Plant N Fluxes and Modulation by Nitrogen, Heat and Water Stresses: A Review Based on Comparison of Legumes and Non Legume Plants

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Modelling Memory: do crop models need to become nostalgic?

International audienceIncreased frequency of stress events such as heat waves has been observed for the last decades. Based on the last IPCC report, they are expected to be more frequent, to last longer and to increase in intensity during the reproductive phase of economically important crops. Many recent studies pointed out induced memory effects of stressing events when plants are challenged several times with similar stresses throughout the crop season. These memory effects were shown to be potentially beneficial since the plants are 'primed' and thus more prepared to develop an earlier, more rapid, intense and/or sensitive response when the stress recurs [1]. Therefore, the new climatic patterns prompts to take into account stress memory into predictive crop modelling approaches so as to estimate the effects of repeated stresses and their consequences on crop yield, quality of harvested products. During the last decades, the use of crop models have been enlarged to climate change driven predictions [2]. While evidence for improving crop climate models and especially the temperature response functions in order to reduce uncertainty in yield simulations before any decision making in agriculture, no modelling studies have attempted to decipher and interpret simulation bias in the light of stress memory nor they focused on methodologies to take into account stress memory effects

Unraveling the role of transient starch in the response of Arabidopsis to elevated CO2 under long-day conditions

Previous studies on Arabidopsis under long-term exposure to elevated CO2 have been conducted using starch synthesis and breakdown mutants cultured under short day conditions. These studies showed that starch synthesis can ameliorate the photosynthetic reduction caused by soluble sugar-mediated feedback regulation. In this work we characterized the effect of long-term exposure to elevated CO2 (800 ppm) on growth, photosynthesis and content of primary photosynthates in long-day grown wild type plants as well as the near starch-less (aps1) and the starch-excess (gwd) mutants. Notably, elevated CO2 promoted growth of both wild type and aps1 plants but had no effect on gwd plants. Growth promotion by elevated CO2 was accompanied by an increased net photosynthesis in WT and aps1 plants. However, the plants with the highest starch content (wild type at elevated CO2, gwd at ambient CO2, and gwd at elevated CO2) were the ones that suffered decreased in in vivo maximum carboxylation rate of Rubisco, and therefore, photosynthetic down-regulation. Further, the photosynthetic rates of wild type at elevated CO2 and gwd at elevated CO2 were acclimated to elevated CO2. Notably, elevated CO2 promoted the accumulation of stress-responsive and senescence-associated amino acid markers in gwd plants. The results presented in this work provide evidence that under long-day conditions, temporary storage of overflow photosynthate as starch negatively affect Rubisco performance. These data are consistent with earlier hypothesis that photosynthetic acclimation can be caused by accelerated senescence and hindrance of CO2 diffusion to the stroma due to accumulation of large starch granules

Concerted changes in N and C primary metabolism in alfalfa (Medicago sativa) under water restriction

Although the mechanisms of nodule N2 fixation in legumes are now well documented, some uncertainty remains on the metabolic consequences of water deficit. In most cases, little consideration is given to other organs and, therefore, the coordinated changes in metabolism in leaves, roots, and nodules are not well known. Here, the effect of water restriction on exclusively N2-fixing alfalfa (Medicago sativa L.) plants was investigated, and proteomic, metabolomic, and physiological analyses were carried out. It is shown that the inhibition of nitrogenase activity caused by water restriction was accompanied by concerted alterations in metabolic pathways in nodules, leaves, and roots. The data suggest that nodule metabolism and metabolic exchange between plant organs nearly reached homeostasis in asparagine synthesis and partitioning, as well as the N demand from leaves. Typically, there was (i) a stimulation of the anaplerotic pathway to sustain the provision of C skeletons for amino acid (e.g. glutamate and proline) synthesis; (ii) re-allocation of glycolytic products to alanine and serine/glycine; and (iii) subtle changes in redox metabolites suggesting the implication of a slight oxidative stress. Furthermore, water restriction caused little change in both photosynthetic efficiency and respiratory cost of N2 fixation by nodules. In other words, the results suggest that under water stress, nodule metabolism follows a compromise between physiological imperatives (N demand, oxidative stress) and the lower input to sustain catabolism

How does sulphur availability modify N acquisition of white clover (Trifolium repens L.)?

