Chemical fixation of ammonia to soil organic matter after application of urea

Chemical fixation of NH₃ to soil organic matter was studied in two Swedish soils with different contents of organic matter: a clay soil with 2.3% C and an organic soil with 36.6% C. ¹⁵N‐labelled urea was applied at different rates to both sterilized and non‐sterilized soils. After 10 days, the soils were extracted and washed with K₂SO₄ and determined for total N and atom% ¹⁵N excess. Urea N was recovered as non‐extractable N in sterilized soil corresponding to 9.7% of supplied ^l5N‐labelled urea in the organic soil and 2.2% in the clay soil. Since no biological immobilization is thought to occur in the sterile soil, this non‐extractable N is suggested to be chemically fixed to soil organic matter. Owing to urea hydrolysis in the clay soil, pH increased from 6.3 to 9.3 and in the organic soil from 5.7 to 6.9 and 8.8, respectively, at the low and high urea supply.     

Mineralization of soil organic nitrogen solubilized by aqua ammonia fertilizer

Organic N solubilized by NH_3(aq) was extracted from ¹⁵N-labelled or unlabelled soil, concentrated and added to non-extracted soil, which was incubated under aerobic conditions at 27±1°C. Gross N mineralization, gross N immobilization, and nitrification in soils with or without addition of unlabelled soluble organic N were estimated by models based on the dilution of the NH ₄ ⁺ or NO _inf3 ^sup- pools, which were labelled with ¹⁵N at the beginning of incubation. Mineralization of labelled organic N was measured by the appearance of label in the mineral N pool. Although gross N mineralization and gross N immobilization were increased in two soils between day 0 and day 7 following addition of unlabelled organic N solubilized by NH_3(aq), there was no increase in net N mineralization. Solubilization of ¹⁵N-labelled organic N increased and the ¹⁵N enrichment of the soluble organic N decereased as the concentration of NH_3(aq) added increased. A constant proportion of approximately one-quarter of the labelled organic N added at different rates to non-extracted soil was recovered in the mineral N pool after an incubation period of 14 days, and the availability ratios calculated from net N mineralization data were 1.1:1 and 2.1:1 for 111 and 186 mg added organic-N kg^-1 soil, respectively, indicating that the mineralization of organic N was increased by solubilization.     

Estimating N2 fixation by field-grown lupins (Lupinus angustifolius L.) using soil and plant 15N enrichment

Summary Biological N₂ fixation was estimated in a field experiment following the addition of NH₄Cl or KNO₃ to unconfined microplots (1.5 m²) at 2.5 g N m^-2 (10 atom% ¹⁵N). A model of total N and ¹⁵N accumulation in lupins and decreasing ¹⁵N enrichment in the KCl-extractable soil-N pool (0–0.15 m depth) was used to estimate the proportion of N in lupins derived from biological N₂ fixation. Estimates of N₂ fixation derived from the model were compared with ¹⁵N isotope-dilution estimates obtained using canola, annual ryegrass, and wheat as nonfixing reference plants. Biomass, total N accumulation, or ¹⁵N enrichment in the lupin and reference crops did not differ whether NH _inf4 ^sup+ or NO _inf3 ^sup- was added as the labelled inorganic-N source. The decrease in soil ¹⁵N enrichment was described by first-order kinetics, whereas total N and ¹⁵N accumulation in the lupins were described by logistical equations. Using these equations, the uptake of soil N by lupins was estimated and was then used to calculate fixed N₂. Estimates of N₂ fixation derived from the model increased from 0 at 50 days after sowing to a maximum of 0.79 at 190 days after sowing. Those based on the ¹⁵N enrichment of the NO _inf3 ^sup- pool were 10% higher than those based on the mineral-N pool. ¹⁵N isotope-dilution estimates of N₂ fixation ranged from 0.37 to 0.55 at 68 days after sowing and from 0.71 to 0.77 at 190 days after sowing. Reference plant-derived values of N₂ fixation were all higher than modelled estimates during the early states of growth, but were similar to modelled estimates at physiological maturity. The use of the model to estimate N₂ derived from the atmosphere has the intrinsic advantage that the need for a non-fixing reference plant is avoided.     

