Uptake and Transport of Calcium and Phosphorus in Lolium perenne in Response to N Supplied to Halves of a Divided Root System

The uptake and transport of Ca²⁺ and HPO₄^2? from roots of Lolium perenne L. was studied using variable N nutrition supplied to halves of a divided root system. Plants were grown for 4 weeks in solution containing 0.11 mM NO₃^?–N; then one-half of the root system was supplied with either 4.0 mM NO₃^?–N or 0.28 mM NH₄⁺–N while the other half of the root system remained in low-N solution. Uptake and transport of Ca²⁺ increased and uptake of HPO₄^2? declined in root halves supplied with high NO₃^?–N for 16 h. After supply of high NO₃^?–N or NH₄⁺–N to half the root system for 6 days, the roots supplied with high-N exhibited significantly higher rates of uptake and percentage transport to shoots of both Ca²⁺ and HPO₄^2?–. However, in neither the 16-h nor 6-day treatment did Ca²⁺ or HPO₄^2? uptake of the root half supplied with low N differ significantly from the control (low N supplied to both halves of the root). Significantly higher N concentrations were found in low-N supplied roots (compared to the control) as a result of internal translocation of N from high-N supplied roots to low-N supplied roots. Although N concentration in the low-N supplied roots increased, uptake rates of Ca²⁺ or HPO₄^2? did not change implying that external N concentration may be the important factor which influences or governs N mediated uptake responses. This would further suggest that the site of uptake regulation for Ca²⁺ and HPO₄^2? exists on the outer plasma membrane which is in direct contact with the external solution. Transport of Ca²⁺ and HPO₄^2? to the shoot was generally increased in low-N root halves after 6 days of high-N supply to the other half of the root. This implies that plant growth demand may be a major factor in regulating rates of Ca²⁺ and HPO₄^2? transport from roots to the shoot. It also reinforces the hypothesis that uptake and transport of ions out of the root are separately controlled or regulated in the plant.     

Nitrate transport processes in Fagus- Laccaria-Mycorrhizae

The contribution of influx and efflux of NO₃ ^- on NO₃ ^- net uptake has been studied in excised mycorrhizae of 18–20 week old beech (Fagus sylvatica L.) trees. Net uptake rates of NO₃ ^- followed uniphasic Michaelis-Menten kinetics in the concentration range between 10 μM and 1.0 mM external NO₃ ^-, with an apparent K_m of 88±7 μM, and a V_max of 110±7 nmol g^-1 root f.wt. h^-1. The relative xylem loading of N, i.e. the portion of NO₃ ^- taken up that was loaded into the xylem vessels as NO₃ ^- plus reduced N, was constant over the concentration range tested (4.6–7.7%). NO₃ ^- influx proceeded linearly with increasing external NO₃ ^- supply. When the assumed regulators of net NO₃ ^- uptake, i.e. NH₄ ⁺ or L-glutamate, were applied together with NO₃ ^-, net uptake rates of NO₃ ^- decreased. This inhibitory effect was caused by a reduction of NO₃ ^- influx rather than an enhanced efflux. The comparison of the present data with a recent study with non-mycorrhizal beech roots (Kreuzwieser et al., 1997; J. Exp. Bot. 48, 1431–1438) revealed that mycorrhization leads to reduced rates of NO₃ ^- net uptake. This effect is caused by reduced influx plus enhanced efflux of NO₃ ^- as compared with non-mycorrhizal beech roots. This revised version was published online in June 2006 with corrections to the Cover Date.     

Transport and Distribution of 14CO2 Assimilate in Lolium perenne in Response to Varying Nitrogen Supply to Halves of a Divided Root System

