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International audience; Aim: Nitrogen is a major element conditioning grapevine growth, yield and aromatic profiles of berries and wines. Different tools can be used in order to detect differences in N status of the plant, including direct measurements of soil, plant nitrogen status (eg. petiole; must), or indirect observations of plant nutritional status such as leaf transmittance or reflectance (eg. SPAD; NDVI). However, the relationships between these indicators of nitrogen status and the overall plant functioning over vintages remain poorly known. The present study aimed at quantifying key vegetative and reproductive responses to plant nitrogen status over two successive seasons under different nitrogen supply levels. Methods and results: Potted plants of Sauvignon Blanc grafted onto SO4 were grown outdoors in 2017 and 2018 with no water limitation. Four mineral nitrogen fertilization levels (equivalent to 0 kg of N ha-1 or 0U, 20U, 40U, 80U) and one organic nitrogen fertilization level (40U) were imposed in 2017. These treatments were doubled in 2018 to increase the degree of nitrogen supply and consequently, the range of observed effects on plant growth and yield. Plant nitrogen status (SPAD) was monitored weekly during both growing cycles. Yield components were determined over the two seasons. Lastly, plant carbon status was addressed through dynamic measurement of plant development and photosynthesis, and destructive measurement of dry matter accumulation and carbon storage in annual and perennial organs at flowering, veraison and harvest. The SPAD values progressively decreased under lower N supply (0N) during the first year (from 31 to 16) and they were more than halved between the maximum and the minimum N treatments straight after budburst in year two (40 for 160N and 19 for 0N). Then, the differences in SPAD values among treatments were maintained up to harvest (2018). The gradient of N status resulted in a gradient of berry numbers per inflorescence (from 180 to 34 berries/inflorescence for 80N and 0N, respectively in 2018) and of individual berry dry matter at harvest (from 0.13 to 0.41 g for 160N and 0N, respectively in 2018). Quantitative relationships between N status and the relative reductions (% of reduction per %SPAD decrease) in terms of C gain (leaf area, photosynthesis), C growth (shoot, berry, trunk and root dry matter) and C storage (trunk and root) were fitted at flowering, veraison and harvest. The reduction in C gain under lower N supply was mainly related to the decrease in total leaf area before flowering (-1.64%). Although the photosynthesis rate tended to decrease under N deficiency over the season, it only poorly contributed to the reduction in C gain. The whole plant C growth was inhibited when N status decreased (-1.13% at harvest), due to the inhibition of shoot dry matter before veraison (-1.81%) and to a lower extent, to the lower dry matter in berries (-0.80%), trunks (-0.42%) and roots (-0.84%) at harvest. Part of the reduction in root dry matter was related to the lower starch reserves (-0.31%) at harvest. Interestingly, starch reserves tended to be higher under organic N supply than mineral N supply. Conclusion: The present results provided a general framework of carbon gain and use over time (within and between seasons) as impacted by N supply levels and form. Such a framework will be useful when building a model of the pluri-annual dynamics of carbon balance related to yield elaboration in grapevines.

Background After uptake from soil into the root tissue, distribution and allocation of nitrate throughout the whole plant body, is a critical step of nitrogen use efficiency (NUE) and for modulation of plant growth in response to various environmental conditions. In legume plants nitrate distribution is also important for the regulation of the nodulation process that allows to fix atmospheric N (N2) through the symbiotic interaction with rhizobia (symbiotic nitrogen fixation, SNF). Results Here we report the functional characterization of the Lotus japonicus gene LjNPF2.9, which is expressed mainly in the root vascular structures, a key localization for the control of nitrate allocation throughout the plant body. LjNPF2.9 expression in Xenopus laevis oocytes induces 15NO3 accumulation indicating that it functions as a nitrate importer. The phenotypic characterization of three independent knock out mutants indicates an increased shoot biomass in the mutant backgrounds. This phenotype is associated to an increased/decreased nitrate content detected in the shoots/roots. Furthermore, our analysis indicates that the accumulation of nitrate in the shoot does not affect the nodulation and N-Fixation capacities of the knock out mutants. Conclusions This study shows that LjNPF2.9 plays a crucial role in the downward transport of nitrate to roots, occurring likely through a xylem-to-phloem loading-mediated activity. The increase of the shoot biomass and nitrate accumulation might represent a relevant phenotype in the perspective of an improved NUE and this is further reinforced in legume plants by the reported lack of effects on the SNF efficiency. Electronic supplementary material The online version of this article (10.1186/s12870-019-1978-5) contains supplementary material, which is available to authorized users.

International audience; Senescence associated gene (SAG) 12, which encodes a cysteine protease isconsidered to be important in nitrogen (N) allocation to Arabidopsis thaliana seeds.A decrease in the yield and N content of the seeds was observed in the ArabidopsisSAG12 knockout mutants (sag12) relative to the wild type (Col0) under limited nitrogennutrition. However, leaf senescence was similar in both lines. To test whether SAG12is involved in N remobilization from organs other than the leaves, we tested whetherroot N could be used in N mobilization to the seeds. Root architecture, N uptakecapacity and 15N partitioning were compared in the wild type and sag12 under eitherhigh nitrogen (HN) or low nitrogen (LN) conditions. No differences in root architectureor root N uptake capacity were observed between the lines under HN or LN. However,under LN conditions, there was an accumulation of 15N in the sag12 roots comparedto the wild type with lower allocation of 15N to the seeds. This was accompanied by anincrease in root N protein contents and a significant decrease in root cysteine proteaseactivity. SAG12 is expressed in the root stele of the plants at the reproductive stage,particularly under conditions of LN nutrition. Taken together, these results suggest anew role for SAG12. This cysteine protease plays a crucial role in root N remobilizationthat ensures seed filling and sustains yields when nitrogen availability is low.

