Effect of low static nitrate concentrations on mineral nitrogen uptake,nodulation, and nitrogen fixation in field pea

Abstract

Combined nitrogen [nitrate (NO₃‐), ammonium (NH₄+), and urea] will inhibit all components of symbiotic nitrogen (N₂) fixation if present in sufficient concentrations. It is generally accepted that nitrate is particularly inhibitory to nodule growth and nitrogenase activity, and somewhat less inhibitory to the infection process. This project examined whether providing low (0.1 ‐ 0.5 mM) static concentrations of NO₃‐ to pea (Pisum sativum L. cv. Express), seedlings could avoid the period of N hunger experienced prior to the establishment of N₂ fixation, without delaying or reducing the symbiotic N₂ fixation. All concentrations of NO₃ ^? tested significantly inhibited all measured components of N₂ fixation. The nodulation process as measured by nodule number was inhibited to a similar degree as the other parameters. A concentration dependent response was evident, with 0.1 mM NO₃ ^? causing less inhibition than the 0.2 or 0.5 mM concentrations. Our results indicate the within the concentrations of 0.1 mM and 0.5 mM NO₃ ^?, it is not possible to stimulate the growth of pea plants without inhibiting nodulation and N₂ fixation.     

Effect of boron on nitrate reductase activity in young sunflower plants

The effect of variable levels of B supply on the content of total N, nitrate and nitrate reductase activities in roots and shoots of young sunflower (Helianthus annuus L.) plants was investigated. It was found that both a deficiency and high toxic levels of B decreased the total N content and the activity of nitrate reductase and increased the content of nitrates in roots and shoots of the plants. The treatment of the excised segments of roots and leaves of plants supplied with insuficient levels of B and adequate levels of glucose significantly increased the activity of nitrate reductase. The study concluded that one of the factors influencing the primary assimilation of N is B supply to plants. Supported by these findings and corroborated by literature data, the possible effect of B on nitrate reductase activity via the metabolisms of nucleic acids and proteins and on energy turnover is discussed.     

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.     

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。     

Nitrate nitrogen and nitrate reductase activity in Verna lemon tree leaves

The determination of the leaf nitrate concentration, as well as the nitrate reductase activity have been proposed as a parameters for the estimation of the nitrogen requirements of citrus plants. Because this, it is interesting to dispose of a well criteria for their suitable diagnosing. On nutritionally normal Verna lemon trees we study the annual evolution of the leaf nitrate levels as well as the nitrate reductase activities. At the same time, the cause of the nitrogen alterations induced by iron chlorosis are determined. The results show that is the ferredoxin the iron compound responsible of this nitrogen unbalance.     

Effect of ammonium or nitrate nutrition on photosynthesis,growth, and nitrogen assimilation in tomato plants

Nitrogen (N) is taken up by most plant species in the form of nitrate (NO ) or ammonium (NH ). Plant response to continuous NH nutrition is species‐dependent. In this study, we compare the responses of tomato (Solanum lycopersicum L. cv. Rio Grande) plants to N source (NO or NH ). To this end, early plant growth, photosynthesis, chlorophyll, carbohydrate, and N‐compound concentrations as well as the activities of main enzymes involved in N metabolism (nitrate reductase, nitrite reductase, glutamine synthetase, and glutamate dehydrogenase) were analyzed. Early plant growth was remarkably ameliorated under NH ‐ in comparison to NO ‐based nutrition. Concomitantly, photosynthetic activity, total chlorophyll, and carbohydrate concentrations were significantly increased. With increasing external NH concentration, NH accumulated mainly in roots. In addition, root protein concentration was significantly increased, reflecting high NH incorporation into organic nitrogen. Root glutamine synthetase (GS) activity was enhanced by NH for concentrations below 5 mM, whereas root glutamate dehydrogenase (GDH) activity increased in parallel to NH availability. Together with the positive effect of NH on tomato plant cv. Rio Grande growth, these results reveal that GDH could have, in addition to GS, a possible role in NH detoxification and tolerance of NH ‐based nutrition.     

