Développement, nutrition minérale, relations hydriques et métabolisme azoté chez deux plantes fourragères Sulla carnosa et Medicago truncatula en condition de déficit hydrique

Le déficit hydrique est à l’origine de problèmes économiques et écologiques majeurs, dont particulièrement la forte réduction de la production des plantes, à l’instar des espèces fourragères. L’effet de la sécheresse sur la production de biomasse est souvent associé à une perturbation du métabolisme de l’azote, qui est un élément essentiel pour la plante. En se basant sur des approches physiologiques, biochimiques et moléculaires nous avons caractérisé le comportement de Sulla carnosa et de Medicago truncatula vis-à-vis du déficit hydrique seul ou combiné à la salinité, en vue d’identifier parmi ces deux légumineuses la plus tolérante a ces contraintes prises séparément ou en combinaison et d’établir des traits de tolérance liés au métabolisme azoté. Les effets du déficit hydrique sur la régulation de l’expression de gènes codant pour des enzymes impliquées dans la régulation du métabolisme azoté (Glutamine synthétase, GS et la Glutamate déshydrogénase, GDH), la biosynthèse de composés compatibles azotés tel que la proline (Pyrroline-5-carboxylate synthétase, P5CS et Proline déshydrogénase, PDH) et l’expression d’un transporteur du nitrate (MtNRT1.3) ont été étudiés. Les résultats montrent un effet dépressif du déficit hydrique sur la production de biomasse chez les deux espèces, avec cependant un effet plus prononcé sur S. carnosa, espèce halophytique que sur M. truncatula. Le déficit hydrique affecte également les activités GS et GDH impliquées dans la voie d’assimilation de l’ammonium. Chez les deux espèces le métabolisme de glutamate est fortement sollicité pour l’élaboration des métabolites d’intérêt adaptatif tel que la proline. Après la levé du stress, un rétablissement partiel de la croissance est observé, ce comportement est considéré très important chez les plantes fourragères, puisque dans leurs biotopes naturels, ces plantes sont exposées à une alternance de périodes sèches et pluvieuses. L’ajout du sel en condition limitante en eau atténue les effets délétères de la contrainte hydrique sur la croissance seulement chez l’espèce halophytique S. carnosa

Functional expression of PHO1 to the Golgi and trans-Golgi network and its role in export of inorganic phosphate.

Arabidopsis thaliana PHO1 is primarily expressed in the root vascular cylinder and is involved in the transfer of inorganic phosphate (Pi) from roots to shoots. To analyze the role of PHO1 in transport of Pi, we have generated transgenic plants expressing PHO1 in ectopic A. thaliana tissues using an estradiol-inducible promoter. Leaves treated with estradiol showed strong PHO1 expression, leading to detectable accumulation of PHO1 protein. Estradiol-mediated induction of PHO1 in leaves from soil-grown plants, in leaves and roots of plants grown in liquid culture, or in leaf mesophyll protoplasts, was all accompanied by the specific release of Pi to the extracellular medium as early as 2-3 h after addition of estradiol. Net Pi export triggered by PHO1 induction was enhanced by high extracellular Pi and weakly inhibited by the proton-ionophore carbonyl cyanide m-chlorophenylhydrazone. Expression of a PHO1-GFP construct complementing the pho1 mutant revealed GFP expression in punctate structures in the pericycle cells but no fluorescence at the plasma membrane. When expressed in onion epidermal cells or in tobacco mesophyll cells, PHO1-GFP was associated with similar punctate structures that co-localized with the Golgi/trans-Golgi network and uncharacterized vesicles. However, PHO1-GFP could be partially relocated to the plasma membrane in leaves infiltrated with a high-phosphate solution. Together, these results show that PHO1 can trigger Pi export in ectopic plant cells, strongly indicating that PHO1 is itself a Pi exporter. Interestingly, PHO1-mediated Pi export was associated with its localization to the Golgi and trans-Golgi networks, revealing a role for these organelles in Pi transport

Differential performance of two forage species, Medicago truncatula and Sulla carnosa, under water-deficit stress and recovery

