NH4 + and NO3 − requirement for wheat somatic embryogenesis

The influence of three nitrogen salts: NH₄NO₃, KNO₃ and NH₄Cl on wheat in vitro cultures was investigated. Both NO ₃ ⁻ and NH ₄ ⁺ ions were indispensable for proliferation of embryogenic calli and development of wheat somatic embryos. It is possible to obtain wheat somatic embryos when the medium is enriched with NH₄NO₃ only as a source of inorganic nitrogen. The results of the statistical analysis showed that the level of NH₄NO₃ and KNO₃ in the medium had a great influence on the efficiency of somatic embryogenesis. We observed tendency that calli on media containing 50 mM NH₄NO₃ and 0 to 20 mM KNO₃ turned out to be more embryogenic than on control MS medium. High concentrations of KNO₃- 100 mM inhibited somatic embryogenesis, while 100 mM NH₄NO₃ did not. The level of total N did not have significant influence on wheat somatic embryogenesis. Ratio NO ₃ ⁻ :NH ₄ ⁺ also turned out to be not substantial. We observed that mutual connection of concentration levels between NH₄NO₃ and KNO₃ and between NH₄Cl and KNO₃ was more important. The efficiency of somatic embriogenesis obtained in the experiment with NH₄Cl and KNO₃ was significantly lower than in experiment with NH₄NO₃ and KNO₃.     

Role of cytokinin in enhanced productivity of maize supplied with NH4 + and NO3 −

Supplying both N forms (NH₄ ⁺+NO₃ ⁻) to the maize (Zea mays L.) plant can optimize productivity by enhancing reproductive development. However, the physiological factors responsible for this enhancement have not been elucidated, and may include the supply of cytokinin, a growth-regulating substance. Therefore, field and gravel hydroponic studies were conducted to examine the effect of N form (NH₄ ⁺+NO₃ ⁻ versus predominantly NO₃ ⁻) and exogenous cytokinin treatment (six foliar applications of 22 μM 6-benzylaminopurine (BAP) during vegetative growth versus untreated) on productivity and yield of maize. For untreated plants, NH₄ ⁺+NO₃ ⁻ nutrition increased grain yield by 11% and whole shoot N content by 6% compared with predominantly NO₃ ⁻. Cytokinin application to NO₃ ⁻-grown field plants increased grain yield to that of NH₄ ⁺+NO₃ ⁻-grown plants, which was the result of enhanced dry matter partitioning to the grain and decreased kernel abortion. Likewise, hydroponically grown maize supplied with NH₄ ⁺+NO₃ ⁻ doubled anthesis earshoot weight, and enhanced the partitioning of dry matter to the shoot. NH₄ ⁺+NO₃ ⁻ nutrition also increased earshoot N content by 200%, and whole shoot N accumulation by 25%. During vegetative growth, NH₄ ⁺+NO₃ ⁻ plants had higher concentrations of endogenous cytokinins zeatin and zeatin riboside in root tips than NO₃ ⁻-grown plants. Based on these data, we suggest that the enhanced earshoot and grain production of plants supplied with NH₄ ⁺+NO₃ ⁻ may be partly associated with an increased endogenous cytokinin supply.     

Composition of the xylem sap of tomato seedlings cultivated on media with HCO3 − and nitrogen source as NO3 − or NH4 +

