The ability of riparian forests to reduce nutrient flows from crops to streams is well known. However, their role as sources and sinks of greenhouse gases (GHGs) has been less studied, particularly in Mediterranean environments. We assessed the changes in daily soil N2O and CH4 fluxes between 2021 and 2023 in a riparian zone located in Central Spain. We also evaluated if cumulative fluxes of such GHGs depended on soil NO3−, NH4+ and DOC contents, and water-filled pore space (WFPS). Their dependence to different vegetation types and distances to the riverbank, as well as on the existence of wet periods, which include occasional floods, was also analysed. Daily N2O and CH4 fluxes were both low. Those of N2O were positive whereas those of CH4 were mostly negative, but positive fluxes of this GHG were nevertheless observed in late autumn 2021 and in spring 2022. Cumulative fluxes were mainly driven by soil NH4+ contents in the case of N2O and by WFPS in the case of CH4, and influenced by the distance from the riverbank in both cases. The relationships between cumulative fluxes of N2O and its major drivers were positively and significantly influenced by the wet periods. Our results indicate that our riparian zone acted as a net source of N2O and a net sink of CH4, but it became a net source of CH4 in cold and wet periods, where anoxic conditions in which methanogenesis occurs are favoured. Soil N2O emissions mainly originate as a by-product of nitrification but also from incomplete denitrification after heavy rainfall events in warm months. Thus, we advocate for preventive strategies to reduce nitrogen flows from cropping systems to reduce soil N2O emissions in Mediterranean riparian zones.

Plants grown under exclusive ammonium (NH4+) nutrition have high carbon (C) demand to sustain proper nitrogen (N) assimilation and energy required for plant growth, generally impaired when compared to nitrate (NO3-) nutrition. Thereby, the increment of the atmospheric carbon dioxide (CO2) concentration, in the context of climate change, will potentially allow plants to better face ammonium nutrition. In this work, tomato (Solanum lycopersicum L.) plants were grown under ammonium or nitrate nutrition in conditions of ambient (aCO(2), 400 ppm) or elevated CO2 (eCO(2), 800 ppm) atmosphere. Elevated CO2 increased photosynthesis rate and tomato shoot growth regardless of the N source. In the case of NH4+-fed leaves the positive effect of elevated CO2 occurred despite of the high tissue NH4+ accumulation. Under eCO(2) ammonium nutrition triggered, among others, the modulation of genes related to C provision pathways (including carbonic anhydrase and glyoxylate cycle), antioxidant response and cell membranes protection. The enhanced photosynthate production at eCO(2) facilitated C skeleton provision through the TCA cycle and anaplerotic pathways to promote amino acid synthesis. Moreover, photorespiratory activity was stimulated by eCO(2) and contributed to yield serine as additional sink for NH4+ excess. Overall, these changes denote a connection between the respiratory and the photorespiratory pathways linked to ammonium nutrition. This metabolic strategy may allow crops to grow efficiently using ammonium as fertilizer in a future climate change scenario, while mitigating N losses.

This study explores the potential use of frass, the larval excrement residue obtained from mealworm rearing, as organic fertilizer for crops. Its high organic matter content means that its joint application with a biostimulant based on efficient microorganisms, favoring its mineralization, is of interest. An experiment with lettuce plants (Lactuca sativa L.) was conducted with two factors and six replicates under greenhouse conditions. The first factor was frass amendment at 0%, 1%, 2.5%, and 5% of the peat substrate, and the second factor was a Bacillus-based BS at two levels, with and without efficient microorganism application. The results reveal that frass shows great potential as an organic fertilizer, providing macronutrients and increasing lettuce aerial biomass, although its effect is mediated by the application rate. Rates of 2.5% or higher proved negative for lettuce plant growth, especially root development, probably due to an increased incidence of potentially pathogenic fungi. The negative effect of medium–high frass rates was counteracted by the addition of a PGP-based biostimulant, enhancing lettuce plant nutrient uptake, aerial biomass, and quality in terms of succulence, but also favoring microbial diversity in the rhizosphere, increasing the incidence of beneficial microorganisms, and decreasing potentially pathogenic fungi. This positive synergy observed between frass and the PGP-based biostimulant is of interest for the design of new organic fertilization strategies.

