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International audience; The essential oils (EOs) of the aerial parts of Origanum libanoticum and Origanum ehrenbergii, endemic to Lebanon, and Origanum syriacum, endemic to the Levantine, were obtained by distillation with a Clevenger apparatus. GC and GC/MS allowed identification of 96.4%, 93.5%, and 95.2% of their constituents, respectively. Carvacrol was the major component of both O. syriacum EO (79%) and O. ehrenbergii EO (60.8%). This compound was absent in O. libanoticum EO and the major compounds were β-caryophyllene (26.8%), caryophyllene oxide (22.6%), and germacrene D (17.2%). The assessment of their antimicrobial activity against Candida albicans and six pathogenic bacteria revealed that O. libanoticum EO was inactive, while O. syriacum and O. ehrenbergii showed moderate antimicrobial activity with minimal inhibitory concentrations varying from 400 to 1200 μg/ml. These results support the traditional use of these last two species in traditional herbal preparations in Lebanon.

Mineral nutrient availability and in particular nitrogen abundance has a huge impact on plant fitness and yield, so that plants have developed sophisticated adaptive mechanisms to cope with environmental fluctuations. The vast natural variation existing among the individuals of a single species constitutes a great potential to decipher complex traits such as nutrient use efficiency. By using natural accessions of Arabidopsis thaliana that differ for their pattern of adaptation to nitrogen stress, we investigated the plant response to nitrate supplies ranging from 0.01 mM up to 50 mM nitrate. The biomass allocation and the different nitrogen pools in shoot and in roots were monitored to establish the nutrition status of each plant. Analysis of variation for these traits revealed genetic differences between accessions for their sensibility to nitrate availability and for their capacity to produce shoot biomass with the same nitrogen nutrition index. From the correlation matrix of all traits measured, a statistical model was formulated to predict the shoot projected area from the nitrate supply. The proposed model points out the importance of genetic variation with respect to the correlation between root thickness and amino acids content in roots. The model provides potential new targets in plant breeding for nitrogen use efficiency.

International audience; The essential oils obtained from Origanum libanoticum Boiss., a plant endemic to Lebanon, were analyzed by GC/MS. Seventy compounds were identified, covering till 99.8% of the total oil composition. All samples were p-cymene and/or beta-caryophyllene chemotype, with variable percentage of other compounds such as alpha-pinene, myrcene, a-phellandrene, limonene, etc. Compared to traditional drying method, lyophilized samples provided the highest essential oil (EO) yields and yields were higher at flowering stage (Chouwen: 0.33% in 2013 and 0.32% in 2014; Qartaba: 0.27% in 2013 and 0.37% in 2014). According to independent components analysis (ICA), date and site of harvest, altitude and drying technique had no effect on the variation of O. libanoticum EO, chemical composition. An annual variation of EOs composition was observed since a particular variation in some major components concentration was revealed monthly and annually between 2013 and 2014.

The essential oils were extracted by hydrodistillation from Satureja cuneifolia, Satureja thymbra, Coridothymus capitatus, Thymus syriacus and Thymbra spicata growing wild in Lebanon. Their phytochemical analysis performed by GC/MS showed that the aforementioned species are characterized either by carvacrol (60.9%) or thymol (54.3%) or by a more or less equal amounts of these two phenols. Assessment of their in vitro antimicrobial activity against Candida albicans and six pathogenic bacteria using the broth dilution method revealed that the tested oils have a broad activity spectrum with minimum inhibitory concentration ranging from 0.1 to 0.8 mg/ml. Among the tested species, S. thymbra EO showed the highest antimicrobial potential whereas T. syriacus showed the lowest inhibitory activity. These results give scientific evidence for the use of those species in the Lebanese folk medicine and lend support to implement them as natural alternatives for synthetic antimicrobials. This article is protected by copyright. All rights reserved.

