Fractionation of Nitrogen Isotopes during the Uptake and Assimilation of Ammonia by Plants

Two varieties (Nihonbare and Koshihikari) of rice plants (Oryzasativa L.) were grown hydro-ponically with two levels (20 and100 mg N liter ^–1) of ammonia. Variations in levels ofnatural abundance of ¹⁵N (¹⁵N) were analyzed in the ammoniaand organic nitrogen of shoots and roots, as well as in theammonia in the culture solution. There was substantial fractionationof nitrogen isotopes during the uptake of ammonia. When plantsabsorbed a large proportion of ammonia from a solution witha low concentration, less negative ¹⁵N values in plants andhigh positive ¹⁵N values in the ammonia remaining in solutionwere observed. The reverse was found when a smaller fractionof ammonia was absorbed from a solution with a higher concentrationof ammonia. The ^l5N values of ammonia in shoots and roots werehigher than in the respective constituent organic nitrogen,suggesting the fractionation of nitrogen isotopes during theassimilation of ammonia. Wild-type and mutant cells of the cyanobacterium(blue-green alga) Synechococcus PCC 7942 were grown in nitrate-or ammonia-containing medium as the source of nitrogen. Fractionationof nitrogen isotopes during the uptake of nitrate was limited,whereas that during the uptake of ammonia was considerable. ¹ In this report, the term ammonia refers indiscriminately toboth NH₃ or NH₄⁺. (Received June 13, 1991; Accepted September 12, 1991)     

Effect of Salinity and Potassium on the Uptake of Nitrogen and on Nitrogen Metabolism in Young Barley Plants

In a solution culture experiment with 4-week-old barley plants (variety Villa) the influence of NaCl salinization and of KCl additions on the uptake and turnover of labelled N (¹⁵NH₄¹⁵NO₃) was studied. Labelled N was applied for 24 h at the end of the growth period. Salinization impaired growth and uptake of labelled N. The incorporation of labelled N into the protein fraction, however, was improved by NaCl salinization. Additions of KCl to the nutrient solution diminished the negative effect of NaCl salinization on growth. At both NaCl salinization levels (60 and 120 mM) K additions favoured the uptake of labelled N and particularly its incorporation into the protein fraction. It is suggested that the negative influence of the NaCl stress is not primarily due to an impaired protein synthesis, but is possibly caused by a deterimental effect of Na on other metabolic processes.     

Gas Exchange and Water Vapor Uptake in the Atmospheric CAM Bromeliad Tillandsia recurvata L.: The Influence of Trichomes

Tillandsia recurvata is an epiphytic atmospheric CAM bromeliad without functional roots. As an adaptation to very exposed and often dry habitats T. recurvata takes up liquid water via trichomes, which cover the plant. Gas exchange measurements also show an absorption of water vapor at the trichomes simultaneously with an increase in relative humidity at the beginning of the dark period. A similar amount of water is lost again when the relative humidity is decreased. Experiments with heat-killed plants show that this was due exclusively to physical equilibration of the hygroscopic walls of the dead trichome cells. This effect would have prevented an accurate calculation of transpiration and conductance, but can be accounted for by substracting water vapor exchange of heat-killed plants from that of the living tissue which yields the true water vapor conductance of the tissue. This is found to be rather low (3.37 mmol H₂O kg^?1 s^?1) in comparison to other plants, and the proportion of total conductance due to trichomes is relatively high (0.53 mmol H₂O kg^?1 s^?1). This explains the slow rates of photosynthesis and growth found in this plant.     

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

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

Effects of Soil Drought and Atmospheric Humidity on Yield,Gas Exchange,and Stable Carbon Isotope Composition of Barley

The combined effects of water status, vapour pressure deficit (VPD), and elevated temperature from heading to maturity were studied in barley. Plants growing at high VPD, either under well-watered or water deficit conditions, had higher grain yield and grain filling rate than plants growing at low VPD. By contrast, water stress decreased grain yield and individual grain dry matter at any VPD. Water regime and to a lesser extent VPD affected ¹³C of plant parts sampled at mid-grain filling and maturity. The differences between treatments were maximal in mature grains, where high VPD increased ¹³C for both water regimes. However, the total amount of water used by the plant during grain filling did not change as response to a higher VPD whereas transpiration efficiency (TE) decreased. The net photosynthetic rate (P _N) of the flag leaves decreased significantly under water stress at both VPD regimes. However, P _N of the ears was higher at high VPD than at low VPD, and did not decrease as response to water stress. The higher correlation of grain yield with P _N of the ear compared with that of the flag leaf support the role of ear as the main photosynthetic organ during grain filling under water deficit and high VPD. The deleterious effects of combined moderately high temperature and drought on yield were attenuated at high VPD.     

