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

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

Effect of Nitrogen Supply on the Grass and Clover Components of Simulated Mixed Swards Grown under Favourable Environmental Conditions II. Nitrogen Fixation and Nitrate Uptake

Simulated mixed swards of Perennial Ryegrass (Lolium perenneL.) cv. S23 and White clover (Trifolium repens L.) cv. S100were grown from seed under a constant 20 °C day/15 °Cnight temperature regime and harvested at intervals over and88 d growht period. The swards received a nutrient solutiondaily, which was either High (220 mg l^–1) or Low (10 mgl^–1) in nitrate N. The nitrate was labelled with the ¹⁵Nisotope. An acetylene reduction assay was carried out on eachsward just prior to harvest. Rates of acetylene reduction agreed qualitatively with the ^l5Nanalyses but absolute values did not match (assuming a 4:1 C₂H₄:N₂ratio) and errors in the acetylene assay are discussed. In theLow-N swards clover relied almost entirely on symbioticallyfixed N₂, fixing more than ten times as much as the High-N cloverplants. In the Low-N treatment the grass was N-deficient despiteobtaining much more nitrate per unit root dry weight than clover.In the High-N swards, however, clover took up more nitrate perunit root weight than grass. The High-N clover plants also fixedsome N₂ and maintained a higher total-N content than grass throughoutthe period. There was no evidence of transfer of symbioticallyfixed N from the clover to the grass in either treatment. Trifolium repens, Lolium perenne, nitrate, nitrogen fixation, ¹⁵N, acetylene reduction     

Nitrogen Fixation by Subterranean Clover at Varying Stages of Nodule Dehydration: II. EFFICIENCY OF NITROGENASE FUNCTIONING

Changes in nitrogenase activity (C₂H₂ reduction and H₂ production),nodulated root respiration and the efficiency of nitrogenasefunctioning were measured in response to progressive dehydrationof nodules on intact well-watered plants of subterranean clover(Trifolium subterraneum L.) cv. Seaton Park. The nodulated rootsof vegetative plants grown to the 14-leaf stage were incubatedin a gas exchange system through which a continuous dry airstreamwas passed over an 8 d period. The root tips were immersed inan N-free nutrient solution during this time so that water andion uptake was unimpeded. The decline in nodulated root respirationresulting from nodule drying was associated with a continualreduction in respiration coupled to nitrogenase activity. Asnodule water potential (_nod) decreased, the proportion of totalnodulated root respiration which was nitrogenase-linked declinedfrom 50% (day 1) to 33% (day 8). This was accompanied by a 79%reduction in specific nitrogenase activity (from 3.79 to 0.81umol C₂H₄ g^–1 nodule dry weight min^–1). Nodule dehydrationalso induced a decline in hydrogen (H₂) production in air. Therelative decline in hydrogen production exceeded that of acetylenereduction activity and this resulted in an increase in the relativeefficiency of nitrogenase functioning. However, the carbon costof nitrogenase activity progressively increased above 2.0 molCO₂ respired per mol C₂H₄ reduced as rood decreased below –0.4to –0.5 MPa. Consecutive measurements of the rates ofhydrogen evolution, ¹⁵N₂ fixation and acetylene reduction activityon intact unstressed plants resulted in a C₂H₄/N₂ conversionfactor of 4.08 and an electron balance of 1.08. These resultsindicated that the pre-decline rate of acetylene reduction activitymeasured in a flow-through system provided a valid measure ofthe total electron flux through nitrogenase. Key words: Subterranean clover, dehydration, efficiency, nitrogenase activity     

Defoliation in White Clover: Nodule Metabolism, Nodule Growth and Maintenance, and Nitrogenase Functioning During Growth and Regrowth