Effects of 15N-labelled ammonium nitrate and urea on soil nitrogen during growth of wheat (Triticum aestivum L.) under field conditions

We studied the effects of ¹⁵N-labelled ammonium nitrate and urea on the yield and uptake of labelled and unlabelled N by wheat (Triticum aestivum L., cv. Mexi-Pak-65) in a field experiment. The dry matter and N yields were significantly increased with fertilizer N application compared to those from unfertilized soil. The wheat crop used 33.6–51.5 and 30.5–40.9% of the N from ammonium nitrate and urea, respectively. Splitting the fertilizer N application had a significant effect on the uptake of fertilizer N by the wheat. The fertilizer N uptake showed that ammonium nitrate was a more available source of N for wheat than urea. The effective use of fertilizer N (ratio of fertilizer N in grain to fertilizer N in whole plant) was statistically similar for the two N fertilizers. The application of fertilizer N increased the uptake of unlabelled soil N by wheat, a result attributed to a positive added N interaction, which varied according to the fertilizer N split; six split applications gave the highest added N interaction compared to a single application or two split applications for both fertilizers. Ammonium nitrate gave 90.5, 33.5, and 48.5% more added N interaction than urea with one, two, and six split N applications. A values were not significantly correlated with the added N interaction (r=0.557). The observed added N interaction may have been the result of pool substitution, whereby added labelled fertilizer N replaced unlabelled soil N.     

Interaction of 15N-labelled ammonium nitrate,urea and ammonium sulphate with soil N during growth of wheat (Triticum aestivum L.) under field conditions

The effects of ¹⁵N-labelled ammonium nitrate, urea and ammonium sulphate on yield and uptake of labelled and unlabelled N by wheat (Triticum aestivum L. cv. Mexi-Pak-65) were studied in a field experiment. The dry matter and N yields were significantly increased with fertilizer N application compared to those from unfertilized soil. The wheat crop used 64.0–74.8%, 61.5–64.7% and 61.7–63.4% of the N from ammonium nitrate, urea and ammonium sulphate, respectively. The fertilizer N uptake showed that ammonium nitrate was a more available source of N for wheat than urea and ammonium sulphate. The effective use of fertilizer N (ratio of fertilizer N in grain to fertilizer N in whole plant) was statistically similar for the three N fertilizers. The application of fertilizer N increased the uptake of unlabelled soil N by wheat, a result attributed to a positive added N interaction, which varied with the method of application of fertilizer N. Ammonium nitrate, urea and ammonium sulphate gave 59.3%, 42.8% and 26.3% more added N interaction, respectively, when applied by the broadcast/worked-in method than with band placement. A highly significant correlation between soil N and grain yield, dry matter and added N interaction showed that soil N was more important than fertilizer N in wheat production. A values were not significantly correlated with added N interaction (r=0.719). The observed added N interaction may have been the result of pool substitution, whereby added labelled fertilizer N stood proxy for unlabelled soil N.     

Fate of Azolla spp. and urea nitrogen applied to wetland rice (Oryza sativa L.)

Summary Using ¹⁵N, the fate of N applied to wetland rice either as Azolla or urea was studied in a field at the International Rice Research Institute (IRRI). In bigger plots nearby, yield response and N uptake were also determined with unlabelled N sources. Azolla microphylla was labelled by repeated application of labelled ammonium sulfate. Labelled and unlabelled N were used alternately in applications of Azolla or urea 0 and 42 days after transplanting, in order to determine the effect of the time of application on the availability of Azolla N. The quantities of Azolla N incorporated were 23% more than those of urea N (30 kg N ha^–1) in the isotope plots or 7% less in the yield response plots. Grain yield and total N uptake by the rice plants in the yield-response plots were higher in the urea-treated plots than in the Azolla-treated plots, but the physiological effect of Azolla N (grain yield response/increase in N uptake) was higher than that of rea. The labelled N balance was studied after the first and second crops of rice. Losses of labelled N after the first crop were higher from urea (30%–32%) than from Azolla (0%–11 %). Losses in N applied as a side dressing 42 days after transplanting were less than those of N applied basally. No further losses of ¹⁵N occurred after the first crop. The recovery of Azolla ¹⁵N in the first crop of rice was 39% from the basal application and 63% from the side dressing. The recovery of urea ¹⁵N was 27% from the basal application and 48% from the side dressing. Recoveries of residual N from both Azolla and urea during the second rice crop were similar. Laboratory incubation of the Azolla used and the changes in labelled exchangeable N in the soil showed that at least 65% of Azolla N (4.7% N content) was mineralized within 10 days.     