Aphid Acceptance of Barley Exposed to Volatile Phytochemicals Differs Between Plants Exposed in Daylight and Darkness

It is well known that volatile cues from damaged plants may induce resistance in neighboring plants. Much less is known about the effects of volatile interaction between undamaged plants. In this study, barley plants, Hordeum vulgare cv. Kara, were exposed to volatiles from undamaged plants of barley cv. Alva or thistle Cirsium vulgare, and to the volatile phytochemicals, methyl salicylate or methyl jasmonate. Exposures were made either during natural daylight or darkness. Acceptance of exposed plants by the aphid Rhopalosiphum padi was assessed, as well as the expression of putative marker genes for the different treatments. Aphid acceptance of plants exposed to either barley or C. vulgare was significantly reduced, and an effect of the volatiles from undamaged plants was confirmed by the induction of pathogenesis-related protein, PR1a in exposed plants. However the effect on aphid acceptance was seen only when plants were exposed during darkness, whereas PR1a was induced only after treatment during daylight. Aphid acceptance of plants exposed to either methyl salicylate or methyl jasmonate was significantly reduced, but only when plants were exposed to the chemicals during daylight. AOS2 (allene oxide synthase) was induced by methyl jasmonate and BCI-4 (barley chemical inducible gene-4) by methyl salicylate in both daylight and darkness. It is concluded that (a) the effects on aphids of exposing barley to volatile phytochemicals was influenced by the presence or absence of light and (b) the response of barley to methyl salicylate/methyl jasmonate and to volatiles from undamaged plants differed at the gene and herbivore level.Key Words: methyl jasmonate, methyl salicylate, allelobiosis, barley, PR1, allene oxide synthase, Rhopalosiphum padi, light     

Changes in the Leaf Polypeptide Patterns of Barley Plants Exposed to Soil Flooding

Exposure of barley plants (Hordeum vulgare L.) to soil flooding for 72 – 120 h led to decrease in the content of the both subunits of ribulose-1,5-bisphosphate carboxylase/oxygenase. The effect was more pronounced on the small subunit. Further, the changes in protein pattern were observed, mainly proteins with molecular masses 30 – 85 kD were down-regulated. This revised version was published online in July 2006 with corrections to the Cover Date.     

Influence of Root Oxygen Deficiency on Photosynthesis and Antioxidant Status in Barley Plants1

Roots of barley plants (Hordeum vulgare L., cv. Alfa) were subjected to hypoxia for 120 h. By 72 to 120 h of soil flooding, a noticeable decrease in the rate of CO₂ assimilation and transpiration was observed. A drop in the activities of Rubisco and photorespiratory enzymes was found. We examined the changes in the activities of enzymes involved in the antioxidative system and stress markers related to membrane integrity, namely, lipid peroxidation and electrolyte leakage. Catalase and peroxidase activities were increased during the experiment, whereas superoxide dismutase activity drastically decreased.     

Effects of Nitrogen Deficiency on Gas Exchange,Chlorophyll Fluorescence,and Antioxidant Enzymes in Leaves of Rice Plants

Gas exchange, chlorophyll (Chl) fluorescence, and contents of photosynthetic pigments, soluble proteins (ribulose-1,5-bisphosphate carboxylase/oxygenase, RuBPCO), and antioxidant enzymes were characterized in the fully expanded 6^th leaves in rice seedlings grown on either complete (CK) or on nitrogen-deficient nutrient (N-deficiency) solutions during a 20-chase period. Compared with the control plants, the lower photosynthetic capacity at saturation irradiance (P _max) was accompanied by an increase in intercellular CO₂ concentration (C_i), indicating that in N-deficient plants the decline in P _max was not due to stomatal limitation but due to the reduced carboxylation efficiency. The fluorescence parameters _PS2, F_v/F_m, electron transport rate (ETR), and q_P showed the same tendency as P _max in N-deficient plants. Correspondingly, a higher q_N paralleled the rise of the ratio of carotenoid (Car) to Chl contents. However, F_v/F_m was still diminished, suggesting that photoinhibition did occur in the photosystem 2 (PS2) reaction centres. In addition, the activities of antioxidant enzymes on a fresh mass basis were gradually lowered, leading to the aggravation of membrane lipid peroxidation with the proceeding N-deficiency. The accumulation of malonyldialdehyde resulted in the lessening of Chl and soluble protein content. Analyses of regression showed PS2 excitation pressure (1 - q_P) was linearly correlated with the content of Chl and inversely with soluble protein (particularly RuBPCO) content. There was a lag phase in the increase of PS2 excitation pressure compared to the decrease of RuBPCO content. Therefore, the increased excitation pressure under N-deficiency is probably the result of saturation of the electron transport chain due to the limitation of the use of reductants by the Calvin cycle. Rice plants responded to N-deficiency and high irradiance by decreasing light-harvesting capacity and by increasing thermal dissipation of absorbed energy.     