In two experiments, the functioning and metabolism of nodulesof white clover, following a defoliation which removed abouthalf the shoot tissue, were compared with those of undefoliatedplants. In one experiment, the specific respiration rates of nodulesfrom undefoliated plants varied between 1160 and 1830 µmolCO₂ g^–1h^–1, of which nodule ‘growth and maintenance’accounted for 22 ± 2 per cent, or 27 ± 3.6 percent, according to method of calculation. Defoliation reducedspecific nodule respiration and nodule ‘growth and maintenance’respiration by 60–70 per cent, and rate of N₂ fixationby a similar proportion. The original rate of nodule metabolismwas re-established after about 5 d of regrowth; during regrowthnodule respiration was quantitatively related to rate of N₂,fixation: 9.1 µmol CO₂ µmol^–1N₂. With the possible exception of nodules examined 24 h after defoliation,the efficiency of energy utilization in nitrogenase functioningin both experiments was the same in defoliated and undefoliatedplants: 2.0±0.1 µmol CO₂ µmol^–1 C₂H₄;similarly, there was no change in the efficiency of nitrogenasefunctioning as rate of N₂ fixation increased with plant growthfrom 1 to 22 µmol N₂ per plant h^–1. Exposure of nodulated white clover root systems to a 10 percent acetylene gas mixture resulted in a sharp peak in rateof ethylene production after 1.5–2.5 min; subsequently,rate of ethylene production declined rapidly before stabilisingafter 0.5–1 h at a rate about 50 per cent of that initiallyobserved. Regression of ‘peak’ rate of ethyleneproduction on rate of N₂ fixation indicated a value of 2.9 µmolC₂H₄ µmol^–1 N₂, for rates of N₂ fixation between1 and 22 µmol N₂ per plant h^–1. The relationshipsbetween nitrogenase respiration, acetylene reduction rates andN₂ fixation rates are discussed. Trifolium repens, white clover, defoliation, nodule respiration, N₂, fixation, nitrogenase     

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

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

Growth of White Clover, Dependent on N2 Fixation, in Elevated CO2 and Temperature

Clonal plants of white clover (Trifolium repens L ), whollydependent on N₂ fixation, were grown for 6 weeks in controlledenvironments providing either (C680 regime) 23/18 °C day/nighttemperatures and a CO₂, concentration of 680 µmol mol^–1,or (C340 regime) 20/15 °C day/night temperatures and a CO₂,concentration of 340 µmol mol^–1 During the firsthalf of the experimental period the C680 plants grew fasterthan their C340 counterparts so that by week 3 they were twicethe weight this 2 1 superiority in weight persisted until theend of the experiment The faster initial growth of the C680plants was based on an approx 70 % increase in leaf numbersand an approx 30 % increase in their individual area Initially,specific leaf area (cm2 g^–1 leaf) was lower in C680 thanin C340 leaves but became similar in the latter half of theexperiment Shoot organ weights, including petioles and stolons,reflected the C680 plant's better growth in terms of photosyntheticsurface Throughout, C680 plants invested less of their weightin root than C340 plants and this disparity increased with timeAcetylene reduction assays showed that nitrogenase activityper unit nodule weight was the same in both C680 and C340 plantsBoth groups of plants invested about the same fraction of totalweight in nodules Nitrogen contents of plant tissues were similarirrespective of growth regime, but C680 expanded leaves containedslightly less nitrogen and their stolons slightly more nitrogenthan their C340 counterparts However, C680 leaves containedmore non-structural carbohydrate Young, unshaded C680 leavespossessed larger palisade cells, packed more tightly withinthe leaf, than equivalent C340 leaves The reason for the C680regime's loss of superiority in relative growth rate duringthe second half of the experiment was not clear, but more accumulationof non-structural carbohydrate, constriction of root growthand increased self-shading appear to be the most likely causes Trifolium repens, white clover, elevated CO₂, elevated temperature, growth, N₂ fixation, leaf structure     

Comparisons of the Respiratory Effluxes of Nodules and Roots in Six Temperate Legumes

The specific respiration rates of nodulated root systems, ofnodules and of roots were determined during active nitrogenfixation in soya bean, navy bean, pea, lucerne, red clover andwhite clover, by measurements on whole plants before and afterthe removal of nodule populations. Similar measurements weremade on comparable populations of the six legumes, lacking nodulesbut receiving abundant nitrate-nitrogen, to determine the specificrespiration of their roots. All plants were grown in a controlled-environmentclimate which fostered rapid growth. The specific respiration rates of nodulated root systems ofthe three grain and three forage legumes during a 7–14-dayperiod of vegetative growth varied between 10 and 17 mg CO₂g^–1 (dry weight) h^–1. This mean value consistedof two components: a specific root respiration rate of 6–9mg CO₂ g^–1 h^–1 and a specific nodule respirationrate of 22–46 mg CO₂ g^–1 h^–1. Nodule respirationaccounted for 42–70 per cent of nodulated root respiration;nodule weight accounted for 12–40 per cent of nodulatedroot weight. The specific respiration rates of roots lackingnodules and utilizing nitrate nitrogen were generally 20–30per cent greater than the equivalent rates of roots from nodulatedplants. The measured respiratory effluxes are discussed in thecontext of nitrogen nitrogen fixation, nitrate assimilation. Glycine max, Phaseolus vulgaris, Pisum sativum, Medicago sativa, Trifolium pratense, Trifolium repens, soya bean, navy bean, pea, lucerne, red clover, white clover, nodule respiration, root respiration, fixation, nitrate assimilation     

