Diurnal variation of root respiration rates and nitrate uptake as influenced by nitrogen supply

Root respiration rates of Lolium multiflorum supplied with nitrate or ammonium were measured continuously during several days (Exp. A). Net uptake rate of nitrate was similarly measured by an ion selective nitrate electrode in a system of flowing nutrient solution (Exp. B). Diurnal variation of in vitro nitrate reductase activity and nitrate content of tops and roots were determined (Exp. C). Two levels of irradiance were applied throughout, with day:night of 16:8 h. Root respiration rates showed diurnal patterns, most pronounced in the nitrate treatment, with two peaks appearing about 6 and 16 h after commencement of the photoperiod. Respiration rates were highest in the nitrate treatment and at high irradiance. Respiration rates fell after removal of nitrogen, particularly in the nitrate supplied plant and at high irradiance. Net uptake rate of nitrate exhibited diurnal patterns, often with two peaks occurring at the same times as those of respiration rates. In vitro nitrate reductase activity of tops increased steeply 16 h after commencement of the photoperiod and remained at the high level during the following 8 h of darkness. Nitrate content of tops was highest during the 8 h dark period and fell at the start of the photoperiod. Possible controlling systems of the apparent coincidences of diurnal variation rates, net nitrate uptake and nitrate reduction are discussed.     

Relatively large nitrate efflux can account for the high specific respiratory costs for nitrate transport in slow-growing grass species

In this paper we address the question why slow-growing grass species appear to take up nitrate with greater respiratory costs than do fast-growing grasses when all plants are grown with free access to nutrients. Specific costs for nitrate transport, expressed as moles of ATP per net mole of nitrate taken up, were 1.5 to 4 times higher in slow-growing grasses than in fast-growing ones (Scheurwater et al., 1998, Plant, Cell & Environ. 21, 995–1005). The net rate of nitrate uptake is determined by two opposing nitrate fluxes across the plasma membrane: influx and efflux. To test whether differences in specific costs for nitrate transport are due to differences in the ratio of nitrate influx to net rate of nitrate uptake, nitrate influx and the net rate of nitrate uptake were measured in the roots of two fast-growing ( Dactylis glomerata L. and Holcus lanatus L.) and two slow-growing (Deschampsia flexuosa L. and Festuca ovina L.) grass species at four points during the diurnal cycle, using ¹⁵NO₃ ^-. Efflux was calculated by subtraction of net uptake from influx; it was assumed that efflux of nitrogen represents the flux of nitrate. Transfer of the plants to the solution containing the labelled nitrate did not significantly affect nitrate uptake in the present grass species. The net rate of nitrate uptake was highest during the middle of the light period in all species. Diurnal variation in the net rate of nitrate uptake was mostly due to variation in nitrate influx. Variation in nitrate efflux did not occur in all species, but efflux per net mole of nitrate taken up was higher during darkness than in the light in the slow-growing grasses. The two fast-growing species, however, did not show diurnal variation in the ratio of efflux to net nitrate uptake. Integrated over 24 hours, the slow-growing grasses clearly exhibited higher ratios of influx to net uptake than the fast-growing grass species. Our results indicate that the higher ratio of nitrate influx to net nitrate uptake can account for higher specific costs for nitrate transport in slow-growing grass species compared with those in their fast-growing counterparts, possibly in combination with greater activity of the non-phosphorylating alternative respiratory path. Therefore, under our experimental conditions with plants grown at a non-limiting nitrate supply, nitrate uptake is less efficient (from the point of ATP consumption) in slow-growing grasses than in fast-growing grass species. This revised version was published online in June 2006 with corrections to the Cover Date.     