Soil fertility effects on growth,yield, nodulation and nitrogenase activity of Austrian winter pea

Austrian winter pea (Pisum sativum subspecies arvense (L.) Poir) is grown as a cool season annual to produce high protein seed and forage as well as for soil fertility improvement. This legume is grown on a wide range of soil types with many different cropping systems. The objective of these studies was to determine the influence of K levels, with and without P and Ca fertilization, for increased growth, yield, nodulation and nitrogenase activity. Results were from 3 years’ field and greenhouse experiments with a Psammentic Paleustalf (Eufaula series) utilizing Rhizobium leguminosarum (Frank), ATCC 10314 as inoculum. Soil fertility effects on composition and histology of field‐grown nodules are presented.

Available soil P was a limiting plant nutrient in field studies with significant response to K resulting with PK combinations for top growth, tillers, pods, seed yield, nodule mass, and nitrogenase activity levels (C₂H₂, red.). Multiple regression for nitrogenase (umol C₂H₄ h^‐1) = 1.09 tiller number + 3.37 nodule weight + 2.29 pod number, R² = 0.837, C.V. = 29.9%. Results from the greenhouse experiments indicated significant responses with increased K application levels when combined with P and Ca fertilization for top growth, nodule weight, number of nodules and nitro‐genase activity. Highly significant correlations resulted with nitrogenase x nodule weight (r=0.538) and nitrogenase x top growth (r=0.359) with multiple regression of treatment effects for nitrogenase (μmol C₂H₄ h^‐1) = 2.73 P + 1.04 K + 4.92 Ca, R² = 0.797 and C.V. = 48.8%. Soil addition of plant nutrients resulted in significantly increased concentrations of those elements within nodules. Magnesium content was not consistently influenced by P, Ca, and K amendments. Sodium decreased with increased K fertilization. Multiple regression of elemental composition (mg g^‐1 nodule) for nitrogenase (pmol C₂H₄ h^‐1) = 0.21 P + 0.86 K + 2.35 Ca ‐ 2.01 Na, R² = 0.772, C.V. = 55.6%. The proportion of plant nutrients in nodules contained within the nodule cytosol was highest for K (56.2%) and lowest for Ca (21.4%) with intermediate levels of Mg (50.2%), P (45.4%), and Na (37.2%).

Practical application from these data include the requirement of adequate available soil K for increased yield and nitrogen fixation with favorable P and Ca soil levels in Austrian winter pea production.     

Effect of green synthesized molybdenum nanoparticles on nitrate accumulation and nitrate reductase activity in spinach

Abstract

In the present study, the green synthesized Mo nanoparticles (NPs) were firstly prepared using vermicompost extracts and, then, two experiments were separately carried out in a completely randomized design. The first experiment was conducted to investigate the effects of ammonium nitrate (AN) on nitrate (NO₃^?) accumulation rates as well as some other vegetative traits in spinach in four treatments and three replicates and the second experiment was done to investigate the effects of elemental Mo and green synthesized Mo NPs on NO₃^? accumulation, nitrate reductase (NR) activity and some morphological parameters in seven treatments with three replicates. The results of the first experiment indicated that the greatest accumulation of NO₃^? in the aerial parts of the plants was observed in the 3?M AN treatment. That is why the same concentration was utilized in the second experiment to study the effects of elemental Mo and green synthesized Mo NPs on the NR activity, NO₃^? accumulation and the other traits. The results of the second experiment indicated that various concentrations of elemental Mo and green synthesized Mo NPs have significant effects on all measured traits including the fresh and dry weights of the plant, NO₃^? concentration, NR activity, chlorophyll a (Chl a) and chlorophyll b (Chl b) rates, total chlorophyll (Chl a?+?b) and the plant height. Moreover, it was found that the green synthesized Mo NPs, as compared to elemental Mo, have a greater effect on the increase of NR activity and, consequently, significant reduction of NO₃^? accumulation. Abbreviations AN ammonium nitrate

Chl a chlorophyll a

Chl b chlorophyll b

Chl a?+?b the total chlorophylls

M Molar

Mo molybdenum

NPs nanoparticles

NR nitrate reductase

N nitrogen

NO₃^? nitrate

     

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.     