Studies on the Relation Between Net Photosynthesis and Nitrogenase-linked Respiration in Subterranean Clover

The relation between the rate of nitrogenase-linked respirationand net photosynthesis, and the effect of defoliation on thisrelation, was studied in plants of subterranean clover (Trifoliumsubterraneum L. cv. Seaton Park). Nitrogenase-linked respirationwas estimated as the difference between the rate of nodulatedroot respiration at 21% O₂ and at 3% O₂. The level to which the rate of nitrogenase-linked respirationfell several hours after defoliation was directly proportionalto the decline in the rate of net photosynthesis. Approximately9% of net photosynthesis was always expended in nitrogenaseactivity, irrespective of whether or not the plants were defoliated.This proportion was maintained during the first 3 d of regrowth. To determine whether the decline in nitrogenase-linked respirationafter defoliation was due solely to the decline in the rateof photosynthesis, a further experiment was conducted in whichthe pre-defoliation rate of net photosynthesis was restoredimmediately (with supplementary light) or within 5 min (supplementarylight and CO₂) after defoliation. Restoring the rate of netphotosynthesis did not prevent the post-defoliation declinein nitrogenase-linked respiration. However, when photosynthesiswas reduced to zero by the imposition of darkness, and the rateof nitrogenase-linked respiration allowed to decline to a steadyrate after 3 h, a rapid recovery in the rate of nodulated rootrespiration began within 2 h of returning the plants to thelight. It was hypothesized that a ‘shoot factor’,which was affected by defoliation, could override the apparentrelation between nitrogenase-linked respiration and the rateof current photosynthesis. Key words: Defoliation, N₂ fixation, photosynthesis, nitrogenase-linked respiration, subterranean clover     

White Clover N2-Fixation in Response to Root Temperature and Nitrate: II. N2-FIXATION, RESPIRATION, AND NITRATE REDUCTASE ACTIVITY

Established, nodulated white clover plants were transferredto eight tanks of a flowing culture apparatus with solutiontemperatures of 5, 11, 17, and 25 ?C (two tanks per temperature).Shoot temperature and light environment were common to all plants.After 7 d, (10 mmol m^–3) was continuouslysupplied to one tank at each temperature while in the remainingfour tanks (one at each temperature) the plants were completelydependent on nodule N₂-fixation. Plants were randomly selected at intervals during the following14 d period in order to measure root and nodule respirationand acetylene reduction activity (ARA) in a flow-through systemset at the adapted root temperature. Additional plants wereassayed for in vitro nitrate reductase activity in leaves, roots,and nodules. Apparent nitrogenase activity (ARA) and respiration associatedwith it were each markedly affected by temperature in two ways;(1) Activity per unit weight of nodule was reduced at lowertemperatures; (2) Development of the plant, and thus also nodulemass, was restricted at lower temperatures which, in turn, restrictedtotal nodule activity per plant. The presence of nitrate significantly reduced ARA of nodules,particularly at higher temperatures. However, significant discrepancieswere found when N₂-fixation rates, estimated from the acetylenereduction assay, were compared with N₂-fixation rates calculatedfrom curves fitted to N accumulation data (minus the rate of uptake in the case of nitrate-treated plants). Carbon use efficiency (CO² respired per C₂H₄ produced) was notsignificantly affected by temperature or the presence of nitrate. Nitrate reductase activity (NRA) developed in all plant partsat the three highest temperatures, but not at 5 ?C. We calculatethat leaf NRA may account for 82, 75, and 68% of total nitratereduction at 11, 17, and 25 ?C respectively. Key words: Trifolium repens, white clover, N₂ fixation, root temperature, acetylene reduction assay, nitrate, nitrate reductase     

N2 Fixation and Nitrate Uptake by White Clover Swards in Response to Root Temperature in Flowing Solution Culture