The role of S in legume growth, N uptake, and N2 fixation was investigated using white clover (Trifolium repens L.) as a model species. We examined whether the effect of sulphate addition on N fixation resulted from a stimulation of host plant growth, a specific effect of S on nodulation, or a specific effect of S on nodule metabolism. Clones of white clover, inoculated with Rhizobium leguminosarum, were grown for 140 d in a hydroponic system with three levels of sulphate concentration (0 mM, 0.095 mM, and 0.380 mM). Nodule morphological and biochemical traits, such as root length, nodule biomass and volume, nodule protein contents (nitrogenase and leghaemoglobin obtained by an immunological approach), and root amino acid concentrations, were used to analyse the effect of sulphate availability on N2 fixation. The application of sulphate increased whole plant dry mass, root length, and nodule biomass, expressed on a root-length basis. N uptake proved less sensitive than N2 fixation to the effects of S-deficiency, and decreased as a consequence of the lower root length observed in S-deficient plants. N2 fixation was drastically reduced in S-deficient plants as a consequence of a low nodule development, but also due to low nitrogenase and leghaemoglobin production. This effect is likely to be due to down-regulation by a N-feedback mechanism, as, under severe S-deficiency, the high concentration of whole plant N and the accumulation of N-rich amino acids (such as asparagine) indicated that the assimilation of N exceeded the amount required for plant growth

Plant physiology and proteomics reveals the leaf response to drought in alfalfa (Medicago sativa L.)

Despite its relevance, protein regulation, metabolic adjustment, and the physiological status of plants under drought is not well understood in relation to the role of nitrogen fixation in nodules. In this study, nodulated alfalfa plants were exposed to drought conditions. The study determined the physiological, metabolic, and proteomic processes involved in photosynthetic inhibition in relation to the decrease in nitrogenase (Nase) activity. The deleterious effect of drought on alfalfa performance was targeted towards photosynthesis and Nase activity. At the leaf level, photosynthetic inhibition was mainly caused by the inhibition of Rubisco. The proteomic profile and physiological measurements revealed that the reduced carboxylation capacity of droughted plants was related to limitations in Rubisco protein content, activation state, and RuBP regeneration. Drought also decreased amino acid content such as asparagine, and glutamic acid, and Rubisco protein content indicating that N availability limitations were caused by Nase activity inhibition. In this context, drought induced the decrease in Rubisco binding protein content at the leaf level and proteases were up-regulated so as to degrade Rubisco protein. This degradation enabled the reallocation of the Rubisco-derived N to the synthesis of amino acids with osmoregulant capacity. Rubisco degradation under drought conditions was induced so as to remobilize Rubisco-derived N to compensate for the decrease in N associated with Nase inhibition. Metabolic analyses showed that droughted plants increased amino acid (proline, a major compound involved in osmotic regulation) and soluble sugar (D-pinitol) levels to contribute towards the decrease in osmotic potential (Ψs). At the nodule level, drought had an inhibitory effect on Nase activity. This decrease in Nase activity was not induced by substrate shortage, as reflected by an increase in total soluble sugars (TSS) in the nodules. Proline accumulation in the nodule could also be associated with an osmoregulatory response to drought and might function as a protective agent against ROS. In droughted nodules, the decrease in N2 fixation was caused by an increase in oxygen resistance that was induced in the nodule. This was a mechanism to avoid oxidative damage associated with reduced respiration activity and the consequent increase in oxygen content. This study highlighted that even though drought had a direct effect on leaves, the deleterious effects of drought on nodules also conditioned leaf responsiveness

Contribution des réserves azotées de la luzerne (Medicago sativa L.) à la tolérance à la sécheresse

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Modifications physiologique et protéomique associées à la remobilisation de l'azote foliaire au cours de la sénescence séquentielle chez le colza (Brassica napus L.)