The movement of ¹⁴C assimilate from shoots to roots and its subsequent metabolism in the root of Lolium perenne L. was studied using variable N nutrition supplied to halves of a divided root system. Half of the N-deficient root system was supplied with either high NO₃-N or high NH₄-N for 16 hours or 6 days before ¹⁴CO₂ labeling of the shoots. The distribution of ¹⁴C in sugars, ethanol-soluble nitrogen and organic acids in roots appeared to be related to the N content of the tissue. Supply of high NO₃-N for 6 days resulted in significant internal translocation of N into the low N supplied root half. Both root halves also had similar patterns of ¹⁴C distribution among soluble and insoluble metabolites. However, NH₄-N supply for 6 days did not result in a significant increase of N in the low N supplied roots, thus only the high NH₄-N supplied roots displayed stimulated sugar metabolism, similar to that in both root halves in the high NO₃-N supply treatment. Percent transport of ₁₄C assimilates from shoot to root was influenced by form and level of N supplied to root halves. Root halves supplied with either high N source for 6 days accumulated greater amounts of ¹⁴C assimilate than the corresponding low N root half. However NH₄-N supply appeared to make roots stronger sinks since NH₄ supply resulted in significantly greater ¹⁴C accumulation in both the high NH₄ supplied and the low N root halves than did NO₃-N supply in corresponding root halves. The data suggest that factors other than root metabolism, such as N mediated metabolism in the shoot, may also influence the percent transport of assimilates to the root. Internal distribution of the incoming assimilate within the root system could be regulated by the metabolic activity or assimilate demand of the roots.     

Uptake and Assimilation of NO(3) and NH(4) by Nitrogen-Deficient Perennial Ryegrass Turf

Assimilation of NO₃⁻ and NH₄⁺ by perennial ryegrass (Lolium perenne L.) turf, previously deprived of N for 7 days, was examined. Nitrogen uptake rate was increased up to four- to five-fold for both forms of N by N-deprivation as compared to N-sufficient controls, with the deficiency-enhanced N absorption persisting through a 48 hour uptake period. Nitrate, but not NH₄⁺, accumulated in the roots and to a lesser degree in shoots. By 48 hours, 53% of the absorbed NO₃⁻ had been reduced, whereas 97% of the NH₄⁺ had been assimilated. During the early stages (0 to 8 hours) of NO₃⁻ uptake by N-deficient turf, reduction occurred primarily in the roots. Between 8 and 16 hours, however, the site of reduction shifted to the shoots. Nitrogen form did not affect partitioning of the absorbed N between roots (40%) and shoots (60%) but did affect growth. Compared to NO₃⁻, NH₄⁺ uptake inhibited root, but not shoot, growth. Total soluble carbohydrates decreased in both roots and shoots during the uptake period, principally the result of fructan metabolism. Ammonium uptake resulted in greater total depletion of soluble carbohydrates in the root compared to NO₃⁻ uptake. The data indicate that N assimilation by ryegrass turf utilizes stored sugars but is also dependent on current photosynthate.     

The role of N-remobilisation and the uptake of NH4+ and NO3- by Lolium perenne L. in laminae growth following defoliation under field conditions

Several studies have previously shown that shoot removal of forage species, either by cutting or herbivore grazing, results in a large decline in N uptake (60%) and/or N₂ fixation (80%). The source of N used for initial shoot growth following defoliation relies mainly on mobilisation of N reserves from tissues remaining after defoliation. To date, most studies investigating N-mobilisation have been conducted, with isolated plants grown in controlled conditions. The objectives of this study were for Lolium perenne L., grown in a dense canopy in field conditions, to determine: 1) the contribution of N-mobilisation, NH₄ ⁺ uptake and NO₃ ^- uptake to growing shoots after defoliation, and 2) the contribution of the high (HATS) and low (LATS) affinity transport systems to the total plant uptake of NH₄ ⁺ and NO₃ ^-. During the first seven days following defoliation, decreases in biomass and N-content of roots (34% and 47%, respectively) and to a lesser extent stubble (18% and 43%, respectively) were observed, concomitant with mobilisation of N to shoots. The proportion and origin of N used by shoots (derived from reserves or uptake) was similar to data reported for isolated plants. Both HATS and LATS contributed to the total root uptake of NH₄ ⁺ and NO₃ ^-. The V_max of both the NH₄ ⁺ and NO₃ ^- HATS increased as a function of time after defoliation, and both HATS systems were saturated by substrate concentrations in the soil at all times. The capacity of the LATS was reduced as soil NO₃ ^- and NH₄ ⁺ concentrations decreased following defoliation. Data from ¹⁵N uptake by field-grown plants, and uptake rates of NH₄ ⁺ and NO₃ ^- estimated by excised root bioassays, were significantly correlated, though uptake was over-estimated by the later method. The results are discussed in terms of putative mechanisms for regulating N uptake following severe defoliation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Root morphological and proteomic responses to growth restriction in maize plants supplied with sufficient N