Nitrogen fertilization is the most important factor increasing nitrous oxide (N2O) emissions from agriculture, which is a powerful greenhouse gas. These emissions are mainly produced by the soil microbial processes of nitrification and denitrification, and the application of nitrification inhibitors (NIs) together with an ammonium-based fertilizer has been proved as an efficient way to decrease them. In this work the NIs dimethylpyrazole phosphate (DMPP) and dimethylpyrazole succinic acid (DMPSA) were evaluated in a temperate grassland under environmental changing field conditions in terms of their efficiency reducing N2O emissions and their effect on the amount of nitrifying and denitrifying bacterial populations responsible of these emissions. The stimulation of nitrifying bacteria induced by the application of ammonium sulphate as fertilizer was efficiently avoided by the application of both DMPP and DMPSA whatever the soil water content. The denitrifying bacteria population capable of reducing N2O up to N2 was also enhanced by both NIs provided that sufficiently high soil water conditions and low nitrate content were occurring. Therefore, both NIs showed the capacity to promote the denitrification process up to N2 as a mechanism to mitigate N2O emissions. DMPSA proved to be a promising NI, since it showed a more significant effect than DMPP in decreasing N2O emissions and increasing ryegrass yield.

Improving fertilizer nitrogen (N) use efficiency is essential to increase crop productivity and avoid environmental damage. This study was conducted during four crop cycles of winter wheat under humid Mediterranean conditions (Araba, northern Spain). The effects of N-fertilization splitting and the application of the nitrification inhibitors (NIs) 3,4-dimethylpyrazole phosphate (DMPP) and 2-(3,4-dimethyl-1H-pyrazol-1-yl) succinic acid isomeric mixture (DMPSA) as strategies to improve grain quality were examined. The hypothesis of this study was to test if the partial ammonium nutrition and the reduction of fertilizer losses presumably induced by the application of NIs can modify the grain gliadin and glutenin protein contents and the breadmaking quality (dough rheological properties). Among both NIs assayed, only DMPP showed a slight effect of decreasing the omega gliadin fraction, following splitting either two or three times, although this effect was dependent on the year and was not reflected in terms of dough extensibility. The slight decreases observed in grain quality in terms of dough strength and glutenin content induced by DMPP suggest that DMPSA is more promising in terms of maintaining grain quality. Nonetheless, these poor effects exerted by NI application on grain quality parameters did not lead to changes in the quality parameters defining the flour aptitudes for breadmaking.

In forest nurseries, intensive use of non-renewable substrates such as peat and high application rates of chemical synthesis fertilizers lead to environmental problems and high susceptibility to biotic and abiotic stresses. This work aims to seek more sustainable crop management to help mitigate these problems, combining the substitution of peat by compost and the use of growth-promoting microorganisms (PGPs) as a fertilization tool. For this purpose, a trial was carried out to test the effectiveness of an agricultural waste compost and a biostimulant based on PGP microorganisms in the production of Castanea sativa plants in a forest nursery. This trial assessed the growth of plants, with both inputs separately and combined, and then studied the tolerance of chestnut seedlings to water deficit. The results showed that partial substitution of peat by compost is possible, but not complete, as the high levels of conductivity and pH generated by a high proportion of compost negatively affected plant growth. It was also noted that the application of the biostimulant enables the complete substitution of mineral fertilization. Moreover, at the end of the nursery phase, chestnut seedlings treated with the biostimulant showed the same or even better quality than chestnut seedlings obtained with conventional fertilization, also resulting in greater resistance to water deficit, based on the increase in root volume and the improvement of the physiological status. Changes observed in both quantity and composition of microbiota associated with chestnut rhizosphere after inoculation with PGPs were related to the improvement observed. In relation to water deficit resistance, a positive synergy was also observed with the combination of both inputs, since plants with full substitution of peat by compost combined with PGP-based fertilization showed the greatest drought resistance.