National audience; Pea seeds have interesting nutritional properties through their protein, vitamin and mineral contents. However, environmental conditions such as soil water availability during plant growth influence both final yield and seed quality in pea. Soil water conditions not only modulates pea nitrogen acquisition whatever the mode of nitrogen nutrition: mineral nitrogen acquisition by roots and atmospheric di-nitrogen fixation by nodules in interaction with rhizobia, but also the overall plant mineral nutrition. The aim of this study is to get a better understanding of soil water deficit impact on nutrient acquisition, storage and distribution inside the plant according to the mode of nitrogen nutrition. In this study the first experimentation aims at characterizing 13 individual mineral deficiencies on nutrient uptake and remobilisation into different plant organs, allowing to design an ionomic fingerprint specific to each mineral deficiency. A second experimentation consists in implementing single (at vegetative or reproductive stage) or repeated water deficits to pea plants grown under both modes of nitrogen nutrition. This experimentation will allow us to analyse how water deficit influences nutrient uptake, storage and remobilisation via (1) ecophysiological analysis by using a structure-function framework including water and mineral fluxes at the whole plant level, (2) transcriptomic analysis of nodules and roots. We intend to identify the main regulatory genes involved in either single water deficit response or water deficit memory stress response. This study will allow us to provide new insights towards a better management of preventive or/and curative mineral fertilization solution in the field, in order to prevent pea from mineral deficiencies and improve its yield and seed quality under fluctuating water environment.

International audience

article accepté; In natural environments, nitrogen (N) concentration in the soil fluctuates greatly and is often limiting for plant growth and development. The ability of plants to respond to changes in N availability is therefore essential for adaptation. The response of plants to N variations consists in particular of adjusting root N uptake systems and root architecture. To do so, plants integrate local sensing and signaling of external N availability with systemic sensing and signaling of their internal N status, in order to tune the functional and structural properties of the root system in accordance with the N demand for growth of the whole plant. Transcriptional regulation of gene expression is one of the most important processes plants use to adapt the properties of the root system in response to local and long-distance N pathways. This review focuses on the mechanisms that give rise to transcriptional responses in Arabidopsis roots under N fluctuations, with an emphasis on those associated with the regulation of nitrate uptake and transport systems. We discuss the putative long-distance signals triggering the gene expression responses, as well as the molecular players that locally induce transcriptional changes. We also highlight several observations revealing the importance of adopting an integrative approach in the regulation of N signaling.

La culture de Légumineuses présente le double intérêt de permettre une production de graines à haute valeur nutritionnelle sans nécessité d’un apport d’engrais azoté. La nutrition azotée des légumineuses dépend en effet majoritairement de la fixation symbiotique de l’azote atmosphérique réalisée par des bactéries du sol, les rhizobia, au sein des nodosités, et dans une moindre mesure, de l’assimilation de l’azote minéral du sol par les racines. Une meilleure compréhension a été acquise, par ce travail de thèse, sur le contrôle génétique de la mise en place des racines et des nodosités et sur leur impact sur la nutrition azotée. Une grande variabilité génétique pour ces caractères a été mise en évidence, ainsi que l’existence de corrélations génétiques entre eux. Une approche de génétique quantitative a permis d’identifier des régions génomiques pouvant être impliquées dans leurs variations. Deux pistes d’amélioration de la nutrition azotée ont aussi été étudiées : l’amélioration de l’acquisition d’azote par les racines à partir d’une étude détaillée d’un mutant de développement racinaire, et l’amélioration de la symbiose via l’étude de la capacité des pois à favoriser les associations symbiotiques avec les rhizobia les plus performants. Les résultats obtenus apportent des bases de réflexion concernant la conception d’un idéotype de nutrition azotée. Au-delà de la complémentarité indispensable entre les deux voies d’acquisition d’azote, il convient d’optimiser l’interaction entre les deux partenaires, mécanisme complexe mettant en jeu la formation et le fonctionnement des nodosités, en lien avec une signalétique et des interactions trophiques complexes entre partenaires symbiotiques et intra-plante. ABSTRACT Grain legume pulse crops are of great interest to allow a production of seeds high nutritional value without any contribution of nitrate fertilizer. The nitrogen nutrition of legumes depends indeed mainly on the fixation in nodules of atmospheric dinitrogen through the plant-rhizobium symbiosis, and to a lesser extent, absorption by roots of soil mineral nitrogen. A better understanding has been obtained on the genetic control of the development of roots and nodules and on their impact on nitrogen nutrition. High genetic variability of these characters has been detected, and the existence of genetic correlations between them demonstrated. A quantitative genetic approach has identified several genomic regions that may be involved in their variations. The two different ways to improve nitrogen nutrition were also studied: the improvement of nitrogen acquisition by roots through a detailed study of a root architecture mutant, and the improvement of symbiosis via the study of the ability of peas to promote symbiotic associations with the most effective rhizobia. The results provide interesting bases for the design of a pea nitrogen-nutrition ‘ideotype’. Beyond the essential complementarity between the two pathways of nitrogen acquisition, it is necessary to optimize the interaction between the two symbiotic partners, which is a complex mechanism involving nodules formation and functioning in connection with complex signaling and trophic interactions between the partners and intraplant, Grain legume pulse crops are of great interest to allow a production of seeds high nutritional value without any contribution of nitrate fertilizer. The nitrogen nutrition of legumes depends indeed mainly on the fixation in nodules of atmospheric dinitrogen through the plant-rhizobium symbiosis, and to a lesser extent, absorption by roots of soil mineral nitrogen. A better understanding has been obtained on the genetic control of the development of roots and nodules and on their impact on nitrogen nutrition. High genetic variability of these characters has been detected, and the existence of genetic correlations between them demonstrated. A quantitative genetic approach has identified several genomic regions that may be involved in their variations. The two different ways to improve nitrogen nutrition were also studied: the improvement of nitrogen acquisition by roots through a detailed study of a root architecture mutant, and the improvement of symbiosis via the study of the ability of peas to promote symbiotic associations with the most effective rhizobia. The results provide interesting bases for the design of a pea nitrogen-nutrition ‘ideotype’. Beyond the essential complementarity between the two pathways of nitrogen acquisition, it is necessary to optimize the interaction between the two symbiotic partners, which is a complex mechanism involving nodules formation and functioning in connection with complex signaling and trophic interactions between the partners and intraplant.

