phloem whereas Mgmovement was restricted. NH₄⁺ versus NO₄⁺ J nutrition alteredthe distribution of these elements. The two broccoli cultivarstested under the greenhouse environment differed in NH⁺₄ toleranceand in the distribution of K and Cu suggesting there was a geneticbasis for cultivar variation in mineral acquisition and redistribution. Key words: Plant nutrition, phloem mobility, elemental composition.     

Evaluation of the 15N Natural Abundance Method and Xylem Sap Analysis for Assessing N2 Fixation of Understorey Legumes in Jarrah (Eucalyptus marginata Donn ex Sm.) Forest in S.W. Australia

Hansen, A. P. and Pate, J. S. 1987. Evaluation of the ¹⁵N naturalabundance method and xylem sap analysis for assessing N₂ fixationof understorey legumes in jarrah (Eucalyptus marginata Donnex Sm.) forest in S.W. Australia.—J. exp. Bot 38: 1446–1458. Nodulated seedlings of Acacia pulchella, A. alata and A. extensawere grown in glasshouse sand culture under a range of levels(0–16 mol m³) of nitrate, supplied as ¹⁵NO₃, or as unenrichedlaboratory grade nitrate (¹⁵N value 5·5%_o). Nitrate at8·0 mol m ³ or above was highly inhibitory to growthof all species. Using ¹⁵N dilution analysis of the ¹⁵N enrichedcultures to measure symbiotic dependency, it was shown that¹⁵N values of the parallel unenriched cultures increased innear linear fashion from close to zero in fully symbiotic plantsto values close to that of the supplied NO₃ in plants experiencingnitrate levels (4·0 mol m³ or above) inhibiting N₂ fixationby over 90%. Xylem sap analyses (0·4 mol m³ NO₃ treatments)showed asparagine as the major nitrogenous solute, relativelylittle spill-over of free nitrate, and no evidence of majorshifts in balance of amino compounds with increasing dependenceon nitrate. This essentially invalidated use of the techniqueas a field assay for N₂ fixation by the species. ¹⁵N values for total N of soil sampled at 64 widely distributedsites in jarrah forest ranged from – 2·15 to +5·4(mean +2·1). Comparable values for soil mineral N (NH⁺₄and NO₃) were +0·3 to + 14·2 (mean +5·1).¹⁵N values of the total plant N of the legumes and of non-N₂-fixingreference species were also highly variable between sites, withlittle evidence of reference plant N accurately reflecting the¹⁵N abundance of soil nitrogen, or of visibly well nodulatedlegume components showing consistently lower ¹⁵N values thantheir companion reference plants. At one site it was possibleto compare ¹⁵N values of first season seedling legumes withpreviously published estimates of their progressive N₂ fixationusing C₂H₂ reduction assays. It was concluded that heterogeneity in ¹⁵N discrimination ofsoil within the ecosystem precluded effective use of the ¹⁵Nnatural abundance technique for assessing legume N₂ fixation. Key words: Acacia spp., ¹⁵N natural abundance,, xylem sap analysis,, nitrogen fixation.     

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     

Nitrate Reduction by Cassava

Nitrate reductase (NR) was assayed in vivo in cassava (Manihotesculenta Crantz). Activity in the leaves ranged from 0 to 2.51µmole of NO₃^– reduced g^–1 h^–1, withno activity in the younger leaves (leaf 1 on top). NR activitywas localized in the sides and toward the tip of the lobes ofthe leaf. (Received December 10, 1985; Accepted April 8, 1986)     

Effect of Applied Nitrogen on N2 Fixation, Assimilation of Nitrate and Ammonia in Nodules of Field-grown Moong (Vigna radiata)