Elevated CO2, but not defoliation, enhances N cycling and increases short-term soil N immobilization regardless of N addition in a semiarid grassland

Elevated CO₂ and defoliation effects on nitrogen (N) cycling in rangeland soils remain poorly understood. Here we tested whether effects of elevated CO₂ (720 μl L⁻¹) and defoliation (clipping to 2.5 cm height) on N cycling depended on soil N availability (addition of 1 vs. 11 g N m⁻²) in intact mesocosms extracted from a semiarid grassland. Mesocosms were kept inside growth chambers for one growing season, and the experiment was repeated the next year. We added ¹⁵N (1 g m⁻²) to all mesocosms at the start of the growing season. We measured total N and ¹⁵N in plant, soil inorganic, microbial and soil organic pools at different times of the growing season. We combined the plant, soil inorganic, and microbial N pools into one pool (PIM-N pool) to separate biotic + inorganic from abiotic N residing in soil organic matter (SOM). With the ¹⁵N measurements we were then able to calculate transfer rates of N from the active PIM-N pool into SOM (soil N immobilization) and vice versa (soil N mobilization) throughout the growing season. We observed significant interactive effects of elevated CO₂ with N addition and defoliation with N addition on soil N mobilization and immobilization. However, no interactive effects were observed for net transfer rates. Net N transfer from the PIM-N pool into SOM increased under elevated CO₂, but was unaffected by defoliation. Elevated CO₂ and defoliation effects on the net transfer of N into SOM may not depend on soil N availability in semiarid grasslands, but may depend on the balance of root litter production affecting soil N immobilization and root exudation affecting soil N mobilization. We observed no interactive effects of elevated CO₂ with defoliation. We conclude that elevated CO₂, but not defoliation, may limit plant productivity in the long-term through increased soil N immobilization.     

Tillage and dicyandiamide influence on nitrogen fertilizer immobilization,remineralization, and utilization by maize (Zea mays L.)

Summary Both tillage and nitrification inhibitors such as dicyandiamide (DCD) have the potential to influence N availability and thus plant N uptake. A field experiment was conducted to investigate the impact of DCD and tillage (rototillage and no-tillage) on N immobilization and the subsequent impact of residual and fertilizer N on N availability to maize (Zea mays L.) ¹⁵N-labeled urea and urea-DCD were surface applied at the rate of 16 g N m^-2, in either 1987 or 1988, to small plots which had been planted to maize (Zea mays L.). Soil samples were collected four times during the 1988 growing season and analyzed for the ¹⁵N and ¹⁴N components of inorganic N, organic N, and hydrolyzable (6 M HCl) amino acid N, hydrolyzable NH _inf4 ^sup+ -N, and non-hydrolyzable N. Plant samples were collected three times during the 1988 growing season, and analyzed for the ¹⁵N and ¹⁴N components of total N. The total amount of NO _inf3 ^sup- percolating through the profile was less than 15 kg N ha^-1 in 1987 and 1988. N uptake by maize was reduced under notillage and when the urea was treated with DCD. The tillage treatments had no effect on the uptake of N fertilizer applied in 1988 or on N immobilization. However, no-till-age reduced the uptake of residual N fertilizer. The reduced use of N fertilizer was attributed to a reduction in the actual mineralization rates of immobilized residual N. DCD reduced the uptake of N fertilizer applied in 1988. The reduced uptake was attributed to increased N immobilization or to organic matter fixation.     

Characterization of the N benefit of a grain legume (Lupinus angustifolius L.) to a cereal (Hordeum vulgare L.) by an in situ 15N isotope dilution technique

Summary A crop of barley was grown on plots which had previously supported pure stands of lupins, canola, ryegrass, and wheat. The plots were labelled with ¹⁵N-enriched fertilizers at the time of sowing of the antecedent crops. The crop of lupins, which derived 79% of its N from symbiotic N₂ fixation at physiological maturity, conferred an N benefit to barley of 3.4 g N m^-2 when compared to barley following wheat. Lupins used less fertilizer N and less unlabelled soil N compared to the other crops, but the ratios of these sources of N in the plant tops were similar. The apparent sparing of soil+fertilizer N under lupins compared with wheat was 13.6 g N m^-2, which was much larger than the measured N benefit. Barley following lupins was less enriched in ¹⁵N compared to barley following wheat, and the measured isotope dilution was used to estimate the proportion of barley N derived from biologically fixed N in the lupin residues. This in turn enabled the N benefit to be partitioned between the uptake of spared N and the uptake of fixed N derived from the mineralization of legume residues. Spared N and fixed N contributed in approximately equal proportions to the N benefit measured in barley following lupins compared to barley following wheat.     