International audience; AbstractThis study deals with the variation in the yield and composition of Lebanese Origanum syriacum L. essential oil (EO) according to harvesting time, drying methods used, and geographical location. Plant material was harvested twice a month all over 2013 and 2014 from Qartaba and Achkout located at high altitude and from Byblos at low altitude. EOs of the aerial parts were obtained by hydrodistillation. The highest yields were obtained at full flowering stage and slightly reduced after flowering. The GC/MS analysis revealed the presence of 50 components representing 90.49 – 99.82%, 88.79 – 100%, and 95.28 – 100% of the total oil extracted from plants harvested from Qartaba, Achkout, and Byblos, respectively. The major components in the oils were: carvacrol (2.1 – 79.8%), thymol (0.3 – 83.7%), p‐cymene (2.8 – 43.8%), thymoquinone (0.4 – 27.7%), γ‐terpinene (0.4 – 10.0%), octan‐3‐ol (0.3 – 4.9%), caryophyllene oxide (0.2 – 4.7%), oct‐1‐en‐3‐ol (0.3 – 3.7%), β‐caryophyllene (0.7 – 3.2%), cis‐sabinene hydrate (0.1 – 2.8%), terpinen‐4‐ol (0.1 – 2.8%), and α‐terpinene (0.2 – 2.2%). Independent components analysis (ICA) revealed that two groups were discriminated, reflecting compositional differences in the EOs profiles of the Lebanese oregano samples: O. syriacum grown in Qartaba and Achkout belongs to carvacrol chemotype, while O. syriacum grown in Byblos belongs to thymol chemotype. The flowering phase was the most productive period in terms of yield, bringing marked changes in the EO composition by increasing the amounts of carvacrol or thymol, and decreasing those of thymoquinone and p‐cymene.

We followed C and N reserves of grapevines grown in trenches under semi-controlled conditions over a 3-year period after planting. Temporal mobilization of stored C and N and subsequent distribution of reserve materials within the vines were described in parallel with 15N uptake, particularly during the third growing season. Storage C in the perennial tissues (roots, trunk, canes) was mainly made of starch, which accumulated in the ray parenchyma of the wood. In the permanent tissues, starch and total nitrogen contents were found to decrease early in the development (bleeding sap, budbreak) whereas, on a concentration basis, they decreased only after stage 7 (first leaf fully expanded). Starch started to accumulate again in the perennial tissues during flowering. The same observation was made with total nitrogen, although N levels were much lower than those of starch. The 15N study showed that N uptake by the roots started at budbreak and increased with vine development, becoming predominant over reserve mobilization only after the onset of flowering. Taken together, these results indicate that the spring growth period can be divided into three main phases: In the first (dormancy to budbreak), significant losses of C and N proceed mainly via root necrosis. In the second period (first leaf to the onset of bloom), a strong mobilization of starch (and, to a lower extent, of N) occurred for supporting vegetative and reproductive growth. At that point, most of the C and N reserves used on the spring flush were those of the roots, rather than those of the old wood (trunk, canes). In the third period (bloom and early berry development), the mobilization process became low and was relieved by N uptake (and CO2 assimilation) supplying nutrients to the sink structures.

A glutamine synthetase gene (GS15) coding for soybean cytosolic glutamine synthetase (GS1) fused to a constitutive promoter (CaMV 35S), a putative nodule-specific promoter (LBC3) and a putative root-specific promoter (rolD) was transformed into Pisum sativum L. cv. Greenfeast. Four lines with single copies of GS15 (one 35S-GS15 line, one LBC 3 -GS15 line, and two rolD-GS15 lines) were tested for the expression of GS15, levels of GS1, GS activity, N accumulation, N2 fixation, and plant growth at different levels of nitrate. Enhanced levels of GS1 were detected in leaves of three transformed lines (the 35S-GS15 and rolD-GS15 transformants), in nodules of three lines (the LBC 3 -GS15 and rolD-GS15 transformants), and in roots of all four transformants. Despite increased levels of GS1 in leaves and nodules, there were no differences in GS activity in these tissues or in whole-plant N content, N2 fixation, or biomass accumulation among all the transgenic lines and the wild-type control. However, the rolD-GS15 transformants, which displayed the highest levels of GS1 in the roots of all the transformants, had significantly higher GS activity in roots than the wild type. In one of the rolD-GS15 transformed lines (Line 8), increased root GS activity resulted in a lower N content and biomass accumulation, supporting the findings of earlier studies with Lotus japonicus (Limami et al. 1999 ). However, N content and biomass accumulation was not negatively affected in the other rolD-GS15 transformant (Line 9) and, in fact, these parameters were positively affected in the 0.1 mM $$ {\rm NO}_3^ - $$ treatment. These findings indicate that overexpression of GS15 in various tissues of pea does not consistently result in increases in GS activity. The current study also indicates that the increase in root GS activity is not always consistent with decreases in plant N and biomass accumulation and that further investigation of the relationship between root GS activity and growth responses is warranted.