A field experiment was conducted to study the effect of nitrogenapplication at 15, 30 and 45 kg ha^–1 of urea at pre-flowering(PF) and pod initiation (PI) stages on the activity of nitrogenase(N₂ase), nitrate reductase (NR) and other related parametersin the nodules of moong (Vigna radiata). Nitrogen applied atPF or PI stage was found to be inhibitory to N₂ase and glutaminesynthetase (GS) activities except at 15 kg N ha^–1 whenapplied at PF in the case of N₂ase. At both the stages therewas increase in NR and glutamate dehydrogenase (GDH) activitieswith the application of nitrogen. Seed yield increased by 18per cent with the application of 15 kg N ha^–1 at PI stagewhereas nitrogen application at PF stage only increased strawyield significantly. Nitrate reductase, nitrogenase, nitrogen application, ammonia assimilation, Vigna radiata     

The Distribution of Nitrate Assimilation Between Root and Shoot in Vicia faba L.

Nitrate assimilation was examined in two cultivars (Banner Winterand Herz Freya) of Vicia faba L. supplied with a range of nitrateconcentrations. The distribution between root and shoot wasassessed. The cultivars showed responses to increased applied nitrateconcentration. Total plant dry weight and carbon content remainedconstant while shoot: root dry weight ratio, total plant nitrogen,total plant leaf area and specific leaf area (SLA) all increased.The proportion of total plant nitrate and nitrate reductase(NR) activity found in the shoot of both cultivars increasedwith applied nitrate concentrations as did NO₃^–: Kjeldahl-Nratios of xylem sap. The cultivars differed in that a greaterproportion of total plant NR activity occurred in the shootof cv. Herz Freya at all applied nitrate concentrations, andits xylem sap NO₃^–: Kjeldahl-N ratio and SLA were consistentlygreater. It is concluded that the distribution of nitrate assimilationbetween root and shoot of V. faba varies both with cultivarand with external nitrate concentration. Vicia faba L., field bean, nitrate assimilation, nitrate reductase, xylem sap analysis     

Pyridine nucleotide specificity and other properties of purified nitrate reductase from Chlorella variegata

Nitrate reductase (NR) (EC 1.6.6.2) from Chlorella variegata 211/10d has been purified by blue sepharose affinity chromatography. The enzyme can utilise NADH or NADPH for nitrate reduction with apparent K _m values of 11.5 M and 14.5 M, respectively. Apparent K _m values for nitrate are 0.13 mM (NADH-NR) and 0.14 mM (NADPH-NR). The diaphorase activity of the enzyme is inhibited strongly by parachloromercuribenzoic acid; NADH or NADPH protects the enzyme against this inhibition. NR proper activity of the enzyme is partially inactive after extraction and may be activated after the addition of ferricyanide. The addition of NAD(P)H and cyanide causes a reversible inactivation of the NR proper activity although preincubation with either NADH or NADH and ADP has no significant effect.Abbreviations NR Nitrate reductase - FAD Flavin-adenine dinucleotide - FMN Riboflavin 5-phosphate - p-CMB para-Chloromercuribenzoic - BV Benzyl viologen     

Photosynthetic Nitrogen Metabolism in High and Low CO2-adapted Scenedesmus: II. EFFECT OF AMMONIUM AND METHIONINE SULPHOXIMINE ON NITRATE UTILIZATION