Daily changes in nitrate influx,efflux and metabolism in maize and pearl millet

Maize (Zea mays L.) and pearl millet (Pennisetum americanum (L.) Leeke) seedlings were exposed to [¹⁵N]nitrate for 1-h periods at eight times during a 24-h period (16–8 h light-dark for maize; 14–10 h for millet). Influx of [¹⁵N]nitrate as well as its reduction and translocation were determined during each period. The efflux of previously absorbed [¹⁴N]nitrate to the uptake solution was also estimated. No marked diurnal changes in [¹⁴N]nitrate efflux or [¹⁵N]nitrate influx were evident in maize. In contrast, [¹⁴N]nitrate efflux from millet increased and eventually exceeded [¹⁵N]nitrate influx during the late dark and early light periods, resulting in net nitrate efflux from the roots. The dissimilarity of their diurnal patterns indicates that influx and efflux are independently regulated. In both species, [¹⁵N]nitrate reduction and ¹⁵N translocation to shoots were curtailed more by darkness than was [¹⁵N]nitrate influx. In the light, maize reduced 15% and millet 24% of the incoming [¹⁵N]nitrate. In darkness, reduction dropped to 11 and 17%, respectively. Since the accumulation of reduced-¹⁵N in shoots declined abruptly in darkness, whereas that in roots was little affected, it is suggested that in darkness [¹⁵N]nitrate reduction occurred primarily in roots. The decrease in nitrate uptake and reduction in darkness was not related to efflux, which remained constant in maize and did not respond immediately to darkness in pearl millet.Paper No. 6722 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh     

Metabolism of carbon and nitrogen by soybean seedlings in response to vegetative apex removal

Short-term (31-hour diurnal) growth-chamber studies were conducted to determine the effects of removing the vegetative apex (meristem and developing trifoliolate leaves) on net photosynthesis (changes in plant dry weight), on distribution of metabolites among plant parts, and on nitrate metabolism and reduced-N accumulation by soybean [Glycine max (L.) Merr.] seedlings. Roots and stems served as alternate sinks for dry matter accumulation in the absence of the vegetative apex. Sugar concentration in roots increased (42%) within 4 hours of vegetative apex removal, and remained higher than for the controls during the 31-hour experimental period. Nitrate assimilation (nitrate reductase activity and total accumulation of reduced-N) was also enhanced in response to vegetative apex removal. Although dry matter accumulation was similar between treated and control plants (113 versus 116 milligrams per plant) over the 31-hour sampling period, more nitrate (1.31 versus 0.79 milligrams per plant) and more reduced-N (3.96 versus 3.45 milligrams per plant) accumulated in treated plants during the same interval. It was concluded that vegetative apex removal had little effect on overall net photosynthesis of soybean seedlings during the 31-hour treatment period, but did alter partitioning of photosynthate and enhanced uptake, transport, and reduction of nitrate. Implications are that uptake and metabolism of nitrate by soybeans may be limited by flux of carbohydrate to the roots, although hormonal effects due to vegetative apex removal cannot be ruled out.     

Nitrate content and nitrate reductase activity in Rumex obtusifolius L.

Summary With Rumex obtusifolius L., the influence of some environmental conditions on nitrate uptake and reduction were investigated. Nitrate concentrations of plant material were determined by HPLC, the activity of nitrate reductase by an in vivo test. As optimal incubation medium, a buffer containing 0.04 M KNO₃; 0.25 M KH²PO₄; 1.5% propanol (v/v); pH 8.0 was found. Vacuum infiltration caused an increase of enzyme activity of up to 40%.High nitrate concentrations were found in roots and leaf petioles. Nitrate reductase activity of these organs, however, was low. On the other hand, the highest nitrate reductase activity was observed in leaf laminae, which contained lowest nitrate concentrations.In leaves, nitrate content and nitrate reductase activity exhibited inverse diurnal fluctuations. During darkness, decreasing activities of the enzyme were followed by increasing nitrate concentrations, while during light the contrary was true. In petioles diurnal fluctuations in nitrate content were observed, too. No significant correlations with illumination, however, could be found.Our results prove that Rumex obtusifolius is characterized by an intensive nitrate turnover. Theoretically, internal nitrate content of the plant would be exhausted within a few hours, if a supply via the roots would be excluded.     

