Opium poppy capsule growth and alkaloid production is constrained by shade during early floral development

Morphinan alkaloids accumulate in the capsules of Papaver somniferum L. (opium poppy) most likely as defence against herbivory. Thus, capsule size is an important component of alkaloid yield. Shade during early cell division-dominated growth of reproductive structures generally reduces final fruit size more than shade during later cell expansion-dominated growth. The current study aimed to determine whether this response is found in opium poppy and the subsequent impact on alkaloid yield, composition and seed production. First the timing of key reproductive developmental events was resolved relative to macromorphological traits. Plants were then shaded during either (a) floral initiation, (b) early floral development or (c) capsule expansion before being harvested at maturation. Shade during floral initiation dramatically reduced final capsule size, alkaloid yield and seed number, and increased the concentration of precursor compounds relative to morphine, despite plants later returning to full sun. Shade during later capsule growth enhanced capsule size and alkaloid yield but had little effect on alkaloid composition or seed number. Thus, early developmental processes, including morphine biosynthesis, appear to have a relatively greater demand for carbohydrates compared with later processes. Crop management practices and environmental factors that limit carbohydrate availability during early development are thus predicted to have significant negative impacts on alkaloid production and reproductive success.     

Accumulation of Ergopeptide Alkaloids in Symbiotic Tall Fescue Grown under Deficits of Soil Water and Nitrogen Fertilizer

The fungus Acremonium coenophialum is endophytically associated with tall fescue (Festuca arundinacea Schreber). Within this symbiotum the fungus produces ergopeptide alkaloids, which are associated with livestock toxicoses. Environmental effects on the production of ergot alkaloids within the symbiotum are unknown. We conducted a greenhouse study of the effects of flooding, nitrogen rate during fertilization (11, 73, and 220 mg of N per pot weekly), nitrogen form (3.4 and 34 mg of N as NH₄⁺ or NO₃^- per pot), and drought stress (-0.03, -0.05, and -0.50 MPa) on ergopeptide alkaloid concentrations in one genotype of nonsymbiotic and symbiotic tall fescue grown in plastic pots. It was determined that the concentration of ergovaline, the major type of ergopeptide alkaloid, was increased but was not as high as that in nonflooded controls. Total ergopeptide and ergovaline concentrations in plants receiving high (220 mg of N per pot) and low (11 mg of N per pot) levels of NH₄NO₃ fertilization were not affected by flooding. The form of nitrogen was important since all concentrations of NO₃^--N increased ergopeptide alkaloid content, as opposed to the effects of NH₄⁺-N, which was effective only at high concentrations (34 mg of N per pot). Ergopeptide concentrations were highest in drought-stressed plants grown at -0.50 MPa and fertilized at the moderate or high N rate. The results suggest that within this genotype, ergopeptide alkaloid biosynthesis by the fungus is not appreciably affected by flooding but is greatly increased by high rates of N fertilization and moderate water deficit.     

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.     

Effect of nitrogen form and phosphorus source on the growth,nutrient uptake and rhizosphere soil property of Camellia sinensis L.

