The growth and some physiological responses of rice to Cd toxicity as affected by nitrogen form

A hydroponic experiment was conducted to examine the effect of Cd stress on anti-oxidative enzyme activities at heading stage, yield components, root exudation and Cd and N uptake of rice plants grew in different N source i.e. (NH₄)₂SO₄, NH₄NO₃ and Ca(NO₃)₂. The results show that the effect of Cd stress on all measured parameters were N source dependent. Cd stress (1 μM) caused a remarkable reduction in grain yield and shoot biomass, an increase in root exudation, glutathione content, Cd concentration and catalase (CAT) and peroxidase (POD) activities of rice plants. In the plants under the control (without Cd addition) N source had no distinctive effect on the above measured parameters, but the differences among the three N forms in these parameters became significant when plants were exposed to Cd stress. Cd stress significantly increased POD and CAT activities, and gultathione content, with Ca(NO₃)₂-fed plants having the greatest POD and CAT activities and lowest glutathione content, and (NH₄)₂SO₄-fed plants being just opposite. Moreover, organic acid exudation varied also with N form for the Cd-stressed plants. In comparison with other two N forms, (NH₄)₂SO₄,-treated plants had higher grain yield, N concentration and lower Cd concentration in plants. The current results indicated that (NH₄)₂SO₄ is a better fertilizer for use in Cd contaminated soil.     

Toxic Effect of Cadmium on Rice as Affected by Nitrogen Fertilizer Form

A nutrient solution experiment was conducted to determine the influence of N forms on growth, oxidative stress, and Cd and N uptake in rice plants. The treatments were consisted of two Cd levels (0 and 1 μmol) and three N forms (NH₄)₂SO₄, NH₄NO₃ and Ca(NO₃)₂. The results indicated that without Cd addition in the culture solution, the N forms had no significant effect on all measured parameters, including plant growth, photosynthetic traits, malondialdehyde (MDA) concentration, superoxide dismutase (SOD) activity, and Cd and N concentration, while Cd addition in the medium resulted in significant differences in measured parameters among the three forms of N fertilizers. The least inhibition of growth was noted in (NH₄)₂SO₄-fed plants, and the largest in Ca(NO₃)₂-fed plants, when plants were exposed to Cd stress. The highest photosynthetic rate and chlorophyll content was also recorded in (NH₄)₂SO₄-fed plants. Addition of Cd caused a remarkable increase in SOD activity and MDA content in plants, and the extent of increase varied with N form, with (NH₄)₂SO₄-fed plants being smallest. In comparison with the control plants, the N concentration in roots and shoots was not significantly affected in (NH₄)₂SO₄-fed plants, but significant decrease in root N concentration was found for the NH₄NO₃ and Ca(NO₃)₂-fed plants under Cd stress. Moreover, the significant differences were also noted among the three N forms in both root and shoot Cd concentrations, with (NH₄)₂SO₄-fed plants being the lowest. The results indicated that the toxic effect of Cd on rice varied with the form of N fertilizer.     

Foliar applications of nitrogen to slash pine seedlings

Summary The absorption of N from foliar applications of various N sources by pine seedlings was studied under greenhouse conditions. Needles dipped into solutions of 4,000 ppm N from Ca(NO₃)₂ were burned slightly at the tips at two weeks. Although higher concentrations of (NH₂)₂CO and (NH₄)₂SO₄ could be used without plant damage, a uniform concentration of 3,000 ppm was used in all comparative tests of sources. The level of tissue N, brought about by soil fertilization 6 weeks previously, did not significantly influence absorption of foliarly applied N¹⁵. Soil moisture maintained at near 100, 60 to 70, and 25 to 35 per cent of water-holding capacity of the Leon fs did not significantly affect the absorption of tagged N. Greater quantities of N¹⁵ were absorbed as urea than as Ca(NO₃)₂ or (NH₄)₂SO₄. The use of a spreader-sticker increased the N¹⁵ uptake, regardless of the N compound used. However, the magnitude of the increased absorption associated with use as a sticker varied from 490 per cent with (NH₄)₂SO₄ to 260 per cent with urea. It was calculated that approximately 71, 45, and 39 per cent of foliar applied N was absorbed into needles within 24 hours from urea, Ca(NO₃)₂, and (NH₄)₂SO₄, respectively. Journal Paper No3588 of the Florida Agricultural Experiment Station, Gainesville, Florida.     