Daily Changes in CO(2) and Water Vapor Exchange, Chlorophyll Fluorescence, and Leaf Water Relations in the Halophyte Mesembryanthemum crystallinum during the Induction of Crassulacean Acid Metabolism in Response to High NaCl Salinity

Simultaneous measurements of net CO₂ exchange, water vapor exchange, and leaf water relations were performed in Mesembryanthemum crystallinum during the development of crassulacean acid metabolism (CAM) in response to high NaCl salinity in the rooting medium. Determinations of chlorophyll a fluorescence were used to estimate relative changes in electron transport rate. Alterations in leaf mass per unit area, which—on a short-term basis—largely reflect changes in water content, were recorded continuously with a beta-gauge. Turgor pressure of mesophyll cells was determined with a pressure probe. As reported previously (K Winter, DJ von Willert [1972] Z Pflanzenphysiol 67: 166-170), recently expanded leaves of plants grown under nonsaline conditions showed gas-exchange characteristics of a C₃ plant. Although these plants were not exposed to any particular stress treatment, water content and turgor pressure regularly decreased toward the end of the 12 hour light periods and recovered during the following 12 hours of darkness. When the NaCl concentration of the rooting medium was raised to 400 millimolar, in increments of 100 millimolar given at the onset of the photoperiods for 4 consecutive days, leaf water content and turgor pressure decreased by as much as 30 and 60%, respectively, during the course of the photoperiods. These transient decreases probably triggered the induction of the biochemical machinery which is required for CAM to operate. After several days at 400 millimolar NaCl, when leaves showed features typical of CAM, overall turgor pressure and leaf mass per unit area had increased above the levels before onset of the salt treatment, and diurnal alterations in leaf water content were reduced. Net carbon gain during photoperiods and average intercellular CO₂ partial pressures at which net CO₂ uptake occurred, progressively decreased upon salinization. Reversible diurnal depressions in leaf conductance and net CO₂ uptake, with minima recorded in the middle of the photoperiods, preceded the occurrence of nocturnal net CO₂ uptake. During these reductions, intercellular CO₂ partial pressure and rates of photosynthetic electron transport decreased. With advancing age, leaves of plants grown under nonsaline conditions exhibited progressively greater diurnal reductions in turgor pressure and developed a low degree of CAM activity.     

Diurnal variations of intracellular nitrate storage by marine diatoms: effects of nutritional state

Nitrate uptake and accumulation were measured in N-sufficient, N-limited, and 24 h N-starved cells of Phaeodactylum tricornutum Bohlin and Skeletonema costatum Grev., growing under a light-dark cycle. In N-sufficient cells the uptakerate was reduced at night and showed possible variation during the light period. In N-limited and N-starved cells such diurnal changes in uptake were absent, except extremely rapid, but short-lived nitrate uptake was observed early in the morning in N-limited cells. The nitrate accumulation inside the cells reflects a transient uncoupling between uptake and reduction mostly due to the light-dark cycle and strongly influenced by the physiological state of the cells. This accumulation is high during the night and at the beginning of the day, but decreases during the light period in N-sufficient cells. On the other hand, nitrate storage in N-sufficient and N-limited cultures shows a strong diurnal pattern, with maximum accumulation, suggesting the greatest uncoupling between uptake and assimilation, in the morning. In N-starved cells, accumulation is high and constant during the entire light period. Consequently, the uncoupling between nitrate uptake and reduction decreases during the light period but increases with N deficiency. These results indicate the importance of light periodicity and nutritional state of the cells on the nitrate utilization. They reveal the need for more systematic studies on N dynamics in relation to nutrient-light regimes.     

Growth requirement for N as a criterion to assess the effects of physical manipulation on nitrate uptake fluxes in spinach