The effects of nitrogen form and phosphorus source on the growth, nutrient uptake and rhizosphere soil property of tea (Camellia sinensis L.) were investigated in a pot experiment. The experiment was performed with a compartmental cropping device, which enables the collection of rhizosphere soil at defined distances from the root of tea plant. Nitrogen was supplied as nitrate or ammonium in combination with soluble phosphorus as Ca(H₂PO₄)₂ or insoluble P as rock phosphate. The leaf dry matter production of tea was significantly greater in the treatments with NH₄ ⁺ than NO₃ ^-, whereas dry matter production of root and stem was not significantly affected. Addition of phosphorus as either source did not influence the dry matter production. The concentrations of K in root, Mg and Ca in both the shoot and root supplied with NO₃ ^- were significantly higher than in NH₄ ⁺ and influence of P sources was minor. On the contrary, Al and Mn concentrations were significantly larger in NH₄ ^--fed plants which could be attributed to remarkably increased availability of Al and Mn caused by acidification of the rhizosphere soil (the first 1-mm soil section from the root surface) with NH₄–N nutrition. The concentration of N in shoot was also significantly higher in NH₄- than in NO₃-fed plants, indicating higher use efficiency of NH₄–N. Whatever the phosphate source, rhizosphere pH declined in ammonium compared to in nitrate treatment. The pH decrease was much larger when no P or soluble P were applied and reached 0.85–1.30 units which extended to 3–5 mm away from the root surface. Exchangeable acidity, content of exchangeable Al and Mn were also considerably higher in the rhizosphere soils of NH₄ ⁺ fed tea plants. Significant amounts of P dissolved from rock phosphate accumulated in rhizosphere of NH₄ ⁺, not NO₃ ^-, suggesting that the dissolution of rock phosphate was induced by the proton excreted by tea root fed with ammonium. With soluble P addition, shoot and root P concentrations were greater in NH₄ ⁺ than in NO₃ ^- treatment and it appeared that this difference could not be sufficiently explained by the available P content in soil which was only slightly higher in NH₄ ⁺ treatment. With rock phosphate addition, the shoot and root P concentrations were hardly affected by nitrogen form, although the available P content was much higher and accumulated in the rhizosphere soil supplied with ammonium. The reason for this was discussed with regard to the inter-relationship of Al with P uptake. This revised version was published online in June 2006 with corrections to the Cover Date.     

Ammonium under solution culture alleviates aluminum toxicity in rice and reduces aluminum accumulation in roots compared with nitrate

Al stress and ammonium–nitrogen nutrition often coexist in acidic soils due to their low pH and weak nitrification ability. Rice is the most Al-resistant species among small grain cereal crops and prefers NH₄ ⁺ as its major inorganic nitrogen source. This study investigates the effects of NH₄ ⁺ and NO₃ ^? on Al toxicity and Al accumulation in rice, and thereby associates rice Al resistance with its NH₄ ⁺ preference. Two rice subspecies, indica cv. Yangdao6 and japonica cv. Wuyunjing7, were used in this study. After treatment with or without Al under conditions of varying NH₄ ⁺ and NO₃ ^? supply, rice seedlings were harvested for the determination of root elongation, callose content, biomass, Al concentration and medium pH. The results indicated that Wuyunjing7 was more Al-resistant and NH₄ ⁺-preferring than Yangdao6. NH₄ ⁺ alleviated Al toxicity in two cultivars compared with NO₃ ^?. Both NH₄ ⁺-Al supply and pretreatment with NH₄ ⁺ reduced Al accumulation in roots and root tips compared with NO₃ ^?. NH₄ ⁺ decreased but NO₃ ^? increased the medium pH, and root tips accumulated more Al with a pH increase from 3.5 to 5.5. Increasing the NO₃ ^? concentration enhanced Al accumulation in root tips but increasing the NH₄ ⁺ concentration had the opposite effect. These results show NH₄ ⁺ alleviates Al toxicity for rice and reduces Al accumulation in roots compared with NO₃ ^?, possibly through medium pH changes and ionic competitive effects. Making use of the protective effect of NH₄ ⁺, in which the Al resistance increases, is advised for acidic soils, and the hypothesis that rice Al resistance is associated with the preferred utilization of NH₄ ⁺ is suggested.     

Importance of the nitrogen source in the grass species Brachiaria brizantha responses to sulfur limitation

Background and aims

Plant responses to S supply are highly dependent on N nutrition. We investigated the effect of S status on metabolic, nutritional, and production variables in Brachiaria brizantha treated with different N forms. Additionally, ¹⁵N and ³⁴S root influx were determined in plants under short- and long-term S deprivation.

Methods

Plants were submitted to soil fertilization treatments consisted of combinations of N forms [without N, ammonium (NH₄ ⁺), nitrate (NO₃ ^?) or NH₄ ⁺+NO₃ ^?] at S rates (0, 15, 30, or 45 mg dm^?3). N and S influx capacity was determined in hydroponically-grown plants.