Urea fertilization is a widely spread source of nitrogen (N) for agriculture because of its easy accessibility. However, its use is highly inefficient, since a large amount of the applied N is lost to the environment mainly in the form of ammonia (NH3) volatilization, causing serious environmental and economic damages. Although there are simple strategies to reduce these losses, as the subsurface application, this practice cannot be adopted in all types of cultivations and tillage systems. To address this problem, combinations of urease and nitrification inhibitors (double inhibitors) have been developed to close all possible N escape routes, which is not achieved when these inhibitors are applied individually. The objective of this work was to evaluate the potential of a new double inhibitor combining N-(n-butyl) thiophosphoric triamide (NBPT) and 2-(3,4-dimethylpyrazole-1-YL)-succinic acid (DMPSA) to mitigate N losses in a rapeseed crop established in conventional tillage (CT) and no-tillage (NT) soils under Humid Mediterranean conditions. To do so, N losses in form of NH3 volatilization and nitrous oxide (N2O) emissions were determined through the Drager-Tube method (DTM) and closed chamber method respectively. The specific weather conditions present during the experiment promoted great losses, mostly in form of NH3 volatilization. The treatments receiving urea without inhibitors or urea with DMPSA lost up to 57.3 and 59.9 kg N ha(-1) in CT and NT respectively. Nonetheless, the application of the double inhibitor was able to diminish these losses by more than 50%, reducing the losses to a minimum of 18.8 kg N ha(-1) (CT) and 25 kg N ha(-1) (NT). As a whole, a CT system fertilized with a single application of urea with the double inhibitor showed the lowest global warming potential (GWP) (3.93 Mg CO2-eq ha(-1)), at the same level of the unfertilized treatment. (C) 2020 Elsevier Ltd. All rights reserved.

Nitrate and ammonium are the main nitrogen sources in agricultural soils. In the last decade, ammonium (NH 4 + ), a double-sided metabolite, has attracted considerable attention by researchers. Its ubiquitous presence in plant metabolism and its metabolic energy economy for being assimilated contrast with its toxicity when present in high amounts in the external medium. Plant species can adopt different strategies to maintain NH 4 + homeostasis, as the maximization of its compartmentalization and assimilation in organic compounds, primarily as amino acids and proteins. In the present study, we report an integrative metabolic response to ammonium nutrition of seven plant species, belonging to four different families: Gramineae (ryegrass, wheat, Brachypodium distachyon ), Leguminosae (clover), Solanaceae (tomato), and Brassicaceae (oilseed rape, Arabidopsis thaliana ). We use principal component analysis (PCA) and correlations among metabolic and biochemical data from 40 experimental conditions to understand the whole-plant response. The nature of main amino acids is analyzed among species, under the hypothesis that those Asn-accumulating species will show a better response to ammonium nutrition. Given the provision of carbon (C) skeletons is crucial for promotion of the nitrogen assimilation, the role of different anaplerotic enzymes is discussed in relation to ammonium nutrition at a whole-plant level. Among these enzymes, isocitrate dehydrogenase (ICDH) shows to be a good candidate to increase nitrogen assimilation in plants. Overall, metabolic adaptation of different carbon anaplerotic activities is linked with the preference to synthesize Asn or Gln in their organs. Lastly, glutamate dehydrogenase (GDH) reveals as an important enzyme to surpass C limitation during ammonium assimilation in roots, with a disparate collaboration of glutamine synthetase (GS).