Natural genetic variation in Arabidopsis is considerable, but has not yet been used extensively as a source of variants to identify new genes of interest. From the cross between two genetically distant ecotypes, Bay-0 and Shahdara, we generated a Recombinant Inbred Line (RIL) population dedicated to Quantitative Trait Locus (QTL) mapping. A set of 38 physically anchored microsatellite markers was created to construct a robust genetic map from the 420 F6 lines. These markers, evenly distributed throughout the five chromosomes, revealed a remarkable equilibrium in the segregation of parental alleles in the genome. As a model character, we have analysed the genetic basis of variation in flowering time in two different environments. The simultaneous mapping of both large- and small-effect QTLs responsible for this variation explained 90% of the total genotypic variance. Two of the detected QTLs colocalize very precisely with FRIGIDA and FLOWERING LOCUS C genes; we provide information on the polymorphism of genes confirming this hypothesis. Another QTL maps in a region where no QTL had been found previously for this trait. This confirms the accuracy of QTL detection using the Bay-0 x Shahdara RIL population, which constitutes the largest in size available so far in Arabidopsis. As an alternative to mutant analysis, this population represents a powerful tool which is currently being used to undertake the genetic dissection of complex metabolic pathways.

A large number of nitrate transporters ensure uptake and distribution of this main nutrient. Nitrate also acts as a signal, and elements of the signalling pathway have been identified.Plants have developed adaptive responses allowing them to cope with nitrogen (N) fluctuation in the soil and maintain growth despite changes in external N availability. Nitrate is the most important N form in temperate soils. Nitrate uptake by roots and its transport at the whole-plant level involves a large panoply of transporters and impacts plant performance. Four families of nitrate-transporting proteins have been identified so far: nitrate transporter 1/peptide transporter family (NPF), nitrate transporter 2 family (NRT2), the chloride channel family (CLC), and slow anion channel-associated homologues (SLAC/SLAH). Nitrate transporters are also involved in the sensing of nitrate. It is now well established that plants are able to sense external nitrate availability, and hence that nitrate also acts as a signal molecule that regulates many aspects of plant intake, metabolism, and gene expression. This review will focus on a global picture of the nitrate transporters so far identified and the recent advances in the molecular knowledge of the so-called primary nitrate response, the rapid regulation of gene expression in response to nitrate. The recent discovery of the NIN-like proteins as master regulators for nitrate signalling has led to a new understanding of the regulation cascade.

As a part of a global project aimed at comparing the physiology of several grapevine (Vitis vinifera L.) cultivars, we tested plant cultivation in trenches under semi-controlled conditions. Vegetative growth of vines of two cultivars, Pinot Noir and Merlot, from different locations, were followed during a 3-year period. Roots of trench-grown vines grew widely, leading to a root architecture closer to that observed under field conditions than that of potted vines. In addition, the root system could be sampled exhaustively for analyses. Although morphologically different, vines of the two cultivars exhibited similar patterns in dry matter production during the whole period, except at early flowering in year 3. Surprisingly, perennial parts accounted for this difference, though both cultivars were grafted onto the same SO4 rootstock clone. Consistently, vines exhibited root necrosis at any sampling date throughout spring, this process affecting more roots for the Pinot Noir/SO4 combination than for Merlot/SO4.

A soybean cytosolic glutamine synthetase gene (GS15) was fused with the constitutive 35S cauliflower mosaic virus (CaMV) promoter in order to direct overexpression in Lotus corniculatus L. plants. Following transformation with Agrobacterium rhizogenes, eight independent Lotus transformants were obtained which synthesized additional cytosolic glutamine synthetase (GS) in the shoots. To eliminate any interference caused by the T-DNA from the Ri plasmid, three primary transformants were crossed with untransformed plants and progeny devoid of TL- and TR-DNA sequences were chosen for further analyses. These plants had a 50-80% increase in total leaf GS activity. Plants were grown under different nitrogen regimes (4 or 12 mM NH4+) and aspects of carbon and nitrogen metabolism were examined. In roots, an increase in free amino acids and ammonium was accompanied by a decrease in soluble carbohydrates in the transgenic plants cultivated with 12 mM NH4+ in comparison to the wild type grown under the same conditions. Labelling experiments using 15NH4+ were carried out in order to monitor the influx of ammonium and its subsequent incorporation into amino acids. This experiment showed that both ammonium uptake in the roots and the subsequent translocation of amino acids to the shoots was lower in plants overexpressing GS. It was concluded that the build up of ammonium and the increase in amino acid concentration in the roots was the result of shoot protein degradation. Moreover, following three weeks of hydroponic culture early floral development was observed in the transformed plants. As all these properties are characteristic of senescent plants, these findings suggest that expression of cytosolic GS in the shoots may accelerate plant development, leading to early senescence and premature flowering when plants are grown on an ammonium-rich medium.