Results

Shoot production due to S supply increased 53, 145 and 196 % with NH₄ ⁺, NH₄ ⁺+NO₃ ^? and NO₃ ^? treatments, respectively. No or low S impaired protein synthesis and led to high accumulation of N-NO₃ ^? and asparagine in NO₃ ^?-fed plants, both alone and with NH₄ ⁺. Proline accumulation was observed in NH₄ ⁺-fed plants. Short- and long-term S deprivation did not promote considerable changes in ¹⁵N influx. ³⁴S absorption decreased depending on the N form provided: NH₄ ⁺+NO₃ ^? > only NH₄ ⁺ > only NO₃ ^? > low N.

Conclusions

Including both NH₄ ⁺ and NO₃ ^? forms in fertilizer increases N and S intake potential and thereby enhances plant growth, nutritional value and production.     

NO3/NH4 ratios affect the growth and N removal ability of Acorus calamus and Iris pseudacorus in a hydroponic system

Sweet flag (Acorus calamus L.) and yellow flag (Iris pseudacorus L.) have been used increasingly in constructed wetlands (CWs) for treatment of eutrophic wastewater. In order to properly match plant species with the type of wastewater being treated, it is important to know the performance of plant species under different NO₃⁻/NH₄⁺ ratios. We investigated dry matter (DW) production and N content of A. calamus and I. pseudacorus under five NO₃⁻/NH₄⁺ ratios (100/0, 75/25, 50/50, 25/75, and 0/100) in a hydroponic system. Results showed that the two species exhibited different preferences for NO₃⁻ and NH₄⁺. Total DW, shoot DW, and N content were greater with NO₃⁻/NH₄⁺ ratios of 50/50 and 75/25 than otherwise for A. calamus, but these parameters were only higher under the sole NO₃⁻ treatment in I. pseudacorus. We conclude that A. calamus could be best used for treating wastewater in constructed wetlands with NO₃⁻/NH₄⁺ ratios between 50/50 and 75/25, while I. pseudacorus for treating wastewater with NO₃⁻ only to achieve the highest biomass production and efficiency in the removal of N.     

Functional role of DNRA and nitrite reduction in a pristine south Chilean <Emphasis Type="Italic">Nothofagus</Emphasis> forest

Nitrite (NO₂ ⁻) is an intermediate in a variety of soil N cycling processes. However, NO₂ ⁻ dynamics are often not included in studies that explore the N cycle in soil. Within the presented study, nitrite dynamics were investigated in a Nothofagus betuloides forest on an Andisol in southern Chile. We carried out a ¹⁵N tracing study with six ¹⁵N labeling treatments, including combinations of NO₃ ⁻, NH₄ ⁺ and NO₂ ⁻. Gross N transformation rates were quantified with a ¹⁵N tracing model in combination with a Markov chain Monte Carlo optimization routine. Our results indicate the occurrence of functional links between (1) NH₄ ⁺ oxidation, the main process for NO₂ ⁻ production (nitritation), and NO₂ ⁻ reduction, and (2) oxidation of soil organic N, the dominant NO₃ ⁻ production process in this soil, and dissimilatory NO₃ ⁻ reduction to NH₄ ⁺ (DNRA). The production of NH₄ ⁺ via DNRA was approximately ten times higher than direct mineralization from recalcitrant soil organic matter. Moreover, the rate of DNRA was several magnitudes higher than the rate of other NO₃ ⁻ reducing processes, indicating that DNRA is able to outcompete denitrification, which is most likely not an important process in this ecosystem. These functional links are most likely adaptations of the microbial community to the prevailing pedo-climatic conditions of this Nothofagus ecosystem.     

Effect of infection by the endophytic fungus Acremonium lolii on growth and nitrogen uptake by perennial ryegrass (Lolium perenne) in flowing solution culture

Clonal tillers of a genotype of perennial ryegrass (Lolium perenne), either with or without the endophytic fungus Acremonium lolii, were grown under natural light in flowing nutrient solutions with mineral N maintained automatically at concentrations of 3 or 30μm NH₄NO₃ for 28 days. Uptake of N was monitored daily and dry matter production was assessed by sequential harvesting. The presence of endophyte had no significant effect on shoot or root biomass production at either N level, but shoot: root ratios were significantly increased by endophyte infection at both N levels at some harvests. All plants absorbed NH₄⁺ preferentially to NO₃^- and the ratio was not affected by endophyte infection. Also, infection did not affect total N content of plants, which was significantly more in plants at the higher N level than at the lower level. It is concluded that endophyte infection had only minor effects on growth and N economy of the plant, under the conditions imposed in this experiment.     