Plante de grande culture, le colza (Brassica napus L.) présente une faible efficience d utilisation de l azote (EUA) due à une mauvaise remobilisation de l azote (N) au cours de la sénescence foliaire. Afin de caractériser les mécanismes clés susceptibles d améliorer l EUA, les objectifs visaient à identifier les processus physiologiques, protéomiques et moléculaires impliqués dans la remobilisation du N au cours de la sénescence foliaire et de déterminer l impact de bas intrants azotés sur ces processus. Les jeunes feuilles de colza privées en nitrate présentent un ralentissement de la sénescence concomitant à l accumulation d un inhibiteur de protéases correspondant à BnD22 (Brassica napus Drought 22 kDa), une protéine capable de se lier aux chlorophylles. La double fonction de BnD22 interviendrait dans la protection des tissus juvéniles en maintenant l intégrité des protéines et la capacité photosynthétique en réponse aux stress abiotiques. L analyse des événements physiologiques et protéomiques associés à la remobilisation du N et à la sénescence foliaire a montré l implication de protéines du métabolisme énergétique, de réponse aux stress et de protéolyse. Une protéase chloroplastidiale FtsH, une protéase à aspartate, une sous-unité du protéaosome et une protéase à cystéine SAG12 sont successivement induites au cours de la sénescence. La privation en N accélère l entrée en sénescence mais n affecte pas la séquence des événements. Le faible niveau de N résiduel observé dans les feuilles chutées de colzas privés en N s explique par la conjonction d une sénescence précoce, d une protéolyse rapide, et d une augmentation de la durée et de l intensité du recyclage du N.senescence, the faster proteolysis, and the increase of duration as well as intensity of N recycling. Brassica napus L. (oilseed rape) is an important crop plant with low nitrogen use efficiency (NUE) due to a weak remobilization of nitrogen (N) during leaf senescence. In order to characterize the key processes able to improve the NUE, the objectives were to identify the physiological, proteomics and molecular events implied in N remobilization during leaf senescence, and to determine whether low mineral N availability impact on these events. Young leaves of nitrate-deprived plants presented a delay of senescence concomitant with the accumulation of a trypsin inhibitor corresponding to BnD22 (Brassica napus Drought 22 kDa), a protein capable of binding chlorophylls. The dual function of BnD22 could be involved in the protection of younger tissues by maintaining protein integrity and photosynthesis capacity in response to abiotic stresses. The analysis of proteomics and physiological events associated with N remobilization and leaf senescence revealed the involvement of proteins acting in energy metabolism, plant stress response and proteolysis. A chloroplastidial protease FtsH, an aspartic protease, a proteaosome subunit and a cysteine protease SAG12 were successively induced during leaf senescence. The N starvation led to accelerate the onset of leaf senescence but did not affect the sequence of events. The weak level of residual N observed in fallen leaves of oilseed rape deprived in N was explained by the prematureCAEN-BU Sciences et STAPS (141182103) / SudocSudocFranceF

Leaf senescence and nitrogen remobilization efficiency in oilseed rape (<em>Brassica napus</em> L.)