The Influence of Nitrogen (15NO3) Supply to Chicory (Cichorium intybus L.) Plants During Forcing on the Uptake and Remobilization of N Reserves for Chicon Growth

Tuberized tap roots of Witloof chicory (Cichorium intybus L.)were forced by placing in a dark chamber in a hydroponic systemunder high RH to produce an etiolated bud, the chicon. Plantswere fed nutrient solutions with two NO₃concentrations of 1·5or 18 mol m^–3 NO₃, or demineralized water. The nutrientsolutions were labelled with 2% atom excess ¹⁵N. Although thechicon biomass increased with increasing NO₃ concentration inthe nutrient solution, the chicon dry weight remained unchanged.The increased chicon biomass was, therefore, due to more waterin the chicon. The N in the chicon originated from either anendogenous source, the root, and/or an exogenous source, thenutrient solution. Organic N reserves remobilization and transferto the chicon were not been affected by NO₃ supply. At the endof the forcing period 75% of the root N had been remobilized.Differences in the amount of N in the chicons of the three treatmentswere due to the uptake of exogenous N. The flux of exogenousnitrogen to the chicon in high NO₃-plants was 2- to 6-fold higherthan in the low NO₃-plants and, at the end of the forcing period,exogenous nitrogen contributed 30% of total chicon N in highNO₃-plants and 10% in low NO₃-plants. Net uptake of NO₃ by chicory plants during the forcing processwas a function of N influx and N efflux. The increase in N influxwas accompanied by an increase in exogenous N flux to the chiconand probably a shift in root and/or chicon osmotic potentialwhich increased water flux to the chicon. Since NO₃ did notaccumulate in either the chicon or the root, it is proposedthat osmotic solutes, such as organic acids and amino acidsmay be involved in osmotic potential changes in chicory duringthe forcing process. Key words: Cichorium intybus L., efflux, influx, nitrogen (¹⁵NO₃) nutrition, remobilization     

Gas Exchange Characteristics of the Submerged Aquatic Crassulacean Acid Metabolism Plant, Isoetes howellii

The submerged aquatic plant Isoetes howellii Engelmann possesses Crassulacean acid metabolism (CAM) comparable to that known from terrestrial CAM plants. Infrared gas analysis of submerged leaves showed Isoetes was capable of net CO₂ uptake in both light and dark. CO₂ uptake rates were a function of CO₂ levels in the medium. At 2,500 microliters CO₂ per liter (gas phase, equivalent to 1.79 milligrams per liter aqueous phase), Isoetes leaves showed continuous uptake in both the light and dark. At this CO₂ level, photosynthetic rates were light saturated at about 10% full sunlight and were about 3-fold greater than dark CO₂ uptake rates. In the dark, CO₂ uptake rates were also a function of length of time in the night period. Measurements of dark CO₂ uptake showed that, at both 2,500 and 500 microliters CO₂ per liter, rates declined during the night period. At the higher CO₂ level, dark CO₂ uptake rates at 0600 h were 75% less than at 1800 h. At 500 microliters CO₂ per liter, net CO₂ uptake in the dark at 1800 h was replaced by net CO₂ evolution in the dark at 0600 h. At both CO₂ levels, the overnight decline in net CO₂ uptake was marked by periodic bursts of accelerated CO₂ uptake. CO₂ uptake in the light was similar at 1% and 21% O₂, and this held for leaves intact as well as leaves split longitudinally. Estimating the contribution of light versus dark CO₂ uptake to the total carbon gain is complicated by the diurnal flux in CO₂ availability under field conditions.     