Effect of nitrogen forms on growth and chemical changes in the rhizosphere of strawberry plants

The experiment was set up to examine the influence of different nitrogen forms: (NH₄)₂SO₄, Ca(NO₃)₂ or NH₄NO₃ on growth response, root induced pH changes in the rhizosphere, root-borne acid phosphatase activity in strawberry plants cv. Senga Sengana. The plants grown on sandy mineral soil were fertilized with 3 forms of nitrogen, in concentrations of 46 mg N·kg⁻¹ soil. The plants were grown in rhizoboxes with removable plexiglass lids. To ensure the root growth along the plexiglass lids, the rhizoboxes were placed at an angle of about 50° with the lid on the lower side. In case of ammonium supply, the nitrification inhibitor DIDIN was added (10 mg·kg⁻¹ of moist soil) to prevent conversion of ammonium into nitrate. The growth response (roots and shoots) of strawberry plants were determined after 11 weeks of treatment with different N forms. The best development of the root system and shoots (root and shoot dry weight and root length) was obtained, when ammonium nitrate was supplied. It is suggested therefore, that NH₄NO₃ stimulates vegetative growth of strawberry plants cv. Senga Sengana. However, there were no statistical differences in a leaf and flower number of the plants grown under different forms of N-fertilization. Determination of rhizosphere pH, and acid phosphatase activity were executed using non-destructive techniques, which enabled weekly measurement of chemical changes in the rhizosphere. The results revealed that the form of nitrogen supplied had a predominant effect on chemical changes in the rhizosphere of strawberry plants. The highest pH values (average pH 6.8) were measured in the rhizosphere of individual plants supplied with Ca(NO₃)₂. Whereas the lowest pH values (average pH 5.8) were detected in the presence of (NH₄)₂SO₄. The curve of rhizosphere pH measured along individual roots of the plants treated with Ca(NO₃)₂ represents the highest pH values whereas the curve of rhizosphere pH under (NH₄)₂SO₄ treatment had the lowest pH values. The highest activity of acid phosphatase were observed in the rhizosphere of strawberry plants grown in the presence of (NH₄)₂SO₄, at pH 5.8.     

Yield and nitrogen uptake of rapessed (Brassica campestris L.) with ammonium and nitrate

Water culture, growth chamber, greenhouse and field experiments were conducted to compare the effect of NH₄−N and NO₃−N on yield and N uptake of rapeseed (Brassica campestris L.). In water culture, the yields of 28-day old rapeseed plants grown at 14 μg N ml⁻¹ were double with NO₃ compared to NH₄, but N uptake was little affected. There was no such effect when concentration was reduced to 3.5 or 7 μg N ml⁻¹. The yield and N uptake of 26-day old rapeseed grown on six soils (pH 4.6 to 6.5) in pots in a growth chamber were much greater with NO₃ than with NH₄, although N concentration was more in the NH₄- than the NO₃-grown plants. In a greenhouse experiment with rapeseed grown on 12 potted soils, the N uptake of applied N was greater with NO₃ than with NH₄ on all soils. Averages were 63% with NH₄ and 78% with NO₃. However, NH₄-fixation capacities of the soils were only weakly correlated with yield from the two sources of N (r=0.48) and the relation was similar with N uptake. In contrast to the behavior of water culture, growth chamber and greenhouse experiments, the 33 field experiments did not show consistent difference in seed yield with NH₄ and NO₃ applied at time of seeding. In nine field experiments where band application was used for Ca(NO₃)₂, (NH₄)₂ SO₄, NH₄ NO₃, yield tended to be greatest for (NH₄)₂SO₄. However, in 19 experiments on acid soils with and without lime, yields in most cases were similar with (NH₄)₂SO₄ and NH₄ NO₃. Nitrification inhibitors were added to spring banded NH₄-based fertilizers in five experiments, but the yields were not influenced. Scientific Paper No. 558, Lacombe Research Station, Agriculture Canada.     