The primary objective of this study was to better understand how root morphological alteration stimulates N uptake in maize plants after root growth restriction, by investigating the changes in length and number of lateral roots, ¹⁵NO₃⁻ influx, the expression level of the low-affinity Nitrate transporter ZmNrt1.1, and proteomic composition of primary roots. Maize seedlings were hydroponically cultured with three different types of root systems: an intact root system, embryonic roots only, or primary roots only. In spite of sufficient N supply, root growth restriction stimulated compensatory growth of remaining roots, as indicated by the increased lateral root number and root density. On the other hand, there was no significant difference in ¹⁵NO₃⁻ influx between control and primary root plants; neither in ZmNrt1.1 expression levels in primary roots of different treatments. Our data suggested that increased N uptake by maize seedlings experiencing root growth restriction is attributed to root morphological adaptation, rather than explained by the variation in N uptake activity. Eight proteins were differentially accumulated in embryonic and primary root plants compared to control plants. These differentially accumulated proteins were closely related to signal transduction and increased root growth.     

Inter-varietal differences in xylem exudate composition and growth under contrasting forms of N supply in cucumber

Variations in the inorganic and organic composition of xylem exudate, growth and N content under contrasting forms of N supply in three cucumber cultivars (Hyclos, Medusa and Victory) were studied in glasshouse conditions. The plants were grown hydroponically with two NO₃ ^-:NH₄ ⁺ ratios (100:0 and 60:40).The xylem sap of Medusa grown with both N sources displayed an increase of organic N and carboxylate concentrations and a decrease of cations, inorganic anions and carbohydrates compared with that of those grown with NO₃ ^- alone, showing a higher growth and N content in tissues and thus better utilization of N supplied as NO₃ ^- and NH₄ ⁺. Mixed N nutrition in Hyclos caused the greatest amounts of NO₃ ^- and NH₄ ⁺ in xylem sap, lower root weight and N levels in the leaves, while its root was unable to generate an adequate supply of organic N compounds. Despite the levels of cations, inorganic and organic anions were reduced by the NH₄ ⁺ supplied to Victory, the ionic balance in the xylem sap, growth and N content remained similar to that of those supplied with NO₃ ^- alone. Finally, the cucumber cultivars studied here, responded differently to the form of N supplied, it may partly be due to their ability of assimilating N in the roots and partly to the form in which the N is translocated to the shoot.     

Exogenous NO(3) Influx and Endogenous NO(3) Efflux by Two Maize (Zea mays L.) Inbreds during Nitrogen Deprivation

The influence of nitrogen stress on net nitrate uptake resulting from concomitant ¹⁵NO₃⁻ influx and ¹⁴NO₃⁻ efflux was examined in two 12-day-old inbred lines of maize. Plants grown on ¹⁴NO₃⁻ were deprived of nitrogen for up to 72 hours prior to the 12th day and then exposed for 0.5 hour to 0.15 millimolar nitrate containing 98.7 atom% ¹⁵N. The nitrate concentration of the roots declined from approximately 100 to 5 micromolar per gram fresh weight during deprivation, and ¹⁴NO₃⁻ efflux was linearly related to root nitrate concentration. Influx of ¹⁵NO₃⁻ was suppressed in nitrogen-replete plants and increased with nitrogen deprivation up to 24 hours, indicating a dissipation of factors suppressing influx. Longer periods of nitrogen-deprivation resulted in a decline in ¹⁵NO₃⁻ influx from its maximal rate. The two inbreds differed significantly in the onset and extent of this decline, although their patterns during initial release from influx suppression were similar. Except for plants of high endogenous nitrogen status, net nitrate uptake was largely attributable to influx, and genetic variation in the regulation of this process is implied.     

Effects of pH and nitrogen on cadmium uptake in potato

This study investigated the effects of pH and nitrogen form and concentration on cadmium (Cd) uptake by potato (Solanum tuberosum L.) grown in hydroponic culture. Potato plants grown in a pH-buffered nutrient solution for 10 d were exposed for 24 h to 25 nM CdCl₂ labelled with ¹⁰⁹Cd. Plants showed a significantly higher Cd uptake and accumulation at pH 6.5 than at pH 4.5 and 5.5. Nitrogen supplied as nitrate (NO₃ ^?) generally resulted in a higher Cd uptake and accumulation than N supplied as ammonium (NH₄ ⁺). This effect was most pronounced at pH 6.5. The N concentration increasing from 6.5 to 26 mM resulted in a decreased Cd influx when either NO₃ ^? or NH₄ ⁺ was used. Cd translocation to the shoot was increased when NO₃ ^? was used as the sole N source. In conclusion, pH had a strong influence on Cd uptake by roots and N form is especially important for Cd translocation within the potato plant.     