In this paper we discuss the ways in which our understanding of the nature of the molecular controls of nitrogen assimilation has been increased by the use of non-leguminous and leguminous plants with genetically-altered capacities for ammonia assimilation. Using tobacco or Lotus as model plants, Glutamine synthetase (GS) and glutamate synthase (GOGAT) activities have been altered by stimulating or inhibiting in an organ- or tissue-specific manner the expression of the corresponding genes. In a few selected examples, the physiological impact of these genetic manipulations has been studied on plants grown under different nitrogen regimes. The use of such genetically-modified plants will allow us to better understand the molecular control of this metabolic pathway. It is also potentially of great importance in agriculture if such internal and stable modifications are beneficial in terms of nitrogen use efficiency, thus avoiding an excessive utilization of fertilizers or herbicides (GS inhibitors). Our current knowledge and prospects for future development are explored. Manipulation der NH4-Assimilation in transgenen Nicht-Leguminosen und Leguminosen Das Verstandnis der molekularen Steuerungsmechanismen der Stickstoffassimilation ist durch die Verwendung von Pflanzen mit genetisch veranderter Kapazitat der NH4+-Assimilation betrachtlich gewachsen. Bei den Modellpflanzen Tabak (fur Nicht-Leguminosen) und Lotus corniculatus (fur Leguminosen) lassen sich die Aktivitaten der Glutaminsynthetase (GS) und der Glutamatsynthase (GOGAT) auf Organ-und Gewebsebene durch Stimulierung oder Hemmung der Expression der entsprechenden Gene verandern. Die physiologischen Konsequenzen dieser Genmanipulation wurden an Pflanzen unterschiedlicher N-Versorgung untersucht. Sollten sich solche internen und stabilen Modifikationen der Pflanzen als gunstig im Hinblick auf eine Verbesserung der N-Dungereffizienz erweisen, konnte zukunftig der Aufwand fur N-Dunger und (uber die GS-Inhibierung wirkende) Herbizide reduziert werden. Die Chancen hierfur bedurfen jedoch noch weiterer Forschung.

Our understanding of plant growth in response to nitrogen (N) supply is mainly based on studies of mutants and transformants. This study explored the natural variability of Arabidopsis thaliana first to find out its global response to N availability and secondly to characterize the plasticity for growth and N metabolism among 23 genetically distant accessions under normal (N+), limited (N-), and starved (N0) N supplies. Plant growth was estimated by eight morphological traits characterizing shoot and root growth and 10 metabolic parameters that represented N and carbon metabolism. Most of the studied traits showed a large variation linked to genotype and nutrition. Furthermore, Arabidopsis growth was coordinated by master traits such as the shoot to root ratio of nitrate content in N+, root fresh matter and root amino acids in N-, and shoot fresh matter together with root thickness in N0. The 23 accessions could be gathered into four different groups, according to their growth in N+, N-, and N0. Phenotypic profiling characterized four different adaptative responses to N- and N0. Class 1 tolerated N limitation with the smallest decrease in shoot and root biomass compared with N+, while class 2 presented the highest resistance to N starvation by preferential increased root growth, huge starch accumulation, and high shoot nitrate content. In contrast, class 3 plants could tolerate neither N limitation nor N starvation. Small plants of class 4 were different, with shoot biomass barely affected in N- and root biomass unaffected in N0.

To address the questions of whether allocation of carbohydrates to roots is influenced by ionic form of nitrogen absorbed and whether allocation of carbohydrates to roots in turn influences proportionality between NH4+ and NO3- uptake from mixed sources, NH4+ and NO3- were supplied separately to halves of a split-root hydroponic system and were supplied in combination to a whole-root system. Dry matter accumulation in the split-root system was 18% less in the NH4(+)-fed axis than in the NO3(-)-fed axis. This, however, does not indicate that partitioning of carbohydrate between the two axes was different. Most of the reduction in dry matter accumulation in the NH4(+)-fed axis can be accounted for by the retransport of CH2O equivalents from the root back to the shoot with amino acids produced by NH4+ assimilation. Uptake of NH4+ or NO3- by the respective halves of the split-root system was proportional to the estimated allocation of carbohydrate to that half. When NH4+ and NO3- were supplied to separate halves of the split-root system, the cumulative NH4+ to NO3- uptake ratio was 0.81. When supplied in combination to the whole-root system, the cumulative NH4+ to NO3- uptake ratio was 1.67. Thus, while the shoot may affect total nitrogen uptake through the export of carbohydrates to roots, the shoot (common for halves of the split-root system) apparently does not exert a direct effect on proportionality of NH4+ and NO3- uptake by roots. For whole roots supplied with both NH4+ and NO3-, the restriction in uptake of NO3- may involve a stimulation of NO3- efflux rather than an inhibition of NO3- influx. While only the net uptake of NH4+ and NO3- was measured by ion chromatography, monitoring at approximately hourly intervals during the first 3 days of treatment revealed irregularly occurring intervals of both depletion (net influx) and enrichment (net efflux) in solutions. In the case of NH4+, numbers of net efflux events were similar (21 to 24 out of 65 sequential sampling intervals) whether NH4+ was supplied with NO3- to whole-root systems or separately to an axis of the split-root system. In the case of NO3-, however, the number of net efflux events increased from 8 when NO3- was supplied to a separate axis of the split-root system to between 19 and 24 when NO3- was supplied with NH4+ to whole-root systems.