L’azote (N) est un facteur majeur limitant la croissance des plantes. Sa disponibilité peut également avoir un impact sur la résistance des plantes aux pathogènes en régulant leur immunité. Afin de mieux comprendre les liens entre la nutrition azotée et les défenses de la plante, nous avons analysé l’impact de la disponibilité en N sur la résistance de Medicago truncatula à un pathogène racinaire, Aphanomyces euteiches, en prenant en compte la variabilité génétique de la plante. Cet oomycète est considéré comme un des facteurs limitant le plus la production des légumineuses. Deux conditions de nutrition azotée, non limitante ou carencée en N, et dix génotypes de M. truncatula ont été testés in vitro. Les résultats ont montré que la résistance est modulée par les conditions nutritionnelles, dépendament du génotype. Les analyses d’expression de gènes impliqués dans le métabolisme azoté et dans les réponses de défense ainsi que la quantification des teneurs en acides aminés et des composés métaboliques secondaires ont montré des réponses différentes selon les génotypes et la condition nutritive. Elles ont souligné en particulier le rôle potentiellement important de la glutamine dans ce pathosystème. De plus, nous avons mis en évidence l’importance de l’homéostasie du monoxyde d’azote (NO) dans la résistance de M. truncatula à A. euteiches et que la disponibilité en azote impactait l’homéostasie du NO en affectant les niveaux de S-nitrosothiols et l’activité de la S-nitrosoglutathion réductase dans les racines. Ces résultats soulignent l’importance du métabolisme azoté et de son interaction avec le génotype de la plante dans les réactions de défense chez M. truncatula., Nitrogen (N) is a major limiting factor for plant growth. N availability can also impact plant resistance to pathogens by regulating plant immunity. To better understand the links between N nutrition and plant defense, we analyzed the impact of N availability of plant on Medicago truncatula resistance to the root pathogen, Aphanomyces euteiches, taking into account plant genetic variability. This oomycete is considered as the most limiting factor for legume production. Two conditions of N nutrition, non-limiting or deprived in N, and ten plant genotypes were tested in vitro. The results showed that the resistance is modulated by nutritional conditions, depending on plant genotype. Analysis of the expression of genes involved in N metabolism and defense and quantification of different amino-acids contents and secondary metabolic compounds showed different responses of the genotypes and highlighted a potential role of glutamine in this pathosystem. Furthermore, our work underlined the importance of nitric oxide (NO) homeostasis for M. truncatula resistance to A. euteiches and that N availability impacts NO homeostasis by affecting S-nitrosothiol levels and S-nitrosoglutathione reductase activity in roots. These studies highlight, therefore, the importance of N metabolism and its interaction with plant genotype in defense responses in M. truncatula.

Modelling the distribution of leaf nitrogen is central to specify photosynthetic parameters and simulate canopy photosynthesis using leaf gas exchange models. Nitrogen distribution depends both on soil nitrogen availability and within-canopy light distribution. We propose the use of a set of empirical relationships with measurable parameters to dynamically account for it. The model was assessed on alfalfa canopies under contrasting N nutrition levels. It proved able to predict nitrogen distribution and daily patterns of photosynthesis from leaf to whole canopy for a large range of situations., Modelling the spatial and temporal distribution of leaf nitrogen (N) is central to specify photosynthetic parameters and simulate canopy photosynthesis. Leaf photosynthetic parameters depend on both local light availability and whole-plant N status. The interaction between these two levels of integration has generally been modelled by assuming optimal canopy functioning, which is not supported by experiments. During this study, we examined how a set of empirical relationships with measurable parameters could be used instead to predict photosynthesis at the leaf and whole-canopy levels. The distribution of leaf N per unit area (Na) within the canopy was related to leaf light irradiance and to the nitrogen nutrition index (NNI), a whole-plant variable accounting for plant N status. Na was then used to determine the photosynthetic parameters of a leaf gas exchange model. The model was assessed on alfalfa canopies under contrasting N nutrition and with N2-fixing and non-fixing plants. Three experiments were carried out to parameterize the relationships between Na, leaf irradiance, NNI and photosynthetic parameters. An additional independent data set was used for model evaluation. The N distribution model showed that it was able to predict leaf N on the set of leaves tested. The Na at the top of the canopy appeared to be related linearly to the NNI, whereas the coefficient accounting for N allocation remained constant. Photosynthetic parameters were related linearly to Na irrespective of N nutrition and the N acquisition mode. Daily patterns of gas exchange were simulated accurately at the leaf scale. When integrated at the whole-canopy scale, the model predicted that raising N availability above an NNI of 1 did not result in increased net photosynthesis. Overall, the model proposed offered a solution for a dynamic coupling of leaf photosynthesis and canopy N distribution without requiring any optimal functioning hypothesis.

International audience; Oilseed rape, a crop requiring a high level of nitogen (N) fertilizers, is characterized by low N use efficiency. To identify the limiting factors involved in the N use efficiency of winter oilseed rape, the response to low N supply was investigated at the vegetative stage in 10 genotypes by using long-term pulse-chase (15)N labelling and studying the physiological processes of leaf N remobilization. Analysis of growth and components of N use efficiency allowed four profiles to be defined. Group 1 was characterized by an efficient N remobilization under low and high N conditions but by a decrease of leaf growth under N limitation. Group 2 showed a decrease in leaf growth under low N supply that was associated with a low N remobilization efficiency under both N supplies despite a high remobilization of soluble proteins. In response to N limitation, Group 3 is characterized by an increase in N use efficiency and leaf N remobilization compared with high N that is not sufficient to sustain the leaf biomass production at a similar level to non-limited plants. Genotypes of Group 4 subjected to low nitrate were able to maintain leaf growth to the same level as under high N. The profiling approach indicated that enhancement of amino acid export and soluble protein degradation was crucial for N remobilization improvement. At the whole-plant level, N fluxes revealed that Group 4 showed a high N remobilization in source leaves combined with a better N utilization in young leaves. Consequently, an enhanced N remobilization limits N loss in fallen leaves, but this remobilized N needs to be efficiently utilized in young leaves to improve N use efficiency.