叶面追施硝态氮肥抑制菠萝营养生长效应

采用叶面淋施的盆栽试验方法,以我国菠萝主栽品种-巴厘为试材,研究不同形态氮素对盆栽菠萝营养生长和菠萝叶片黄化的影响,为菠萝氮肥合理施用提供参考。试验结果表明,叶面淋施硝态氮处理的菠萝根、茎叶生物量显著低于叶面淋施铵态氮、酰胺态氮,黄叶数显著高于叶面淋施铵态氮、酰胺态氮。与铵态氮相比,硝态氮处理的菠萝总叶数、根数目、根重、茎叶重分别减少18.7%、26.5%、49.7%、43.5%,黄叶数增加192.7%。叶面淋施硝态氮抑制菠萝营养生长主要机理是硝态氮提高了土壤p H值,减少了铁吸收,降低菠萝叶片中全铁、活性铁、叶绿素含量(与铵态氮相比,分别减少25.9%、66.9%、23.2%)。     

Role of Azospirillum brasilense nitrate reductase in nitrate assimilation by wheat plants

Summary The nitrogen metabolism of wheat plants inoculated with various Azospirillum brasilense strains and nitrate reductase negative (NR^–) mutants was studied in two monoxenic test tube experiments. The spontaneous mutants selected with chlorate under anaerobic conditions with nitrite as terminal electron acceptor fixed N₂ in the presence of 10 mM NO₃ ^– and were stable after the plant passage. One strain (Sp 245) isolated from surface-sterilized wheat roots produced significant increases in plant weight at both NO₃ levels (1 and 10 mM) which were not observed with the NR^– mutants or with the two other strains. Similar effects were observed in a pot experiment with soil on dry weight and total N incorporation but only at the higher N fertilizer level. In the monoxenic test tube experiments plants inoculated with the mutants showed lower nitrogenase activities than NR⁺ strains at the low NO₃ ^– level (1 = mM) but maintained the same level of activity with 10 mM NO₃ ^– where the activity of all NR⁺ strains was completely repressed. The nitrate reductase activity of roots increased with the inoculation of the homologous strains and with the mutants at both NO₃ ^– levels. At the low NO₃ ^– level this also resulted in increased activity in the shoots, but at the high NO₃ ^– level the two homologous strains produced significantly lower nitrate reductase activity in shoots while the mutants more than doubled it. The possible role of the bacterial nitrate reductase in NO₃ ^– assimilation by the wheat plant is discussed.     

Impact of soil nitrogen concentration on Glomus spp.-Sinorhizobium interactions as affecting growth, nitrate reductase activity and protein content of Medicago sativa

Our objective was to evaluate how increasing levels of N in the medium (0, 4, 8 and 16 mmol N added kg^-1 soil) affect the interaction between Sinorhizobium and arbuscular mycorrhiza (AM) fungi in the tripartite symbiosis with Medicago sativa. Growth response, nutrient acquisition, protein content, and nitrate reductase (NR) activity were measured both in plant shoots and roots. Results showed that N levels in soil did not affect mycorrhizal colonization but they strongly influenced nodulation, particularly of mycorrhizal plants. Mycorrhizal colonization was required for a proper nodulation when no N was applied to soil. In contrast, the addition of 4 mmol N kg^-1 soil reduced nodulation only in mycorrhizal plants and 8 mmol N added kg^-1 soil allowed nodule formation only in non-mycorrhizal plants. Nodulation was totally inhibited in all treatments with the addition of 16 mmol N added kg^-1 soil. N addition enhanced NR activity in all the treatments, while AM colonization increased the proportion of NR allocated to roots. This effect was more pronounced under the lowest N levels in the medium. The two AM fungal species showed different distribution pattern of enzymatic activities in plant tissues indicating specific physiological traits. Protein content as well as the relative proportion of protein in roots were greatly increased after mycorrhizal colonization. Glomus intraradices-colonized plants had the highest protein content in shoot and root. Mycorrhizal effects on growth, N acquisition and biochemical variables cannot be interpreted as an indirect P-mediated effect since P content was lower in mycorrhizal plants than in those which were P fertilized. Mycorrhizal colonization increased the N content in plants irrespective of the N level, but the effectiveness of AM fungi on plant N acquisition depended on the AM fungus involved, G. intraradices being the most effective, particularly at the highest N rate. N₂ fixation, enhanced by AM colonization, contributed to N acquisition when a moderate N quantity was available in the soil. Nevertheless, under a high N amount the nodulating process and/or fixing capacity by Sinorhizobium was reduced in AM plants. In contrast, the AM fungal mycelium from a particular mycorrhizal fungus may continue to contribute efficiently to the N uptake from the soil even at high N levels. These results demonstrate the particular sensitivity of AM fungal species in terms of their growth and/or function to increasing N amounts in the medium. A selection of AM fungi used to address specific environmental conditions, such as N fertilization regimes comparable to those used in agronomic practices, is required for a better use of N applied to soil.     

