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Proper carbon (C) supply is essential for nitrogen (N) assimilation especially when plants are grown under ammonium (NH4+) nutrition. However, how C and N metabolic fluxes adapt to achieve so remains uncertain. In this work, roots of wheat (Triticum aestivum L.) plants grown under exclusive NH4+ or nitrate (NO3−) supply were incubated with isotope-labelled substrates (15NH4+, 15NO3−, or [13C]Pyruvate) to follow the incorporation of 15N or 13C into amino acids and organic acids. Roots of plants adapted to ammonium nutrition presented higher capacity to incorporate both 15NH4+ and 15NO3− into amino acids, thanks to the previous induction of the NH4+ assimilative machinery. The 15N label was firstly incorporated into [15N]Gln vía glutamine synthetase; ultimately leading to [15N]Asn accumulation as an optimal NH4+ storage. The provision of [13C]Pyruvate led to [13C]Citrate and [13C]Malate accumulation and to rapid [13C]2-OG consumption for amino acid synthesis and highlighted the importance of the anaplerotic routes associated to tricarboxylic acid (TCA) cycle. Taken together, our results indicate that root adaptation to ammonium nutrition allowed efficient assimilation of N thanks to the promotion of TCA cycle open flux modes in order to sustain C skeleton availability for effective NH4+ detoxification into amino acids.

White lupin (Lupinus albus L.) is a legume that produces seeds recognized for their high protein content and good nutritional value (lowest glycemic index of all grains, high dietary fiber content, and zero gluten or starch)1–5. White lupin can form nitrogen-fixing nodules but has lost the ability to form mycorrhizal symbiosis with fungi6. Nevertheless, its root system is well adapted to poor soils: it produces cluster roots, constituted of dozens of determinate lateral roots that improve soil exploration and phosphate remobilization7. As phosphate is a limited resource that comes from rock reserves8, the production of cluster roots is a trait of interest to improve fertilizers efficiency. Using long reads sequencing technologies, we provide a high-quality genome sequence of a modern variety of white lupin (2n=50, 451 Mb), as well as de novo assemblies of a landrace and a wild relative. We describe how domestication impacted soil exploration capacity through the early establishment of lateral and cluster roots. We identify the APETALA2 transcription factor LaPUCHI-1, homolog of the Arabidopsis morphogenesis coordinator9, as a potential regulator of this trait. Our high-quality genome and companion genomic and transcriptomic resources enable the development of modern breeding strategies to increase and stabilize yield and to develop new varieties with reduced allergenic properties (caused by conglutins10), which would favor the deployment of this promising culture.

Trees can be associated with dozens of fungi helping them to acquire resources from forest soils. The most widespread mutualistic association in boreal and temperate forests is the ectomycorrhizal symbiosis. This symbiosis involves mushroom-forming fungi of basidiomycota, ascomycota, and some zygomycota clades and the roots of woody plant species, including oaks, poplars or pines. Although the impact of this association on ecosystem production and tree nutrition is investigated for about a century, our understanding on the molecular mechanisms that control water and nutrient fluxes between plant and fungal partners is still limited. Here, we review the recent knowledge on the ectomycorrhizal contribution to tree nutrition. We specifically highlight the molecular mechanisms driving the acquisition, translocation and release of water and nutrients in ectomycorrhizal systems. We particularly focus on the transport of macronutrients, including nitrogen, phosphorus, potassium, sulphur and calcium, micronutrients, and water by the symbiotic partner. We also provide background on the evolution, diversity, and importance of this symbiosis, identify knowledge gaps, and propose future research directions.

International audience; Rapid adjustment of plant physiology and development to external fluctuations is critical for sessile organism, giving a singular interest to network signaling controlling these mechanisms. Among many adaptation processes, root plasticity is primordial to optimize nutrient acquisition but relies on a complex network integrating local and systemic (root shoot) signaling. Indeed, locally, plants invest resource in soil area where nutrients are available and systemically they adjust nutrient acquisition to the whole plant demand.Our main goal is to decipher systemic signaling underlying the perception of nitrate heterogeneous provision, in Arabidopsis. Using the split-root system, in which physically isolated root systems of the same plant were challenged with different environments, we previously demonstrated that cytokinin biosynthesis constitutes one critical component of root-shoot-root communication. By combining the use of cytokinin mutants with hormone measurements, transcriptomic analysis, nitrate uptake assays, and root growth measurements, we show that root to shoot trans-zeatin (tZ) translocation is likely crucial for long distance signaling controlling rapid sentinel gene regulation and long-term functional acclimation to heterogeneous nitrate supply. Interestingly, shoot transcriptome profiling revealed that glutamate/glutamine metabolism is likely a target of tZ root-to-shoot translocation, prompting an interesting hypothesis regarding shoot-to-root communication. Finally, this study also highlights tZ-independent pathways triggered by variation into nitrogen supply.