Responses of soybean to ammonium and nitrate supplied in combination to the whole root system or separately in a split-root system

To address the questions of whether allocation of carbohydrates to roots is influenced by ionic form of nitrogen absorbed and whether allocation of carbohydrates to roots in turn influences proportionality between NH₄⁺ and NO₃^? uptake from mixed sources, NH₄⁺ and NO₃^? were supplied separately to halves of a split-root hydroponic system and were supplied in combination to a whole-root system. Dry matter accumulation in the split-root system was 18% less in the NH₄⁺-fed axis than in the NO₃^?-fed axis. This, however, does not indicate that partitioning of carbohydrate between the two axes was different. Most of the reduction in dry matter accumulation in the NH₄⁺-fed axis can be accounted for by the retransport of CH₂O equivalents from the root back to the shoot with amino acids produced by NH₄⁺ assimilation. Uptake of NH₄⁺ or NO₃^? by the respective halves of the split-root system was proportional to the estimated allocation of carbohydrate to that half. When NH₄⁺ and NO₃^? were supplied to separate halves of the split-root system, the cumulative NH₄⁺ to NO₃^? uptake ratio was 0.81. When supplied in combination to the whole-root system, the cumulative NH₄⁺ to NO₃^? uptake ratio was 1.67. Thus, while the shoot may affect total nitrogen uptake through the export of carbohydrates to roots, the shoot (common for halves of the split-root system) apparently does not exert a direct effect on proportionality of NH₄⁺ and NO₃^? uptake by roots. For whole roots supplied with both NH₄⁺ and NO₃^?, the restriction in uptake of NO₃^? may involve a stimulation of NO₃^? efflux rather than an inhibition of NO₃^? influx. While only the net uptake of NH₄⁺ and NO₃^? was measured by ion chromatography, monitoring at approximately hourly intervals during the first 3 days of treatment revealed irregularly occurring intervals of both depletion (net influx) and enrichment (net efflux) in solutions. In the case of NH₄⁺, numbers of net efflux events were similar (21 to 24 out of 65 sequential sampling intervals) whether NH₄⁺ was supplied with NO₃^? to whole-root systems or separately to an axis of the split-root system. In the case of NO₃^?, however, the number of net efflux events increased from 8 when NO₃^? was supplied to a separate axis of the split-root system to between 19 and 24 when NO₃^? was supplied with NH₄⁺ to whole-root systems.     

Effect of the nitrogen form used in the growth medium (NO3 −, NH4 +) on alkaloid production in Datura stramonium L.

Plants of Datura stramonium var. tatula L. Torr. were cultivated on vermiculite and received two different mineral solutions. In one treatment only NO₃ ^–-nitrogen was added, while in the other NO₃ ^–-nitrogen was partly (20%) replaced by NH₄ ⁺-nitrogen. Total dose of nitrogen as well as interionic ratios were kept constant in both treatments. With the combined treatment (NO₃ ^–-NH₄ ⁺) a significant higher hyoscyamine content was found at the time when highest biomass was reached. This was apparently the result of an increased alkaloid biosynthesis. Also scopolamine content was positively influenced, but only at a point past maximal biomass yield.No significant differences in amounts of nitrogen bound per plant were found between both treatments.The higher alkaloid content observed with the combined treatment was associated with a higher relative proportion of bound nitrogen present in the alkaloids. It seems that more nitrogen is available for secondary metabolism when NH₄ ⁺-nitrogen is present in the culture medium.     