Nitrogen (N) input from fertilizers modifies the properties of agricultural soils as well as bacterial community diversity, composition and relationships. This can lead to negative impacts such as the deterioration of system multifunctionality, whose maintenance is critical to normal nutrient cycling. Synthetic nitrification inhibitors (NIs) can be combined with fertilizers to improve the efficiency of N use by reducing N losses. However, analysis of their effects on non-target bacteria are scarce. This study aimed to analyze the effect of applying the NIs DMPP and DMPSA on the whole bacterial community. Through 16S rRNA amplicon sequencing we determined the differences between samples in terms of microbial diversity, composition and co-occurrence networks. The application of DMPP and DMPSA exerted little impact on the abundance of the dominant phyla. Nevertheless, several significant shifts were detected in bacterial diversity, co-occurrence networks, and the abundance of particular taxa, where soil water content played a key role. For instance, the application of NIs intensified the negative impact of N fertilization on bacterial diversity under high water-filled pore spaces (WFPS) (>64%), reducing community diversity, whereas alpha-diversity was not affected at low WFPS (<55%). Interestingly, despite NIs are known to inhibit ammonia monooxygenase (AMO) enzyme, both NIs almost exclusively inhibited Nitrosomonas genera among AMO holding nitrifiers. Thus, Nitrosomonas showed abundance reductions of up to 47% (DMPP) and 66% (DMPSA). Nonetheless, non-target bacterial abundances also shifted with NI application. Notably, DMPSA application partially alleviated the negative effect of fertilization on soil multifunctionality. A remarkable increase in populations related to system multifunctionality, such as Armatimonadetes (up to +21%), Cyanobacteria (up to +30%) and Fibrobacteres (up to +25%) was observed when DMPSA was applied. NI application substantially influenced microbial associations by decreasing the complexity of co-occurrence networks, decreasing the total edges and node connectivity, and increasing path distances.

Various studies have established that feedstock choice, pyrolysis temperature, and pyrolysis type influence final biochar physicochemical characteristics. However, overarching analyses of pre-biochar creation choices and correlations to biochar characteristics are severely lacking. Thus, the objective of this work was to help researchers, biochar-stakeholders, and practitioners make more well-informed choices in terms of how these three major parameters influence the final biochar product. Utilizing approximately 5400 peer-reviewed journal articles and over 50,800 individual data points, herein we elucidate the selections that influence final biochar physical and chemical properties, total nutrient content, and perhaps more importantly tools one can use to predict biochar’s nutrient availability. Based on the large dataset collected, it appears that pyrolysis type (fast or slow) plays a minor role in biochar physico- (inorganic) chemical characteristics; few differences were evident between production styles. Pyrolysis temperature, however, affects biochar’s longevity, with pyrolysis temperatures > 500 °C generally leading to longer-term (i.e., > 1000 years) half-lives. Greater pyrolysis temperatures also led to biochars containing greater overall C and specific surface area (SSA), which could promote soil physico-chemical improvements. However, based on the collected data, it appears that feedstock selection has the largest influence on biochar properties. Specific surface area is greatest in wood-based biochars, which in combination with pyrolysis temperature could likely promote greater changes in soil physical characteristics over other feedstock-based biochars. Crop- and other grass-based biochars appear to have cation exchange capacities greater than other biochars, which in combination with pyrolysis temperature could potentially lead to longer-term changes in soil nutrient retention. The collected data also suggest that one can reasonably predict the availability of various biochar nutrients (e.g., N, P, K, Ca, Mg, Fe, and Cu) based on feedstock choice and total nutrient content. Results can be used to create designer biochars to help solve environmental issues and supply a variety of plant-available nutrients for crop growth.