Nodulated white clover plants (Trifolium repens L. cv. Huia)were grown as simulated swards for 71 d in flowing nutrientsolutions with roots at 11 C and shoots at 20/15 C, day/night,under natural illumination. Root temperatures were then changedto 3, 5, 7, 11, 13, 17 or 25 C and the total N₂, fixation over21 d was measured in the absence of a supply mineral N. Alltreatments were subsequently supplied with 10 mmol m^–2NO₂ ^– in the flowing solutions for 14 d, and the relativeuptake of N by N₂, fixation and NO₃ ^– uptake was compared.Net uptake of K⁺ was measured on a daily basis. Root temperature had little effect on root d. wt over the 35-dexperimental period, but shoot d. wt increased by a factor of3.5 between 3 and 25 C, with the sharpest increase occurringat 7–11 C. Shoot: root d. wt ratios increased from 25to 68 with increasing temperature at 7–25 C. N₂-fixationper plant (in the absence of NO₂ ^–) increased with roottemperature at 3–13C, but showed little change above13 C. The ratios of N₂ fixation: NO₂ ^– uptake over 14d (mol N: mol N) were 0.47–0.77 at 3–7 C, 092–154at 11–17 C, and 046 at 25 C, reflecting the dominanceof NO₃ ^– uptake over N₂ fixation at extremes of high andlow root temperature. The total uptake of N varied only slightlyat 11–25 –C (095–110 mmol N plant^–1),the decline in N₂ fixation as root temperature increased above11 C was compensated for by the increase in NO^– ₃ uptake.The % N in shoot dry matter declined with decreasing root temperature,from 32% at 13 C to 15% at 3 C. In contrast, concentrationsof N expressed on a shoot water content basis showed a modestdecrease with increasing temperature, from 345 mol m^–3at 3 C to 290 mol m^–3 at 25 C. Trifolium repens L, white clover, root temperature, N₂ fixation, potassium uptake, nitrate uptake, flowing solution culture     

The Effect of Root Temperature on Growth and Uptake of Ammonium and Nitrate by Brassica napus L. cv. Bien venu in Flowing Solution Culture: II. UPTAKE FROM SOLUTIONS CONTAINING NH4NO3

Macduff, J. H., Hopper, M. J. and Wild, A. 1987. The effectof root temperature on growth and uptake of ammonium and nitrateby Brassica napus L. CV. Bien venu in flowing solution culture.II. Uptake from solutions containing NH₄NO₃.—J. exp. Bot.38: 53–66 The effects of root temperature on uptake and assimilation ofNH₄⁺ and NO₃^– by oilseed rape (Brassica napus L. CV. Bienvenu) were examined. Plants were grown for 49 d in flowing nutrientsolution at pH 6?0 with root temperature decrementally reducedfrom 20?C to 5?C; and then exposed to different root temperatures(3, 5, 7, 9, 11, 13, 17 or 25?C) held constant for 14 d. Theair temperature was 20/15?C day/night and nitrogen was suppliedautomatically to maintain 10 mmol m^–3 NH₄NO₃ in solution.Total uptake of nitrogen over 14 d increased threefold between3–13?C but was constant above 13?C. Net uptake of NH₄⁺exceeded that of NO₃^– at all temperatures except 17?C,and represented 47–65% of the total uptake of nitrogen.Unit absorption rates of NH₄⁺ and of 1?5–2?7 for NO₃^–suggested that NO₃^– absorption was more sensitive thanNH₄⁺ absorption to temperature. Rates of absorption were relativelystable at 3?C and 5?C compared with those at 17?C and 25?C whichincreased sharply after 10 d. Tissue concentration of N in theshoot, expressed on a fresh weight basis, was independent ofroot temperature throughout, but doubled between 3–25?Cwhen expressed on a dry weight basis. The apparent proportionof net uptake of NO₃^– that was assimilated was inverselyrelated to root temperature. The results are used to examinethe relation between unit absorption rate adn shoot:root ratioin the context of short and long term responses to change ofroot temperature Key words: Brassica napus, oilseed rape, root temperature, nitrogen uptake     

Components of Growth and Dark Respiration of Kikuyu (Pennisetum clandestinum Chiov.) at Various Temperatures