Culture medium optimization for camptothecin production in cell suspension cultures of Nothapodytes nimmoniana (J. Grah.) Mabberley

The effect of culture medium nutrients on growth and alkaloid production by plant cell cultures of Nothapodytes nimmoniana (J. Grah.) Mabberley (Icacinaceae) was studied with a view to increasing the production of the alkaloid camptothecin, a key therapeutic drug used for its anticancer properties. Amongst the various sugars tested with Murashige and Skoog (MS) medium, such as glucose, fructose, maltose, and sucrose, maximum accumulation of camptothecin was observed with sucrose. High nitrate in the media supports the biomass, while high ammonium enhances the camptothecin content. Selective feeding of 60 mM total nitrogen with a NH₄ ⁺/NO₃ ^? balance of 5/1 on day 15 of the culture cycle results in a 2.4-fold enhancement in the camptothecin content over the control culture (28.5 μg/g DW). Furthermore, the sucrose feeding strategy greatly stimulated cell biomass and camptothecin production. A modified MS medium was developed in the present study, which contained 0.5 mM phosphate, a nitrogen source feeding ratio of 50/10 mM NH₄ ⁺/NO₃ ^? and 3 % sucrose with additional 2 % sucrose feeding (added on day 12 of the cell culture cycle) with 10.74 μM naphthaleneacetic acid and 0.93 μM kinetin. Finally, the selective medium has 1.7- and 2.3-fold higher intracellular and extracellular camptothecin content over the control culture (29.2 and 8.2 μg/g DW), respectively.     

Nitrogen Acquisition from Atmospheric NH3 by Lolium perenne

Ammonia (NH₃) is the third most abundant N species in the atmosphere and, due to various natural and anthropogenic sources, can reach high concentrations in some areas. While some plants show effects of toxicity, others are capable of using this N-form and grow well without any utilization of soil-N. Acquisition of atmospheric NH₃ will affect the acid-base balance of the plants as absorption and dissolution causes an alkalinisation (production of OH^?) and assimilation of NH₃ results in an acidification (generation of H⁺). As there is only a limited capacity for biochemical disposal of excess H⁺ in shoots, pH regulation may involve H⁺/OH^? extrusion into the media via roots and transport of (in)organic ions between roots and above-ground parts of the plant. Our aim therefore was to assess NH₃ acquisition by Lolium perenne and to study the effects of gas phase NH₃ on growth, acid-base balance and mineral composition of the plants. The experiments therefore included application of a range of ¹⁴NH₃ to the shoots and of ¹⁵N as NO₃^?, NH₄⁺ or NH₄NO₃ to the roots, from which the amount of gas phase NH₃ acquisition could be quantified. Analysis of the mineral composition provided data for calculation of acid-base balance as well as for water use efficiencies of the plants. The results indicate that over the range of NH₃ supplied, plants from all treatments could utilize gas-phase NH₃ as demonstrated by increases in growth and in N and C use efficiencies. Plants receiving NO₃^? via their roots had a higher capacity to use gaseous NH₃ than those growing with NH₄⁺. NH₃ assimilation in shoots reduced both the acid load with NH₄⁺ nutrition and the alkaline load with NO₃^? supply to the roots. The results of the experiments are discussed in relation to possible acid-base regulation mechanisms of the whole plant.     

Identification of Heterotrophic Nitrification in a Sierran Forest Soil

A potential for heterotrophic nitrification was identified in soil from a mature conifer forest and from a clear-cut site. Potential rates of NO₂⁻ production were determined separately from those of NO₃⁻ by using acetylene to block autotrophic NH₄⁺ oxidation and chlorate to block NO₂⁻ oxidation to NO₃⁻ in soil slurries. Rates of NO₂⁻ production were similar in soil from the forest and the clear-cut site and were strongly inhibited by acetylene. The rate of NO₃⁻ production was much greater than that of NO₂⁻ production, and NO₃⁻ production was not significantly affected by acetylene or chlorate. Nitrate production was partially inhibited by cycloheximide, but was not significantly reduced by streptomycin. Neither the addition of ammonium nor the addition of peptone stimulated NO₃⁻ production. ¹⁵N labeling of the NH₄⁺ pool demonstrated that NO₃⁻ was not coming from NH₄⁺. The potential for heterotrophic nitrification in these forest soils was greater than that for autotrophic nitrification.     