Larsson, C.-M., Larsson, M. and Guerrero, M. G. 1985. Photosyntheticnitrogen metabolism in high and low CO²-adapted Scenedesmus.II. Effect of ammonium and methionine sulphoximine on nitrateutilization.—J. exp. Bot. 36: 1387–1395 In 3% CO²-grown Scenedesmus obtusiusculus Chod. utilizing NO₃^–J as the N source, NH₄⁺ addition caused a prompt inhibitionof NO₃^– utilization. Nitrate reductase (NR) activity declinedrapidly in response to the presence of NO₄⁺, but the cessationof NO₃^– utilization was too rapid to be accounted forby the loss in NR activity. The first site of NO₄⁺ inhibitionin these cells seems to be the entrance of NO₃^– into thecells. Upon exhaustion of NO₄⁺ from the medium, NO₃^– utilizationwas rapidly restored and NR activity increased. Air-grown cellswere much less sensitive to the effect of NO₄⁺, more than 30min being required for added NO₄⁺ to cause complete inhibitionof NO₃^– utilization. In these cells, NO₃^– uptakeand NR activity decreased in parallel in response to NO₄⁺ addition.In 3% CO²-grown cells simultaneously subjected to NO₄⁺ and air-levelof CO², NO₄⁺ initially inhibited NO₃^– utilization completely,but a slight recovery took place after approximately 20 min The glutamine synthetase (GS) inhibitor L-methionine D, L-sulphoximine(MSO) behaved as a potent inhibitor of NO₃^– uptake in3% CO²-grown cells, but had considerably less effect in air-growncells, although the time-course of the MSO-induced inhibitionof GS was the same in both cases Key words: Ammonium, nitrate utilization, Scenedesmus     

Dimethipin (2,3-Dihydro-5,6-dimethyl-1,4-dithiin 1,1,4,4-tetraoxide) Effects on Soybean Seedling Growth and Metabolism

Three-day-old dark-grown soybean [Glycine max (L.) Merr.] seedlingswere transferred to 2 mM CaSO₄ or 10^–5 M dimethipin in2 nM CaSO₄ and root-fed via liquid culture. Plants were placedin continuous darkness or in continuous white light (200 µE.m^–2?s^–11,PAR) at 25?C. Dimethipin inhibited root and shoot elongationin dark-grown plants after 24 h and 48 h, respectively. In thelight, root elongation was inhibited also after 24 h, but hypocotylelongation was not significantly affected. Extractable phenylalanineammonia-lyase (PAL) activity per axis in dimethipin-treateddark-grown axes was not generally affected but, in the lightdimethipin caused a significant decrease in PAL activity (24to 96 h). Total soluble hydroxyphenolics in axes were not affectedby dimethipin in light- or dark-grown plants. Anthocyanin andchlorophyll levels were lowered in hypocotyls of dimethipin-treatedplants after 48 to 96 h. Soluble protein in hypocotyls of light-or dark-grown seedlings was not substantially affected by dimethipin.Nitrate reductase (NR) activity (per organ) was generally notaffected by dimethipin in light-grown cotyledons, but in theroots of these seedlings, NR activity was significantly decreased.Proteolytic enzyme activity using three substrates (leucine-p-nitroanilide,LPNA; proline-p-nitroanilide, PPNA; and benzoylarginine-p-nitroanilide,BAPA) indicated little effect on enzyme activities per organin roots and hypocotyls. These data suggest that dimethipinat low concentrations can cause significant growth inhibitionin soybean seedlings grown in either light or darkness and thatfurthermore, extractable activities of some enzymes associatedwith nitrogen metabolism and secondary metabolism are alteredby this chemical. Light also plays a role in the activity ofthis chemical. (Received November 29, 1983; Accepted January 25, 1984)     

Incorporation of radioactive molybdenum into protein during nitrate reductase formation and effect of molybdenum on nitrate reductase and diaphorase activities of spinach (Spinacea oleracea L.)