Growth and dark respiration were measured in dense, miniatureswards of kikuyu grass grown at constant temperatures of 15,20, 25 and 30 °C. Total respiration over the first 12 hof darkness was very high and CO² efflux per unit surface areavaried from 2.4 to 3.9 g CO₂ m^–2 h^–1 at 15 and 30°C respectively. Such rates were consistent with the correspondinglyhigh net growth rates of 24 and 63 g d. wt m^–2 d^–1and the heavy yields of herbage. When plants were kept in thedark, CO₂ efflux subsequently declined rapidly to a lower, constantrate which was taken to be the maintenance respiration rate.The half-life of the declining phase of respiration averaged10.9 and 6.0 h at 15 and 30 °C respectively, and was curvilinearlyrelated to the specific maintenance respiration rate (m). Therapid decline in respiration was consistent with the low concentrationsof total soluble carbohydrate and starch in the herbage. Valuesof m for lamina and top growth increased with temperature witha Q₁₀ of 2.6 and 1.42 respectively, but m of stems alone wasnot affected by temperature. Using results from this study forkikuyu and from McCree (1974) for sorghum and white clover,it was noted that all three species have similar m when grownat temperatures which are near their respective optimums forgrowth. Kikuyu, Pennisetum clandestinum, growth, respiration, temperature     

Concurrent Rates of N2 Fixation, Nitrate and Ammonium Uptake by White Clover in Response to Growth at Different Root Temperatures

Nodulated white clover plants (Trifolium repens L. cv. Huia)were grown for 71 d in flowing nutrient solutions containingN as 10 mmol m_–3 NH₄NO₃, under artificial illumination,with shoots at 20/15°C day/night temperatures and root temperaturereduced decrementally from 20 to 5°C. Root temperatureswere then changed to 3, 7, 9, 11, 13, 17 or 25°C, and theacquisition of N by N₂ fixation, NH₄+ and NO₃^– uptakewas measured over 14 d. Shoot specific growth rates (d. wt)doubled with increasing temperature between 7 and 17°C,whilst root specific growth rates showed little response; shoot:root ratios increased with root temperature, and over time at11°C. Net uptake of total N per plant (N₂ fixation + NH₄++ NO₃^–) over 14 d increased three-fold between 3 and 17°C.The proportion contributed by N₂ fixation decreased with increasingtemperature from 51% at 5°C to 18% at 25°C. Uptake ofNH₄+ as a proportion of NH₄+ + NO₃^– uptake over 14 d variedlittle (55–62%) with root temperature between 3 and 25°C,although it increased with time at most temperatures. Mean ratesof total N uptake per unit shoot f. wt over 14 d changed littlebetween 9 and 25°C, but decreased progressively with temperaturebelow 9°C, due to the decline in the rates of NH₄+ and NO₃^–uptake, even though N₂ fixation increased. The results suggestthat N₂ fixation in the presence of sustained low concentrationsof NH₄+ and NO₄^– is less sensitive to low root temperaturethan are either NH₄+ or NO₃^– uptake systems. White clover, Trifolium repens L. cv. Huia, root temperature, nitrogen fixation, ammonium, nitrate     

Effect of Temperature and Nitrogen Supply on the Growth of Perennial Ryegrass and White Clover. 2. A Comparison of Monocultures and Mixed Swards

White clover (Trifolium repens L.) and Perennial ryegrass (Loliumperenne L.) plants were grown, in Perlite, in simulated swardsas either monocultures or mixtures of equal plant numbers. Theywere supplied with a nutrient solution either high (220 µgg^–1) or low (40 µg g^–1) in ¹⁵N-labelled nitrateand grown to ceiling yield at either high (20°C day/15°Cnight) or low (10°C day/8°C night) temperature. Temperature had little effect on the maximum rates of grosscanopy photosynthesis which were similar in High-N grass andHigh-N and Low-N clover monocultures. However these maxima werereached more slowly in clover than grass, and more slowly atlow rather than high temperature. Nitrogen supply increasedphotosynthesis in grass but not in clover. Clover had higherN contents than grass in all four treatments, although in anygiven treatment its N content was lower, and contribution ofN₂-fixation relative to nitrate uptake higher, in mixture thanin monoculture. Conversely, grass had higher N contents in mixturethan monoculture, because more nitrate was available per plantand not because of transfer of biologically fixed N from clover. Under Low-N, clover outyielded grass in mixture, particularlyat high temperature. The grass plants in the Low-N mixtureshad higher N contents and higher SLA, LAR and shoot: root ratiosthan those in monoculture. It is proposed that competition forlight is the cause of the low relative yield and negative aggressivityof grass in these swards. Under High-N, grass outyielded cloverin monoculture and mixture, at both temperatures but particularlyat low temperature when grass had a high aggressivity. Nitrogenand yield component analyses shed no light on clover's apparentlylow competitive ability and evidence is drawn from the previouspaper to demonstrate that grass grew faster than clover onlyas spaced individuals during non-com petitive growth. The relativemerits of measures of competitive ability based on final harvestdata and physiological data taken over a growth period are discussed. Trifolium repens L., white clover, Lolium perenne, perennial ryegrass, competition, temperature, nitrogen     