Application of nitrogen in NO 3 − form increases rhizosphere alkalisation in the subsurface soil layers in an acid soil

Leaching of NO ₃ ^? derived from ammoniacal fertilizers in the topsoil and subsequent uptake of NO ₃ ^? by plants from deeper layers may be used as a method of biological amelioration of subsurface soil acidity. This paper reports a glasshouse column experiment testing the above concept. Nitrogen with labelled ¹⁵N was supplied with and without lime to the surface soil (0–10 cm) as urea, (NH₄)₂SO₄ or Ca(NO₃)₂ at a rate equivalent to 120 kg N ha^?1. Soil columns were regularly watered from the top to facilitate NO ₃ ^? leaching. An aluminium-tolerant wheat genotype was grown for 38 days. The application of lime with nitrogen fertilizers increased growth of shoot and roots in all soil layers. The application of Ca(NO₃)₂ resulted in about 66% of recovery efficiency irrespective of whether lime was applied in the surface. This in turn resulted in about 0.2 units increase in rhizosphere pH in the subsurface (10–15 cm) soil layer compared to the same layer of the unlimed control. The supply of urea and (NH₄)₂SO₄ alone or with lime did not increase rhizosphere pH in the subsurface soil layers. Importantly, this study indicates that it is possible to exploit the process of nitrate uptake by wheat to increase pH in acidic subsurface soil.     

Multiple stress and growth of barley: Effect of salinity and temperature shock

The effect of preconditioning to NaCl salinity (0 to 135 mmol L^-1) on the subsequent response of barley (Hordeum vulgare L.) to two days of low (5°C) temperature shock (LTS) was investigated. Both salinity and LTS reduced the final growth of barley tops and roots. The effect of LTS on growth of tops and roots depended on the level of salinity stress imposed. At salinity level of 45 mmol L^-1, for example, exposing the plants to LTS reduced top growth by an additional 34%; at 135 mmol L^-1 salinity, however, LTS reduced the top growth by only 2%. Salinity increased the concentration of Na, Cl, total P, PO₄, and Zn, reduced the concentration of K, Ca, total N, NO₃, and SO₄, but did not affect the concentration of total S in the barley tops. LTS increased the concentration of Ca and Zn in the tops; the concentrations of other elements (cations and anions) were not changed by the temperature treatment. In the tops of the control plants, NO₃, PO₄, and SO₄ accounted for 15%, 72% and 93% of the total N, P, and S, respectively. In the plants grown at 135 mmol L^-1 NaCl, however, the above values were 8%, 84%, and 70%, respectively, which indicates that salinity had altered the incorporation of N, P, and S into organic compounds. We suggest that salinity and low temperature affect growth and nutrient uptake and incorporation into organic matter by different mechanisms. Although barley subjected to low salinity becomes more sensitive to subsequent low temperature stress, preconditioning of barley to higher salinity stress seems to reduce the plant's sensitivity to subsequent low temperature.     

Toxicity of nitrapyrin to sunflower under different N nutrition regimes

Summary The purpose of this study was to investigate the phytotoxicity of nitrapyrin 2-chloro-6-(trichloromethyl)pyridine to sunflower (Helianthus annuus L.) under different N regimes and to see if N forms affect the phytotoxicity of nitrapyrin. Sunflower was grown in pot culture for 21 days and was fertilized with (NH₄)₂SO₄, NH₄NO₃ and NaNO₃ to provide 0, 100 and 200 ppm N and with nitrapyrin application of 0 and 20 ppm. All N-treated sunflower plants in all N regimes and regardless of titrapyrin treatment produced more root and shoot dry weights and contained a significantly higher N than untreated check. Nitrapyrin toxicity appeared as a curling of leaf margin and a tendril type of stem growth, the visible toxicity symptoms decreased in the order: (NH₄)₂SO₄>NH₄NO₃>NaNO₃. Furthermore nitrapyrin addition suppressed sunflower growth in each N regime, the suppressing effect being greater with (NH₄)₂SO₄ and NH₄NO₃ than as with NaNO₃. Although, shoot growth from plants receiving nitrapyrin was not significantly affected by any N regime, root growth of nitrapyrin-treated plants was somewhat restricted by NH₄ ⁺−N nutrition relative to NO₃ ⁻−N nutrition.     