Absorption of nitrate and ammonium ions by Lolium perenne from flowing solution cultures at low root temperatures

Lolium perenne L. cv. 23 (perennial ryegrass) plants were grown in flowing solution culture and acclimatized over 49 d to low root temperature (5°C) prior to treatment at root temperatures of 3, 5, 7 and 9°C for 41 d with common air temperature of 20/15°C day/night and solution pH 5·0. The effects of root temperature on growth, uptake and assimilation of N were compared with N supplied as either NH₄ or NO3 at 10 mmol m^?3. At any given temperature, the relative growth rate (RGR) of roots exceeded that of shoots, thus the root fraction (Rf) increased with time. These effects were found in plants grown with the two N sources. Plants grown at 3 and 5°C had very high dry matter contents as reflected by the fresh weight: freeze-dried weight ratio. This ratio increased sharply, especially in roots at 7 and 9°C. Expressed on a fresh weight basis, there was no major effect of root temperature on the [N] of plants receiving NHJ but at any given temperature, the [N] in plants grown with NHJ was significantly greater than in those grown with NO₃. The specific absorption rate (SAR) of NH⁺₄ was greater at all temperatures than SAR-NO₃. In plants grown with NH⁺, 3–5% of the total N was recovered as NH⁺₄, whereas in those grown with NO^?₃ the unassimilated NO^?₃ rose sharply between 7 and 9°C to become 14 and 28% of the total N in shoots and roots, respectively. The greater assimilation of NH⁺₄ lead to concentrations of insoluble reduced N (= protein) which were 125 and 20% greater, in roots and shoots, respectively, than in NO^?₃-grown plants. Plants grown with NH⁺₄ had very much greater glutamine and asparagine concentrations in both roots and shoots, although other amino acids were more similar in Concentration to those in NO^?₃ grown plants. It is concluded that slow growth at low root temperature is not caused by restriction of the absorption or assimilation of either NH⁺₄ or NO^?₃. The additional residual N (protein) in NH⁺₄ grown plants may serve as a labile store of N which could support growth when external N supply becomes deficient.     

Nitrogen nutrition of Myriophyllum spicatum: uptake and translocation of 15N by shoots and roots

Intact Myriophyllum spicatum plants were grown in compartmentalized containers in a growth room so that the roots were separated from the shoots by a watertight partition. Nitrogen ¹⁵N was added to the water or sediment to trace the uptake of inorganic N by the plant shoots or roots. Myriophyllum spicatum is capable of taking up inorganic N through both roots and shoots. Plant N requirements can apparently be met by root uptake alone. However, when about 0·1 mg/l of NH₄-N were present in the water, foliar uptake supplied more N to the plants than did root uptake. Foliar uptake of NH₄-N was found to be several times faster than that of NO₃-N when both forms of N were present in the water. Only about 1% of the N taken up by the roots was subsequently released to the water through the foliage.     

Regulation of nitrogen uptake on the whole plant level

The largest part of nitrogen requirements of crops is mostly covered by nitrate. The uptake of this ion is thermodynamically uphill and thus dependent on metabolism. This article considers regulation of N uptake in higher plants putting emphasis on NO₃ ^- and the whole plant level.In field conditions the transport rate depends on the concentration at the root surface in Michaelis-Menten-Kinetics. Maximum net influx of NO₃ ^- (I_max) was often reported at concentrations of 100 M NO₃ ^- and even lower. There are indications that for unrestricted growth the NO₃ ^- concentration at root surface has to be in the order of magnitude allowing I_max if plants are not able to compensate for lower NO₃ ^- concentrations by increasing root surface per unit of shoot.I_max is not a constant but depends for a given variety on N status of plants, the availability of NO₃ ^- and plant age. The decrease of I_max with increasing plant age is closely related to relative growth rate as long as the relationship between N demand and new growth is linear and the root:shoot ratio keeps constant. It seems that I_max is a meaningful physiological characteristic of NO₃ ^- uptake reflecting absolute N demand. There is evidence that shoot demand is linked to NO₃ ^- uptake of the root through an amino acid transport pool cycling in the plant via phloem and xylem.The N demand of a crop depends on increase of dry mass and might not be linear if the critical level of nitrogen in plant dry matter changes during crop development or if retranslocation of nitrogen from older leaves to meristematic tissue occurs. Radiation and temperature drive plant growth and thus N demand of crops. These relationships can be described by mathematical models.     