Plant nitrophily/oligotrophy provides information on the nitrogen richness of their natural habitat. It plays a key role in structuring weed communities. Based on an ecophysiological analysis of the ecological character of nitrophily, our main goal was to better understand the success of nitrophilic weed species and the decline of oligotrophic ones in conventional cropping systems which are characterized by an abundant use of nitrogen fertilizers. A greenhouse experiment was conducted using a range eleven weed and crop species contrasted for their nitrophily status and grown at two different soil-nitrogen concentrations. We show that, compared to an oligotrophic species, a nitrophilic species has a growth highly responsive to soil nitrogen. This is due to higher nitrogen uptake or use efficiency for nitrophilic species when soil-nitrogen availability is high. Our results provide ecophysiological clues to better understand weed community assembly. Applications to weed management in cropping systems are proposed., Le caractère nitrophile/oligotrophe des espèces végétales, qui renseigne sur la richesse en azote de leur habitat préférentiel, joue un rôle déterminant dans la structuration des communautés adventices. L’objectif a été de mieux comprendre, sur la base d’une analyse écophysiologique de ce caractère écologique, le succès des adventices nitrophiles et le déclin des adventices oligotrophes dans les systèmes de cultures conventionnels caractérisés notamment par une utilisation importante d’engrais azotés. Une expérimentation a été conduite en serre sur onze espèces contrastées pour le caractère écologique de nitrophilie et à deux niveaux d’azote du sol. Nous montrons que, par rapport à une espèce oligotrophe, une espèce nitrophile a une croissance fortement stimulée par l’azote en raison d’une meilleure efficience de prélèvement ou de valorisation de l’azote lorsque la disponibilité en azote est élevée. Ces résultats fournissent des clefs de compréhension de l’assemblage des communautés adventices. Des applications pour la gestion des adventices sont proposées.

The microbial loop based on the grazing of bacteria by bacterial-feeding nematodes is thought to play a major role in the mineralization of nutrients such as nitrogen (N) and phosphorus (P) in terrestrial ecosystems. This study was undertaken to quantify the effect of bacterial grazing nematodes on root architecture, growth and mineral nutrition (N and P) in the mycorrhizosphere of woody species (Pinus pinaster). Thirty five days simplified sterile experiment was conducted with young mycorrhizal P. pinaster seedlings (inoculated with Hebeloma cylindrosporum) along with bacteria (Bacillus subtilis) and bacterivorous nematodes (Rhabditis sp.). To know better the role of microbial complexity on N nutrition, two N sources, bacterial N labeled with N-15 and nitrate were supplied in the medium. Phosphorus was supplied as insoluble inorganic tri-calcium phosphate (TCP). The results showed that the N-15 flow (4 mu g plant(-1)) from the bacteria to the plant shoots was only significant when nematodes were present. Plants cultivated with bacteria or nematodes also accumulated higher total N and P in their shoots than mycorrihzal ones. These results revealed that the presence of bacterial-feeding nematodes significantly enhanced N and P availability to mycorrihzal P. pinaster seedlings, probably by improving plant use of nitrate and insoluble P supplied in the medium.

Si l'accroissement de la température et du taux de CO2 a un effet globalement positif sur la photosynthèse, la production fourragère et la fixation symbiotique, les sécheresses et, dans une moindre mesure, une fertilisation azotée limitante des graminées, pourraient annuler cette amélioration. Les interactions entre les différents effets sont analysés, selon le type de couvert (graminée / légumineuse, espèce annuelle ou pérenne, tempérée ou méditerranéenne, plante en C3 ou en C4 (maïs, sorgho)..) ainsi que les phénomènes d'adaptation des espèces et populations aux différents types de climats. Dans le cas de températures estivales fréquemment >30° et/ou de déficit hydrique avril-aoüt >500mm, la survie des populations d'espèces prairiales d'origine tempérée est compromise. un changement de variétés cultivées est donc à anticiper, en poursuivant l'amélioration génétique des espèces., Climatic variables (average and variability) will be undergoing a significant change in the 21st century. What will be the impact on the physiology, production and perennity of forage species? Higher temperatures and increased CO2 levels will have a globally positive effect on photosynthesis, production and symbiotic fixation, but droughts (as well as limited nitrogen supplies) could cancel this. The interactions between these different effects are analyzed, per type of cover (forage grass/legumes, annual and perennial species, temperate or Mediterranean, C3 and C4 plants (maize, sorghum)...) and the way in which species adapt to different types of climate. In extreme conditions (high summer temperatures and/or hydric deficit in April-August > 500 mm), the survival of temperate grassland species is compromised. The genetic improvement of species must anticipate the change in cultivated varieties.

National audience; Climatic variables (average and variability) will be undergoing a significant change in the 21st century. What will be the impact on the physiology, production and perennity of forage species? Higher temperatures and increased CO2 levels will have a globally positive effect on photosynthesis, production and symbiotic fixation, but droughts (as well as limited nitrogen supplies) could cancel this. The interactions between these different effects are analyzed, per type of cover (forage grass/legumes, annual and perennial species, temperate or Mediterranean, C3 and C4 plants (maize, sorghum)...) and the way in which species adapt to different types of climate. In extreme conditions (high summer temperatures and/or hydric deficit in April-August > 500 mm), the survival of temperate grassland species is compromised. The genetic improvement of species must anticipate the change in cultivated varieties.; Si l'accroissement de la température et du taux de CO2 a un effet globalement positif sur la photosynthèse, la production fourragère et la fixation symbiotique, les sécheresses et, dans une moindre mesure, une fertilisation azotée limitante des graminées, pourraient annuler cette amélioration. Les interactions entre les différents effets sont analysés, selon le type de couvert (graminée / légumineuse, espèce annuelle ou pérenne, tempérée ou méditerranéenne, plante en C3 ou en C4 (maïs, sorgho)..) ainsi que les phénomènes d'adaptation des espèces et populations aux différents types de climats. Dans le cas de températures estivales fréquemment >30° et/ou de déficit hydrique avril-aoüt >500mm, la survie des populations d'espèces prairiales d'origine tempérée est compromise. un changement de variétés cultivées est donc à anticiper, en poursuivant l'amélioration génétique des espèces.