Effects of parathion and malathion on transformations of urea and ammonium sulfate nitrogen in soils

Summary A study of the effects of malathion and parathion applied at 10 and 50 g/g of soil on transformations of urea and (NH₄)₂SO₄–N in a sandy loam showed that the insecticides retarded urea hydrolysis as well as nitrification of urea and (NH₄)₂SO₄–N. At 50 parts/10⁶ rate of the insecticides, inhibition of urea hydrolysis ranged from 44 to 61% after 0.5 week and from 7 to 21% after 3 weeks of application. The insecticides inhibited the conversion of NH₄ ⁺ to NO₂ ^– without appreciably affecting the subsequent oxidation of NO₂ ^– to NO₃ ^– –N. This resulted in accumulation of higher amounts of NH₄ ⁺–N in soil samples treated with ammonium sulfate or urea N.The results suggest that transformations of urea and NH₄ ⁺ fertilizers in soils may be influenced by the amount of organophosphorus insecticide present and this may affect plant nutrition and fertilizer use.     

Effect of different concentrations of some ammonium fertilizers on the germination ofPennisetum typhoides Stapf & Hubb. andSorghum vulgare Pers

Summary Germination experiments were carried out with varying but equivalent amounts of NH₄Cl, (NH₄)₂SO₄, and NH₄NO₃, while in one experiment urea was included. It could be stated that the inhibitory effect on germination was ionic in nature, the effect of urea being rather small, while it was safer to use NH₄Cl or (NH₄)₂SO₄ than NH₄NO₃, especially under dry conditions.     

Assay of molybdenum cofactor of barley

Conditions for assay of molybdenum cofactor in barley shoot extracts in the presence of molybdate (25 mM N₂MoO₄) and the sulphydryl-group protector, reduced glutathione (5 mM) were optimized. Both total Mo-cofactor (assayed after heat-treatment of cell-free extracts) and ‘free’ Mo-cofactor (assayed in untreated cell-free extracts) were assayed. Compared to control plants grown in the absence of an exogenous nitrogen source total Mo-cofactor levels increased around 70 % when plants were grown for 4 days in the presence of either 15 mM KNO₃ or 15 mM NH₄NO₃. Growth in the presence of 15 mM (NH₄)₂SO₄ did not affect the Mo-cofactor level. Very similar results were seen when plants were transferred to these nitrogen sources for 24 hr after previous growth in the absence of an exogenous nitrogen source. In contrast ‘free’ Mo-cofactor levels of both KNO₃ and NH₄NO₃-treated plants were increased 2-3-fold over untreated controls. Growth in the presence of (NH₄)₂SO₄ did not affect the ‘free’ Mo-cofactor level.     

Comparison of the efficiency of urea,aqueous ammonia and ammonium sulphate as nitrogen fertilizers

Summary Nitrogen-15 labelled urea, aqueous NH₃ and (NH₄)₂SO₄ were applied to soils contained in pots. The fertilizers were injected in 5 cm³ of solution, 3.5 cm below the soil surface, to simulate a fertilizer band in the field. Ryegrass (Lolium perenne) was planted, and several cuttings and roots were harvested. Efficiency was determined as the recovery of fertilizer-N in the plant tissues and soil.Total recovery varied from 94 to 100%. There was no significant difference between the total recovery of the 3 fertilizer forms, although recovery in the soil component was lower for (NH₄)₂SO₄ than for urea or NH₃. There was a significant difference in total recovery between soils due to the soil component. Only small amounts of¹⁵N were not recovered, whereas laboratory experiments reported elsewhere had demonstrated that substantial gaseous losses of N as N₂, N₂O and NO +NO₂ occurred in these soils during nitrification of added NH₃ fertilizer.     