Nitrogen (N) applied with fertilizers is not efficiently used in agriculture. In the soil, this N is transformed into different compounds by means of several biological processes. As a result, there is a negative economic and environmental impact due to water contamination, via nitrate (NO3-) leaching, and greenhouse gasses emission, via nitrous oxide (N2O). To reverse this situation, nitrification inhibitors (NI) such as dicyandiamide (DCD), nytrapirin and 3,4-dimethylpyrazole phosphate (DMPP) are widely applied to agricultural soils in order to delay ammonium (NH4+) transformation. A new NI, 3,4-dimethylpyrazole-succinic acid (DMPSA), has been recently developed with the aim of deploying a specific action on ammonium-oxidizing bacteria (AOB). However, previous studies have demonstrated that DMPSA application increases nosZl gene abundance. Thus, non-target populations involved in N-cycle are also affected by its application. For better understanding the effects of DMPSA addition, this NI was applied with ammonium sulfate (AS) fertilizer in a winter wheat crop soil under Humid Mediterranean conditions, in two different soil tillage managements (conventional tillage, CT; and notillage, NT). Soil samples were then analyzed by 16S rRNA amplicon sequencing. DMPSA application induced a decrease in bacterial alpha-diversity under the NT management, which showed higher water-filled pore space (WFPS) than the CT management. This suggests that water content played a key role in DMPSA effects. Even at the phyla level, the abundances of several non-target organisms, either involved or not in the N-cycle, were affected by DMPSA application. Within them, the biggest changes were found in Cyanobacteria (+ 48%) phylum (considered promising bio-agents for sustainable agriculture), which may have also triggered the increase of Bacteroidetes (+ 20%) and the decrease of certain phytopathogens. This decrease of phytopathogens may have also been helped by the great increase observed after DMPSA application in the genus Vermamoeba vermiformis, a protist known to control/regulate several soil-borne pathogens. Altogether, the results showed that DMPSA may lead to a reduction of the environmental impacts derived not only from the loss of reactive N, but also from the maintenance of a safer microbial community for plant health. However, further studies would be necessary to analyze the persistence and the consequences of all these effects in the long-term.

In higher plants ammonium (NH4+) assimilation occurs mainly through the glutamine synthetase/glutamate synthase (GS/GOGAT) pathway. Nevertheless, when plants are exposed to stress conditions, such as excess of ammonium, the contribution of alternative routes of ammonium assimilation such as glutamate dehydrogenase (GDH) and asparagine synthetase (AS) activities might serve as detoxification mechanisms. In this work, the in vivo functions of these pathways were studied after supplying an excess of ammonium to tomato (Solanum lycopersicum L. cv. Agora Hybrid F1) roots previously adapted to grow under either nitrate or ammonium nutrition. The short-term incorporation of labelled ammonium (15NH4+) into the main amino acids was determined by GC-MS in the presence or absence of methionine sulphoximine (MSX) and azaserine (AZA), inhibitors of GS and GOGAT activities, respectively. Tomato roots were able to respond rapidly to excess ammonium by enhancing ammonium assimilation regardless of the previous nutritional regime to which the plant was adapted to grow. The assimilation of 15NH4+ could take place through pathways other than GS/GOGAT, since the inhibition of GS and GOGAT did not completely impede the incorporation of the labelled nitrogen into major amino acids. The in vivo formation of Asn by AS was shown to be exclusively Gln-dependent since the root was unable to incorporate 15NH4+ directly into Asn. On the other hand, an in vivo aminating capacity was revealed for GDH, since newly labelled Glu synthesis occurred even when GS and/or GOGAT activities were inhibited. The aminating GDH activity in tomato roots responded to an excess ammonium supply independently of the previous nutritional regime to which the plant had been subjected.