Relationships between Root Temperature and the Transport of Ammonium and Nitrate Ions by Italian and Perennial Ryegrass (Lolium multiflorum and Lolium perenne)

At root temperature below 14 C the absorption of ¹⁵N from NH₄⁺ greatly exceeded that from NO₂⁻ by tillers of Lolium multiflorum and Lolium perenne under conditions where pH, external concentration, plant N status, and pretreatment temperature were varied. There was a marked increase in the temperature sensitivity of NO₃⁻ transport below 14 C, irrespective of the temperature at which plants were grown previously. A marked increase in the temperature sensitivity was also seen for NH₄⁺ transport, but this occurred at the lower temperature of 10 C. Pretreatment of roots at 8 C lowered this still further to 5 C. Above and below these transition temperatures the Q₁₀ values for NO₃⁻ and NH₄⁺ transport were similar. Thus, the increased absorption of NH₄⁺ relative to NO₃⁻ at low temperatures seems to be related primarily to the difference in transition temperatures.     

Growth of Salvinia molesta as affected by water temperature and nutrition I. Effects of nitrogen level and nitrogen compounds

The growth of Salvinia molesta D.S. Mitchell was studied in a greenhouse using controlled-temperature water-baths at 16, 19 and 22°C and 4 different nitrogen compounds (NO₃^?, NH₄⁺, NH₄NO₃ and urea) at levels up to 60 mg N l^?1. Little growth occurred at 16°C even if 20 mg N l^?1 was supplied together with other nutrients including phosphorus (2 mg H₂PO₄-P l^?1). The highest relative growth rate and total dry matter production occurred at 22°C when plants were supplied with 20 mg NH₄-N l^?1. At this temperature, the NH₄⁺ ion was superior to the NO₃^? ion or urea as a nitrogen source (almost doubling the biomass), but was not significantly better than NH₄NO₃. Over a period of 19 days for plants receiving 0.02 mg NH₄-N l^?, biomass increased 4-fold at 16°C, 9-fold at 19°C and 10-fold at 22°C. In contrast, for plants receiving 20 mg NH₄-N l^?1, biomass increased 4-fold at 16°C, 18-fold at 19°C and 38-fold at 22°C.     

Dissimilatory Reduction of NO2− to NH4+ and N2O by a Soil Citrobacter sp.

Dissimilatory reduction of NO₂⁻ to N₂O and NH₄⁺ by a soil Citrobacter sp. was studied in an attempt to elucidate the physiological and ecological significance of N₂O production by this mechanism. In batch cultures with defined media, NO₂⁻ reduction to NH₄⁺ was favored by high glucose and low NO₃⁻ concentrations. Nitrous oxide production was greatest at high glucose and intermediate NO₃⁻ concentrations. With succinate as the energy source, little or no NO₂⁻ was reduced to NH₄⁺ but N₂O was produced. Resting cell suspensions reduced NO₂⁻ simultaneously to N₂O and free extracellular NH₄⁺. Chloramphenicol prevented the induction of N₂O-producing activity. The K_m for NO₂⁻ reduction to N₂O was estimated to be 0.9 mM NO₂⁻, yet the apparent K_m for overall NO₂⁻ reduction was considerably lower, no greater than 0.04 mM NO₂⁻. Activities for N₂O and NH₄⁺ production increased markedly after depletion of NO₃⁻ from the media. Amendment with NO₃⁻ inhibited N₂O and NH₄⁺ production by molybdate-grown cells but not by tungstate-grown cells. Sulfite inhibited production of NH₄⁺ but not of N₂O. In a related experiment, three Escherichia coli mutants lacking NADH-dependent nitrite reductase produced N₂O at rates equal to the wild type. These observations suggest that N₂O is produced enzymatically but not by the same enzyme system responsible for dissimilatory reduction of NO₂⁻ to NH₄⁺.     

Role of nitrite reductase in the ammonia-oxidizing pathway of <Emphasis Type="Italic">Nitrosomonas europaea</Emphasis>