NaHSO3对盐胁迫下小麦幼苗氮同化酶及脯氨酸含量的影响

以普通小麦(Triticum aestivumL.)为材料,研究了NaHSO3对不同盐度胁迫下小麦幼苗氮素同化酶和脯氨酸含量的调节。结果表明,盐胁迫降低了叶片中硝酸还原酸(NR)的活性,加入NaHSO3之后,NR活性表现出进一步的降低。谷氨酰胺合成酶(GS)在低浓度盐胁迫下活性增加,在高浓度盐胁迫下活性降低;NaHSO3加入时,即便在低盐浓度下GS活性也降低。依赖于NADH的谷氨酸脱氢酶(NADH-GDH)和依赖于NADP的异柠檬酸脱氢酶(NADP-ICDH)的变化趋势一致,在盐胁迫下它们的活性都明显增加;NaHSO3加入促进了它们活性的进一步增加,尤其对NADH-GDH活性的促进更为明显。游离脯氨酸在高浓度盐胁迫下大量积累,在低浓度盐胁迫下含量增加不明显;NaHSO3促进了盐胁迫下脯氨酸的积累,提示了NaHSO3促进了盐胁迫下小麦幼苗碳氮营养元素的贮存。     

Transformation of Lotus corniculatus Plants with Escherichia coli Asparagine Synthetase A: Effect on Nitrogen Assimilation and Plant Development

Asparagine and glutamine are major forms of nitrogen in the phloem sap of many higher plants. In vascular plants, glutamine-dependent asparagine synthetase (AS) is the primary source of asparagine. In Escherichia coli, asparagine is synthesized by the action of two distinct enzymes, AS-A which utilizes ammonia as a nitrogen donor, and AS-B which utilizes both glutamine and ammonia as substrates, but with a preference for glutamine. In this study, the possibility to endow plants with ammonia-dependent AS activity was investigated by heterologous expression of E. coli asnA gene with the aim to introduce a new ammonium assimilation pathway in plants. The bacterial gene is placed under the control of light-dependent promoters, and introduced by transformation into Lotus corniculatus plants. Analysis of transgenic plants has revealed a phenomenon of transgene silencing which has prevented asnA expression in several transgenics. The asnA-expressing plants are characterized by premature flowering and reduced growth. A significant reduction of the total free amino acid accumulation in transgenic plants is observed. Surprisingly, the content of asparagine in wild-type plants is about 2.5-fold higher than that of transgenic plants. While glutamine levels in transgenic plants are about 3–4-fold higher than those in wild-type plants, aspartate levels are significantly lower. Transformation with asnA also induced a significant reduction of photosynthesis when measured under saturated light and ambient CO₂ conditions.     

稀土元囊对小麦幼苗氮素吸收及同化的影响

稀土元素浸种能够促进小麦(Triticum aestivum L.)幼苗对No3-的吸收,提高硝酸还原酶活力。这些效应与稀土浓度有关,低浓度有促进作用,高浓度则有抑制作用。稀土元素处理还能促进小麦幼苗体内No3-的同化还原,使硝态氮含量降低,氨基氮含量增加,促进了氮素代谢过程。     

Carbon Assimilation Characteristics of the Aquatic CAM Plant, Isoetes howellii

The relationship between malic acid production and carbon assimilation was examined in the submerged aquatic Crassulacean acid metabolism (CAM) plant, Isoetes howellii Engelmann. Under natural conditions free-CO₂ level in the water was highest at 0600 hours and ¹⁴CO₂ assimilation rates in I. howellii were also highest at this time. After 0900 hours there was a similar pattern in (a) rate of free-CO₂ depletion from the water, (b) reduction of carbon assimilation rates, and (c) rate of deacidification in leaves. Rates of daytime deacidification increased under CO₂-free conditions and as irradiance intensity increased. Nighttime CO₂ uptake was estimated to contribute one-third to one-half of the total daily gross carbon assimilation. CO₂ uptake, however, accounted for only one-third to one-half of the overnight malic acid accumulation. Internal respiratory CO₂ may be a substrate for a large portion of overnight acid accumulation as leaves incubated overnight without CO₂ accumulated substantial levels of malic acid. Loss of CAM occurred in emergent leaf tips even though submerged bases continued CAM. Associated with loss of CAM in aerial leaves was an increase in total chlorophyll, a/b ratio, and carotenoids, and a decrease in leaf succulence. δ¹³C values of I. howellii were not clearly distinguishable from those for associated non-CAM submerged macrophytes.     

