In this review, we present the recent developments and future prospects of improving nitrogen use efficiency (NUE) in crops using various complementary approaches. These include conventional breeding and molecular genetics, in addition to alternative farming techniques based on no-till continuous cover cropping cultures and/or organic nitrogen (N) nutrition. Whatever the mode of N fertilization, an increased knowledge of the mechanisms controlling plant N economy is essential for improving NUE and for reducing excessive input of fertilizers, while maintaining an acceptable yield and sufficient profit margin for the farmers. Using plants grown under agronomic conditions, with different tillage conditions, in pure or associated cultures, at low and high N mineral fertilizer input, or using organic fertilization, it is now possible to develop further whole plant agronomic and physiological studies. These can be combined with gene, protein and metabolite profiling to build up a comprehensive picture depicting the different steps of N uptake, assimilation and recycling to produce either biomass in vegetative organs or proteins in storage organs. We provide a critical overview as to how our understanding of the agro-ecophysiological, physiological and molecular controls of N assimilation in crops, under varying environmental conditions, has been improved. We have used combined approaches, based on agronomic studies, whole plant physiology, quantitative genetics, forward and reverse genetics and the emerging systems biology. Long-term sustainability may require a gradual transition from synthetic N inputs to legume-based crop rotation, including continuous cover cropping systems, where these may be possible in certain areas of the world, depending on climatic conditions. Current knowledge and prospects for future agronomic development and application for breeding crops adapted to lower mineral fertilizer input and to alternative farming techniques are explored, whilst taking into account the constraints of both the current world economic situation and the environment.

A field study was conducted in northern France over two consecutive years to evaluate the combined effect of conventional tillage (CT) vs no till (NT) with or without cover crops (cc) and nitrogen (N) fertilization on various agronomic traits related to N use efficiency in winter wheat. Five years after conversion of CT to NT, significant increases in N use efficiency, N utilization efficiency, N agronomic efficiency, N partial factor productivity, N apparent recovery fraction and N remobilization were observed under three N fertilization regimes (0, 161, 215 kg ha⁻¹). It was also observed that grain yield and grain N content were similar under CT and NT. The N nutrition index was higher under NT at the three rates of N fertilization. Moreover, N use efficiency related traits were increased in the presence of cc both under NT and CT. Thus, agronomic practices based on continuous NT in the presence of cc, appear to be promising strategies to increase N use efficiency in wheat, while reducing both the use and the loss of N-based fertilizers.

Salinity is one of the most important stresses that reduces plant growth and productivity in several parts of the world. Nine Tunisian durum wheat genotypes grown under hydroponic conditions were subjected to two levels of salt stress (100 and 170 mM NaCl) for 21 days. An integrative analysis revealing the impact of salinity on key phenotypic and physiological marker traits was then conducted. Principal component analysis grouped these traits into three different clusters corresponding to the absence of salt stress and the two levels of salt stress. This analysis also allowed the identification of genotypes exhibiting various levels of tolerance to NaCl. Among the nine genotypes of Triticum durum Desf., cultivar Om Rabiaa was the most tolerant whereas cultivar Mahmoudi genotype was the most sensitive. Following the multivariate analysis of the examined phenotypic and physiological traits, we found that shoot length, shoot fresh weight, leaf area, the whole-plant stable isotope ratios of nitrogen (δ¹⁵N), shoot ammonium and proline contents, and shoot glutamine synthetase activity could be used as markers for the selection of salt-tolerant wheat genotypes.