The response patterns during water deficit stress and subsequent recovery of two forage species, Medicago truncatula and Sulla carnosa, were studied. After germination and pre-treatment, seedlings were individually cultivated for two months under two irrigation modes: 100% and 33% of field capacity. Measured parameters were plant growth, water relations, leaf osmotic potential, lipid peroxidation, and leaf inorganic (Na+ and K+) and organic (proline and soluble sugars) solute contents, as well as delta-1-pyrroline-5-carboxylate synthase (P5CS) and proline dehydrogenase (PDH) activities. Our results showed that under control conditions, and in contrast to roots, no significant differences were observed in shoot biomass production between the two species. However, when subjected to water-deficit stress, M. truncatula appeared to be more tolerant than S. carnosa (reduction by 50 and 70%, respectively). In the two studied species, water-deficit stress led to an increase in root/shoot ratio and leaf proline and soluble sugar contents, and a decrease in leaf osmotic potential. Enzymatic assay revealed that in the two species, P5CS activity was stimulated whereas that of PDH was inhibited under stress conditions. Despite greater accumulation of proline, sugar, and potassium in leaves of S. carnosa, M. truncatula was more tolerant to water deficit. This was essentially due to its capacity to control tissue hydration and water-use efficiency, in addition to its greater ability to protect membrane integrity. Following stress relief, M. truncatula and S. carnosa showed partial re-establishment of growth capacity

Comparative responses to water deficit stress and subsequent recovery in the cultivated beet Beta vulgaris and its wild relative B. macrocarpa

The effects of water deficit stress and recovery on growth, photosynthesis, physiological and biochemical parameters were investigated in the cultivated Beta vulgaris and in two Tunisian provenances (Soliman and Enfidha) of its wild relative B. macrocarpa. Seedlings were cultivated for 4 weeks under optimal or limiting water supply (respectively, 100% and 25% of field capacity, FC). After 2 weeks of treatment, a lot of stressed plants were rehydrated to 100% FC. In the Control, B. vulgaris was more productive than B. macrocarpa, whereas Enfidha provenance showed the highest biomass production (1.6- and 3-fold compared with B. vulgaris and Soliman, respectively), under water deficit stress. A partial re-establishment of growth occurred in both species upon recovery at 100% FC. The sensitivity of B. vulgaris and Soliman provenance to drought was associated with the disturbance of leaf water status and the sharp decrease in net CO2 assimilation (–66% and –82% as compared with the Control, respectively). On the contrary, the better behaviour of Enfidha provenance was related to its better photosynthetic capacity and leaf relative water content, along with a higher accumulation of amino acids (proline, glycine, and glutamine) implied in the osmotic adjustment. Leaf hexose concentration increased significantly under drought stress in both species whereas leaf sucrose concentration declined only in drought-stressed B. vulgaris and Soliman provenance. Leaf glutamate dehydrogenase activity increased under water deficit in both species despite to a higher extent in B. vulgaris. As glutamate dehydrogenase is implied in catabolism of glutamate to oxoglutarate, it might contribute to provide stressed plants with carbon skeletons. Enfidha provenance of the spontaneous species B. macrocarpa could be used in the marginal arid ecosystems in order to limit the deficit in fodder and to improve the pastoral value of these regions. In addition, this species could serve as a source of genes for genetic improvement to water deficit stress

Detecting the Effects of Changes on the Compliance of Cross-organizational Business Processes

An emerging challenge for collaborating business partners is to properly define and evolve their cross-organizational processes with respect to imposed global compliance rules. Since compliance verification is known to be very costly, reducing the number of compliance rules to be rechecked in the context of process changes will be crucial. Opposed to intra-organizational processes, however, change effects cannot be easily assessed in such distributed scenarios, where partners only provide restricted public views and assertions on their private processes. Even if local process changes are invisible to partners, they might affect the compliance of the cross-organizational process with the mentioned rules. This paper provides an approach for ensuring compliance when evolving a cross-organizational process. For this purpose, we construct qualified dependency graphs expressing relationships between process activities, process assertions, and compliance rules. Based on such graphs, we are able to determine the subset of compliance rules that might be affected by a particular change. Altogether, our approach increases the efficiency of compliance checking in cross-organizational settings

