Differences in nitrogen‐assimilating enzyme activity in halophyte species are habitat‐related

The chemical form and content of available nitrogen (N) in salt marsh substrates varies considerably. On the western coast of Ireland, habitats designated as Ombrogenic Atlantic salt marshes were formed on ombrogenic peat substrate. The peat substrate in these systems has three times more ammonium than substrate from adjacent salt marsh habitats on sand and mud substrate. This study examined the extent to which the high concentration of ammonium in peat salt marsh substrate influences the N‐ assimilating enzyme activity of halophytes and the extent to which N metabolism differs between species. Specifically, this work investigated whether plants from peat salt marshes are more likely to assimilate ammonium than plants from non‐peat substrates. Four halophyte plant species—Armeria maritima, Aster tripolium, Plantago maritime, and Triglochin maritime—were sampled from various saltmarsh habitats including three sites on peat substrate and three on non‐peat substrate, comprising sand, mud and sand/mud. The activities of N‐metabolising enzymes—glutamine synthetase (GS), glutamate synthase, glutamate dehydrogenase (GDH), and nitrate reductase (NR)—were quantified in shoot and root parts. Root GS activity in Armeria maritima and shoot GS activity in Triglochin maritima were positively correlated with increasing soil ammonium levels. Root NR activity in Aster tripolium and shoot NR activity in Plantago maritima were significantly higher in plants grown on non‐peat substrates than peat substrates. The shoot : root GS activity ratio in Triglochin maritima on peat substrate was more than double the ratio on non‐peat substrates. It is concluded that all species tested displayed differences in N‐metabolising activities depending on the chemical form and/or concentration of N in the substrate, while three out of the four species were capable of taking advantage of the high levels of ammonium in peat substrates.     

Effect of potassium on growth and the electrophoretic pattern of leaf proteins of sunflower supplied with ammonium or nitrate

Sunflower (Helianthus annuus L.) was grown on liquid medium containing either nitrate, ammonium or ammonium + nitrate, with or without potassium. The growth of plants supplied with nitrate or ammonium + nitrate in the presence of potassium (5 mmol/l) was comparable. Plants grown on ammonium‐N only showed significantly lower growth. Dry matter and organic nitrogen content in plants supplied with ammonium + nitrate (total nitrogen 10 mmol/1) in the presence of potassium (5 mmol/l) was higher than in nitrate grown plants. Leaf protein pattern on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE) was not affected by the nitrogen source provided in the medium.

Potassium deficiency led to reduced growth, necrotic spots on the leaves and specific alterations in leaf protein pattern, expressed by an increase or decrease in several polypeptides. This was common to all the nitrogen forms tested. A most pronounced change was the increased expression of polypeptides of molecular weight 60 and 62 kilodaltons.     

Diurnal patterns of nitrate assimilation in Kentucky bluegrass

Kentucky bluegrass (Poapratensis L.) is a major C₃‐type forage and turfgrass, but it is less efficient than many grasses in utilizing nitrogen(N). To determine how this grass can accommodate its greater N need, diurnal patterns of nitrate reductase activity (NRA) and nitrite reductase activity (NiRA) in its leaves and roots were examined and compared with those in barley (Hordeum vulgare L.). Plants were grown under greenhouse or growth room conditions and assayed for NRA and NiRA by optimized in vivo methods. The diurnal patterns of NRA and NiRA indicated that Kentucky bluegrass could assimilate nitrate during the night at rates greater than or similar to those during the day. Leaf NRA of Kentucky bluegrass was minimal approximately 4 and 10 h after illumination commenced and increased at night. The diurnal pattern of leaf NRA among Kentucky bluegrass cultivars did not differ significantly. In roots, NRA of Kentucky bluegrass was high in the morning and decreased sharply during the afternoon and evening, but increased again late at night. Unlike Kentucky bluegrass, barley exhibited greater leaf NRA during the day than during the night and exhibited the greatest activity 6 or 10 h after illumination commenced. In both species, the equilibrium leaf nitrate pool was 20 to 30 times larger than the ammonium pool and 3, 000 to 13, 000 times larger than the nitrite pool. Leaf nitrate pool size showed a diurnal pattern complementary to that of leaf NRA. Our results suggest that a nighttime N use strategy might exist in Kentucky bluegrass.     