Spinach plants grown without molybdenum lack nitrate reductaseand when plants are deprived of nitrate existing activity islost. Transfer of molybdenum-deficient plants to a solutioncontaining (NH₄)₂⁹⁹MoO₄) or nitrate-starved plants to NaNO₃solution induced enzyme activity in 24 hr. After purificationby selective adsorption, precipitation and disc electrophoresis,the protein from molybdenum-deficient plants given ⁹⁹Mo showedradioactivity only where nitrate reductase was revealed on theacrylamide gel. Molybdenum was similarly selectively concentratedinto the enzyme as a result of induction by nitrate in plantsgrown with sub-optimal molybdenum supply in order to minimizeeffects of isotope dilution on measurement of ⁹⁹Mo incorporation. There was no exchange in vitro between ⁹⁹Mo and purified activeenzyme in the resting state over 18 hr at 4°C, or with functioningenzyme held at room temperature for 24 hr. There was evidenceeither for possible in vivo exchange of ⁹⁹Mo andenzyme boundMo or for slight synthesis of fresh enzyme under conditionsof net loss of enzyme in nitrate starved plants. Five NADH₂ and two NADPH₂ reactive diaphorases which could beseparated by electrophoresis were present in extracts. Onlyone of these having strong NADH₂ and weak NADPH₂ activity wasdirectly associated with nitrate reductase. The same complexalso showed the only benzyl viologen (BV.) reactive nitratereductase. Nitrate reductase in spinach is therefore considered to be amolybdenum-dependant and molybdenum-containing protein in whichNADH₂ (with weak NADPH₂) and BVelectron donor functions anddiaphorase/reductase activities remain closely associated duringpurification and electrophoresis. The techniques provide a simple means for the production andpurification of enzyme containing radioactively labelled Moapplicable to investigations on the structure of the enzyme. (Received January 16, 1971; )     

Action of Thiol Proteinase on Nitrate Reductase in Leaves of Hordeum distichum L.

Nitrate reductase (NR) from the leaves of Hordeum distichumwas very susceptible to inactivation by barley leaf thiol proteinase,trypsin, and papain. A cytochrome c reductase species with amolecular weight of about 40,000 was derived from the NR complexby the proteolytic actions. The barley proteinase seemed toattack the Mo⁺-containing component of NR, just like trypsinand papain. It showed a preference for the alanine and tryptophanesters among the carbobenzoxyamino acid-nitrophenylesters tested. In vivo NR activity in the presence of leupeptin was fairlyhigher than that in its absence. Leupeptin also protected NRfrom its cleavage to small cytochrome c reductase species, suggestingthat the barley proteinase may play a role in the in vivo changein NR activity. (Received May 21, 1984; Accepted September 10, 1984)     

Short-term Effects of Ferulic Acid on Ion Uptake and Water Relations in Cucumber Seedlings

Ferulic acid (FA) is commonly found in soils and is consideredan allelochemical. Studies have suggested that FA and otherphenolic acids decrease plant growth in part by decreasing theabsorption of mineral nutrients and water. However, no studieshave examined these parameters in a single experimental systemto investigate how FA affected both ion uptake and plant-waterrelations in whole plants. Using intact cucumber (Cucumis sativusL. cv. Early Green Cluster) seedlings, we examined short-termeffects of FA on ion uptake kinetics, transport promoters andinhibitors, and water relations as indicated by a pressure-volumeanalysis. We found that after 3 h of treatment, 200 µMFA inhibited net ion uptake, particularly NO₃, and promotednet K⁺ efflux from seedling roots. The addition of fusicoccin,a K⁺ transport promoter, counteracted the inhibitory effectof FA on net K⁺ uptake. Concurrent treatment of seedlings withFA and tetraethylammonium, a channel-blocking salt, reducedaverage K⁺ efflux by 66%. Treatment of seedlings with FA alsodecreased leaf water potential (₁ and turgor pressure (P_T).However, decreased ₁ and P_T were not caused by changes in theosmotic properties of the symplast or stomatal conductance.A decrease in water absorption is a likely explanation for theloss of P_T observed. The results of our experiments indicatethat both ion uptake and plant-water relations can be adverselyaffected by FA. Key words: Cucumis sativus, ferulic acid, allelopathy, ion uptake, water relations     

Characterization of a rice (Oryza sativa L.) mutant deficient in the heme domain of nitrate reductase