Arbuscular mycorrhizal fungi (AMF) play major roles in nutrient acquisition by crops and are key actors of agroecosystems productivity. However, agricultural practices can have deleterious effects on plant–fungi symbiosis establishment in soils, thus inhibiting its potential benefits on plant growth and development. Therefore, we have studied the impact of different soil management techniques, including conventional moldboard ploughing and no-till under an optimal nitrogen (N) fertilization regime and in the absence of N fertilization, on AMF spore density and soil chemical, physical, and biological indicators in the top 20 cm of the soil horizon. A field experiment conducted over six years revealed that AMF spore density was significantly lower under conventional tillage (CT) combined with intensive synthetic N fertilization. Under no-till (NT) conditions, the density of AMF spore was at least two-fold higher, even under intensive N fertilization conditions. We also observed that there were positive correlations between spore density, soil dehydrogenase enzyme activity, and soil penetration resistance and negative correlations with soil phosphorus and mineral N contents. Therefore, soil dehydrogenase activity and soil penetration resistance can be considered as good indicators of soil quality in agrosystems. Furthermore, the high nitrate content of ploughed soils appears to be detrimental both for the dehydrogenase enzyme activity and the production of AMF spores. It can be concluded that no-till, by preventing soil from structural and chemical disturbances, is a farming system that preserves the entire fungal life cycle and as such the production of viable spores of AMF, even under intensive N fertilization.

Arbuscular mycorrhizal fungi (AMF) play a major role in the uptake of nutrients by agricultural plants. Nevertheless, some agricultural practices can interrupt fungal-plant signaling and thus impede the establishment of the mycorrhizal symbiosis. A field experiment performed over a 5-year period demonstrated that both the absence of tillage and of nitrogen (N) fertilization improved AMF colonization of wheat roots. Moreover, under no-till conditions, N uptake and aboveground biomass production did not vary significantly between N-fertilized and N-unfertilized plots. In contrast, both N uptake and above ground biomass were much lower when N fertilizer was not added during conventional tillage. This finding strongly suggests that for wheat, no-till farming is a sustainable agricultural system that allows a gradual reduction in N fertilizer use by promoting AMF functionality and at the same time increasing N uptake.

The use of nitrogen (N) fertilizer and glyphosate-based herbicides is increasing worldwide, with agriculture holding the largest market share. The agronomic and socioeconomic utilities of glyphosate are well established; however, our knowledge of the potential effects of glyphosate applied in the presence or absence of long-term N fertilization on microbial functional activities and the availability of soil nutrients remains limited. Using an ex situ approach with soils that did (N+) or did not (N0) receive synthetic N fertilization for 6 years, we assessed the impact of different rates (no glyphosate, CK; field rate, FR; 100 × field rate, 100FR) of glyphosate application on biological and chemical parameters. We observed that, after immediate application (1 day), the highest dose of glyphosate (100FR) negatively affected the alkaline phosphatase (AlP) activity in soils without N fertilization history and decreased the cation exchange capacity (CEC) in N0 compared to CK and FR treatments with N+. Conversely, the 100FR application increased nitrate (NO₃^-) and available phosphorus (PO₄^3-) regardless of N fertilization history. Then, after 8 and 15 days, the N+\100FR and N+\FR treatments exhibited the lowest values for dehydrogenase (DH) and AlP activities, respectively, while urease (URE) activity was mainly affected by N fertilization. After 15 days and irrespective of N fertilization history, the FR glyphosate application negatively affected the degradation of carbon substrates by microbial communities (expressed as the average well color development, AWCD). By contrast, the 100FR treatment positively affected AWCD, increasing PO₄^3- by 5 and 16% and NO₃^- by 126 and 119% in the N+ and N0 treatments, respectively. In addition, the 100FR treatment resulted in an increase in the average net nitrification rate. Principal component analysis revealed that the 100FR glyphosate treatment selected microbial communities that were able to metabolize amine substrates. Overall, the lack of N fertilization in the 6 past years combined with the highest glyphosate application rate (100FR) induced the highest values of AWCD, functional diversity, NO₃^-, PO₄^3- and nitrification. We concluded that the intensive use of N fertilization for 6 years may change the non-target effects of glyphosate application on enzyme activities. The functional activities, nitrification and nutrient contents were increased by glyphosate only when applied at 100 times the field application rate.