The results of the experiments discussed here present changes in the chemical composition of xylem sap of tomato seedlings cultivated in hydroponics on media containing 5 mmol HCO₃ ^– and an N-source given as NO₃ ^–, NH₄ ⁺ or these two forms in different proportions. The occurrence of free NH₄ ⁺ in the xylem sap of NH₄ ⁺-seedlings and in NO₃ ^–-seedlings indicates that the process of N-assimilation was not only confined to roots. The application of HCO₃ ^– to the medium effected a decrease in the concentration of NH₄ ⁺ in the xylem sap of NH₄ ⁺-seedlings, having no effect on changes in the concentration of NO₃ ^– or NH₄ ⁺ in NO₃ ^–-seedlings. Malate, citrate, fumarate, and succinate were identified in the xylem sap. The concentration of carboxylates in NO₃ ^–-seedlings exceeded by about 50% that recorded in NH₄ ⁺-seedlings. The highest concentration of malate constituting from 80% to 93.5% of this fraction, was determined in this group of compounds. The enrichment of the medium with HCO₃ ^– ions induced an increase in the content of carboxylates, chiefly of malate. In these experimental conditions an increase in the malate concentration in the xylem sap of NO₃ ^– and NH₄ ⁺-seedlings reached relative values of 100% and 36%, respectively. The total concentration of amides and amino acids was about 2.6 times higher in the xylem sap of NH₄ ⁺-seedlings than in NO₃ ^–-seedlings. Amide glutamine was the main component of this fraction in xylem sap and its total concentration was about 3.3 times higher in NH₄ ⁺-seedlings than that determined in NO₃ ^–-seedlings. Glutamine, glutamate, aspargine, and aspartate constituted from 69% to 77% of this fraction. The concentration of the remaining amino acids varied from 0.6% to 7%. The enrichment of the medium with HCO₃ ^– ions also effected an increase in the concentration of amides and amino acids in the xylem sap by about 17% and 56% in the case of NO₃ ^– and NH₄ ⁺-seedlings, respectively, in comparison with the respective controls (without HCO₃ ^–). Abbreviations: DAG – days after germination; DIC – dissolved inorganic carbon; GOGAT – glutamine:2-oxoglutarate aminotransferase; GS – glutamine synthetase; PAR – photosynthetically active radiation; PEPc – phosphoenolpyruvate carboxylase     

Effects of clipping and shading on 15NO3− and 15NH4+ recovery by plants in grazed and ungrazed temperate grasslands

Plant and Soil - In natural ecosystems, plants generally promote the acquisition of nitrogen (N) through the input of carbon (C) into the soil. The present study aimed to clarify how changes in C...     

Influence of nitrogen form and NH4 +-N/:NO3 −-N ratios on adventitious shoot formation from pear (Pyrus communis) leaf explants in vitro

The effect of various basal salts media, containing different nitrogen levels on in vitro adventitious shoot regeneration from leaf explants of Louise Bonne Panachee and Seckel pear (Pyrus communis L.) were investigated. Among the different basal salt formulae tested, Nitsch (1969) gave significantly better regeneration in most of the experiments. Shoot regeneration was altered with different NH₄ ⁺-N/NO₃ ^–-N ratios. The best regeneration was obtained when NH₄ ⁺:NO₃ ^– was either 1:2 or 1:3 regardless of overall N concentration. In addition, these data show that NH₄ ⁺ was essential for adventitious shoot regeneration from pear leaf explants on White's (1943) medium.     

Effect of NH4+/NO3− availability on nitrate reductase activity and nitrogen accumulation in wetland helophytes Phragmites australis and Glyceria maxima

The effect of NH₄⁺/NO₃⁻ availability on nitrate reductase (NR) activity in Phragmites australis and Glyceria maxima was studied in sand and water cultures with the goal to characterise the relationship between NR activity and NO₃⁻ availability in the rhizosphere and to describe the extent to which NH₄⁺ suppresses the utilization of NO₃⁻ in wetland plants.The NR activity data showed that both wetland helophytes are able to utilize NO₃⁻. This finding was further supported by sufficient growth observed under the strict NO₃⁻ nutrition. The distribution of NR activity within plant tissues differed between species. Phragmites was proved to be preferential leaf NO₃⁻ reducer with high NR activity in leaves (NR_max > 6.5 μmol NO₂⁻ g dry wt⁻¹ h⁻¹) under all N treatments, and therefore Phragmites seems to be good indicator of NO₃⁻ availability in flooded sediment. In the case of Glyceria the contribution of roots to plant NO₃⁻ reduction was higher, especially in sand culture. Glyceria also tended to accumulate NO₃⁻ in non-reduced form, showing generally lower leaf NR activity levels. Thus, the NR activity does not necessarily correspond with plant ability to take up NO₃⁻ and grow under NO₃⁻-N source. Moreover, the species differed significantly in the content of compounds interfering with NR activity estimation. Glyceria, but not Phragmites, contained cyanogenic glycosides releasing cyanide, the potent NR inhibitor. It clearly shows that the use of NR activity as a marker of NO₃⁻ utilization in individual plant species is impossible without the precise method optimisation.     