L'article original est publié par The American Society of Plant Biologists; International audience; In Arabidopsis (Arabidopsis thaliana), the NRT2.1 gene codes for the main component of the root nitrate (NO(3)(-)) high-affinity transport system (HATS). Due to the strong correlation generally found between high-affinity root NO(3)(-) influx and NRT2.1 mRNA level, it has been postulated that transcriptional regulation of NRT2.1 is a key mechanism for modulation of the HATS activity. However, this hypothesis has never been demonstrated, and is challenged by studies suggesting the occurrence of posttranscriptional regulation at the NRT2.1 protein level. To unambiguously clarify the respective roles of transcriptional and posttranscriptional regulations of NRT2.1, we generated transgenic lines expressing a functional 35S::NRT2.1 transgene in an atnrt2.1 mutant background. Despite a high and constitutive NRT2.1 transcript accumulation in the roots, the HATS activity was still down-regulated in the 35S::NRT2.1 transformants in response to repressive nitrogen or dark treatments that strongly reduce NRT2.1 transcription and NO(3)(-) HATS activity in the wild type. In some treatments, this was associated with a decline of NRT2.1 protein abundance, indicating posttranscriptional regulation of NRT2.1. However, in other instances, NRT2.1 protein level remained constant. Changes in abundance of NAR2.1, a partner protein of NRT2.1, closely followed those of NRT2.1, and thus could not explain the close-to-normal regulation of the HATS in the 35S::NRT2.1 transformants. Even if in certain conditions the transcriptional regulation of NRT2.1 contributes to a limited extent to the control of the HATS, we conclude from this study that posttranscriptional regulation of NRT2.1 and/or NAR2.1 plays a predominant role in the control of the NO(3)(-) HATS in Arabidopsis.

Cereal-legume intercrops are gaining increasing interest in Europe because of low-input preoccupations, preservation of environment and biodiversity. Previous studies had shown that intercropping performances are highly dependant on soil N availability. This practice could provide interesting applications to develop crops for multi-service outcomes along with lower inputs (particularly N). However, very few references are nowadays available to allow N management of intercrops for contrasted production targets. The objectives of this work was (i) to use current knowledge on winter pea-wheat intercrop in order to study whether N fertilization could be a helpful tool to manage such intercrop towards production targets under a conventional agriculture; (ii) to improve knowledge on the response of intercrop to different N-availability dynamics (N resources sharing; inhibition and reversibility of Symbiotic Nitrogen Fixation (SNF)); (iii) to give propositions towards decision rules about Nmanagement of pea-wheat intercrops for different production goals. Our field experiments demonstrated that N-fertilization is an efficient tool to manage pea-wheat intercrop, notably contribution of cereal in total intercrop yield, which is nowadays a badly controlled criterion. N-supply favoured cereal growth to the detriment to the legume. Cereal is more competitive than the legume for mineral N resources if N-application occurs before pea beginning of seed filling. However, the intensity of the response to date of N-fertilizer application is variable according to gap between intercrops species in growth dynamics and stage of development. This is determinant for N-sharing and SNF response. Experiments under greenhouse conditions investigated inhibition effect of nitrate and ability of pea SNF to recover through a structure –function analysis according to different stages of nitrate exposure. Nitrate reduced rate of nodule establishment when nodulated root were exposed to nitrate during vegetative phases while it entailed damage on existing nodules when applied during flowering and seed filling. Nitrate exposure always decreased specific activity of nodules. Moreover, the ability to recover of SNF after nitrate removal is partly dependant on level of C availability to nodules. Thus, pea SNF can recover when a short-term inhibition by nitrate occurs before seed filling stages, which is in agreement with our own field observations. Crop model AZODYN-IC was built to simulate pea-wheat intercrop. It is based on two sole crop models (AZODYN and AFISOL for wheat and pea, respectively). Its main interests are (i) to be able to satisfactorily simulate response of such intercrops to various soil mineral N availability dynamics in order to be used as decision support tool; (ii) to use simple formalisms to simulate resources sharing (light, water, and nitrogen), tightly linking light sharing and Nacquisition, (iii) to run with the sole-crop parameters. This model was used to extend field experimentations by simulating wider range of N-management strategies (sowing densities x Nsupply rates x N-supply dates) using numerous climatic datasets. This work was able to give propositions towards decision rules for N-management of intercrop according to soil N mineral content at the end of winter, contribution of wheat in intercrop biomass at the end of winter, estimation of N-mineralization from end of winter to harvest, and rate and date of N-application.; Avec l'émergence des préoccupations d'économie d'intrants, de préservation de l'environnement et de la biodiversité, les associations, qui consistent en la culture d'au moins deux espèces différentes sur la même surface pendant une période significative de leur développement, présentent un regain d'intérêt en Europe. Ces associations pourraient avoir des applications intéressantes pour le développement de cultures « multiservices » à moindre niveau d'intrants (azote particulièrement). Des travaux antérieurs ont montré que les performances des associations céréale-légumineuse dépendent fortement du niveau d'azote minéral du sol Cependant on ne dispose pas à l'heure actuelle de références suffisantes pour piloter la fertilisation azotée de l'association en fonction de différents objectifs de production. Les objectifs de la thèse étaient (i) d'utiliser les connaissances précédemment acquises sur le fonctionnement dynamique d'une association pois-blé d'hiver pour étudier la pertinence de la fertilisation azotée comme levier pour orienter les performances des associations pois-blé vers différents objectifs de production en agriculture conventionnelle ; (ii) d'approfondir les connaissances sur le fonctionnement de l'association en réponse à différentes dynamiques de disponibilité en azote (partage des ressources azotées ; inhibition et réversibilité de la fixation symbiotique) ; (iii) d'apporter des pistes vers des règles de décision pour la gestion la fertilisation azotée de ces associations pour différents objectifs de production. Nos expérimentations de plein champ démontrent que la fertilisation azotée est un levier efficace pour orienter les performances finales notamment la proportion de chaque espèce dans le mélange, critère aujourd'hui mal maîtrisé. Un apport d'azote favorise la croissance de la céréale et pénalise celle de la légumineuse. La céréale apparaît plus compétitive que la légumineuse pour les ressources d'azote minéral pour une date d'apport intervenant avant début du remplissage des grains du pois. Cependant, l'intensité de la réponse à la date de fertilisation varie en fonction des écarts de dynamiques de croissance et de phénologie de chaque espèce avant l'apport, facteurs qui apparaissent déterminants dans le partage de l'N minéral et le comportement de la fixation symbiotique. Des expérimentations en conditions contrôlées ont permis d'approfondir l'effet inhibiteur des nitrates et la réversibilité de la fixation chez le pois en analysant séparément l'impact sur la structure et sur l'activité de l'appareil fixateur en fonction du stade phénologique et de la disponibilité en carbone. Une exposition aux nitrates pendant la phase végétative réduit la vitesse d'apparition des nodosités alors qu'une exposition durant la floraison et le remplissage des grains réduit la croissance des nodosités existantes. Les nitrates réduisent fortement l'activité fixatrice quelle que soit la date d'exposition aux nitrates. De plus, il a été démontré que la réversibilité de la fixation symbiotique après courte exposition aux nitrates était fonction de l'allocation carbonée aux nodosités. Ainsi, la réversibilité de la fixation symbiotique est possible chez le pois si une courte inhibition due nitrates survient avant les stades de remplissage du grain, ce qui est confirmé en situations de plein champ. Notre démarche de modélisation a par ailleurs aboutit au développement, à partir de deux modèles de culture pures (AZODYN pour le blé et AFISOL pour le pois), d'un modèle dynamique du fonctionnement de ces associations (AZODYN-IC) dont l'intérêt et l'originalité se situent dans (i) sa capacité à bien simuler la réponse à des disponibilités en azote variées permettant ainsi d'être directement opérationnel et utilisé comme outil d'aide à la gestion de la fertilisation azotée (ii) des formalismes relativement simples de partage des ressources (lumière, eau, azote) et comprenant un lien très étroit entre le partage de la lumière et l'acquisition de l'azote, déterminant dans le fonctionnement du peuplement, (iii) l'absence de paramétrage spécifique pour simuler le fonctionnement de l'association. Le modèle a permis de prolonger les expérimentations en simulant une gamme de stratégies de fertilisation plus large (combinaisons de proportions de semis x doses x dates de fertilisation) et ceci pour une gamme importante d'années climatiques. Ce travail a également permis de proposer des pistes vers des règles de décision de conduites azotées en fonction des reliquats d'azote observés sortie hiver, de la proportion de blé dans la biomasse de l'association observée sortie hiver, et de l'estimation de la minéralisation depuis la sortie hiver jusqu'à la récolte.