Plants need to cope with strong variations in the nitrogen content of the soil solution. Although many molecular actors are being discovered concerning how plants perceive NO3- provision, it is less clear how plants recognize a lack of Nitrogen. Indeed, following N removal plants activate their Nitrogen Starvation Response (NSR) being characterized in particular by the activation of very high affinity nitrate transport systems (NRT2.4, NRT2.5) and other sentinel genes such as GDH3. Here we show using a combination of functional genomics (via TF perturbation) and molecular physiology studies, that the GARP Transcription Factors (TFs) belonging the HHO sub-family are important regulators of the NSR through two potential mechanisms. First, HHOs directly repress NRT2.4 and NRT2.5 high-affinity nitrate transporters. Genotypes affected in HHO genes (mutants and overexpressors) display modified high-affinity nitrate transport activities opening interesting perspectives in biotechnology applications. Second, we show that Reactive Oxygen Species (ROS) are important to control NSR in wild type plants and that HRS1 and HHO1 overexpressors are affected in their ROS content, defining a potential feedforward branch of the signaling pathway. Taken together our results define two new classes of molecular actors in the control of NSR including ROS and the first transcription factors to date. This work (i) opens perspectives on a poorly understood nutrient related signaling pathway, and (ii) defines targets for molecular breeding of plants with enhanced NO3- uptake.

Plants are subjected to variable nitrogen (N) availability including frequent spatial nitrate (NO3-) heterogeneity in soil. Thus, plants constantly adapt their genome expression and root physiology in order to optimize N acquisition from this heterogeneous source. These adaptations rely on a complex and long-distance root-shoot-root signaling network that is still largely unknown. Here, we used a combination of reverse genetics, transcriptomic analysis, NO3- uptake experiments and hormone profiling under conditions of homogeneous or heterogeneous NO3- availability to characterize the systemic signaling involved. We demonstrate the important role of the trans-zeatin form of cytokinin (CK) in shoots, in particular using a mutant altered for ABCG14-mediated trans-zeatin-translocation from the root to the shoot, in mediating: (i) rapid long distance N-demand signaling and (ii) long term functional adaptations to heterogeneous NO3- supply, including changes in NO3- transport capacity and root growth modifications. We also provide insights into the potential CK-dependent and independent shoot-to-root signals involved in root adaptation to heterogeneous N availability.

Plants face temporal and spatial variation in nitrogen (N) availability. This includes heterogeneity in soil nitrate (NO3-) content. To overcome these constraints, plants modify their gene expression and physiological processes to optimize N acquisition. This plasticity relies on a complex long-distance root-shoot-root signaling network that remains poorly understood. We previously showed that cytokinin (CK) biosynthesis is required to trigger systemic N signaling. Here, we performed split-root experiments and used a combination of CK-related mutant analyses, hormone profiling, transcriptomic analysis, NO3- uptake assays, and root growth measurements to gain insight into systemic N signaling in Arabidopsis thaliana. By comparing wild-type plants and mutants affected in CK biosynthesis and ABCG14-dependent root-to-shoot translocation of CK, we revealed an important role for active trans-Zeatin (tZ) in systemic N signaling. Both rapid sentinel gene regulation and long-term functional acclimation to heterogeneous NO3- supply, including NO3- transport and root growth regulation, are likely mediated by the integration of tZ content in shoots. Furthermore, shoot transcriptome profiling revealed that glutamate/glutamine metabolism is likely a target of tZ root-to-shoot translocation, prompting an interesting hypothesis regarding shoot-to-root communication. Finally, this study highlights tZ-independent pathways regulating gene expression in shoots as well as NO3- uptake activity in response to total N-deprivation.