The effects of physical manipulation of hydroponically grown plants of spinach (Spinacia oleracea L., cvs Subito and Glares) on nitrate uptake fluxes were studied in a long-term experiment (3 days), and in short-term label experiments (2 h) with ¹³N-nitrate and ¹⁵N-nitrate. In the long-term experiment, net nitrate uptake rate (NNUR) was measured by following the nitrate depletion in the uptake solution, which was replaced at regular intervals. In the short-term experiments, NNUR and nitrate influx were measured by simultaneous application of ¹³N-nitrate and ¹⁵N-nitrate. Plants were gently transferred into the labelled uptake solution, as is usually done in nutrient uptake studies. In addition, a more severe physical manipulation was carried out, including blotting of the roots, to mimic pretreatments which involve more handling of the plants prior to uptake measurements. Nitrate influx was measured immediately after physical manipulation and after 2 h of recovery. To assess the impact of the physical manipulation the experimentally determined nitrate uptake fluxes were compared with the N demand for growth, defined as relative growth rate (RGR) times plant nitrogen concentration (PNC) of parallel plants, which were left undisturbed. Nitrate influx and efflux were both subject to changes after physical manipulation of the plants. Physical handling, however, did not always result in an alteration of NNUR, which complicates the determination of the length of the recovery period. The impact of the handling and the time course of the recovery depended on the severity of the disturbance and were independent of the light conditions during the experiments. Even after a gentle transfer of the plants, recovery, in most cases, was not complete within 2 h. The data emphasise the need for minimal disturbance of plants during the last hours prior to nutrient uptake measurements.     

Modulation of NO 3 - uptake by water-extractable humic substances: involvement of root plasma membrane H+ATPase

The effect of a water extractable humic substances fraction (WEHS) on nitrate uptake and plasma membrane (pm) H⁺-ATPase activity of maize roots was investigated. Four days old maize root seedlings were exposed for 4 to 24 h to a nutrient solution containing 200 μ M nitrate in the absence or presence of 5 mg org. C { L ^-1 WEHS. Plants exposed to nitrate developed a higher capacity to absorb the anion (induction): the net uptake rate progressively increased up to 12 h of contact with the solution; thereafter, a decline was observed. When WEHS was present together with nitrate in the nutrient solution, the induction of nitrate uptake was evident and maximal already 4 h after starting the treatment. The rate of net nitrate uptake decreased only slightly during the remaining period (4-24 h). Stimulation of net nitrate uptake rate was also observed when WEHS was added to a nitrogen- or nitrate-free nutrient solution or to a 5 mM CaSO₄ solution. The activity of pmH⁺-ATPase raised upon exposure of the roots to nitrate with the same pattern observed for nitrate uptake. The contemporary presence of nitrate and WEHS caused a further stimulation of the pmH⁺-ATPase activity after 4 h treatment. An increase in the enzyme activity was also observed when plants were treated for 4 h in the presence of WEHS in CaSO₄, nitrogen- or nitrate-free solutions. However, when nitrate was present the enhancement was even greater. Results support the idea that the plasma membrane proton pump might be one of the primary targets of the action of humic substances on plant nutrient acquisition. A role of WEHS in the modulation of nitrate uptake via an interaction with the pm H⁺-ATPase is also discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

STIMULATION OF NITRATE NET UPTAKE AND REDUCTION BY RED AND BLUE LIGHT AND REVERSION BY FAR-RED LIGHT IN THE GREEN ALGA ULVA RIGIDA1

The steady-state levels of nitrate, nitrite, and ammonium were estimated in the green alga Ulva rigida C. Agardh in darkness after addition of 0.5 mM KNO₃ and irradiation with red (R) and blue (B) light pulses of different duration (5 and 30 min). The net uptake of nitrate was very rapid. Seventy-five percent of the nitrate added was consumed after 60 min in darkness. Although uptake was stable after R or B, efflux of nitrate occurred within 3 h in the dark control and when R or B were followed by far-red (FR) irradiation. The internal nitrate concentration after 3 h in darkness was similar after R and B light pulses; however, the intracellular ammonium was higher after R than after B. The intracellular nitrate and ammonium decreased when FR tight pulses were applied immediately after R or B. Thus, the involvement of phytochrome in the transport of nitrate and ammonium is proposed. Nitrate reductase activity, measured by the in situ method, was increased by both R and B light pulses. The effect was partially reversed by FR light. Nitrate reductase activity was higher after 5 min of R light than after 5 min of B. However, after 30-min light pulses, the relative increase in activity was reversed for R and B. We propose that phytochrome and a blue-light photoreceptor are involved in regulation of nitrogen metabolism. Nitrate uptake and reduction correlates with previously detected light-regulated accumulation of protein in Ulva rigida under the same experimental conditions.     