New insights into carbon acquisition and exchanges within the coral–dinoflagellate symbiosis under NH4+ and NO3? supply

Anthropogenic nutrient enrichment affects the biogeochemical cycles and nutrient stoichiometry of coastal ecosystems and is often associated with coral reef decline. However, the mechanisms by which dissolved inorganic nutrients, and especially nitrogen forms (ammonium versus nitrate) can disturb the association between corals and their symbiotic algae are subject to controversial debate. Here, we investigated the coral response to varying N : P ratios, with nitrate or ammonium as a nitrogen source. We showed significant differences in the carbon acquisition by the symbionts and its allocation within the symbiosis according to nutrient abundance, type and stoichiometry. In particular, under low phosphate concentration (0.05 µM), a 3 µM nitrate enrichment induced a significant decrease in carbon fixation rate and low values of carbon translocation, compared with control conditions (N : P = 0.5 : 0.05), while these processes were significantly enhanced when nitrate was replaced by ammonium. A combined enrichment in ammonium and phosphorus (N : P = 3 : 1) induced a shift in nutrient allocation to the symbionts, at the detriment of the host. Altogether, these results shed light into the effect of nutrient enrichment on reef corals. More broadly, they improve our understanding of the consequences of nutrient loading on reef ecosystems, which is urgently required to refine risk management strategies.     

Effects of Al on nitrogen (NH 4 + and NO 3 − ) uptake,nitrate reductase activity and proton release in two sorghum cultivars differing in Al tolerance

After growth for 17 to 36 days on nutrient solutions with NH₄NO₃ as nitrogen source (pH 4.2) dry matter of sorghum genotype SC0283 was much less affected by Al (1.5 and 3.0 ppm) than that of genotype NB9040. In the absence of Al both cultivars released protons into the nutrient solution as a result of an excess of cationic nutrients taken up. When Al was present, this proton efflux per unit dry weight increased drastically, especially with the sensitive genotype NB9040. Chemical analysis of plant material and continuous analyses of NO ₃ ⁻ and NH ₄ ⁺ in the nutrient solution indicated, that the Al-induced shift in H⁺-balance of both genotypes could almost completely be attributed to a decreased NO ₃ ⁻ /NH ₄ ⁺ uptake ratio. In vivo nitrate reductase activity (NRA) was reduced in the shoot of NB9040 and to a lesser degree in SC0283. Al-induced decrease in NRA was accompanied by similar percentual decreases in NO ₃ ⁻ tissue concentrations. Therefore this decrease is interpreted as being indirect,i.e., the consequence of the reduced NO ₃ ⁻ uptake of the plants. A direct repression of NRA by Al seems also unlikely because nitrate reductase activity of the roots (where cellular Al-concentrations should be higher than in shoots) was not affected in Al-treated plants of either genotype.     

Short-term studies of NO 3 − uptake in Pisum using 13NO 3 −

Influx, efflux and net uptake of NO ₃ ^– was studied in Pisum sativum L. cv. Marma in short-term experiments where ¹³NO ₃ ^– was used to trace influx. The influx rate in N-limited plants was similar both during net uptake at external concentrations of around 50 M, and at low external NO ₃ ^– concentrations (4–6 M) when net uptake was practically zero. Efflux could be inferred from discrepancies between influx and net uptake but was never very high in the N-limited plants during net uptake. Close to the threshold concentration for not NO ₃ ^– uptake, efflux was high and equalled influx. Thus, the threshold concentration can be regarded as a NO ₃ ^– compensation point. The inclusion of NH ₄ ⁺ in the outer medium decreased influx by about 40% but did not significantly affect efflux. The roles of NO ₃ ^– fluxes and nitrate-reductase activity in regulating/limiting NO ₃ ^– utilization are discussed.Abbreviations DW dry weight - FW fresh weight - R_N relative nitrogen addition rate     

Alteration in enrichment of NO 3 − and reduced-N in xylem exudate during and after extended plant exposure to15NO 3 −