Agricultural sustainability is compromised by nitrogen (N) losses caused by soil microbial activity. Nitrous oxide (N2O) is a potent greenhouse gas (GHG) produced as consequence of nitrification and denitrification processes in soils. Nitrification inhibitors (NI) as 3,4-dimethylpyrazole-succinic acid (DMPSA) are useful tools to reduce these N losses from fertilization. The objective of this work was to test the efficiency of DMPSA in two different tillage management systems, conventional tillage (CT) and no-tillage (NT), in a winter wheat crop under Humid Mediterranean conditions. N fertilizer was applied as ammonium sulphate (AS) with or without DMPSA in a single or split application, including an unfertilized treatment. GHG fluxes (N2O, CO2 and CH4) were measured by the closed chamber method. amoA and nosZI genes were quantified by qPCR as indicators of nitrifying and denitrifying populations. Nitrification was inhibited by DMPSA in both CT and NT, while the higher water filled pore space (WFPS) in NT promoted a better efficiency of DMPSA in this system. This higher efficiency might be due to a greater N2O reduction to N2 as result of the nosZI gene induction. Consequently, DMPSA was able to reduce N2O emissions down to the unfertilized levels in NT. Provided that NT reduced CO2 emissions and maintained crop yield compared to CT, the application DMPSA under NT management is a promising strategy to increase agro-systems sustainability under Humid Mediterranean conditions.

Biochar can reduce both nitrous oxide (N2O) emissions and nitrate (NO3-) leaching, but refining biochar's use for estimating these types of losses remains elusive. For example, biochar properties such as ash content and labile organic compounds may induce transient effects that alter N-based losses. Thus, the aim of this meta-analysis was to assess interactions between biochar-induced effects on N2O emissions and NO3- retention, regarding the duration of experiments as well as soil and land use properties. Data were compiled from 88 peer-reviewed publications resulting in 608 observations up to May 2016 and corresponding response ratios were used to perform a random effects meta-analysis, testing biochar's impact on cumulative N2O emissions, soil NO3- concentrations and leaching in temperate, semi-arid, sub-tropical, and tropical climate. The overall N2O emissions reduction was 38%, but N2O emission reductions tended to be negligible after one year. Overall, soil NO3- concentrations remained unaffected while NO3- leaching was reduced by 13% with biochar; greater leaching reductions (>26%) occurred over longer experimental times (i.e. >30 days). Biochar had the strongest N2O emission reducing effect in paddy soils (Anthrosols) and sandy soils (Arenosols). The use of biochar reduced both N2O emissions and NO3- leaching in arable farming and horticulture, but it did not affect these losses in grasslands and perennial crops. In conclusion, the time-dependent impact on N2O emissions and NO3- leaching is a crucial factor that needs to be considered in order to develop and test resilient and sustainable biochar-based N loss mitigation strategies. Our results provide a valuable starting point for future biochar-based N loss mitigation studies. (C) 2018 Elsevier B.V. All rights reserved.

Grasslands are subject to a wide range of land management practices that influence the exchange of the three main agricultural greenhouse gases (GHGs) that are related to agriculture: carbon dioxide (CO2), nitrous oxide (N2O), and methane (CH4). Improving nitrogen fertilization management practices through the use of nitrification inhibitors (NIs) can reduce GHGs emissions. We conducted a field experiment at the Colombian Agricultural Research Corporation with four fertilization treatments: urea (typical fertilizer used in this region), ammonium sulfate nitrate (ASN), ASN plus the NI 3,4-dimethylpyrazole phosphate (ASN+DMPP), and an unfertilized control. The highest grassland yields (1956 and 2057 kg DM ha−1, respectively) and apparent fertilizer nitrogen recoveries (34% and 33%, respectively) were generated by the conventional urea fertilizer and ASN+DMPP. Furthermore, the use of ASN+DMPP reduced the N2O emissions that were related to N fertilization to the level of the unfertilized treatment (ca. 1.5 g N2O-N ha−1), with a significant reduction of N-yield-scaled N2O emissions (ca. 20 g N2O-N kg N uptake−1). These results support the application of DMPP as an alternative strategy to increase grassland yield while simultaneously reducing the environmental impact of N fertilization.