水分胁迫下氮素对分蘖期小麦某些生理特性的影响

本研究结果表明 :水分胁迫且充足供氮时 ,叶片硝酸还原酶活性降幅较小 ;水分胁迫且不供氮时 ,硝酸还原酶活性几乎检测不出。水分胁迫下叶片可溶性蛋白含量和叶绿素含量均低于对照 ;根系活力在水分胁迫下均升高 ,根系还原活性在水分胁迫下显著降低。在小麦分蘖期 ,水分胁迫使根系的分布范围增大 ;氮素供给充足时 ,水分胁迫使根系干物质积累增多 ;但水分胁迫下使地上干物质积累降低 ,不供氮处理降幅更大。分蘖期的水分胁迫导致小麦收获期株高显著低于对照处理 ,不供氮处理更是明显 ;根系、地上干物质积累低于对照水平 ;不供氮处理降幅极大。分蘖期水分胁迫处理影响小麦的产量     

Effect of combined nitrogen supply and nodulation on nitrate reductase activity and growth of pea plants

Nitrate reductase activity (NRA; EC 1.6.6.1) was measured in leaves, stems and roots of Pisum sativum cv. Lincoln supplied with different nitrate concentrations and inoculated with selected Rhizobium leguminosarum strains. As a control, noninoculated plants were grown in the same nutrient medium. NRA was determined by an in vivo‐nitrate assay. Although differences in tissue NRA were mainly related to nitrate concentration in the growing medium, nodulation much affected the NRA in the different plant organs, especially in root. An increased proportion of total plant NRA occurred in the leaves and stems as nitrate concentration was increased. Nitrate accumulation in leaves, stems and roots was correlated with the concentration of this ion in the nutrient solution. Nodulation also affects the nitrate accumulation in the different plant organs.     

Nitrogen uptake efficiency of finger millet grown in hydroponic culture

The optimum nitrogen requirement along with nitrogen uptake efficiency of finger millet (Eleusine coracana) for obtaining plants of a high phenotypic quality was studied in this research using hydroponic culture. 20 days old seedlings were transferred in nutrient solution containing all the essential macro and micro nutrients. Plants were subjected to three treatments of nitrate viz. 0.05 mM, 0.1 mM and 0.5 mM by dissolving it in the nutrient solution. Plants grown in nutrient solution devoid of nitrate were treated as control. Plants grown in 0.5 mM nitrate conc. attained more height compared to other low nitrogen treatments (0.1 mM and 0.05 mM). Similarly plants of 0.5 mM nitrate treatments possessed more number of lateral roots, surface area of leaves, dry weight of plants and chlorophyll content compared to other low nitrogen treatments. Specific activity of nitrate reductase in plants treated with 0.5 mM concentration was found nearly 3 times higher than that of control plants; similarly, specific activity of nitrite reductase in 0.5 mM treated plants was also high compared to other low nitrogen treatments followed by control. The results of the present investigation therefore indicate that nitrogen uptake efficiency as revealed by the activities of nitrate reductase and nitrite reductase was high in 0.5 mM nitrate treatment. It also shows that 0.5 mM nitrate is optimum nitrogen concentration for the growth of finger millet in hydroponic condition.     

Diurnal variations in uptake,transport and assimilation of NO3 ‐and efflux of OH‐ in maize plants

Maize plants, grown for 7 and 21 days on a nutrient solution with NO₃ ^‐ as the sole nitrogen source showed a clear diurnal pattern with respect to the in vivo NRA. Especially in roots dark/light fluctuations of the enzyme activity were high. Also in NO₃ ^‐ uptake, OH^‐ efflux and endogenous content of water soluble carbohydrates a diurnal variation was found. The plant age did not significantly affect the daily rhythm.