International audienceDespite its worldwide economic importance for food (oil, meal) and non-food (green energy and chemistry) uses, oilseed rape has a low nitrogen (N) use efficiency (NUE), mainly due to the low N remobilization efficiency (NRE) observed during the vegetative phase when sequential leaf senescence occurs. Assuming that improvement of NRE is the main lever for NUE optimization, unravelling the cellular mechanisms responsible for the recycling of proteins (the main N source in leaf) during sequential senescence is a prerequisite for identifying the physiological and molecular determinants that are associated with high NRE. The development of a relevant molecular indicator (SAG12/Cab) of leaf senescence progression in combination with a N-15-labelling method were used to decipher the N remobilization associated with sequential senescence and to determine modulation of this process by abiotic factors especially N deficiency. Interestingly, in young leaves, N starvation delayed senescence and induced BnD22, a water-soluble chlorophyll-binding protein that acts against oxidative alterations of chlorophylls and exhibits a protease inhibitor activity. Through its dual function, BnD22 may help to sustain sink growth of stressed plants and contribute to a better utilization of N recycled from senescent leaves, a physiological trait that could improve NUE. Proteomics approaches have revealed that proteolysis involves chloroplastic FtsH protease in the early stages of senescence, aspartic protease during the course of leaf senescence, and the proteasome beta 1 subunit, mitochondria processing protease and SAG12 (cysteine protease) during the later senescence phases. Overall, the results constitute interesting pathways for screening genotypes with high NRE and NUE

Impacts de la disponibilité en sulfate sur la physiologie de la feuille et sur la qualité, le métabolisme soufré et la germination de la graine de colza (Brassica napus L.)

Le colza est une oléagineuse très exigeante en soufre (S). L étude des impacts de limitations en S sur la physiologie du colza et sa qualité grainière revêt un intérêt majeur dans un contexte de baisse des dépôts atmosphériques entrainant un appauvrissement des sols en S. Les objectifs étaient donc d étudier l incidence d une limitation en S sur la physiologie de jeunes feuilles, et sur la qualité, le métabolisme soufré et la vigueur germinative des graines. L analyse physiologique (photosynthèse, flux de S par utilisation de traceur 34S-sulfate), protéomique et biochimique (métabolites S, espèces réactives de l O2) a démontré qu une limitation en S provoque des perturbations du métabolisme carboné et soufré de la feuille et de la graine, pouvant affecter la qualité grainière. Ainsi, une restriction en S au stade rosette se traduit par la chute de l activité photosynthétique des jeunes feuilles et conduit à un stress oxydatif. Des restrictions en S à différents stades reproducteurs altèrent la qualité protéique et lipidique de la graine aboutissant à une accumulation amoindrie des acides oléique, linoléique, linolénique et des protéines de stockage (SSPs) riches en S. Une accumulation accrue de SSPs pauvres en S permet un maintien de la teneur en protéines de la graine en cas de restriction survenant en fin de cycle. L accumulation de S dans les protéines de la graine apparaît principalement contrôlée par la synthèse protéique. La vigueur germinative des graines produites est réduite en cas de restriction précoce en S. Ces travaux ont également permis de démontrer que le péricarpe et la graine en développement sont capables d assimiler le sulfate par la voie réductrice.Oilseed rape is a high sulphur (S) demanding crop. Study the impacts of S limitations on oilseed rape physiology and seed quality is a major issue in a context of declining S atmospheric depositions on soil. The objectives were therefore to study the impact of S limitation on the physiology of young leaves, and on quality, sulphur metabolism and germination vigour of seeds. The physiological (photosynthesis, S flows using 34S labelling), proteomics and biochemical (S metabolites, reactive oxygen species) analyses showed that S limitation leads to disturbances of carbon and sulphur metabolisms in leaf and seeds, which may affect seed quality. An S restriction applied at vegetative stage results in a lower photosynthetic activity of young leaves and leads to oxidative stress. S restrictions applied at different reproductive stages reduce seed protein and lipid quality resulting in a lower accumulation of oleic, linoleic, linolenic acids and of S-rich seed storage proteins (SSPs). A higher accumulation of S-poor SSPs allows maintenance of the seed protein content if S restriction occurs at the end of the plant growth cycle. S accumulation in seed proteins appears mainly controlled by protein synthesis. Early S restriction leads to reduce seed germination capacity and vigour. These studies have also demonstrated that developing seeds and pod walls are able to assimilate sulphate by the reductive pathway.CAEN-BU Sciences et STAPS (141182103) / SudocSudocFranceF