Cereal-legume intercrops are gaining increasing interest in Europe because of low-input preoccupations, preservation of environment and biodiversity. Previous studies had shown that intercropping performances are highly dependant on soil N availability. This practice could provide interesting applications to develop crops for multi-service outcomes along with lower inputs (particularly N). However, very few references are nowadays available to allow N management of intercrops for contrasted production targets. The objectives of this work was (i) to use current knowledge on winter pea-wheat intercrop in order to study whether N fertilization could be a helpful tool to manage such intercrop towards production targets under a conventional agriculture; (ii) to improve knowledge on the response of intercrop to different N-availability dynamics (N resources sharing; inhibition and reversibility of Symbiotic Nitrogen Fixation (SNF)); (iii) to give propositions towards decision rules about Nmanagement of pea-wheat intercrops for different production goals. Our field experiments demonstrated that N-fertilization is an efficient tool to manage pea-wheat intercrop, notably contribution of cereal in total intercrop yield, which is nowadays a badly controlled criterion. N-supply favoured cereal growth to the detriment to the legume. Cereal is more competitive than the legume for mineral N resources if N-application occurs before pea beginning of seed filling. However, the intensity of the response to date of N-fertilizer application is variable according to gap between intercrops species in growth dynamics and stage of development. This is determinant for N-sharing and SNF response. Experiments under greenhouse conditions investigated inhibition effect of nitrate and ability of pea SNF to recover through a structure –function analysis according to different stages of nitrate exposure. Nitrate reduced rate of nodule establishment when nodulated root were exposed to nitrate during vegetative phases while it entailed damage on existing nodules when applied during flowering and seed filling. Nitrate exposure always decreased specific activity of nodules. Moreover, the ability to recover of SNF after nitrate removal is partly dependant on level of C availability to nodules. Thus, pea SNF can recover when a short-term inhibition by nitrate occurs before seed filling stages, which is in agreement with our own field observations. Crop model AZODYN-IC was built to simulate pea-wheat intercrop. It is based on two sole crop models (AZODYN and AFISOL for wheat and pea, respectively). Its main interests are (i) to be able to satisfactorily simulate response of such intercrops to various soil mineral N availability dynamics in order to be used as decision support tool; (ii) to use simple formalisms to simulate resources sharing (light, water, and nitrogen), tightly linking light sharing and Nacquisition, (iii) to run with the sole-crop parameters. This model was used to extend field experimentations by simulating wider range of N-management strategies (sowing densities x Nsupply rates x N-supply dates) using numerous climatic datasets. This work was able to give propositions towards decision rules for N-management of intercrop according to soil N mineral content at the end of winter, contribution of wheat in intercrop biomass at the end of winter, estimation of N-mineralization from end of winter to harvest, and rate and date of N-application.; Avec l'émergence des préoccupations d'économie d'intrants, de préservation de l'environnement et de la biodiversité, les associations, qui consistent en la culture d'au moins deux espèces différentes sur la même surface pendant une période significative de leur développement, présentent un regain d'intérêt en Europe. Ces associations pourraient avoir des applications intéressantes pour le développement de cultures « multiservices » à moindre niveau d'intrants (azote particulièrement). Des travaux antérieurs ont montré que les performances des associations céréale-légumineuse dépendent fortement du niveau d'azote minéral du sol Cependant on ne dispose pas à l'heure actuelle de références suffisantes pour piloter la fertilisation azotée de l'association en fonction de différents objectifs de production. Les objectifs de la thèse étaient (i) d'utiliser les connaissances précédemment acquises sur le fonctionnement dynamique d'une association pois-blé d'hiver pour étudier la pertinence de la fertilisation azotée comme levier pour orienter les performances des associations pois-blé vers différents objectifs de production en agriculture conventionnelle ; (ii) d'approfondir les connaissances sur le fonctionnement de l'association en réponse à différentes dynamiques de disponibilité en azote (partage des ressources azotées ; inhibition et réversibilité de la fixation symbiotique) ; (iii) d'apporter des pistes vers des règles de décision pour la gestion la fertilisation azotée de ces associations pour différents objectifs de production. Nos expérimentations de plein champ démontrent que la fertilisation azotée est un levier efficace pour orienter les performances finales notamment la proportion de chaque espèce dans le mélange, critère aujourd'hui mal maîtrisé. Un apport d'azote favorise la croissance de la céréale et pénalise celle de la légumineuse. La céréale apparaît plus compétitive que la légumineuse pour les ressources d'azote minéral pour une date d'apport intervenant avant début du remplissage des grains du pois. Cependant, l'intensité de la réponse à la date de fertilisation varie en fonction des écarts de dynamiques de croissance et de phénologie de chaque espèce avant l'apport, facteurs qui apparaissent déterminants dans le partage de l'N minéral et le comportement de la fixation symbiotique. Des expérimentations en conditions contrôlées ont permis d'approfondir l'effet inhibiteur des nitrates et la réversibilité de la fixation chez le pois en analysant séparément l'impact sur la structure et sur l'activité de l'appareil fixateur en fonction du stade phénologique et de la disponibilité en carbone. Une exposition aux nitrates pendant la phase végétative réduit la vitesse d'apparition des nodosités alors qu'une exposition durant la floraison et le remplissage des grains réduit la croissance des nodosités existantes. Les nitrates réduisent fortement l'activité fixatrice quelle que soit la date d'exposition aux nitrates. De plus, il a été démontré que la réversibilité de la fixation symbiotique après courte exposition aux nitrates était fonction de l'allocation carbonée aux nodosités. Ainsi, la réversibilité de la fixation symbiotique est possible chez le pois si une courte inhibition due nitrates survient avant les stades de remplissage du grain, ce qui est confirmé en situations de plein champ. Notre démarche de modélisation a par ailleurs aboutit au développement, à partir de deux modèles de culture pures (AZODYN pour le blé et AFISOL pour le pois), d'un modèle dynamique du fonctionnement de ces associations (AZODYN-IC) dont l'intérêt et l'originalité se situent dans (i) sa capacité à bien simuler la réponse à des disponibilités en azote variées permettant ainsi d'être directement opérationnel et utilisé comme outil d'aide à la gestion de la fertilisation azotée (ii) des formalismes relativement simples de partage des ressources (lumière, eau, azote) et comprenant un lien très étroit entre le partage de la lumière et l'acquisition de l'azote, déterminant dans le fonctionnement du peuplement, (iii) l'absence de paramétrage spécifique pour simuler le fonctionnement de l'association. Le modèle a permis de prolonger les expérimentations en simulant une gamme de stratégies de fertilisation plus large (combinaisons de proportions de semis x doses x dates de fertilisation) et ceci pour une gamme importante d'années climatiques. Ce travail a également permis de proposer des pistes vers des règles de décision de conduites azotées en fonction des reliquats d'azote observés sortie hiver, de la proportion de blé dans la biomasse de l'association observée sortie hiver, et de l'estimation de la minéralisation depuis la sortie hiver jusqu'à la récolte.