Summary Distinctly different patterns of¹⁵N enrichment were observed in the nitrate and reduced-N fractions of xylem exudate from soybean plants during and after 5 to 6 days of exposure to¹⁵NO ₃ ^– . Within 1 d after changes in solution NO ₃ ^– label, more than 90% of the exudate NO ₃ ^– originated from the exogenous supply. Alterations in the enrichment of exudate reduced-N were much slower, however, and the enrichment reached only 40% even after 5 d of continuous exposure to¹⁵NO ₃ ^– . Taking into account possible reduction of endogenous NO ₃ ^– and delayed translocation of NO ₃ ^– reduction products, it was concluded that root reduction could have contributed only 30 to 42% of the reduced-N found in the exudate.     

The role of gross and net N transformation processes and NH4 + and NO3 − immobilization in controlling the mineral N pool of a temperate mixed deciduous forest soil

In many forests of Europe and north-eastern North America elevated N deposition has opened the forest N cycle, resulting in NO₃ ^? leaching. On the other hand, despite this elevated N deposition, the dominant fate of NO₃ ^? and NH₄ ⁺ in some of these forests is biotic or abiotic immobilization in the soil organic matter pool, preventing N losses. The environmental properties controlling mineral N immobilization and the variation and extent of mineral N immobilization in forest soils are not yet fully understood. In this study we investigated a temperate mixed deciduous forest, which is subjected to an average N deposition of 36.5 kg N ha^?1 yr^?1, but at the same time shows low NO₃ ^? concentrations in the groundwater. The aim of this study was to investigate whether the turnover rate of the mineral N pool could explain these low N leaching losses. A laboratory ¹⁵N pool dilution experiment was conducted to study gross and net N mineralization and nitrification and mineral N immobilization in the organic and uppermost (0–10 cm) mineral layer of the forest soil. Two locations, one at the forest edge (GE) and another one 145 m inside the forest (GF1), were selected. In the organic layers of GE and GF1, the gross N mineralization averaged 10.9 and 11.1 mg N kg^?1 d^?1, the net N mineralization averaged 6.1 and 6.8 mg N kg^?1 d^?1 and NH₄ ⁺ immobilization rates averaged 3.8 and 3.6 mg N kg^?1 d^?1. In the organic layer of GE and GF1, the average gross nitrification was 3.8 and 4.6 mg N kg^?1 d^?1, the average net nitrification was ?25.2 and ?31.3 mg N kg^?1 d^?1 and the NO₃ ^? immobilization rates averaged 29.0 and 35.9 mg N kg^?1 d^?1. For the mineral (0–10 cm) layer the same trend could be observed, but the N transformation rates were much lower for the NH₄ ⁺ pool and not significantly different from zero for the NO₃ ^? pool. Except for the turnover of the NH₄ ⁺ pool in the mineral layer, no significant differences were observed between location GE and GF1. The ratio of NH₄ ⁺ immobilization to gross N mineralization, gross N mineralization to gross nitrification, and NO₃ ^? immobilisation to gross nitrification led to the following observations. The NH₄ ⁺ pool of the forest soil was controlled by N mineralization and NO₃ ^? immobilization was importantly controlling the forest NO₃ ^? pool. Therefore it was concluded that this process is most probably responsible for the limited NO₃ ^? leaching from the forest ecosystem, despite the chronically high N deposition rates.     

Hg2+-induced leakage of electrolytes and inhibition of NO3 − utilization inNostoc calcicola

Summary The effect of mercury (Hg²⁺) in the absence and presence of methylmercury (CH₃Hg⁺), cadmium (Cd²⁺), copper (Cu²⁺), nickel (Ni²⁺) and calcium (Ca²⁺) on Nostoc calcicola Bréb. has been studied in terms of electrolyte leakage, NO₃ ^– uptake and in vivo nitrate reductase (NR) activity to discover any possible correlation among such parameters under Hg²⁺ stress. Leakage of electrolytes from Hg²⁺-treated cyanobacterial cells was directly proportional to Hg²⁺ concentrations and exposure time. In comparison to NO₃ ^– uptake, an about 60-fold slower rate of NR activity was observed in the untreated cultures, the former being five times more Hg²⁺-sensitive. A non-competitive synergistic interaction of Hg²⁺ with CH₃Hg⁺ or Cd²⁺ and antagonistic with that of Ni²⁺ or Ca²⁺ has been observed for both the processes of NO₃ ^– utilization. The antagonistic interaction of Cu²⁺ with Hg²⁺ in terms of NO₃ ^– uptake and synergistic with respect to NR activity, has been attributed to the dual bonding preference of Cu²⁺ for cellular ligands. These findings suggest that (a) a statistically significant correlation exists among such parameters; (b) Hg²⁺ predominantly attacks the cyanobacterial cell membrane; (c) Hg²⁺ inhibits NO₃ ^– utilization; (d) the presence of other cations increases or decreases the inhibitory actions of Hg²⁺.     