Transformations of nitrogen fertilizers in soil

The effects of ¹⁵N-labelled urea, (NH₄)₂SO₄ and KNO₃ on immobilization, mineralization, nitrification and ammonium fixation were examined under aerobic conditions in an acid tropical soil (pH 4.0) and in a neutral temperate soil (pH 6.8). Urea, (NH₄)₂SO₄ and KNO₃ slightly increased net mineralization of soil organic nitrogen in both soils. There was also an apparent Added Nitrogen Interaction (ANI) i.e. added labelled NH₄-N stood proxy for unlabelled NH₄-N that would otherwise have been immobilized. So far as immobilization and nitrification were concerned, urea and (NH₄)₂SO₄ behaved very similarly in each soil. Immobilization of NO₃-N was negligible in both soils. Some of the added labelled NH₄-N was rapidly fixed, more by the temperate soil than by the tropical soil. This labelled fixed NH₄-N decreased during incubation, in contrast to labelled organic N, which did not decline.     

Priming effects of 15N-labelled ammonium nitrate on uptake of soil N by wheat (Triticum aestivum L.) under field conditions

Summary The uptake of labelled and unlabelled N by wheat was measured in a field experiment using ¹⁵N-labelled ammonium nitrate fertilizer. The dry matter yield and N yields were significantly increased with fertilizer N application compared to those from unfertilized soil. The uptake of applied N by wheat ranged between 25 and 34%. Fertilizer N application increased the uptake of unlabelled soil N which was attributed to a positive priming effect or added N interaction. The added N interaction observed by applying 20, 60, and 120 kg fertilizer N was 11.4, 19.1, and 27.9 kg, corresponding to 26, 44 and 64%, respectively of the N taken up from unfertilized soil. The A values did not alter with the increase in fertilizer N application. The observed added N interaction may have been the result of pool substitution whereby added labelled fertilizer N stood proxy for unlabelled soil N. A significant correlation coefficient (r=0.996^**) between the uptake of soil N and the dry matter yield showed that soil N was more important than fertilizer N in wheat production.     

脲酶抑制剂/硝化抑制剂对土壤中尿素氮转化及形态分布的影响

运用^15N尿素示踪技术,以壤质草甸棕壤和春小分别作供试土壤和作物进行盆栽试验,结果表明,与单^15N尿素或^15N尿素＋氢素＋氢醌（HQ）相比,配施双氰胺（DCD）尤其与HQ组合可使土壤保持较高的^15N回收率,其中有^15N占相当大的比例,在小麦孕穗期前,上述两处理可有效地保持尿素解后土壤中NH^ 4 -^15N含量,并显著降低（NO^ 3 NO^-2)-^15N的富集;促进肥料^15N固持－矿化的周转,HQ与DCD配合施用对尿素施用后土壤中残留^15N量及有机^15N的再矿化和NH^ 4-^15N含量具有一定的协同作用。     

Mineralization of carbon and nitrogen from fresh and anaerobically stored sheep manure in soils of different texture

A sandy loam soil was mixed with three different amounts of quartz sand and incubated with (¹⁵NH₄)₂SO₄ (60 g N g^-1 soil) and fresh or anaerobically stored sheep manure (60 g g^-1 soil). The mineralization-immobilization of N and the mineralization of C were studied during 84 days of incubation at 20°C. After 7 days, the amount of unlabelled inorganic N in the manure-treated soils was 6–10 g N g^-1 soil higher than in soils amended with only (¹⁵NH₄)₂SO₄. However, due to immobilization of labelled inorganic N, the resulting net mineralization of N from manure was insignificant or slightly negative in the three soil-sand mixtures (100% soil+0% quartz sand; 50% soil+50% quartz sand; 25% soil+75% quartz sand). After 84 days, the cumulative CO₂ evolution and the net mineralization of N from the fresh manure were highest in the soil-sand mixutre with the lowest clay content (4% clay); 28% fo the manure C and 18% of the manure N were net mineralized. There was no significant difference between the soil-sand mixtures containing 8% and 16% clay, in which 24% of the manure C and -1% to 4% of the manure N were net mineralized. The higher net mineralization of N in the soil-sand mixture with the lowest clay content was probably caused by a higher remineralization of immobilized N in this soil-sand mixture. Anaerobic storage of the manure reduced the CO₂ evolution rates from the manure C in the three soil-sand mixtures during the initial weeks of decomposition. However, there was no effect of storage on net mineralization of N at the end of the incubation period. Hence, there was no apparent relationship between net mineralization of manure N and C.     