Remobilization of leaf S compounds and senescence in response to restricted sulphate supply during the vegetative stage of oilseed rape are affected by mineral N availability

The impact of sulphur limitation on the remobilization of endogenous S compounds during the rosette stage of oilseed rape, and the interactions with N availability on these processes, were examined using a long-term 34SO42− labelling method combined with a study of leaf senescence progression (using SAG12/Cab as a molecular indicator) and gene expression of the transporters, BnSultr4;1 and BnSultr4;2, involved in vacuolar sulphate efflux. After 51 d on hydroponic culture at 0.3 mM 34SO42− (1 atom% excess), the labelling was stopped and plants were subject for 28 d to High S-High N (HS-HN, control), Low S-High N (LS-HN) or Low S-Low N (LS-LN) conditions. Compared with the control, LS-HN plants showed delayed leaf senescence and, whilst the shoot growth and the foliar soluble protein amounts were not affected, S, 34S, and SO42− amounts in the old leaves declined rapidly and were associated with the up-regulation of BnSultr4;1. In LS-LN plants, shoot growth was reduced, leaf senescence was accelerated, and the rapid S mobilization in old leaves was accompanied by decreased 34S and SO42−, higher protein mobilization, and up-regulation of BnSultr4;2, but without any change of expression of BnSultr4;1. The data suggest that to sustain the S demand for growth under S restriction (i) vacuolar SO42− is specifically remobilized in LS-HN conditions without any acceleration of leaf senescence, (ii) SO42− mobilization is related to an up-regulation of BnSultr4;1 and/or BnSultr4;2 expression, and (iii) the relationship between sulphate mobilization and up-regulation of expression of BnSultr4 genes is specifically dependent on the N availability

Regulatory feedback response mechanisms to phosphate starvation in rice

Phosphorus is a growth-limiting nutrient for plants. The growing scarcity of phosphate stocks threatens global food security. Phosphate-uptake regulation is so complex and incompletely known that attempts to improve phosphorus use efficiency have had extremely limited success. This study improves our understanding of the molecular mechanisms underlying phosphate uptake by investigating the transcriptional dynamics of two regulators: the Ubiquitin ligase PHO2 and the long non-coding RNA IPS1. Temporal measurements of RNA levels have been integrated into mechanistic mathematical models using advanced statistical techniques. Models based solely on current knowledge could not adequately explain the temporal expression profiles. Further modeling and bioinformatics analysis have led to the prediction of three regulatory features: the PHO2 protein mediates the degradation of its own transcriptional activator to maintain constant PHO2 mRNA levels; the binding affinity of the transcriptional activator of PHO2 is impaired by a phosphate-sensitive transcriptional repressor/inhibitor; and the extremely high levels of IPS1 and its rapid disappearance upon Pi re-supply are best explained by Pi-sensitive RNA protection. This work offers both new opportunities for plant phosphate research that will be essential for informing the development of phosphate efficient crop varieties, and a foundation for the development of models integrating phosphate with other stress responses

Nuclear localised more sulphur accumulation1 epigenetically regulates sulphur homeostasis in Arabidopsis thaliana

Sulphur (S) is an essential element for all living organisms. The uptake, assimilation and metabolism of S in plants are well studied. However, the regulation of S homeostasis remains largely unknown. Here, we report on the identification and characterisation of the more sulphur accumulation1 (msa1-1) mutant. The MSA1 protein is localized to the nucleus and is required for both S adenosylmethionine (SAM) production and DNA methylation. Loss of function of the nuclear localised MSA1 leads to a reduction in SAM in roots and a strong S-deficiency response even at ample S supply, causing an over- accumulation of sulphate, sulphite, cysteine and glutathione. Supplementation with SAM suppresses this high S phenotype. Furthermore, mutation of MSA1 affects genome-wide DNA methylation, including the methylation of S-deficiency responsive genes. Elevated S accumulation in msa1-1 requires the increased expression of the sulphate transporter genes SULTR1;1 and SULTR1;2 which are also differentially methylated in msa1-1. Our results suggest a novel function for MSA1 in the nucleus in regulating SAM biosynthesis and maintaining S homeostasis epigenetically via DNA methylation