White Clover N2-Fixation in Response to Root Temperature and Nitrate: I GROWTH AND UPTAKE OF NITRATE FROM FLOWING NUTRIENT SOLUTIONS

Established, nodulated white clover plants (Trifolium repensL. cv. Blanca) were transferred to eight plant culture unitsof a system of flowing solution culture. Solution temperatureswere 5, 11, 17, and 25 ?C (two units per temperature), withshoot temperature of 25/15 ?C day/night and light regime commonto all plants. After 7 d, was supplied at 10 mmol m^–3 to one of each pair of culture units and thenet uptake of was monitored over 14 d. The remaining four culture units served as controls in which plantswere entirely dependent on N₂ fixation, as estimated by sequentialharvesting and the change in the amount of N in plants. Totalfresh and dry weights increased exponentially with time andwith increasing root temperature, between 5–25 ?C in nitrateplants and 5–17 ?C in control plants, respectively, byfactors of 2.9 and 1.8. Nodule dry weight of nitrate plantsshowed little increase after 6 d compared with control plants.Dry weight shoot: root ratios increased with time in all treatmentsexcept 5 ?C control plants. Total net uptake of over 13 d was 0.41, 4.27, 11.92, and 12.77 mmol plant-1, respectively,at 5, 11, 17, and 25 ?C. At all temperatures except 5 ?C, plantsaccumulated high concentrations of (10–40 mol m^–3) in leaflets and roots, within 2 or 3 days ofsupplying . Daily mean unit absorption rates of increased 12-fold with increasing temperature between 5 and 25 ?C, and showed little change with time at 5,17, and 25 ?C. The total N₂ fixed by nitrate plants over 14d increased 5-fold with temperature between 5 and 17 ?C, butwas always less than the amount fixed by control plants. Thepercentage contributed by N2 fixation to total N acquisitionby nitrate plants over 14 d decreased with increasing temperature,from 77% at 5 ?C to 11 % at 25 ?C. Mean daily rates of N₂ fixationper unit dry weight of nodule were lower in nitrate plants thanin control plants throughout treatment at 5 ?C and 25 ?C, butat 11 ?C and 17 ?C the rates for nitrate plants increased progressivelywith time and exceeded the rates for control plants after 8d. In both nitrate and control plants the effect of temperatureon N₂ fixation per nodule dry weight was proportionately lessthan that on unit absorption rate of . The results are discussed in terms of the overall regulation ofN accumulation by white clover and the adaptive significanceof differences in the sensitivities of uptake and N₂ fixation to root temperature. Key words: Trifolium repens, white clover, root temperature, N₂ fixation, nitrate uptake     

Growth and Preferences for Ammonium or Nitrate Uptake by Barley in Relation to Root Termperature

Barley plants (Hordewn vulgare L. cv. Atem) were grown fromseed for 28 d in flowing solution culture, during which timeroot temperature was lowered decrementally to 5?C. Plants werethen subjected to root temperatures of 3, 5, 7, 9, 11, 13, 17or 25 ?C, with common air temperature of 25/15 ?C (day/night).Changes in growth, plant total N, and NO₃^– levels, andnet uptake of NH₄⁺ and NO₃^– from a maintained concentrationof 10 mmol m^–3 NH₄NO₃ were measured over 14 d. Dry matterproduction increased 6-fold with increasing root temperaturebetween 3–25 ?C. The growth response was biphasic followingan increase in root temperature. Phase I, lasting about 5 d,was characterized by high root specific growth rates relativeto those of the shoot, particularly on a fresh weight basis.During Phase I the shoot dry weight specific growth rates wereinversely related to root temperature between 3–13 ?C.Phase 2, from 5–14 d, was characterized by the approachtowards, and/or attainment of, balanced exponential growth betweenshoots and roots. Concentrations of total N in plant dry matterincreased with root temperature between 3–25 ?C, moreso in the shoots than roots and most acutely in the youngestfully expanded leaf (2?l–6?9% N). When N contents wereexpressed on a tissue fresh weight basis the variation withtemperature lessened and the highest concentration in the shootwas at 11 ?C. Uptake of N increased with root temperature, andat all temperatures uptake of NH₄⁺, exceeded that of NO₃^–,irrespective of time. The proportions of total N uptake over14 d absorbed in the form of NH₄⁺ were (%): 86, 91, 75, 77,76, 73, 77, and 80, respectively, at 3, 5, 7, 9, Il, 13, 17,and 25 ?C. At all temperatures the preference for NH₄⁺ overNO₃^– uptake increased with time. An inverse relationshipbetween root temperature (3–11 ?C) and the uptake of NH₄⁺as a proportion of total N uptake was apparent during PhaseI. The possible mechanisms by which root temperature limitsgrowth and influences N uptake are discussed. Key words: Hordeum vulgare, root temperature, ammonium, nitrate, ion uptake, growth rate     