Daily changes in uptake, reduction and storage of nitrate in spinach grown at low light intensity

Under poor light conditions, as normally used during winter production of greenhouse vegetables, the nitrate concentration in the shoot of spinach ( Spinacia oleracea L. cv. Vroeg Reuzenblad) showed a diurnal rhythm. This rhythm was mainly caused by a decrease during the day, followed by an increase during the night in the leaf blade nitrate concentration. Nitrate was mainly located in the vacuoles of the leaf blades. A strong correlation was found between net uptake of nitrate by the roots and the nitrate concentration in the leaf blade vacuoles. The nitrate concentration in the leaf blades increased during the initial hours of the night. This increase was caused by a marked increase in the net uptake rate of nitrate by the roots during the first hours of the dark period. During the second part of the night both net uptake rate of nitrate by the roots and the vacuolar nitrate concentration in the leaf blades remained constant.
We conclude that nitrate is taken up for osmotic purposes when light conditions are poor because of a lack of organic solutes. During the night, nitrate influx into the vacuole is needed for replacement of organic solutes, which are metabolized during the night, and possibly also for leaf elongation growth. During the day, vacuolar nitrate may be exchanged for newly synthesized organic solutes and be metabolized in the cytoplasm. A strong diurnal rhythm in nitrate reductase (NR; EC 1.6.6.1.) activity was absent, due to the poor light conditions, and in vitro NR activity was not correlated with nitrate flux from the roots. In vivo NR activity also lacked a strong diurnal rhythm, but it was calculated that in situ nitrate reduction was much lower during the night, so that the major nitrate assimilation took place during the day.     

Nitrate Uptake during Recovery from Nitrogen Deficiency

Two-week-old nitrogen-deficient wheat plants attained a high rate of nitrate uptake on the first day of exposure to nutrient solutions supplemented with KNO₃. Ammonium uptake from similar solutions supplemented with NH₄NO₃ was also high during the first day of exposure, but nitrate uptake from this solution was lower than from the KNO₃ treatment. During the next two to three days there was a progressive decrease in uptake of both nitrogen ions. A steady increase in uptake then occurred as the plants fully recovered from the nitrogen-deficient state. The transient low nitrate uptake after three or four days of exposure to KNO₃ was not due to an excessive accumulation of nitrate in the tissue, nor to a failure in nitrate reduction as indicated by the rate of nitrate accumulation relative to the uptake rate. Nitrogen supplied as ¹⁵N-nitrite during the low uptake period was effectively incorporated into organic forms and effectively translocated to the shoots. Failure of the root tissue to increase in soluble carbohydrates during illumination was characteristic of the low uptake period. This contrasted with an increase in root soluble carbohydrates in the light during rapid uptake associated with full recovery from the nitrogen-deficient state. It is concluded that carbohydrate translocation to the root system was insufficient during the intermediate recovery period for optimal nitrate uptake, although it was sufficient for effective reduction and translocation of nitrate and reduced nitrogen. Ammonium uptake from NH₄NO₃ was restricted during darkness by the third day whereas there was little difference between light and dark periods in nitrate uptake from KNO₃ until about the sixth day of recovery. The extent to which ammonium restricted nitrate uptake increased progressively for two or three days following which a lessening influence seemed evident, and the effects were not directly associated with the rate of ammonium uptake.     

Nitrogen Utilization in Lemna: II. Studies of Nitrate Uptake Using NO(3)