Because day/night changes of the in vivo root NRA and nitrate uptake were proportional, the relative content of reduced N in the xylem sap of the plants was constant during a day/night interval. At both day 7 and day 21 about 40–50% of the N was transported via the xylem as amino N. As a result of non‐synchronous variation of the specific root and shoot NRA, root reduction capacity showed a great within‐day variation. It varied between 20 and 40% of the whole plant reduction capacity. Since the ratio N‐organic to N‐total in the xylem sap was about 0.5, cycling of organic nitrogen was very likely in these maize plants.     

不同氮素水平对甜菜硝酸还原酶和亚硝酸还原酶活性的影响

以当前甜菜（Beta Vulgaris L.）生产主栽品种KWS0143为试材,设立4个氮素水平（N 0、60、120、180 kg/hm2）,研究硝态氮肥对甜菜硝酸还原酶（nitrate reductase, NR）和亚硝酸还原酶（nitrite reductase, NiR）活性以及光合速率与叶绿素总含量的影响,探讨了氮素水平与NR及NiRA之间的关系。结果表明:在甜菜生育期间,光合速率呈单峰曲线变化,NR、NiR活性及叶绿素总含量基本呈双峰曲线变化,叶绿素变化曲线的高峰期早于NR活性,NR活性早于NiR活性。氮肥用量在N 120 kg/hm2时显著提高了甜菜光合速率、叶绿素总含量、NR活性和NiR活性。相比于N 120 kg/hm2,N 180 kg/hm2时光合速率及叶绿素总含量没有明显变化,NR与NiR活性则有一定的提高。本试验中,甜菜产量随氮水平的增加而提高;含糖率则相反,随氮水平的增加而降低,N 120 kg/hm2水平下产糖量最高。     

菠菜硝态氮累积和还原与植株生长的关系

通过盆栽试验,以两个硝态氮含量差异显著的菠菜品种为供试材料,在不同生长时期,测定了叶柄、叶片干重、水分含量、硝态氮含量及叶片内源和外源硝酸还原酶活性,研究菠菜硝态氮累积和硝酸还原酶活性的动态变化及其与植株生长变化的关系。结果表明,随生长期后移,叶柄、叶片及地上部干重和水分含量先增加而后降低,硝态氮含量则持续降低,低硝态氮累积品种S9的下降更为明显,出苗后52d和62d地上部分别降低了100%和89.7%;叶片内源和外源酶活性则随植株生长量增加而增加,高硝态氮累积品种S4增加（379%和199%）更明显,之后该品种酶活性随植株生长量降低而显著下降,品种S9却显著增加,分别为121%和288%。生长前期,品种S4硝态氮含量、干重增长速率及内源、外源酶活性均显著高于品种S9,内源/外源酶活性比值却明显低于后者;生长后期,除外源酶活性和内源/外源酶活性比值外,品种间差异均不明显。因此,生长前期高累积品种硝态氮含量降低较少,主要原因可能是其内源/外源酶活性比值（70.7%）较低,生长后期该品种的内源/外源酶活性比值（98.2%）显著增加后,硝态氮含量迅速下降进一步证明了这一推测。综合上述结果可知,内源/外源酶活性比值更能揭示植株生长变化引起的品种间硝态氮含量变化差异。     

Interaction of salinity,nitrogen, and phosphorus fertilization on wheat

This study determined whether the application of nitrogen (N) and phosphorus (P) could ameliorate salt‐induced reduction in wheat production. Saline irrigation water (0.5, 4.0, 8.2, and 12.5 dS/m) and N and P fertilizers (150 kg N/ha and 37.5 kg P₂O₅/ha) were applied to wheat (Triticum aestivum L. ‘Saka 92') grown on a calcareous soil in a greenhouse experiment. Plants received equal amounts of each fertilizer, but the time and frequency of application differed. All salinity levels reduced straw and grain yields, leaf soluble proteins, nitrate (NO₃) content, actual and potential nitrate reductase activity (NRA), and grain protein content. The delay in pollen meiotic cell division increased with salinity. Under saline conditions, applying N and P fertilizers at the end of the grain filling stage improved yield and metabolic performance of the plants compared to other fertilizer treatments.     