浅谈NH4NO3和植物根部环境pH的关系

1897年在灭菌条件下证明铵盐不经转变成硝酸盐也能为植物吸收,此后,植物既能吸收NO_3~--N,又能吸收NH_4~_ N已为人所共知。为此,有些学者认为,植物吸收NH_4NO_3的阴阳离子的量很     

The Uptake of NO3?, NO2?, and NH4+ by Intact Wheat (Triticum aestivum) Seedlings : I. Induction and Kinetics of Transport Systems

The inducibility and kinetics of the NO₃⁻, NO₂⁻, and NH₄⁺ transporters in roots of wheat seedlings (Triticum aestivum cv Yercora Rojo) were characterized using precise methods approaching constant analysis of the substrate solutions. A microcomputer-controlled automated high performance liquid chromatography system was used to determine the depletion of each N species (initially at 1 millimolar) from complete nutrient solutions. Uptake rate analyses were performed using computerized curve-fitting techniques. More precise estimates were obtained for the time required for and the extent of the induction of each transporter. Up to 10 and 6 hours, respectively, were required to achieve apparent full induction of the NO₃⁻ and NO₂⁻ transporters. Evidence for substrate inducibility of the NH₄⁺ transporters requiring 5 hours is presented. The transport of NO₃⁻ was mediated by a dual system (or dual phasic), whereas only single systems were found for transport of NO₂⁻ and NH₄⁺. The K_m values for NO₃⁻, NO₂⁻, and NH₄⁺ were, respectively, 0.027, 0.054, and 0.05 millimolar. The K_m for mechanism II of NO₃⁻ transport could not be defined in this study as it exhibited only apparent first order kinetics up to 1 millimolar.     

High-Affinity NO− 3-H+ Cotransport in the Fungus Neurospora: Induction and Control by pH and Membrane Voltage

High-affinity nitrate transport was examined in intact hyphae of Neurospora crassa using electrophysiological recordings to characterize the response of the plasma membrane to NO⁻ ₃ challenge and to quantify transport activity. The NO⁻ ₃-associated membrane current was determined using a three electrode voltage clamp to bring membrane voltage under experimental control and to compensate for current dissipation along the longitudinal cell axis. Nitrate transport was evident in hyphae transferred to NO⁻ ₃-free, N-limited medium for 15 hr, and in hyphae grown in the absence of a nitrogen source after a single 2-min exposure to 100 μm NO⁻ ₃. In the latter, induction showed a latency of 40–80 min and rose in scalar fashion with full transport activity measurable approx. 100 min after first exposure to NO⁻ ₃; it was marked by the appearance of a pronounced sensitivity of membrane voltage to extracellular NO⁻ ₃ additions which, after induction, resulted in reversible membrane depolarizations of (+)54–85 mV in the presence of 50 μm NO⁻ ₃; and it was suppressed when NH₄ ⁺ was present during the first, inductive exposure to NO⁻ ₃. Voltage clamp measurements carried out immediately before and following NO⁻ ₃ additions showed that the NO⁻ ₃-evoked depolarizations were the consequence of an inward-directed current that appeared in parallel with the depolarizations across the entire range of accessible voltages (−400 to +100 mV). Measurements of NO⁻ ₃ uptake using NO⁻ ₃-selective macroelectrodes indicated a charge stoichiometry for NO⁻ ₃ transport of 1(+):1(NO⁻ ₃) with common K _m and J _max values around 25 μm and 75 pmol NO⁻ ₃ cm⁻²sec⁻¹, respectively, and combined measurements of pH _o and [NO⁻ ₃] _o showed a net uptake of approx. 1 H⁺ with each NO⁻ ₃ anion. Analysis of the NO⁻ ₃ current demonstrated a pronounced voltage sensitivity within the normal physiological range between −300 and −100 mV as well as interactions between the kinetic parameters of membrane voltage, pH _o and [NO⁻ ₃] _o . Increasing the bathing pH from 5.5 to 8.0 reduced the current and the associated membrane depolarizations 2- to 4-fold. At a constant pH _o of 6.1, driving the membrane voltage from −350 to −150 mV resulted in an approx. 3-fold reduction in the maximum current and a 5-fold rise in the apparent affinity for NO⁻ ₃. By contrast, the same depolarization effected an approx. 20% fall in the K _m for transport as a function in [H⁺] _o . These, and additional results are consistent with a charge-coupling stoichiometry of 2(H⁺) per NO⁻ ₃ anion transported across the membrane, and implicate a carrier cycle in which NO⁻ ₃ binding is kinetically adjacent to the rate-limiting step of membrane charge transit. The data concur with previous studies demonstrating a pronounced voltage-dependence to high-affinity NO⁻ ₃ transport system in Arabidopsis, and underline the importance of voltage as a kinetic factor controlling NO⁻ ₃ transport; finally, they distinguish metabolite repression of NO⁻ ₃ transport induction from its sensitivity to metabolic blockade and competition with the uptake of other substrates that draw on membrane voltage as a kinetic substrate. Received: 17 March 1997/Revised: 20 June 1997     