Effects of Nitrogen Deficiency on the Absorption of Nitrate and Ammonium by Barley Plants

When young barley plants which had been supplied with nitratewere deprived of this source of N, an enhanced capacity forabsorption of either nitrate or ammonium ions developed, reachinga maximum in about 3 d under the particular experimental conditionsused. The net uptake rate of either nutrient was then approximatelythree times that in plants which had received nitrate throughout.Likewise, withholding external N from plants previously growingwith ammonium caused a 2.4-fold increase in their subsequentcapacity to absorb that ion, compared with control plants grownwith an uninterrupted ammonium supply. Accelerated nitrate uptakein N-starved plants was not accompanied by additional phosphateor sulphate absorption, but the plants had the capacity to absorbmore potassium, whether or not ammonium was also present inthe solution. Indirect evidence from analyses of root tissuesuggests that these responses to mild N-stress may depend onsome property of an N fraction which does not include nitrateor ammonium. Hordeum vulgare, barley, nitrogen, ammonium, nitrate, N-deficiency, absorption     

Influence of Nitrate and Ammonium Nutrition on the Uptake, Assimilation, and Distribution of Nutrients in Ricinus communis

Ricinus communis L. plants were grown in nutrient solutions in which N was supplied as NO₃⁻ or NH₄⁺, the solutions being maintained at pH 5.5. In NO₃⁻-fed plants excess nutrient anion over cation uptake was equivalent to net OH⁻ efflux, and the total charge from NO₃⁻ and SO₄²⁻ reduction equated to the sum of organic anion accumulation plus net OH⁻ efflux. In NH₄⁺-fed plants a large H⁺ efflux was recorded in close agreement with excess cation over anion uptake. This H⁺ efflux equated to the sum of net cation (NH₄⁺ minus SO₄²⁻) assimilation plus organic anion accumulation. In vivo nitrate reductase assays revealed that the roots may have the capacity to reduce just under half of the total NO₃⁻ that is taken up and reduced in NO₃⁻-fed plants. Organic anion concentration in these plants was much higher in the shoots than in the roots. In NH₄⁺-fed plants absorbed NH₄⁺ was almost exclusively assimilated in the roots. These plants were considerably lower in organic anions than NO₃⁻-fed plants, but had equal concentrations in shoots and roots. Xylem and phloem saps were collected from plants exposed to both N sources and analyzed for all major contributing ionic and nitrogenous compounds. The results obtained were used to assist in interpreting the ion uptake, assimilation, and accumulation data in terms of shoot/root pH regulation and cycling of nutrients.     

Effect of Nitrogen Supply on the Photosynthetic Performance of Leaves from Coffee Plants Exposed to Bright Light

Although Coffea arabica L. grows naturally in shaded habitats,it can be cultivated under high light intensity, but not withoutsevere photoinhibition mainly during the period of transferfrom the nursery into the field. The present work examines someof the changes in the photosynthetic performance induced byexposure to high light and the possibility of using enhancednitrogen levels to overcome photoinhibition. For that purpose,young plants of Coffea arabica L. (cv. Catuai) grown in a shadedgreenhouse were treated with 0, 1 and 2 mmol of nitrogen and4 weeks later exposed to full solar irradiation, outside. Visible damage due to exposure to full sunlight appeared within2 d in all plants, resulting in a reduced photosynthetic leafarea and drastic shedding of leaves in the unfertilized plants.These effects were considerably less in plants with the highestN dose. After 130 d of exposure, there was 100% mortality inplants receiving no extra nitrogen, compared with 30% in theplants treated with 2 mmol nitrogen. Photosynthesis rates, leafconductance and transpiration presented minimum values after4 d of light stress. Large changes in the photosynthetic capacity(measured at high CO₂ concentration and high light intensity),quantum efficiency and fluorescence yield (F_v/F_m) indicate thatnet photosynthesis rate in the air had been reduced by bothstomatal closure and by changes at the photochemical level.All indicators show that N-fertilized plants were less affectedby photoinhibition. Key words: Coffee plant, nitrogen, photoinhibition, photosynthesis     

The Influence of Removing Tubers on Dry-matter Production and Net Assimilation Rate of Potato Plants

To study the effect of removing tubers on growth and net assimilationrate (E) of potato, plants were grown in pots partly filledwith soil with the shoot growing through a polythene cover.Tubers developed in the space between the cover and the soilsurface. Removing tubers immediately they began to form had little effecton E at the beginning of the experiment but later greatly reducedit. Shading reduced E more at the beginning of the experimentthan later. Removing tubers decreased total dry weight, butmuch of the material that would have moved to tubers accumulatedin leaves and stems. In intact plants the loss of weight byshading was mainly from the tubers; in plants without tubersit was mainly from stems and leaves. Removing tubers increasedleaves on lateral stems. Increasing the amount of nitrogen supplieddiminished the effect on E of removing tubers, presumably becausethe extra allowed other sinks for carbohydrate to develop. Thegrowth of some buds of the potato plant is so strongly inhibitedthat they cannot grow and act as sinks for excess carbohydratewhen tubers are removed. Such internal inhibition of growthmay sometimes suffice to influence the magnitude of E of normalplants. Removing tubers usually increased sugar and starch contentand protein N content of stems and leaves.