Nitrogen‐use efficiency traits of mini‐watermelon in response to grafting and nitrogen‐fertilization doses

Two experiments were conducted to study the effect of grafting on nitrogen‐use efficiency (NUE) in mini‐watermelon plants. In the first study, mini‐watermelon plants (Citrullus lanatus [Thumb.] Matsum. and Nakai cv. Minirossa) either ungrafted or grafted onto Macis, Vita (Lagenaria siceraria [Mol.] Standl.), PS1313, and RP15 (Cucurbita maxima Duchesne × Cucurbita moschata Duchesne) rootstocks grown in hydroponics were compared in terms of shoot dry biomass, leaf area, root‐to‐shoot ratio, SPAD index, shoot N uptake, and nitrate reductase (NR) activity 40 d after transplantation in response to nitrate concentration in the nutrient solution (0.5, 2.5, 5, 10, 15, or 20 mM of NO$ _3^- $ ). In the second experiment, the suitability of a selected rootstock with high NUE (Vita) to improve crop performance and NUE of grafted mini‐watermelon plants was evaluated under field conditions. In the hydroponic experiment mini‐watermelon grafted onto Vita rootstock needed the lowest nitrate concentration (1.31 mM of NO₃) in the nutrient solution to reach half maximum shoot dry weight. Total leaf area, SPAD index, and shoot N uptake increased in response to an increase of N concentration in the nutrient solution. At 2.5 mM NO$ _3^- $ , mini‐watermelon grafted on either Vita or RP15 had the highest NR activity whereas no significant difference was observed at 10 mM NO$ _3^- $ . The open‐field study indicated that increasing N‐fertilization rates from 0 to 100 kg ha^–1 improved total and marketable yields of mini‐watermelon plants while decreasing NUE. When averaged over N levels, the marketable yield, NUE, N‐uptake efficiency, and N‐utilization efficiency were significantly higher by 39%, 38%, 21%, and 17%, respectively, in Minirossa grafted onto Vita compared to ungrafted Minirossa plants. Therefore, grafting mini‐watermelon plants onto selected rootstocks can be used as a quick and effective method for improving productivity and NUE.     

Physiological studies on salinity and nitrogen interaction in alfalfa plants: III. nitrate reductase activity

Nitrate reductase activity (NRA) was determined to investigate the effect of salinity and nitrogen (N) interactions on alfalfa [Medicago sativa (L) cv. Gilboa] during its vegetative growth. Increasing levels of sodium chloride (NaCl) (0, 30, 65, and 100 mM) decreased NRA in both plant parts, i.e., root and leaf, however to a lesser extent in leaves. The inclusion of Neither as nitrate (NO₃) or ammonium (NH₄) (0, 3, and 6 mM) to the nutrient medium resulted in a substantial enhancement of NR activity in salinized and non‐salinized plants as well.     

Effects of supplemental nitrogen on nitrogen‐assimilation enzymes,free amino nitrogen,soluble sugars,and crude protein of rice

Abstract

An upland rice variety IAC‐47 was grown in a greenhouse to determine the effect of foliar nitrogen (N) supplementation during grain development on the activity of the N assimilation enzymes, nitrate reductase (NR) and glutamine synthetase (GS), on free amino‐N content and leaf soluble sugars, and on grain crude protein content. At 10 and 20 days after anthesis (DAA), the leaves were fertilized with a liquid fertilizer containing 32% N as 12.8% urea, 9.6% ammonium (NH₄), and 9.6% nitrate (NO₃) in increasing rates corresponding to 0,20+20, 40+40, and 60+60 kg N ha^‐1. Leaves were collected twice (at 12 DAA and 14 DAA for GS activity, sugar and amino‐N content, and at 11 and 13 DAA for NRA) after each application of leaf N. The late foliar application of N increased significantly grain crude protein without a corresponding decrease in grain weight. The NR activity (NRA) increased after the foliar application of N. In the flag leaf, 60+60 kg N ha^‐1 (21 DAA) resulted in higher NRA (20x over the control), while GS activity was smaller than the control. At 22 DAA there was an increase in GS activity in the flag leaf at 20+20 N level. However, the GS activity decreased as applied N levels increased. Also at the 20+20 level, there were increases in free amino‐N in the flag leaf and second leaf at the final harvest. Throughout the experiment, plants at the 60+60 N level had the lowest levels of soluble sugars. Increases in crude protein were highest at 40+40 N level (27.9%), followed by 60+60 (18.7%).     