Effect of nitrogen form on maize response to drought stress

Two hybrids of maize (Zea mays L.) differing in resistance to drought, were grown in chernozem soil in a greenhouse and were fertilized with two different forms of nitrogen: Ca(NO₃)₂ and (NH₄)₂SO₄ in concentrations corresponding to 100 kg of N ha^-1. After emergence of the 4th leaf, plants were exposed to drought. During the drought period, the parameters of plant water status (water potential, osmotic potential, turgor pressure and relative water content) and chlorophyll a+b concentration were monitored every two days. N and K concentration and accumulation over the drought period were also monitored.Next to differences in adaptability of the two hybrids to drought, the results demonstrate different adaptability of NH₄ and NO₃-treated plants within each hybrid. NH₄-plants of each hybrid maintain higher turgor pressure during the drought by better osmotic adaptation. Especially significant differences appear between chlorophyll (a+b) values of NH₄ and NO₃-treated plants and as affected by drought. Chlorophyll concentrations of NH₄-plants are higher than those of NO₃-plants both in control and droughted plants. NH₄ plants show a characteristic initial chlorophyll increase at the beginning of the drought period while in NO₃ plants chlorophyll constantly decreases throughout the whole drought period. The influence of the nitrogen form on chlorophyll concentration changes during drought does not appear to be affected by regulation of the K concentration.     

Effect of some nitrogenous salts on nitrogen transfer and protease activity in germinatingZea mays L. seeds

Maize seeds were allowed to germinate in the presence of different nitrogenous salts for 72 h. Changes in the ethanol soluble and insoluble nitrogen were studied in the embryo and in the endosperm. Supply of Ca(NC₃)₂ enhanced germination and protease activity in the endosperm resulting in greater solubilisation of protein to soluble nitrogen in the seeds. NH₄NO₃ and (NH₄)₂SO₄ were less effective as compared to Ca(NO₃)₂. Cycloheximide inhibited germination and protease activity. Pretreatment also resulted in increase in growth, soluble and insoluble nitrogen, and nitrate reductase activity in the primary leaves. Ca(NO₃)₂ was more effective than NH₄NO₃ and (NH₄)₂SO₄.     

Metabolic regulation and gene expression of root phosphoenolpyruvate carboxylase by different nitrogen sources

Alfalfa (Medicago sativa L.) N-sufficient plants were fed 1·5 mM N in the form of NO₃⁻, NH₄⁺ or NO₃⁻ in conjunction with NH₄⁺, or were N-deprived for 2 weeks. The specific activity of phosphoenolpyruvate carboxylase (PEPC) from the non-nodulated roots of N-sufficient plants was increased in comparison with that of N-deprived plants. The PEPC value was highest with NO₃⁻ nutrition, lowest with NH₄⁺ and intermediate in plants that were fed mixed salts. The protein was more abundant in NO₃⁻-fed plants than in either NH₄⁺- or N mixed-fed plants. Nitrogen starvation decreased the level of PEPC mRNA, and nitrate was the N form that most stimulated PEPC gene expression. The malate content was significantly lower in NO₃⁻-deprived than in NO₃⁻-sufficient plants. Root malate accumulation was high in NO₃⁻-fed plants, but decreased significantly in plants that were fed with NH₄⁺. The effect of malate on the desalted enzyme was also investigated. Root PEPC was not very sensitive to malate and PEPC activity was inhibited only by very high concentrations of malate. Asparagine and glutamine enhanced PEPC activity markedly in NO₃⁻-fed plants, but failed to affect plants that were either treated with other N types or N starved. Glutamate and citrate inhibited PEPC activity only at optimal pH. N-nutrition also influenced root nitrate and ammonium accumulation. Nitrate accumulated in the roots of NO₃⁻- and (NO₃⁻ + NH₄⁺)-fed plants, but was undetectable in those administered NH₄⁺. Both the nitrate and the ammonium contents were significantly reduced in NO₃⁻- and (NO₃⁻ + NH₄⁺)-starved plants. Root accumulation of free amino acids was strongly influenced by the type of N administered. It was highest in NH₄⁺-fed plants and the most abundant amides were asparagine and glutamine. It was concluded that root PEPC from alfalfa plants is N regulated and that nitrate exerts a strong influence on the PEPC enzyme by enhancing both PEPC gene expression and activity.     