Soybean yield response to residual NO3−N and applied N

Summary During 1976 through 1978, 10N treatments (combinations of N application times and rates) were used in a corn study. Those treatments created different levels of soil NO ₃ ^– –N content that were well-suited to a study of the influence of residual NO ₃ ^– –N and applied N on soybean yield. In April 1979 we applied ammonium nitrate at rates of 0, 75, or 150 kg N/ha to three subplots formed from each of the whole plots (previous N treatment plots). With N fertilization in 1979, seed yield increased where the residual NO ₃ ^– –N amount was less than 190 kg/ha but decreased where the residual amount was greater than 190 kg/ha. As the NO ₃ ^– –N content in the soil increased by 1 kg/ha, the soybean yield increase due to N fertilization in 1979 decreased by approximately 4 kg/ha.Contribution no. 82-368-J, Dep. of Agronomy, Kansas Agric. Exp. Stn., Manhattan, KS 66506, USA     

The interrelationship between HCO 3 − , NO 3 − , and Cl− as effective anions in the photosynthetic electron transport

Thylakoids were isolated from the leaves of three different plants (Pisum sativium L., Lactuca sativa L., and Raphanus sativus L.). The addition of HCO ₃ ^? to a suspension of salt-and HCO ₃ ^? -epleted thylakoids (suspended in salt-free medium) raised the rate of O₂ evolution up to fourfold. This stimulation could be partially replaced by the addition of chloride or nitrate ions. However, the addition of HCO ₃ ^? in the presence of Cl^? or NO ₃ ^? resulted in a higher stimulation of O₂ evolution (sixfold in the presence of nitrate and sevenfold in the presence of chloride). On the other hand, the addition of HCO ₃ ^? to the thylakoids depleted from salt only raised the rate of O₂ evolution by 10–15%, whereas 40–70% was obtained by the addition of nitrate or chloride ions. The fluorescence induction studies indicated a significant decrease in the yield of the variable fluorescence of the salt- and HCO ₃ ^? -depleted thylakoids. A partial increase in the fluorescence yield was obtained by the addition of HCO ₃ ^? alone. A typical fluorescence induction curves were obtained by the addition of HCO ₃ ^? in the presence of Cl^? or NO ₃ ^? ions. The data obtained suggest a similar role for chloride and nitrate ions in O₂ evolution in the Hill-reaction, which is restricted at the donor side of photosystem II, whereas bicarbonate plays its role at both sides (acceptor and donor sides). The presented data are those obtained for the thylakoids of P. sativium, which were more or less similar to those obtained for L. sativa and R. sativus.