International audience; Background and Aims: Understanding interactions between water and nitrate fluxes in response to nitrate availability and transpiration rate is crucial to select more efficient plants for the use of water and nitrate. Methods: Some of these interactions were investigated in intact Brassica napus plants by combining a non-destructive gravimetric device with (NO3-)-N-15 labeling. The set-up allowed high-resolution measurement of the effects of a cross-combination of two concentrations of KNO3 or KCl (0.5 and 5 mM) with two different rates of transpiration controlled by the relative humidity during a day-night cycle. Key Results: Results show that (1) high external nitrate concentrations increased root water uptake significantly whatever the transpiration rate, (2) nitrate translocation depended both on the rate of nitrate uptake and loading into xylem (3) dilution-concentration effect of nitrate in the xylem was mainly modulated by both external nitrate availability and transpiration rate, (4) dynamic changes in N-15 translocation in the xylem modified shoot growth and capacitance, and (5) variations in tissue concentrations of NO3- induced by the experimental conditions were balanced by changes in concentrations of chloride and sulfate ions. These effects were even more amplified under low transpiration condition and 0.5 mM external nitrate concentration. Conclusion: Taken together, these results highlight the fine and rapid adjustment of anion contents, nitrate and water flows to changes in transpiration rate and nitrate availability during a day-night cycle. The use of this non-invasive gravimetric device is therefore a powerful tool to assess candidates genes involved in nitrogen and water use efficiency.

Plants are subjected to variable nitrogen (N) availability including frequent spatial nitrate (NO3-) heterogeneity in soil. Thus, plants constantly adapt their genome expression and root physiology in order to optimize N acquisition from this heterogeneous source. These adaptations rely on a complex and long distance root-shoot-root signaling network that is still largely unknown. Here, we used a combination of reverse genetics, transcriptomic analysis, NO3- uptake experiments and hormone profiling under conditions of homogeneous or heterogeneous NO3- availability to characterize the systemic signaling involved. We demonstrate the important role of the trans-zeatin form of cytokinin (CK) in shoots, in particular using a mutant altered for ABCG14-mediated trans-zeatin-translocation from the root to the shoot, in mediating: (i) rapid long distance N-demand signaling and (ii) long term functional adaptations to heterogeneous NO3- supply, including changes in NO3- transport capacity and root growthmodifications. We also provide insights into the potential CK-dependent and independent shoot-to-root signals involved in root adaptation to heterogeneous N availability.

Plants need to cope with strong variations in the nitrogen content of the soil solution. Although many molecular actors are being discovered concerning how plants perceive NO3- provision, it is less clear how plants recognize a lack of Nitrogen. Indeed, following N removal plants activate their Nitrogen Starvation Response (NSR) being characterized in particular by the activation of very high affinity nitrate transport systems (NRT2.4, NRT2.5) and other sentinel genes such as GDH3. Here we show using a combination of functional genomics (via TF perturbation) and molecular physiology studies, that the GARP Transcription Factors (TFs) belonging the HHO sub-family are important regulators of the NSR through two potential mechanisms. First, HHOs directly repress NRT2.4 and NRT2.5 high-affinity nitrate transporters. Genotypes affected in HHO genes (mutants and overexpressors) display modified high-affinity nitrate transport activities opening interesting perspectives in biotechnology applications. Second, we show that Reactive Oxygen Species (ROS) are important to control NSR in wild type plants and that HRS1 and HHO1 overexpressors are affected in their ROS content, defining a potential feedforward branch of the signaling pathway. Taken together our results define two new classes of molecular actors in the control of NSR including ROS and the first transcription factors to date. This work (i) opens perspectives on a poorly understood nutrient related signaling pathway, and (ii) defines targets for molecular breeding of plants with enhanced NO3- uptake.

Most plants use nitrate (NO3-) as their major nitrogen (N) source. The NO3- uptake capacity of a plant is determined by three interdependent factors that are sensitive to NO3- availability: (i) the functional properties of the transporters in the roots that contribute to the acquisition of NO3- from the external medium, (ii) the density of functional transporters at the plasma membrane of root cells, and (iii) the surface and architecture of the root system. The identification of factors that regulate the NO3--sensing systems is important for both fundamental and applied science, because these factors control the capacity of plants to use the available NO3-, a process known as the “nitrate use efficiency”. The molecular component of the transporters involved in uptake and sensing mechanism in Arabidopsis roots are presented and their relative contribution discussed.

Nitrogen fertilizer is the most used nutrient source in modern agriculture and represents significant environmental and production costs. In the meantime, the demand for grain increases and production per area has to increase as new cultivated areas are scarce. In this context, breeding for an efficient use of nitrogen became a major objective. In wheat, nitrogen is required to maintain a photosynthetically active canopy ensuring grain yield and to produce grain storage proteins that are generally needed to maintain a high end-use quality. This review presents current knowledge of physiological, metabolic and genetic factors influencing nitrogen uptake and utilization in the context of different nitrogen management systems. This includes the role of root system and its interactions with microorganisms, nitrate assimilation and its relationship with photosynthesis as postanthesis remobilization and nitrogen partitioning. Regarding nitrogen-use efficiency complexity, several physiological avenues for increasing it were discussed and their phenotyping methods were reviewed. Phenotypic and molecular breeding strategies were also reviewed and discussed regarding nitrogen regimes and genetic diversity.