Nitrate Transformation and N2O Emission in a Typical Intensively Managed Calcareous Fluvaquent Soil: A 15-Nitrogen Tracer Incubation Study

A 56-day aerobic incubation experiment was performed with 15-nitrogen (N) tracer techniques after application of wheat straw to investigate nitrate-N (NO₃-N) immobilization in a typical intensively managed calcareous Fluvaquent soil. The dynamics of concentration and isotopic abundance of soil N pools and nitrous oxide (N₂O) emission were determined. As the amount of straw increased, the concentration and isotopic abundance of total soil organic N and newly formed labeled particulate organic matter (POM-N) increased while NO₃-N decreased. When ¹⁵NO₃-N was applied combined with a large amount of straw at 5000 mg carbon (C) kg^?1 only 1.1 ± 0.4 mg kg^?1 NO₃-N remained on day 56. The soil microbial biomass N (SMBN) concentration and newly formed labeled SMBN increased significantly (P < 0.05) with increasing amount of straw. Total N₂O-N emissions were at levels of only micrograms kg^?1 soil. The results indicate that application of straw can promote the immobilization of excessive nitrate with little emission of N₂O.     

Availability of soil and fertilizer nitrogen to wetland rice following wheat straw amendment

Summary A pot experiment was conducted to study the availability of soil and fertilizer N to wetland rice as influenced by wheat straw amendment (organic amendment) and to establish the relative significance of the two sources in affecting crop yield. Straw was incorporated in soil at 0.1, 0.2, and 0.3% before transplanting rice. Inorganic N as ¹⁵N-ammonium sulphate was applied at 30, 60, and 90 g g^-1 soil either alone or together with wheat straw in different combinations. After harvesting the rice, the plant and soil samples were analyzed for total N and ¹⁵N. Straw incorporation significantly decreased the dry matter and N yield of rice, the decrease being greater with higher rates of straw. The reduction in crop yield following the straw incorporation was attributed mainly to a decrease in the uptake of soil N rather than fertilizer N. The harmful effects of organic matter amendment were mitigated by higher levels of mineral N addition. The uptake of applied N increased and its losses decreased due to the straw incorporation. Mineral N applied alone or together with organic amendment substantially increased the uptake of unlabelled soil N. The increase was attributed to a real added N interaction.     

Alder-poplar associations: Determination of plant nitrogen sources by isotope techniques

Summary Three¹⁵N isotopic dilution methods (¹⁵N natural abundance, labelled mineral fertilizer, and organic matter) were used to determine the proportion of N derived from different available sources in seedlines ofAlnus glutinosa andPopulus nigra planted together or in monoculture under natural climatic conditions. The proportion of N derived from N₂ fixation in associated alders was appreciably higher than that determined in monoculture. The reduction of soil N uptake by associated alders contributed to an increase in total plant N and biomass production in associated poplars. When slightly N-labelled organic matter (alder leaf litter) was incorporated into the soil, 10–15% of its initial N content was recovered in poplar tissues, showing that this N source makes an important contribution to the N yield of associated non-fixing plants. There were no significant differences between the results obtained by¹⁵N natural abundance and those obtained by labelled fertilizer methods, suggesting that the ¹⁵N method could be used to evaluate annual N budgets in natural ecosystems.     