Effect of varied soil nitrogen supply on growth and nutrient uptake of young Norway spruce plants grown in a shaded environment

To understand the effect of increased soil N supply on tree growth and nutrient uptake, three-year-old Norway spruce seedlings were grown in pots on low-nutrient mineral forest soil supplemented with N in mineral or organic form. Outdoor shaded growth conditions were used, to test the hypothesis that shaded plants are particularly susceptible to high soil N supply. Plants were harvested eleven months after planting. Shoot growth was not affected by the N supply, but N concentrations in needles and roots were increased in plants supplied with mineral N (150 or 300 mg N [kg soil]^—1). Root growth was drastically reduced and root/shoot ratios were decreased in plants with higher N uptake. A high supply of mineral N to soil also decreased the concentrations of other essential elements (P, K) in the needles and thus had effects on plant growth which may impair the stress resistance of trees. Organic N in the form of keratin (150 mg N [kg soil]^—1) did not influence plant growth significantly. The adverse effects of high mineral N supply were particularly pronounced under shaded conditions in comparison to results from other experiments using higher light intensity and temperature conditions.     

Effect of organic amendments on nodulation and nitrogen fixation by cowpea

Cornstover, garbage compost, and cowdung were added to an Alfisol (Iwo series) at the rates of 0, 23, 69, and 115 Mg ha^‐1 and incubated for 4 weeks in the greenhouse prior to sowing cowpea [Vigna unguiculata (L.) var. Ife Brown]. Two successive plantings were made per pot. Harvesting of the plants was at onset of flowering, 42 days after planting. Nitrogen (N) fixed was assessed using the N₂ difference method. Soil reaction (pH), organic carbon (C), and tissue N contents were increased by all the organic amendments. At both harvests, dry matter yields were significantly enhanced by garbage compost and cowdung. While garbage compost and cowdung increased the amounts of N₂ fixed, the efficiency of N₂ fixation was higher in the cornstover treatments. The cornstover additions also significantly increased nodule size and number. These results suggest that organic amendments would benefit N₂ fixation especially in soils low in indigenous organic matter. The beneficial effects were in the order: cowdung > garbage compost > cornstover. To enhance N₂ fixation in cowpea, a rate of 23 Mg ha^‐1 of these organic amendments is suggested.     

Effect of ammonium-, nitrite- and nitrate nitrogen on anaerobic nitrogenase activity in soil

Sandy loam soil, with added glucose, was incubated anaerobically under N₂ and subjected to repeated 1-h C₂H₂ reduction assays. In the presence of 1% glucose the addition of 50 μg NH₄⁺ ?N/g or of 20 μg NO^?₃ N/g (untreated soil contained 1.2 μg NH⁺₄?N and 7.10 μg NO^?₃-N/g) caused at least some suppression of nitrogenase activity. Activity developed when the KCl-extractable soil inorganic nitrogen concentration dropped below 35 μg/g. In the presence of 0.1 or 0.05% glucose the addition of 5 μg NH⁺₄?N/g caused some suppression of nitrogenase activity. However, activity developed when the soil NH₄⁺-N concentration dropped below about 4 μg/g. With 0.1% glucose and 5 μg added NO^?₂ N/g, activity did not develop until the soil NO^?₂ -N concentration dropped to zero. Added NO^?₃ N was rapidly reduced and denitrified to NO^?₂- N, N₂O-N and NH⁺₄ N and furthermore caused some inhibition of CO₂ evolution. The data from NH₄^?-addition experiments are consistent with a nitrogenase repression/ derepression threshold of 4 and 35μg NH⁺₄-N/g at 0.05 and 1% glucose concentrations, respectively. The data from NO^?₂- and NO^?₃-addition experiments suggest a combination of repression and toxicity effects in the presence of added NO^?₃ N.     