International audience; The strong negative correlation between grain protein concentration (GPC) and grain yield (GY) in bread wheat complicates the simultaneous improvement of these traits. However, earlier studies have concluded that the deviation from this relationship (grain protein deviation or GPD) has strong genetic basis. Genotypes with positive GPD have an increased ability to uptake nitrogen (N) during the post-flowering period independently of the amount of N taken up before flowering, suggesting that genetic variability for N satiety could enable the breakage of the negative relationship. This study is based on two genotypes markedly contrasted for GPD grown under semi-hydroponic conditions differentiated for nitrate availability both before and after flowering. This allows exploration of the genetic determinants of post-flowering N uptake (PANU) by combining whole plant sampling and targeted gene expression approaches. The results highlights the correlation (r(2) = 0.81) with GPC of PANU occurring early during grain development (flowering-flowering + 250 degree-days) independently of GY. Early PANU was in turn correlated (r(2) = 0.80) to the stem-biomass increment after flowering through its effect on N sink activity. Differences in early PANU between genotypes, despite comparable N statuses at flowering, suggest that genetic differences in N satiety could be involved in the establishment of the GPC. Through its strong negative correlation with genes implied in N assimilation, root nitrate concentration appears to be a good marker for evaluating instantaneous plant N demand, and may provide valuable information on the genotypic N satiety level. This trait may help breeders to identify genotypes having high GPC independently of their GY.

International audience; No abstract available

In the past decade, horizontal gene transfer (HGT) has emerged as a major evolutionary process that has shaped the genome of Saccharomyces cerevisiae wine yeasts. We recently showed that a large Torulaspora microellipsoides genomic island carrying two oligopeptide transporters encoded by FOT genes increases the fitness of wine yeast during fermentation of grape must. However, the impact of these genes on the metabolic network of S. cerevisiae remained uncharacterized. Here we show that Fot-mediated peptide uptake substantially affects the glutamate node and the NADPH/NADP(+) balance, resulting in the delayed uptake of free amino acids and altered profiles of metabolites and volatile compounds. Transcriptome analysis revealed that cells using a higher amount of oligopeptides from grape must are less stressed and display substantial variation in the expression of genes in the central pathways of carbon and nitrogen metabolism, amino acid and protein biosynthesis, and the oxidative stress response. These regulations shed light on the molecular and metabolic mechanisms involved in the higher performance and fitness conferred by the HGT-acquired FOT genes, pinpointing metabolic effects that can positively affect the organoleptic balance of wines.

The present work investigates the interactions between soil content, rootstock and scion by focusing on the effects of roostocks and nitrogen supply on grape berry content. Scions of Cabernet Sauvignon (CS) and Pinot Noir (PN) varieties were grafted either on Riparia Gloire de Montpellier (RGM) or 110 Richter (110R) rootstock. The 4 rooststock/scion combinations were fertilized with 3 different levels of nitrogen after fruit set. Both in 2013 and 2014, N supply increased N uptake by the plants, and N content both in vegetative and reproductory organs. Rootstock, variety and year affected berry weight at harvest, while nitrogen did not affect significantly this parameter. Grafting on RGM consistently increased berry weight compared to 110R. PN consistently produced bigger berries than CS. CS berries were heavier in 2014 than in 2013, but the year effect was less marked for PN berries. The berries were collected between veraison and maturity, separated in skin and pulp, and their content was analyzed by conventional analytical procedures and untargeted metabolomics. For anthocyanins, the relative quantitation was fairly comparable with both LC-MS determination and HPLC-DAD, which is a fully quantitative technique. The data show complex responses of the metabolite content (sugars, organic acids, amino acids, anthocyanins, flavonols, flavan-3-ols/procyanidins, stilbenes, hydroxycinnamic and hydroxybenzoic acids.) that depend on the rootstock, the scion, the vintage, the nitrogen level, the berry compartment. This opens a wide range of possibilities to adjust the content of these compounds through the choice of the roostock, variety and nitrogen fertilization.

In wheat, remobilization of nitrogen absorbed before anthesis and regulation of monocarpic senescence is a major issue in breeding for nutrient use efficiency. We identified natural variants of NAM-A1, a gene having the same role as its well-characterized homoeolog NAM-B1, a NAC transcription factor associated with senescence kinetics and nutrient remobilization to the grain. Differences in haplotype frequencies between a worldwide core collection and a panel of European elite varieties were assessed and discussed. Moreover, hypotheses for the loss of function of the most common haplotype in elite European germplasm are discussed.