¹³N-labeled nitrate was used to trace short-term nitrate influx into Lemna gibba L. G3 in experiments where disappearance of both radioactivity and total nitrate from the incubation medium was measured continuously and simultaneously. In plants performing net nitrate uptake from an initial nitrate concentration of 40 to 60 micromolar, there was no discrepancy between net uptake and influx, irrespective of the N status of the plants, indicating that concomitant nitrate efflux was low or nil. Plants treated with tungstate to inactivate nitrate reductase were able to take up nitrate following induction of the uptake system by exposure to a low amount of nitrate. Also, in this case, net uptake was equivalent to influx. In tungstate-treated plants preloaded with nitrate, both net uptake and influx were nil. In contrast to these observations, a clear discrepancy between net uptake and influx was observed when the plants were incubated at an initial nitrate concentration of approximately 5 micromolar, where net uptake is low and eventually ceases. It is concluded that plasmalemma nitrate transport is essentially unidirectional in plants performing net uptake at a concentration of 40 to 60 micromolar, and that transport is nil when internal nitrate sinks (vacuole, metabolism) are eliminated. The efflux component becomes increasingly important when the external concentration approaches the threshold value for net nitrate uptake (the nitrate compensation point) where considerable exchange between internal and external nitrate occurs.     

Nitrogen Utilization in Lemna: II. Studies of Nitrate Uptake Using 13NO3?

¹³N-labeled nitrate was used to trace short-term nitrate influx into Lemna gibba L. G3 in experiments where disappearance of both radioactivity and total nitrate from the incubation medium was measured continuously and simultaneously. In plants performing net nitrate uptake from an initial nitrate concentration of 40 to 60 micromolar, there was no discrepancy between net uptake and influx, irrespective of the N status of the plants, indicating that concomitant nitrate efflux was low or nil. Plants treated with tungstate to inactivate nitrate reductase were able to take up nitrate following induction of the uptake system by exposure to a low amount of nitrate. Also, in this case, net uptake was equivalent to influx. In tungstate-treated plants preloaded with nitrate, both net uptake and influx were nil. In contrast to these observations, a clear discrepancy between net uptake and influx was observed when the plants were incubated at an initial nitrate concentration of approximately 5 micromolar, where net uptake is low and eventually ceases. It is concluded that plasmalemma nitrate transport is essentially unidirectional in plants performing net uptake at a concentration of 40 to 60 micromolar, and that transport is nil when internal nitrate sinks (vacuole, metabolism) are eliminated. The efflux component becomes increasingly important when the external concentration approaches the threshold value for net nitrate uptake (the nitrate compensation point) where considerable exchange between internal and external nitrate occurs.     

Effects of benomyl and drought on the mycorrhizal development and daily net CO2 uptake of a wild platyopuntia in a rocky semi-arid environment

The effects of drought and the fungicide benomyl on a wild platyopuntia, Opuntia robusta Wendl., growing in a rocky semi-arid environment were assessed. Cladode phosphorus content, cladode water potential and daily net CO2 uptake were measured monthly in 2000 and 2001 before, during and after the summer rainy period. During 2000, the formation of new roots and new cladodes was severely suppressed in response to a prolonged drought, impairing the development of the symbiotic relationship between the arbuscular mycorrhizal (AM) fungi and the roots. Hence no effect of benomyl application was observed on daily carbon assimilation by this Crassulacean acid metabolism plant. During 2001, drought was interrupted, and new cladodes and roots were formed in response to rainfall. Benomyl was highly effective in suppressing root colonization by AM-fungi; however, daily C assimilation was reduced by benomyl application only in October. Thus, the inhibition of AM-fungal colonization by benomyl did not affect photosynthesis, water uptake and P uptake under prolonged drought.     

Growth and development of seminal and crown root systems in N-limited barley,and their contributions to nitrate acquisition during vegetative and generative growth