Relationships between Acetylene Reduction Activity, Hydrogen Evolution and Nitrogen Fixation in Nodules of Acacia spp.: Experimental Background to Assaying Fixation by Acetylene Reduction under Field Conditions

Hansen, A. P., Pate, J. S. and Atkins, C. A. 1987. Relationshipsbetween acetylene reduction activity, hydrogen evolution andnitrogen fixation in nodules of Acacia spp.: Experimental backgroundto assaying fixation by acetylene reduction under field conditions.—J.exp. Bot. 38: 1–12 Glasshouse grown, symbiotically-dependent seedlings of Acaciaalata R.Br., .A. extensa Lindl., and A. pulchella R.Br. wereexamined for acetylene reduction in closed assay systems usingundisturbed potted plants, excavated whole plants, nodulatedroots or detached nodules. Nitrogenase activity declined sharplyover the first hour after exposure of detached nodules to acetylene(10% v/v in air), less steeply or not at all over a 3 h periodin assays involving attached nodules. Using detached nodules,rates of acetylene reduction, nitrogen (¹⁵N₂) fixation, andhydrogen evolution in air (¹⁵N₂) and acetylene-containing atmosphereswere measured in comparable 30 min assays. Total electron flowthrough nitrogenase in air was determined from rates of nitrogen(¹⁵N₂) fixation ( ? 3) plus hydrogen evolution, that in thepresence of acetylene from rates of acetylene reduction andhydrogen evolution in air: acetylene. Values for the ratio ofelectron flow in air: acetylene to that in air ranged from 0?43to 0?83 in A. pulcheila, from 0?44 to 0?66 in A. alala and from0?37 to 0?70 in A. extensa, indicating substantial inhibitionof electron flow through nitrogenase of detached nodules byacetylene. Relative efficiencies of nitrogenase functioningbased on hydrogen evolution and acetylene reduction were from0?15 to 0?79, those based on nitrogen (¹⁵N₂) fixation and hydrogenevolution from 0?53 to 0?87. Molar ratios of acetylene reducedto nitrogen (¹⁵N₂) fixed were 2?82 ? 0?24, 201 ? 0?15, and 1?91? 0?11 (?s.e.; n = 7) for A. pulcheila,A. extensa and A. alata respectively A standard 5–10 min acetylene reduction assay, conductedon freshly detached unwashed nodules in daytime (12.00–14.00h), was calibrated for field use by comparing total N accumulationof seedlings with estimated cumulative acetylene reduction overa 7-week period of glasshouse culture. Molar ratios for acetylenereduced: nitrogen fixed using this arbitrary method were 3?58for A. alata, 4?82 for A. extensa and 1?60 for A. pulchella.The significance of the data is discussed. Key words: Acacia spp, nitrogenase functioning     

Occurrence of Inducible Crassulacean Acid Metabolism in Leaves of Talimum triangulare (Portulacaceae)