In Northern Spain, winter wheat is usually fertilized with 180-220kg N/ha split in two applications. To reduce this rate, a strategy was tested which comprised three nitrogen (N)-applications, the latter being tuned to the nutritional needs of the crop using a chlorophyll meter. N-fertilization experiments were performed on winter wheat (Soissons variety). Treatments consisted of three N-broadcasts of 40, 60, and either 0, 40, 60, or 80kg N/ha respectively applied at Zadoks scale growth stages 21, 30, and 37. An over-fertilized treatment of 220kg N/ha was included for normalization of chlorophyll-meter values. Chlorophyll-meter readings were performed at growth stage 37, before fertilization. When normalized chlorophyll-meter values were below 92%, yield and grain protein contents attained with 40+60+0kg N/ha could be respectively enhanced with third applications of 60 or 80kg N/ha and 40kg N/ha.

The increase of the global demand for dairy products is reflected in a rise of animal feed and forage productivity. In the coastal Atlantic climate conditions of northern Spain the maize-ryegrass rotation is a common management used to satisfy this forage demand. With the aim of mitigating greenhouse gases (GHG) emissions associated with fertilization in this type of intensive management, the use of nitrification inhibitors (NI) such as 3,4-dimethylpyrazol phosphate (DMPP) or the isomeric mixture of 2-(3,4-dimethyl-1H-pyrazol-1-yl) succinic acid and 2-(4,5-dimethy1-1H-pyrazol-1-y1) succinic acid (DMPSA) could be a useful strategy. Until now, the new NI DMPSA has only been evaluated under Mediterranean conditions. The objective of this study was to compare the efficiency of DMPSA with respect to DMPP reducing GHG emissions when applied in a maize-ryegrass rotation under Atlantic climate conditions. Nitrogen fertilizer was applied as ammonium sulphate, with and without both NIs, split into two applications of 80 and 100 kg N ha(-1) in the case of maize and in three applications of 80, 60 and 60 kg N ha(-1) in the case of ryegrass. An unfertilized control treatment was also included. Nitrous oxide (N2O) and methane (CH4) fluxes were measured using the closed chamber technique. The new NI DMPSA showed a similar behaviour to DMPP, mitigating N2O emissions down to the levels of the unfertilized soil. The effect of DMPSA reducing N2O losses lasted for the same time as for DMPP (1 < four months). CH4 fluxes were not affected by the application of any of both NIs. In terms of yield and quality, DMPSA and DMPP maintained the yield and the forage crude protein content of both maize and ryegrass.

This work explores whether the natural abundance of N isotopes technique could be used to understand the movement of N within the plant during vegetative and grain filling phases in wheat crop (Triticum aestivum L.) under different fertilizer management strategies. We focus on the effect of splitting the same N dose through a third late amendment at flag leaf stage (GS37) under humid Mediterranean conditions, where high spring precipitations can guarantee the incorporation of the lately applied N to the soil-plant system in an efficient way. The results are discussed in the context of agronomic parameters as N content, grain yield and quality, and show that further splitting the same N dose improves the wheat quality and induces a better nitrogen use efficiency. The nitrogen isotopic natural abundance technique shows that N remobilization is a discriminating process that leads to an impoverishment in 15N of senescent leaves and grain itself. This technique also reflects the more efficient use of N resources (fertilizer and native soil-N) when plants receive a late N amendment.

Nitrogen is the element with the greatest influence on plant production and on protein content in the case of grain crops. Nevertheless, nitrogen over-fertilization produces environmental problems such as water pollution and global warming, which has led to the declaration of vulnerable zones to nitrate pollution in the European Union and to the adhesion of many countries to the Kyoto protocol. In the case of wheat there is a demand for producing quality grain, which is primed with a bonus price. Under these both economical and environmental circumstances, arose the need for a rational system of nitrogen fertilization which enabled the optimization of nitrogen use under the specific edaphoclimatic of Northern Spain. In order to cope with this objective a net of nitrogen fertilization assays was established by means of which a series of fertilization strategies together with some associated diagnosis tools were evaluated. Thus, N losses occurring both by nitrate leaching and by N2O emissions to the atmosphere were quantified, as well as plant N extractions regarding the different nitrogen fertilizer treatments applied.