不同硝响应型水稻品种苗期根系生长对增硝营养的响应

利用控制条件下的水培试验方法,研究了两种铵硝配比(NH4+/NO3-为100/0和75/25)营养条件对4种不同硝响应型水稻品种苗期根系生长的影响。结果表明,在增硝营养(NH4+/NO3-为75/25)条件下,不同水稻品种NO3-的反应差异明显。与全NH4+营养条件相比,增硝营养条件下对NO3-强响应的水稻品种南光的根系干重和氮积累量显著增加,增幅达50%和79%;同时南光的根系总根长、总不定根长和总侧根长增幅均达到显著水平;不定根数、新根数和侧根数亦显著增加;平均不定根长和平均侧根长差异不显著;对硝弱响应型的水稻品种上海97、辽粳和Elio在增硝营养培养下的根系不定根、新根和侧根的长度和数量差异均不显著。这表明增NO3-营养仅仅促进了对NO3-强响应型水稻南光根系的不定根和侧根的发生,进而促进根系对氮素的吸收,并没有促进不定根和侧根的伸长。从本试验的结果可推论,水稻根系对硝态氮的响应度强弱可能是水稻品种氮素效率差异性的因子之一。     

Influence of nitrogen nutrition on growth,nitrate reductase and nitrite reductase of seedlings of maize (Zea mays L. cv LG12)

The effects of three N levels on etiolated and green leaves of Zea mays were studied. It was found that the activity of both nitrate reductase (NR) and nitrite reductase (NiR) was affected, although differently, and the former showed higher sensitivity. NR activity rose with N increase, but different values in the three groups of seedlings appeared only after the second day in light/darkness; both nitrate and light were necessary to obtain a high level of activity. The interference of the N supply on NiR was observed only in seedlings grown in light. Seedling growth was also affected and the leaf weight appeared to correspond with the level of N supplied. In seedlings grown in darkness, the nitrate accumulated in the leaf increased with the decrease of the supplied N, whereas in light/darkness the opposite occurred. Different values of the leaf protein in the three seedling groups appeared only under light/dark and after the second day and by SDS‐PAGE they were found to be quantitative.     

Effects of changes in applied nitrogen concentrations on nodulation,nitrogen fixation and nitrogen accumulation during the soybean growth period

ABSTRACT

The specific mechanism by which nitrogen application affects nodulation and nitrogen fixation in legume crops remains uncertain. To further study the effects of nitrogen application on soybean nodulation and nitrogen accumulation, three consecutive tests were performed during the VC-V4, V4-R1 (10 days), and R1-R2 (10 days) growth periods of soybean. In a dual-root soybean system, seedlings on one side were watered with a nutrient solution containing NH₄⁺ or NO₃^? as the N source (N+ side), and those on the other side were watered with a nitrogen-free nutrient solution (N- side). During the VC-R2 period, on the N+ side, high nitrogen treatment inhibited nodule growth and nitrogenase activity (EC 1.18.6.1), and the inhibition was significantly increased with increasing high nitrogen supply time (10 days, 20 days). When the high nitrogen treatment time reached 20 days, the specific nitrogenase activity (C₂H₄ μmol^?1 g^?1 nodule dry mass h^?1) was similar to that in the low nitrogen treatment, indicating that the nitrogen fixation capacity per gram of dry mass nodules was almost the same. Therefore, it is assumed that long-term high nitrogen treatment mainly reduces nitrogen fixation by reducing the nodule number. The effect of nitrogen concentration on the roots on the N+ side was greater than that on the N- side. Taken together, these results indicate that nitrogen application affects a contact-dependent local inhibition of root nodule growth, nitrogenase activity, and nitrogen accumulation. The whole plant systematically regulates specific nitrogenase activity, and high nitrogen inhibition is recoverable.