Fate and interaction with native soil N of ammonium N applied to wetland rice (Oryza sativa L.) grown under saline and non-saline conditions

Summary The effect of salts on the balance of fertilizer N applied as ¹⁵N-labelled ammonium sulphate and its interaction with native soil N was studied in a pot experiment using rice (Oryza sativa L.) as a test crop. The rice crop used 26%–40% of the applied N, the level of applied N and salts showing no significant bearing on the uptake of fertilizer N. Losses of fertilizer N ranged between 54% and 68% and only 5%–8% of the N was immobilized in soil organic matter. Neither the salts nor the rate of N application had any significant effect on fertilizer N immobilization. The effective use of fertilizer N (fertilizer N in grain/fertilizer N in whole plant) was, however, better in the non-saline soil. The uptake of unlabelled N (N mineralized from soil organic matter and that originating from biological N₂ fixation in thes rhizosphere) was inhibited in the presence of the salts. However, in fertilized soil, the uptake of unlabelled N was significantly enhanced, leading to increased A values [(1-% Ndff/% Ndff)x N fertilizer applied, where Ndff is N derived from fertilizer], an index of interaction with the added N. This added N interaction increased with increasing levels of added N. Since the extra unlabelled N taken up by fertilized plants was greater than the fertilizer N immobilized, and the root biomass increased with increasing levels of added N, a greater part of the added N interaction was considered to be real, any contribution by an apparent N interaction (pool substitution or isotopic displacement) to the total calculated N interaction being fairly small. Under saline conditions, for the same level of fertilizer N addition, the added N interaction was lower, and this was attributed to a lower level of microbial activity, including mineralization of native soil N, rootdriven immobilization of applied N, and N₂ fixation.     

有机、无机肥料施用后土壤生物量C、N、P的变化及N素转化

研究结果表明,有机、无机肥施用后,土壤微生物量C、N、P开始增加很快,随着时间的推移,土壤微生物量C又有所降低,但生物量N和P则基本保持稳定。硫铵施入土壤后,微生物对肥料¹⁵N的生物固持10天后达到最高峰,以后被固持在体内的¹⁵N有一部分被逐渐释放出来,但一个月后仍有17%左右的¹⁵N被固持在微生物体内。硫铵与有机肥配合施用时,微生物对硫铵¹⁵N固持比例有所增加。有机肥中的¹⁵N被微生物固持的比例也较大,在肥料施入20天左右达到最大值,一个月后仍有19-25%存在于微生物体内。硫铵施用一个月后¹⁵N损失高达18%,有机肥中的N也有少量被损失。     

北京地区潮土表层中NO3--N的转化积累及其淋洗损失

本试验利用渗滤池设施，采用化学分析和同位素技术相结合的方法研究了北京地区潮土表层中施用氮肥后ＮＯ３＾－－Ｎ转化积累及其１３０ｃｍ土体的淋洗状况。常规分析结果表明，在春小麦和夏玉米的生育前期可以观察到氮素明显地向ＮＯ３＾－－Ｎ的转化积累，其强度随尿素施用量的增加而明显增加，而尿素、硝铵、硫铵等不同氮肥品种处理之间有差异但大多不显著。同时夏玉米期间转化积累作用比春小麦期间强烈。＾１５Ｎ标记试验结果表明     

Influence of mycorrhiza vs. soluble phosphate on growth,nodulation, and N2 fixation (15N) in alfalfa under different levels of water potential

Summary The legume Medicago sativa (+Rhizobium melilott) was grown under controlled conditions to study the interactions between soluble P in soil (four levels), or a mycorrhizal inoculum, and the degree of water potential (four levels) in relation to plant development and N₂ fixation. ¹⁵N-labelled ammonium sulphate was added to each pot for a qualitative estimate of N₂ fixation, in order to rank the effects of the different treatments.Dry-matter yield, nutrient content and nodulation increased with the amount of plant-available P in the soil, and decreased as the water stress increased, for each P-level. The mycorrhizal effect on dry matter, N yield, and on nodulation was little affected by the water potential. Since P uptake was affected by the water content in mycorrhizal plants, additional mechanisms, other than those mediated by P, must be involved in the mycorrhizal activity.There was a positive correlation between N yield and nodulation for the different P levels and the mycorrhizal treatment at all water levels. A high correlation between plant unlabelled N content and atom% ¹⁵N excess was also found for all levels of P. In mycorrhizal plants, however, the correlation between unlabelled N yield and ¹⁵N was lower. This suggests that mycorrhiza supply plants with other N sources in addition to those derived from the improvement on N₂ fixation.