Metabolism of ammonia (NH₃) and hydroxylamine (NH₂OH) by wild-type and a nitrite reductase (nirK) deficient mutant of Nitrosomonas europaea was investigated to clarify the role of NirK in the NH₃ oxidation pathway. NirK-deficient N. europaea grew more slowly, consumed less NH₃, had a lower rate of nitrite (NO₂ ⁻) production, and a significantly higher rate of nitrous oxide (N₂O) production than the wild-type when incubated with NH₃ under high O₂ tension. In incubations with NH₃ under low O₂ tension, NirK-deficient N. europaea grew more slowly, but had only modest differences in NH₃ oxidation and product formation rates relative to the wild-type. In contrast, the nirK mutant oxidized NH₂OH to NO₂ ⁻ at consistently slower rates than the wild-type, especially under low O₂ tension, and lost a significant pool of NH₂OH–N to products other than NO₂ ⁻ and N₂O. The rate of N₂O production by the nirK mutant was ca. three times higher than the wild-type during hydrazine-dependent NO₂ ⁻ reduction under both high and low O₂ tension. Together, the results indicate that NirK activity supports growth of N. europaea by supporting the oxidation of NH₃ to NO₂ ⁻ via NH₂OH, and stimulation of hydrazine-dependent NO₂ ⁻ reduction by NirK-deficient N. europaea indicated the presence of an alternative, enzymatic pathway for N₂O production.     

Growth responses of the perennial legume Sesbania sesban to NH4 and NO3 nutrition and effects on root nodulation

Preference for NH₄⁺ or NO₃⁻ nutrition by the perennial legume Sesbania sesban (L.) Merr. was assessed by supplying plants with NH₄⁺ and NO₃⁻ alone or mixed at equal concentrations (0.5 mM) in hydroponic culture. In addition, growth responses of S. sesban to NH₄⁺ and NO₃⁻ nutrition and the effects on root nodulation and nutrient and mineral composition of the plant tissues were evaluated in a hydroponic setup at a range of external concentration of NH₄⁺ and NO₃⁻ (0, 0.1, 0.2, 0.5, 2 and 5 mM). Seedlings of S. sesban grew equally well when supplied with either NH₄⁺ or NO₃⁻ alone or mixed and had high relative growth rates (RGRs) ranging between 0.19 and 0.21 d⁻¹. When larger plants of S. sesban were supplied with NH₄⁺ or NO₃⁻ alone, the RGRs and shoot elongation rates were not affected by the external concentration of inorganic N. At external N concentrations up to 0.5 mM nodulation occurred and contributed to the N nutrition through fixation of gaseous N₂ from the atmosphere. For both NH₄⁺ and NO₃⁻-fed plants the N concentration in the plant tissues, particularly water-extractable NO₃⁻, increased at high supply concentrations, and concentrations of mineral cations generally decreased. It is concluded that S. sesban can grow without an external inorganic N supply by fixing atmospheric N₂ gas via root nodules. Also, S. sesban grows well on both NH₄⁺ and NO₃⁻ as the external N source and the plant can tolerate relatively high concentrations of NH₄⁺. This wide ecological amplitude concerning N nutrition makes S. sesban very useful as a N₂-fixing fallow crop in N deficient areas and also a candidate species for use in constructed wetland systems for the treatment of NH₄⁺ rich waters.     

A ground water nitrogen budget for a headwater swamp in an area of permanent ground water discharge

Ground water inputs and outputs of N were studied for a small ground water discharge swamp situated in a headwater drainage basin in southern Ontario, Canada. Darcy's equation with data for piezometers was used to measure inputs of shallow local ground water at the swamp margin and deep regional ground water beneath the swamp. Ground water flux was also quantified by measuring ground water discharge to the outlet stream draining the swamp in combination with a chemical mixing model to separate shallow and deep ground water components based on chloride differences. Estimates of shallow ground water flux determined by these two approaches agreed closely however, the piezometer data seriously underestimated the deep ground water input to the swamp. An average ground water input-output budget of total N (TN) total organic nitrogen (TON) ammonium (NH₄ ⁺-N) and nitrate (NO₃ ^--N) was estimated for stream base flow periods which occurred on an average of 328 days each year during 1986–1990. Approximately 90% of the annual NO₃ ^--N input was contributed by shallow ground water at the swamp margin. Deep ground water represented about 65% of the total ground water input and a similar proportion of TON and NH₄ ⁺-N inputs. Annual ground water NO₃ ^--N inputs and outputs were similar whereas NH₄ ⁺-N retention was 4 kg ha^-1 representing about 68% of annual ground water input. Annual TON inputs in ground water exceeded outputs by 7.7 kg ha (27%). The capacity of the swamp to regulate ground water N fluxes was influenced by the N chemistry of ground water inputs and the hydrologic pathways of transport within the swamp.