The effect of C/N ratio on lipid production byRhodosporidium toruloides ATCC 10788

Summary An optimum initial molar C/N ratio of about 77 was found for maximum lipid production from glucose or fructose with a nitrogen source from either (NH₄)₂SO₄, NH₄NO₃ or urea. The fatty acid composition of the lipids was affected by the initial C/N ratio and by the type of sugar but not significantly by the type of nitrogen.     

Nitrate reductase activity and nitrogen compounds in xylem exudate of Juglans nigra seedlings: relation to nitrogen source and supply

Nitrogen (N) limits plant productivity and its uptake and assimilation may be regulated by N source, N availability, and nitrate reductase activity (NRA). Knowledge of how these factors interact to affect N uptake and assimilation processes in woody angiosperms is limited. We fertilized 1-year-old, half-sib black walnut (Juglans nigra L.) seedlings with ammonium (NH₄ ⁺) [as (NH₄)₂SO₄], nitrate (NO₃ ⁻) (as NaNO₃), or a mixed N source (NH₄NO₃) at 0, 800, or 1,600 mg N plant⁻¹ season⁻¹. Two months following final fertilization, growth, in vivo NRA, plant N status, and xylem exudate N composition were assessed. Specific leaf NRA was higher in NO₃ ⁻-fed and NH₄NO₃-fed plants compared to observed responses in NH₄ ⁺-fed seedlings. Regardless of N source, N addition increased the proportion of amino acids (AA) in xylem exudate, inferring greater NRA in roots, which suggests higher energy cost to plants. Root total NRA was 37% higher in NO₃ ⁻-fed than in NH₄ ⁺-fed plants. Exogenous NO₃ ⁻ was assimilated in roots or stored, so no difference was observed in NO₃ ⁻ levels transported in xylem. Black walnut seedling growth and physiology were generally favored by the mixed N source over NO₃ ⁻ or NH₄ ⁺ alone, suggesting NH₄NO₃ is required to maximize productivity in black walnut. Our findings indicate that black walnut seedling responses to N source and level contrast markedly with results noted for woody gymnosperms or herbaceous angiosperms.     

Effects of Ca(NO<Subscript>3</Subscript>)<Subscript>2</Subscript> stress on oxidative damage,antioxidant enzymes activities and polyamine contents in roots of grafted and non-grafted tomato plants

The effects of Ca(NO₃)₂ stress on biomass production, oxidative damage, antioxidant enzymes activities and polyamine contents in roots of grafted and non-grafted tomato plants were investigated. Results showed that when exposed to 80 mM Ca(NO₃)₂ stress, the biomass production reduction in non-grafted plants was more significant than that of grafted plants. Under Ca(NO₃)₂ stress, superoxide anion radical (O₂•⁻) producing rate, hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) contents of non-grafted plants roots were significantly higher than those of grafted plants, however, nitrate (NO₃ ⁻), ammonium (NH₄ ⁺) and proline contents, superoxide dismutase (SOD, EC1.15.1.1), peroxidase (POD, EC1.11.1.7), catalase (CAT, EC1.11.1.6) and arginine decarboxylase (ADC, EC 4.1.1.19) activities of grafted plants roots were significantly higher than those of non-grafted plants. Regardless of stress, free, conjugated and bound polyamine contents in roots of grafted plants were significantly higher than those of non-grafted plants. The possible roles of antioxidant enzymes, prolines and polyamines in adaptive mechanism of tomato roots to Ca(NO₃)₂ stress were discussed. Gu-Wen Zhang and Zheng-Lu Liu contributed equally to this work.