Responses of soybean to ammonium and nitrate supplied in combination to the whole root system or separately in a split-root system

To address the questions of whether allocation of carbohydrates to roots is influenced by ionic form of nitrogen absorbed and whether allocation of carbohydrates to roots in turn influences proportionality between NH₄⁺ and NO₃^? uptake from mixed sources, NH₄⁺ and NO₃^? were supplied separately to halves of a split-root hydroponic system and were supplied in combination to a whole-root system. Dry matter accumulation in the split-root system was 18% less in the NH₄⁺-fed axis than in the NO₃^?-fed axis. This, however, does not indicate that partitioning of carbohydrate between the two axes was different. Most of the reduction in dry matter accumulation in the NH₄⁺-fed axis can be accounted for by the retransport of CH₂O equivalents from the root back to the shoot with amino acids produced by NH₄⁺ assimilation. Uptake of NH₄⁺ or NO₃^? by the respective halves of the split-root system was proportional to the estimated allocation of carbohydrate to that half. When NH₄⁺ and NO₃^? were supplied to separate halves of the split-root system, the cumulative NH₄⁺ to NO₃^? uptake ratio was 0.81. When supplied in combination to the whole-root system, the cumulative NH₄⁺ to NO₃^? uptake ratio was 1.67. Thus, while the shoot may affect total nitrogen uptake through the export of carbohydrates to roots, the shoot (common for halves of the split-root system) apparently does not exert a direct effect on proportionality of NH₄⁺ and NO₃^? uptake by roots. For whole roots supplied with both NH₄⁺ and NO₃^?, the restriction in uptake of NO₃^? may involve a stimulation of NO₃^? efflux rather than an inhibition of NO₃^? influx. While only the net uptake of NH₄⁺ and NO₃^? was measured by ion chromatography, monitoring at approximately hourly intervals during the first 3 days of treatment revealed irregularly occurring intervals of both depletion (net influx) and enrichment (net efflux) in solutions. In the case of NH₄⁺, numbers of net efflux events were similar (21 to 24 out of 65 sequential sampling intervals) whether NH₄⁺ was supplied with NO₃^? to whole-root systems or separately to an axis of the split-root system. In the case of NO₃^?, however, the number of net efflux events increased from 8 when NO₃^? was supplied to a separate axis of the split-root system to between 19 and 24 when NO₃^? was supplied with NH₄⁺ to whole-root systems.     

Are phloem amino acids involved in the shoot to root control of NO3- uptake in Ricinus communis plants?

The putative role of phloem amino acids as negative feedback signals for root NO3^- uptake was investigated in Ricinus communis L. The NO3^--grown plants were subjected to N-deficiency due either to complete N-deprivation, or to localized N-deprivation on one side of a split-root system. In comparison with controls, complete N-deprivation resulted in a transient increase in ¹⁵NO3^- influx, and in profound changes in downward phloem transport of amino acids. Total amino acid concentration in the phloem sap decreased by 40%, but responses markedly differed between the individual amino acids. Concentrations of Gln and Ser were rapidly lowered by 50%, while those of Val, Phe, Leu, and Ile displayed a marked increase. Localized N-deprivation on one side of the split root system also resulted in the up-regulation of ¹⁵NO3^- influx in the roots still supplied with NO3^-. However, the amino acid composition of the phloem sap directed to these roots was not modified by the treatment, and remained similar to that in N-sufficient control plants. Only amino acid transport to the N-deprived roots was affected as observed in response to complete N-deprivation. The results from split-root plants indicate that the response of root NO3^- influx to N-deficiency is controlled by shoot-borne regulatory signals, and provide a case study where these signals are not related to a qualitative change or a significant decrease in downward phloem transport of amino acids.     