Background and Aims[br/] Both rootstocks and mineral nutrition, particularly nitrogen (N), are known to affect many aspects of plant development including the control of scion vigour. In this study, we investigated the role of the rootstock genotype in grafted grapevine responses to N supply in terms of the root and leaf ionomes, scion vigour and whole plant biomass.[br/] [br/] Methods and Results[br/] Vitis vinifera cv. Cabernet Sauvignon was grafted on two rootstock genotypes known to confer high and low vigour. Plants were grown for 60 days under three levels of nitrate supply. Stem, leaf, trunk and root biomass, and the concentration of 13 macroelements and microelements in roots and leaves were measured. High scion vigour was associated with a high concentration of elements in the leaves. The concentration of some elements in the leaves, such as phosphorus, was affected by the N supply differently in the two scion/rootstock combinations.[br/] [br/] Conclusion[br/] Differences in rootstock conferred vigour were associated with particular shoot and root ionome profiles, and these responses were dependent on N supply.[br/] [br/] Significance of the Study[br/] These results demonstrate that rootstocks alter scion growth and the leaf ionome in response to N supply.

cf. thèse de doctorat (PhD thesis); Below-ground residue of agroforestry trees is an important N source for associated crops. Several studies have shown that its isotopic signature (δ15N) may change after tree pruning, which makes it difficult to study below-ground N inputs from pruned trees by isotopic techniques. We studied how temporal variation of legume root residue δ15N could be explained by considering differential decomposition kinetics and 15N content of residue fractions. A mathematical model on the isotopic patterns of soil and a N recipient plant during root decomposition was developed and applied for testing assumptions about residue characteristics against two experimental datasets. Observed 15N patterns of the recipient plants could be satisfactorily simulated only when the residue was assumed to consist of at least two fractions with distinct δ15N and decomposition rates depending on their C : N ratio. Assuming δ15N of residue constant over time resulted in substantial underestimates of N derived from low-quality residue (%Ndfr) by the recipient plant when compared with experimental data. Results of this study suggest that residue fractionation can help improve estimation of %Ndfr in isotopic studies, as an alternative or complementary method to assuming or aiming at homogenous isotopic composition of N sources.

International audience; Nitrogen (N) is an essential plant nutrient at the centre of plant metabolism being present in numerous vital organic compounds including amino acids, proteins, nucleic acids and phytohormones. In highly fertile arable soils supplied with N fertilisers, NO3 - and NH4 + ions are usually considered to be the main N forms in soil solution taken up by crop plants, the ratio of NO3-N to NH4-N often being in the order of 10-20:1. Soluble organic N is receiving increasing attention. The form of N uptake modifies rhizosphere pH which affects uptake of other soil nutrients. Acquisition of the various N forms is regulated not only by their chemical and spatial availability in the soil, but also by transport systems in the plasma membrane of root cells, root system architecture and mechanisms that regulate the activity of N transport systems and root growth, depending on plant requirements. In most fertile soils well supplied with mineral nutrients, neither N transport from the bulk soil to crop roots nor the efficient processes of NO3 - and NH4 + ion uptake by roots limit N acquisition. Uptake of N is regulated by N demand in which shoot-root interactions play an important role. Nitrate acts as a signal for metabolism and plant development which is associated with cytokinin transport. The biochemistry of N assimilation is well understood but control at molecular level much less so. Nitrogen status of plants can be characterised by the concept of critical N concentration (Nc) in relation to the long-term fall in N concentration in the dry matter as plants age. Nc represents the optimum for growth for a given crop and can be related to actual crop N concentration. Fertilisation is needed when [N] / [Nc], i.e. NNI (nitrogen nutritional index) < 1. By determining N status during crop growth, fertilisation can be carried out more efficiently as exemplified in melon cultivation. Examples are presented to demonstrate practical effects of different N forms including NO3-, NH4- and urea-N. These examples include the value of placed stabilised NH4- N in depressing rhizosphere pH on high-pH soils to increase the availability and uptake of sparingly soluble phosphate and micronutrients iron, manganese, copper, zinc and boron. These latter nutrients play a vital role, protecting plants against both biotic and abiotic stresses. The acidifying effect in the rhizosphere of symbiotically grown legumes can be of similar benefit on high pH soils. The use of NH4-N fertilisers to depress rhizosphere pH which is also associated with increased Si and Mn uptake is regularly practiced to control some plant diseases such as mildew and take-all in wheat. Water use efficiency (WUE) also relates to the form of N supply. Foliar application of urea is discussed. Signaling effects of stabilised N forms can be used to influence tiller number and yield components in wheat and barley crops. The use of NH4-N in restricting lateral shoot development in tomato production is discussed