Summary Biochemical and genetical characterization of a rice nitrate reductase (NR)-deficient mutant, M819, which had been isolated as a chlorate-resistant mutant, was carried out. In M819, leaf NADH-NR activity was found to be about 10% of that of the wild-type cv Norin 8, while NADPH-NR activity was higher than that in the wild-type; FMNH₂-NR and MV-NR activities were also 10% of those of the wild type; BPB-NR activity was higher than that of the wild type; and xanthine dehydrogenase activity was revealed to be present in both. These results suggest that the mutant line M819 lacks the functional heme domain of the NADH-NR polypeptide due to a point mutation or a small deletion within the coding region of the structural gene. Chlorate resistance in M819 was transmitted by a single recessive nuclear gene.Abbreviations NR Nitrate reductase - NiR nitrite reductase - FMNH₂ reduced flavin nucleotide - MV reduced methyl viologen - BPB reduced bromphenol blue - XDH xanthine dehydrogenase     

NO2 Suppression of Light-Induced Nitrate Reductase in Squash Cotyledons

NADH-nitrate reductase (NR) (EC 1.6.6.1 [EC] ) activity in the cotyledonsof squash (Cucurbita maxima Duch.) seedlings showed daily variationwhen the seedlings were subjected to an alternating light-darkcycle. When the seedlings were transferred into continuous darkness,NR activity rose at first and then decreased continuously. Irradiationafter continuous darkness induced a rapid increase in NR activity;this light induction of NR activity was inhibited completelyby fumigation with 4 ppm nitrogen dioxide (NO₂). This inhibitoryeffect of NO₂ was prominent even at 1 ppm and became more pronouncedas the concentration of NO₂ increased. NO₂ fumigation did notremarkably affect the content of reductant (NADH) in the cotyledons.The results of immunoblotting using anti-NR serum indicatedthat irradiation induced the increase in the NR-polypeptidecontent and NO₂ fumigation inhibited the increase, suggestingthat NO₂ put an inhibitory effect on the synthesis of NR inducedby irradiation. ⁴ Present address: College of Environmental Health, Azabu University,Fuchinobe, Sagamihara, Kanagawa 229, Japan ⁵ Present address: Faculty of Home Economics, Otuma Women'sUniversity, Sanban-cho, Chiyoda, Tokyo 102, Japan (Received October 21, 1987; Accepted January 13, 1988)     

Stomatal Control of Photosynthesis of Eucalyptus globulus Labill. Trees under Field Conditions in Portugal

Pereira, J. S., Tenhunen, J. D. and Lange, O. L. 1987. Stomatalcontrol of photosynthesis of Eucalyptus globulus Labill. treesunder field conditions in Portugal.—J. exp. Bot. 38: 1678–1688. Stomatal behaviour of adult leaves of Eucalyptus globulus treeswas studied under field conditions in Portugal. In the absenceof severe plant water stress stomata were open when the summedtotal of photosynthetically active photon flux density incidenton both leaf surfaces was above 100 µmol m²s¹ and leafconductance to water vapour reached 245 mmol m ² s¹ on a total(both epidermes) leaf area basis. The stomata of both leaf epidermesresponded similarly to changes in solar radiation and waterstress. Water stress resulted in decreasing daily maxima inleaf conductance as predawn leaf water potential decreased.Maximal leaf conductance decreased to less than 50 mmol m ²s ¹ when predawn leaf water potential decreased below —1·0MPa. At similar values of predawn leaf water potential stomatawere more closed as the leaf to air water vapour partial pressuredifference increased. The effect of increasing air dryness onstomata was greatest at high predawn leaf water potential. Dailymaxima in photosynthetic rates and in leaf conductance werelinearly related to one another in spring and summer. Both decreasedwith increase in leaf water stress. In autumn and winter, increasesin leaf conductance occurring under natural conditions duringthe course of the day were not necessarily accompanied by increasesin net photosynthesis. Stomata were more closed in the afternoonthan in the morning at the same rates of net photosynthesis,temperature or leaf to air water vapour partial pressure difference. Key words: Eucalyptus globulus,, photosynthesis, stomata, water stress.