Barley (Hordeum vulgare L., cvs Golf and Laevigatum) was grown under nitrogen limitation, controlled by the relative rate of nitrate-N addition (RA), in solution culture. The seminal and crown root systems were kept apart, but in contact with the same nutrient solution throughout culturing. Growth, nitrate uptake, and in vitro nitrate reductase (NR) activity in the different root parts were studied at plant ages from 40 (late vegetative stage) to 110 (mid grain-filling) days. The RA was during this time interval stepwise decreased from 0.08 day^–1 to 0.005 day^–1. The ratio between seminal root dry weight and total plant dry weight decreased drastically during post-anthesis growth, whereas the contribution by crown roots remained unchanged. Tissue nitrogen concentrations in seminal roots did not change with time, but decreased in crown roots after day 80. The NR activity decreased with age in both seminal and crown roots. The V_max for net nitrate uptake decreased throughout the experiment in the seminal root system, but not in the crown root system. The kinetic properties (V_max and K_M) were used to calculate the nitrate concentration required to maintain a relative rate of nitrate-N uptake that equals the relative addition rate. These concentrations (2 to 5 mmol m^–3) were found to closely match actually measured nitrate concentrations in the nutrient solution (1 to 6 mmol m^–3). From uptake kinetics, it was deduced that the contribution by seminal roots to total nitrate uptake at these concentrations decreased from more than 50% in vegetative plants, to about 20% just after main shoot anthesis, and to less than 5% during grain-filling. ei]Section editor: H Lambers     

Increase in nitrate uptake by soybean plants during interruption of the dark period with low intensity light

Diurnal patterns of net NO⁻₃ uptake by nonnodulated soybean [ Glycine max (L.) Merr. cv. Ransom] plants growing in flowing hydroponic culture at 26 and 16°C root temperatures were measured at hourly intervals during alternate days of a 12-day growth period. Ion chromatography was used to determine removal of NO⁻₃ from the culture solution. Day and night periods of 9 and 15 h were used during growth. The night period included two 6-h dark periods and an intervening 3-h period of night interruption by incandescent lamps to effect a long-day photoperiod and repress floral initiation. At both root temperatures, the average specific rates of NO⁻₃ uptake were twice as great during the night interruption period as during the day period; they were greater during the day period than during the dark periods; and they were greater during the dark period immediately following the day period than during the later dark period that followed the night interruption. While these average patterns were repetitious among days, measured rates of uptake varied hourly and included intervals of net efflux scattered through the day period and more frequently through the 2 dark periods. Root temperature did not affect the average daily specific rates of uptake or the qualitative relationships among day, dark and night interruption periods of the diurnal cycle.     

Comparative uptake of nitrate by intact seedlings of C3 (barley) and C4 (corn) plants: Effect of light and exogenously supplied sucrose

The effect of light and exogenously supplied sucrose on NO₃ ⁻ uptake was studied in 9-day-old intact C₃ (barley) and C₄ (corn) seedlings. The seedlings used were uninduced for nitrate uptake system (i.e. had never seen nitrogen during germination and growth) and were exposed to continuous light for 3 days to avoid any diurnal variation and to load the seedlings fully with photosynthates. The uptake assay was conducted either in light or in darkness. Prior to assay, seedlings were treated with darkness or light for 24 h. Accordingly, four sets of seedlings, i.e. pretreated with light and assayed in light (LL); pretreated and assayed in darkness (DD); pretreated with light and assayed in darkness (LD); and pretreated with darkness and assayed in light (DL) were formed. Barley exhibited 55% higher NO₃ ⁻ uptake than corn during light (LL) and 91% higher during darkness (DD). Shifting barley seedlings from light to dark (LD) or dark to light (DL) for uptake assay, did not affect NO₃ ⁻ uptake, i.e. in LD the uptake was similar to LL and in DL it was similar to DD. However, in corn, the light conditions during the assay determined the uptake regardless of the conditions during the period preceding the assay. One percent sucrose in the medium increased NO₃ ⁻ uptake by 31% in barley and 70% in corn during light (LL). The corresponding increase during darkness (DD) was 38% in both barley and corn. Removal of the corn residual endosperm decreased NO₃ ⁻ uptake by 40% during darkness. Etiolated seedlings (those having never seen light) of both barley and corn were able to take up significant amount of NO₃ ⁻ during darkness. Externally supplied sucrose in the assay medium of etiolated seedlings increased the NO₃ ⁻ uptake to about 4 and 2 fold in barley and corn, respectively. The data presented here provide evidence that: 1. In intact seedlings, light per se is not obligatory for NO₃ ⁻ uptake and that the carbohydrate supply may mimic light. 2. Light affected the NO₃ ⁻ uptake differently in barley and corn. This revised version was published online in June 2006 with corrections to the Cover Date.