The aim of this study was to improve knowledge on physiological patterns of oak trees' tolerance to waterlogging (or flooding), in order to discriminate useful settings to assess their tolerance potential. In plants the main constraint caused by flooding is the lack of oxygen (root hypoxia) leading to an energy crisis. The responses to this stress were investigated under controlled conditions in two sympatric oak species known to display different ecological requirements, particularly relative to waterlogging. The tolerant pedunculate oak and the more sensitive sessile oak were studied at the early stage of seedling development, in presence of their cotyledons. Potted seedlings were submitted during two flush of growth to a waterlogging or a control treatment. A drainage stage following the waterlogging was included in a third treatment to mimic a temporary soil flooding (most usual cases on the field). One part of the study was related to seedling growth development, especially root distribution. A second part deals with nitrogen nutrition (assimilation and allocation), that was previously assessed by monitoring changes in mineral nitrogen composition of the rhizosphere. Finally, a third part examines the pattern of carbohydrate management (starch and soluble carbohydrates). In both species, the first flush of growth was not inhibited by waterlogging whereas the second flush was severely disrupted, while the root growth was drastically reduced. Nevertheless, pedunculate oak showed a stronger root system than sessile oak, especially by maintaining root colonization in the shallow waterlogged soil layer (adventitious roots). Moreover, pedunculate oak showed the ability to regenerate a strong root system quickly after drainage. The growth recovery of pedunculate oak seedlings was greater than that of sessile oak, allowing its better establishment on hydromorphic soils since the early stage of seedlings. The nitrogen monitoring in the rhizospheric flooded soil showed a switch from nitrate to ammonia, reversible after drainage. The nitrogen metabolism was similar in the two oak species. The key enzymatic activities of the nitrogen assimilation pathway (nitrate reductase and glutamine synthetase) were not clearly affected in the roots neither in the leaves. However, under waterlogging the two oak species showed nitrogen deficiency in the leaves. Overall, nitrogen nutrition is not a feature of waterlogging tolerance which can discriminate these two oak species. The total starch content of waterlogged seedlings was significantly decreased in both species but starch accumulation process remained active, especially in the first flush stem and in the upper taproot section. The overall management of carbohydrates was significantly different between the two oak species when subjected to waterlogging. Under waterlogging, pedunculate oak showed both starch accumulation and a growth rate higher than sessile oak, while carbohydrate mobilization from cotyledons was delayed by waterlogging especially in pedunculate oak seedlings. The α-amylase enzyme activity in cotyledons was corroborated at the rate of starch mobilization. Unlike crop plants (e.g. rice, wheat), tolerance to waterlogging of oak seedlings was not related to higher rates of carbohydrate mobilization from cotyledons. Pedunculate oak seems able to save its carbohydrate reserves until the growth conditions are more favourable and therefore it was able to better sustain a fast and strong recovery after drainage. Over the duration of the experiments, despite a growth limitation, waterlogged seedlings did not seem under carbon limitation because soluble carbohydrate availability was maintained in roots. However, the obtained results showed a carbon imbalance in sessile oak seedlings. Thus, carbon limitation and then carbohydrate starvation could take place. The features of carbohydrate management, related to the whole plant's carbon metabolism, and root growth appear crucial in tolerance to waterlogging. These physiological patterns are usable at the seedling stage to assess their waterlogging tolerance., Ce travail a pour objectif d'améliorer la connaissance des mécanismes physiologiques qui président à la tolérance à l'ennoyage du chêne. La principale contrainte de l'ennoyage est un déficit en oxygène (hypoxie). Les réponses à ce stress sont étudiées chez le chêne pédonculé (réputé tolérant) et le chêne sessile (plus sensible), à des stades précoces de développement des semis, en présence des cotylédons. Une période de drainage est incluse pour mimer un ennoyage temporaire. La croissance des semis a été suivie, notamment au niveau de l'architecture racinaire. La contribution de la nutrition azotée (assimilation et allocation) a été évaluée en suivant au préalable le devenir des éléments minéraux azotés dans la rhizosphère. La gestion des réserves glucidiques (amidon et sucres solubles) des organes de réserve (cotylédons) et des organes en croissance lors du développement des semis a été mise en relation avec la tolérance à l'ennoyage des chênes. Il ressort des paramètres de croissance étudiés que l'ennoyage a peu d'effets sur la première vague de croissance foliaire mais inhibe la deuxième vague, alors que la croissance racinaire est particulièrement inhibée. Néanmoins, le chêne pédonculé est capable d'une forte colonisation racinaire des horizons superficiels du sol ennoyé et, après drainage, son aptitude à régénérer des racines est plus efficace que chez le chêne sessile. Dans la rhizosphère, l'ennoyage entraîne un basculement réversible des formes azotées nitrates / ammonium. Les modifications du métabolisme azoté sont globalement similaires entre les deux espèces. L'assimilation de l'azote, via la nitrate réductase et la glutamine synthétase, n'est pas réellement perturbée mais, les deux espèces présentent une carence azotée au niveau foliaire. L'assimilation et l'allocation azotée n'apparaissent pas comme des traits discriminants de la tolérance à l'ennoyage chez ces espèces. Sous ennoyage, l'accumulation totale d'amidon est réduite mais elle reste active dans la partie basale du pivot et dans la tige des deux espèces. Elle est cependant plus élevée chez le chêne pédonculé. L'ennoyage restreint la mobilisation des réserves cotylédonaires d'amidon, notamment chez le chêne pédonculé. L'activité des α amylases cotylédonaires corrobore le taux de mobilisation de l'amidon. La tolérance à l'ennoyage des semis de chêne n'est pas associée à une stimulation de la mobilisation des réserves glucidiques cotylédonaires. Le chêne pédonculé serrait plus économe en glucide que le chêne sessile (faible mobilisation cotylédonaire / stockage d'amidon élevé / meilleure croissance). Cette particularité pourrait constituer un marqueur physiologique important de la tolérance à l'ennoyage chez les chênes.