Effects of gypsum on the uptake,assimilation and cycling of 15N-labelled ammonium and nitrate-N by perennial ryegrass

Solution culture studies have shown that plant uptake of NH₄ ⁺ and NO₃ ^- can be improved by increasing the concentration of Ca²⁺ in the root environment: the same may be true for grass grown in soil culture. An experiment was set up to see whether gypsum (CaSO₄ 2H₂O) increased the rate at which perennial ryegrass absorbed ¹⁵NH₄ ⁺ and ¹⁵NO₃ ^- from soil.The results demonstrated that gypsum increases the rates of uptake of both NH₄ ⁺ and NO₃ ^- by perennial ryegrass. However because there was little potential for mineral-N loss from the experimental system, either by gaseous emission or by N immobilization, long term improvements in fertilizer efficiency were not observed. Nitrogen cycling from shoots to roots commenced once net uptake of N into plants had ceased. Labelled N transferred thus to roots underwent isotopic exchange with unlabelled soil N. It was suggested that this exchange of N might constitute an energy drain from the plant, if plant organic N was exchanged for soil inorganic N. The fact that the exchange occurred at all cast doubt on the suitability of the ¹⁵N-isotope dilution technique for assessing fertilizer efficiency in medium to long term experiments. There was evidence that the extra NO₃ ^--N taken up by plants on the all-nitrate treatments as a result of gypsum application, was reduced in root tissue rather than in shoots, but to the detriment of subsequent root growth and N uptake.     

Positive feedback between acidification and organic phosphate mineralization in the rhizosphere of maize (Zea mays L.)

To test the hypothesis that rhizosphere acidification would enhance the hydrolyzation of organic phosphates by increasing phosphatase activity. A Petri dish experiment with sterile agar and a pot experiment with a low P soil were used. In the Petri dish experiment, roots of each plant were cultured in two compartments, each of which contained agar with one of three nitrogen combinations: NH ₄ ⁺ /N0 (N0 = nitrogen free), NH ₄ ⁺ /NO ₃ ^- , and NO ₃ ^- /N0. Phytin was supplied as the sole phosphorus (P) source to all compartments. In the pot experiment, the soil in each pot was treated with N0, KNO₃, or (NH₄)₂SO₄) together with 0 or 75 mg kg^?1 phytin-P. Dry weight, P concentration, and P content of roots were highest in the NH ₄ ⁺ compartments in the Petri dish experiment. In the pot experiment, dry weight, P concentration, and P content of both shoots and roots were higher with NH ₄ ⁺ than with NO ₃ ^- . NH ₄ ⁺ treatments reduced rhizosphere pH, promoted the hydrolization of phytin, enhanced acid phosphatase activity in the rhizosphere, and increased phytin-P utilization relative to N0 and NO ₃ ^- treatments. Phosphatase activity was negatively correlated with rhizosphere pH but was positively correlated with plant P content in both experiments. Rhizosphere acidification optimized the activity of acid phosphatase excreted by maize roots and promoted phytin mineralization. NH ₄ ⁺ -induced acidification in the maize rhizosphere improved the growth of maize roots by improving P uptake from phytin; the improved growth, in turn, increased NH ₄ ⁺ uptake and acidification.     

Nitrate fluxes in soybean seedling roots and their response to amino acids: an approach using 15N