In this paper we report for the first time the occurrence ofan inducible weak CAM in leaves of Talinwn triangulare (Jacq.)Willd. This plant is a terrestrial perennial deciduous herbwith woody stems and succulent leaves which grows under fullexposure and in the shade in northern Venezuela. Plants grownin a greenhouse (‘sun’ plants) and a growth cabinet(‘shade’ plants) with daily irrigation showed CO₂uptake only during the daytime (maximum rate, 4?0 µmolm^–2 s^–1) and a small acid accumulation during thenight (6?0 µmol H⁺g^–1 FW). Twenty-four hours aftercessation of irrigation, no CO₂ exchange was observed duringpart of the night. Dark fixation reached a maximum (1?0 µmolCO₂ m^–2 s^–1, 100 µmol H⁺ g^–1 FW) onday 9 of drought. By day 30 almost no gas exchange was observed,while acid accumulation was still 10 µmol H⁺ g^–1FW. Rewatering reverted the pattern of CO₂ exchange to thatof a C₃ plant within 24 h. Daytime and night-time phosphoenolpyruvatecarboxylase activity increased up to 100% (shade) and 62% (sun)of control values after 10 and 15 d of drought, respectively.Light compensation point and saturating irradiance were similarin well-watered sun and shade plants, values being characteristicof sun plants. CAM seems to be important for the tolerance ofplants of this species to moderately prolonged (up to 2 months)periods of drought in conditions of full exposure as well asshade, and also for regaining high photosynthetic rates shortlyafter irrigation. Key words: Talinum triwigulare, inducible CAM, PEP-C activity, recycling     

Respiratory Efflux in Relation to Temperature of Simulated Swards of Perennial Ryegrass with Contrasting Soluble Carbohydrate Contents

Fully light-intercepting simulated swards of S24 perennial ryegrasswere exposed to contrasting environmental conditions in a growthroom for 4 days. Half experienced 20 h days of 120 Wm^–2(400–700. nm) and 5 °C, and came to have a WSC (watersoluble carbohydrate) content of 235 mg g^–1 and half 4h days of 20 Wm^–2 and 25 °C leading to a WSC of 25mg g^–1. Their rates of CO₂ efflux were monitored at anumber of temperatures during an 8 h dark period; half experiencedincreasing (5–30 °C) and half decreasing (30–5°C) temperatures. The ‘high’ WSC swards hadrespiration rates of 3.7 mg CO₂ g^–1 (d. wt) h^–1at 15 °C, and the ‘low’ swards 0.8 mg CO₂ g^–1h^–1. The order in which the temperatures were experiencedwas immaterial. Even the ‘low’ WSC swards showedno evidence of a respiratory decline during the dark periodthat could be attributed to substrate shortage. The relationshipbetween temperature and CO₂ efflux was best represented by logisticcurves. Even so, a Q₁₀ of 2 fitted the data reasonably well,at least up to 20 °C, and has practical advantages wheninterpolating estimated between measured values of respirationin the construction of a carbon balance sheet. Lohum perenne L., ryegrass, respiration, temperature, Q₁₀, soluble carbohydrate content, simulated sward     

Fruit Number in Relation to Pollen Production and Viability in Groundnut Exposed to Short Episodes of Heat Stress

Hot days and warm nights are important environmental factorslimiting fruit yields of groundnuts in the semi-arid tropics.The objective of the present research was to quantify the effectsof short episodes of heat stress on pollen production and viability,and fruit yield. Plants of cultivar ‘ICGV 86015’were grown at a day/night temperature of 28/22 °C from sowinguntil 9 d after flowering. Cohorts of plants were then exposedto a factorial combination of four day (28, 34, 42 and 48 °C)and two night (22 and 28 °C) temperatures for 6 d. Thereafter,all plants were maintained at 28/22 °C until final harvest9 d later. Number of flowers per plant (FN), the proportionof flowers setting pegs (fruit-set), the number of pegs andpods per plant (reproductive number, RN_t), pollen productionper flower and pollen viability were determined during the 6d stress period. There were strong negative linear relationsbetween day temperature over the range of 28 to 48 °C andFN (slope, -1.1 °C^-1), fruit-set (-2.8% °C^-1), RN_t(-0.90°C^-1), and pollen production (-390 °C^-1) and viability(-1.9% °C^-1). Warmer night temperature (28 vs. 22 °C)had no effect on FN, but reduced fruit-set (31 to 19%), RN_t(8to 5), and pollen production (4389 to 2800) and viability (49to 40%). There were no significant interactions between dayand night temperature. Reduced fruit-set was a consequence offewer pollen grains and reduced pollen viability. The thresholdday temperature for pollen production and viability was 34 °Cand there were strong negative linear relations between bothpollen production and pollen viability and accumulated temperature>34 °C. Copyright 1999 Annals of Botany Company Arachis hypogaea L., fruit-set, groundnut, heat-stress, peanut, pollen viability, pollen production, temperature.