Nitrogen represents the major element in plants. This element is present in most of the basic constituents: purines, pyrimidines, and amino acids, and is important for the autotrophic capacity of the plants, being part of the light harvesting molecules, e.g., chlorophyll. De facto, N deficiency in plants leads rapidly to chlorosis and stunted growth. Plants being sessile organisms, they are completely dependent on the N availability in the soil solution for their growth and productivity. In soil, a harsh competition exists for N acquisition between the microorganisms (mainly bacteria and fungi) and the plants. However, plant species have developed various strategies of N acquisition. These strategies are including microorganism partners into a symbiotic association. First, legumes are associated with bacteria to assimilate aerial N. A second form of mutualism is the association between plants and mycorrhizal fungi. The fungus and the plant are associated underground forming a close bound between the hyphae and the root cells, the so-called mycorrhizal root. These mycorrhizal roots are characterized by the presence of an extensive network of mycelium developing out of the root and exploring the soil. Therefore, the fungal partner considerably increases the volume of soil that is exploited compared to the nonmycorrhizal root system (Rousseau et al. 19994). The mycelium also develops between the root cells inside the root cortex. All mineral and nutrient exchanges between the host cell (carbohydrate release) and the fungal cells (phosphorus and nitrogen release) are thought to take place in this fungal‐plant interface (Smith and Read 1997). Melin and Nilsson (1952) were the first to demonstrate the N uptake by the extra radical mycelium and its transfer to the host plant. Since this pioneering study, a considerable amount of work has been devoted to the capacities of mycorrhizal symbiosis to modify the access to the various N-forms from the soil, including the uptake and assimilation of N by the partners. The main purpose of