This study was undertaken to determine which of the two NO₃^? fluxes (influx or efflux) across plasma membranes of root cells is the target of those amino acids which have been shown to inhibit net NO₃^? uptake (Muller & Touraine 1992, Journal of Experimental Botany 43 , 617–623). Parallel experiments were performed to mea-sure either the time course of ¹⁵NO₃^? release from roots of soybean seedlings previously labelled with this isotope into non-labelled solution, or the time course of ¹⁵N accumulation from labelled ¹⁵NO₃^? solution in non-labelled seedlings. Focusing on the fate of ¹⁵NO₃^? in the cytoplasmic compartment, a model is developed to describe the time courses of the accumulation and release of tracer across the plasma membranes of root cells. Both time courses can be described by the sum of an exponential plus a linear term. In our material, the linear part of the accumulation time course is obscured by the NO₃^? fluxes exiting the cytoplasm, and the curve thus appears to be quasilinear over several minutes. However, we show that the use of the net tracer accumulation rate during this time period as an estimate of NO₃^? influx does not provide accurate estimates of influx and efflux. By contrast, ¹⁵NO₃^? efflux analysis permits calculation of the unidirectional fluxes across plasma membranes of root cells and the kinetic parameters of the cytoplasmic NO₃^? pool. Under our experimental conditions, efflux accounted for 30 to 50% of influx, and the cytoplasmic NO₃^? content was found to be in the 70–400nmol g^?1 fw range. Using this methodology, the effect of amino acid accumulation on unidirectional fluxes of nitrate was then examined. Pretreatments of the seedlings with an amino acid which has been shown to inhibit net NO₃^? uptake led to concomitant decreases in net accumulation rates of ¹⁵NO₃^? and of reduced ¹⁵N in roots and total ¹⁵N in cotyledons. NO₃^? influx was markedly inhibited by these treatments, while NO₃^? efflux remained essentially unaffected, or even decreased. It is concluded that the target of the regulation of NO₃^? uptake by phloemtranslocated amino acids is the influx system.     

Influx,efflux and net uptake of nitrate in Quercus suber seedlings

Nitrate influx, efflux and net nitrate uptake were measured for the slow-growing Quercus suber L. (cork-oak) to estimate the N-uptake efficiency of its seedlings when grown with free access to nitrate. We hypothesise that nitrate influx, an energetically costly process, is not very efficiently controlled so as to avoid losses through efflux, because Q. suber has relatively high respiratory costs for ion uptake. Q. suber seedlings were grown in a growth room in hydroponics with 1 mM NO₃ ^-. Seedlings were labelled with ¹⁵NO₃ ^- in nutrient solution for 5 min to measure influx and for 2 h for net uptake. Efflux was calculated as the difference between influx and net uptake. Measurements were made in the morning, afternoon and night. The site of nitrate reduction was estimated from the ratio of NO₃ ^- to amino acids in the xylem sap; the observed ratio indicated that nitrate reduction occurred predominantly in the roots. Nitrate influx was always much higher than net acquisition and both tended to be lower at night. High efflux occurred both during the day and at night, although the proportion of ¹⁵NO₃ ^- taken up that was loss through efflux was proportionally higher during the night. Efflux was a significant fraction of influx. We concluded that the acquisition system is energetically inefficient under the conditions tested. This revised version was published online in June 2006 with corrections to the Cover Date.     

Differential effects of mineral and organic N sources,and of ectomycorrhizal infection by Hebeloma cylindrosporum,on growth and N utilization in Pinus pinaster

The effect of ectomycorrhizal association of Pinus pinaster with Hebeloma cylindrosporum was investigated in relation to the nitrogen source supplied as mineral (NH₄⁺ or NO₃^?) or organic N (L ‐glutamate) and at 5 mol m^?3. Plants were grown for 14 and 16 weeks with mineral and organic N, respectively, and samples were collected during the last 6 weeks of culture. Total fungal biomass was estimated using glucosamine amount and its viability was assessed using the glucosamine to ergosterol ratio. Non‐mycorrhizal plants grew better with NH₄⁺ than with NO₃^? and grew very slowly when supplied with L ‐glutamate. The presence of the fungus decreased the growth of the host plant with mineral N whereas it increased it with L ‐glutamate. Whatever the N source, most of the living fungal biomass was associated with the roots, whereas the main part of the total biomass was assayed outside the root. The form of mineral N did not significantly affect N accumulation rates over the 42 d in control plants. In mycorrhizal plants grown on either N source, the fungal tissues developing outside of the root were always the main N sink. The ectomycorrhizal association did not change ¹⁵NH₄⁺ uptake rate by roots, suggesting that the growth decrease of the host‐plant was related to the carbon cost for fungal growth and N assimilation rather than to a direct effect on NH₄⁺ acquisition. In contrast, in NO₃^?‐grown plants, in addition to draining carbon for NO₃^? reduction the fungus competed with the root for NO₃^? uptake. With NH₄⁺ or NO₃^? feeding, although mycorrhizal association improved N accumulation in shoots, we concluded that it was unlikely that the fungus had supplied the plant with N. In L ‐glutamate‐grown plants, the presence of the fungus increased the proportion of glutamine in the xylem sap and improved both N nutrition and the growth rate of the host plant.