Nitrogen Metabolism and Tomato Yield in Response to Organic Fertilization

We assessed the effects of organic fertilization on the response of biochemical and physiological indicators and the yield of saladette-type tomato (Solanum lycopersicum L.) grown under greenhouse conditions. Five fertilization forms [sand + inorganic nutrient solution (F1); sand + vermicompost tea (F2); a mixture of sand, compost, + vermicompost tea (F3); a mixture of sand, vermicompost, + vermicompost tea (F4); and a mixture of sand, compost, vermicompost, + vermicompost (F5)] and two genotypes (Cuauhtémoc and El Cid) were evaluated. The parameters analyzed were leaf pigments, enzymatic activity of nitrate reductase (NR) in vivo, and yield. A fertilizer source of sand + vermicompost tea resulted in the best assimilation of nitrate (NO₃-), the greatest NR endogenous activity, the second highest foliar concentration of organic nitrogen (N), and the second best yield. In conclusion, for improved tomato cultivation during organic production, treatment F2 produced the maximum organic yield and resulted in more efficient N utilization.     

小白菜适当增铵下硝酸盐累积机理研究

利用NO3--N/NH44+-N为100∶0和75∶25的营养液对两个硝酸盐累积能力显著不同的小白菜品种(上海青和亮白叶1号)进行培养,测定了小白菜叶片、叶柄及根系硝酸盐含量、硝态氮和铵态氮吸收量及各部位硝酸还原酶活性,以探讨适当增铵降低小白菜硝酸盐含量以及小白菜不同品种和不同器官累积硝酸盐能力差异的机理。结果表明,适当增铵使叶片、叶柄和根系硝酸盐含量分别降低了22%、15%和22%,而硝态氮吸收量则降低了7.5%。小白菜各器官硝酸盐含量为叶柄叶片根系。叶片硝酸还原酶活性分别是叶柄和根系的27和9倍,呈现叶片根系叶柄,叶片是硝态氮的主要还原器官。亮白叶1号叶片、叶柄及根系硝酸盐含量分别较上海青高3%、38%和34%,硝态氮吸收量仅较上海青高11%;而叶片、叶柄及根系硝酸还原酶活性则分别较后者降低44%、56%和38%。适当增铵减少硝态氮吸收量是增铵降低硝酸盐含量的主要原因。不同器官的功能与结构的不同决定其累积硝酸盐能力的不同;不同品种硝酸盐累积的差异取决于还原硝态氮能力的差异。     

Influence of photosynthetic irradiance on nitrate reductase activity,nutrient uptake and partitioning in tomato plants

The objective of the present work was to study the nutritional behavior of tomato plants (Lvcopersicon esculentum Mill.) subjected to high pressure sodium (HPS) supplementary lighting in relation to nitrate reductase activity (NRA). Tomato plants were grown with or without HPS supplementary lighting at 2 different root‐zone temperatures (RZT). Supplementary lighting combined with low RZT promoted NRA and cation uptake. Magnesium uptake appeared particularly related to the NRA daily pattern. Effects of photosynthetic irradiance (PI) at two growth stages on partitioning of 45Ca and 86Rb were also investigated. Low light level stimulated 45Ca uptake in fruiting plants but depressed 86Rb uptake. A hypothetical mechanism involving the influence of NRA and K cycling on HCO₃‐ excretion by root is proposed to explain the effects of treatments on mineral uptake.     

Effect of nitrate and humic substances of different molecular size on kinetic parameters of nitrate uptake in wheat seedlings

The kinetic parameters of nitrate uptake (I_max, Km and C_min) were evaluated in young seedlings of Triticum durum L., cv. Appulo, exposed to nitrate and/or to soil‐extracted humic acids (HAs) of different molecular weight. The uptake was enhanced after induction at low levels of nitrate (50 μM KNO₃), while it was inhibited after induction at higher concentrations (2000 μM). The kinetic parameters of uptake were selectively influenced by pre‐treatment with HAs: total (TE) and, at a greater extent, low (LMS, < 3500 Da) molecular size humic fraction increased either the nitrate uptake rate (I_max) and the efficiency of the whole transport system (low Km and C_min), while an opposite result was evidenced in high molecular size (HMS, > 3500 Da)‐treated plants. An additive effect was shown when nitrate and humic substances were provided simultaneously: the uptake rate was enhanced in TE‐ and LMS‐treated plants, but was strongly delayed in HMS‐treated plants. Removal of nitrate and/or humic fractions de‐induced the system and NO₃^— uptake rate decreased. Exposure to HAs was not able to induce nitrate reductase activity in root and leaf tissues. Inhibitors of protein synthesis p‐fluorophenylalanine and cycloheximide reversed the positive effect of LMS fraction on nitrate uptake. This would support the hypothesis of a promoting effect of HAs on the molecular expression of proteins of the nitrate transport system.     

Nitrogen metabolism is related to improved water‐use efficiency of nodulated alfalfa grown with sewage sludge under drought

Leguminous plants grown in sewage sludge–amended soils can acquire nitrogen by assimilation of nitrate and ammonium from the soil solution or from atmospheric‐dinitrogen (N₂) fixation through association with N₂‐fixing bacteria. We proposed that operation of both metabolic processes could contribute to alleviate the impact of drought in sludge‐treated plants. A greenhouse experiment was conducted to evaluate the involvement of nodule metabolism in the use efficiency of water and N in sludge‐treated plants. Treatments comprised (1) plants inoculated with rhizobia and amended with sewage sludge; (2) plants inoculated with rhizobia without any amendment; and (3) noninoculated plants supplied with ammonium nitrate, each under well‐watered and drought conditions. Under drought, sludge‐treated plants had increased plant growth and higher photosynthetic and water‐use efficiencies than untreated plants. Drought stimulated nitrate reductase and GS/GOGAT activities but did not affect the activities of phosphoenolpyruvate carboxylase and malate dehydrogenase or the leghemoglobin concentration. The results suggest that under drought conditions, both N₂ fixation and nitrate assimilation in nodules of sludge‐treated plants contributed to improve plant N supply and to increase the drought tolerance of alfalfa.     

Sensitivity of soft red winter wheat cultivars to chlorate‐induced toxicity

The use of chlorate as a nitrate analogue to screen soft red winter wheat (Triticum aestivum L.) cultivars for differences in nitrate reductase activity (NRA) was studied by adding potassium chlorate to a hydroponic nutrient solution in which wheat seedlings were growing. After 14 days, leaf symptoms indicating chlorate‐induced toxicity were rated. It was hypothesized that wheat plants which were susceptible to chlorate‐induced toxicity reduced chlorate and nitrate more rapidly than did resistant plants. In experiments testing the potential of this assay, wheat and barley (Hordeum vulgare L.) cultivars previously reported to have low NRA were less susceptible to chlorate‐induced toxicity than were cultivars reported to have high NRA. The assay was used to screen 15 soft red winter wheat cultivars for differences in sensitivity to chlorate‐induced toxicity. Variable toxic reactions were observed both among and within the cultivars. To determine whether the within‐cultivar variation was environmental or genetic, single plant selections for contrasting chlorate response were made, and bulked progeny were rescreened. In eight of 15 cultivars, the contrasting selections were different for chlorate‐induced toxic response, indicating heterogeneity for this trait within these eight cultivars. These chlorate‐selected lines may also be near‐isogenic lines for NRA. Seedling screening of wheat for chlorate response may be useful for identification of high NRA breeding lines.