Soybean [Glycine max (L.) Merrill] plants that had been subjected to 15 d of nitrogen deprivation were resupplied for 10 d with 1.0 mol m-3 nitrogen provided as NO3-, NH4+, or NH4(+) + NO3- in flowing hydroponic culture. Plants in a fourth hydroponic system received 1.0 mol m-3 NO3- during both stress and resupply periods. Concentrations of soluble carbohydrates and organic acids in roots increased 210 and 370%, respectively, during stress. For the first day of resupply, however, specific uptake rates of nitrogen, determined by ion chromatography as depletion from solution, were lower for stressed than for non-stressed plants by 43% for NO3- resupply, by 32% for NH4(+) + NO3- resupply, and 86% for NH4+ resupply. When specific uptake of nitrogen for stressed plants recovered to rates for non-stressed plants at 6 to 8 d after nitrogen resupply, carbohydrates and organic acids in their roots had declined to concentrations lower than those of non-stressed plants. Recovery of nitrogen uptake capacity of roots thus does not appear to be regulated simply by the content of soluble carbon compounds within roots. Solution concentrations of NH4+ and NO3- were monitored at 62.5 min intervals during the first 3 d of resupply. Intermittent 'hourly' intervals of net influx and net efflux occurred. Rates of uptake during influx intervals were greater for the NH4(+)-resupplied than for the NO3(-)-resupplied plants. For NH4(+)-resupplied plants, however, the hourly intervals of efflux were more numerous than for NO3(-)-resupplied plants. It thus is possible that, instead of repressing NH4+ influx, increased accumulation of amino acids and NH4+ in NH4(+)-resupplied plants inhibited net uptake by stimulation of efflux on NH4+ absorbed in excess of availability of carbon skeletons for assimilation. Entry of NH4+ into root cytoplasm appeared to be less restricted than translocation of amino acids from the cytoplasm into the xylem.

In non-nodulated soybean [Glycine max (L.) Merrill cv. Ransom] plants that were subjected to 15 d of nitrogen deprivation in flowing hydroponic culture, concentrations of nitrogen declined to 1.0 and 1.4 mmol Ng −1 dry weight in shoots and roots, respectively, and the concentration of soluble amino acids (determined as primary amines) declined to 40 μmol g −1 dry weight in both shoots and roots. In one experiment, nitrogen was resupplied for 10 d to one set of nitrogen-depleted plants as 1.0 mol m −3 NH 4 + to the whole root system, to a second set as 0.5 mol m −3 NH 4 + plus 0.5 mol m −3 NO 3 − to the whole root system, and to a third set as 1.0 mol m −3 NH 4 + to one-half of a split-root system and 1.0 mol m −3 NO 3 − to the other half

International audience; In a low-input agricultural context, plants facing temporal nutrient deficiencies need to be efficient. By comparing the effects of NOFormula-starvation in two lines of Arabidopsis thaliana (RIL282 and 432 from the Bay-0xShahdara population), this study aimed to screen the physiological mechanisms allowing one genotype to withstand NOFormula-deprivation better than another and to rate the relative importance of processes such as nitrate uptake, storage, and recycling. These two lines, chosen because of their contrasted shoot N contents for identical shoot biomass under N-replete conditions, underwent a 10 d nitrate starvation after 28 d of culture at 5 mM NOFormula. It was demonstrated that line 432 coped better with NOFormula-starvation, producing higher shoot and root biomass and sustaining maximal growth for a longer time. However, both lines exhibited similar features under NOFormula-starvation conditions. In particular, the nitrate pool underwent the same drastic and early depletion, whereas the protein pool was increased to a similar extent. Nitrate remobilization rate was identical too. It was proportional to nitrate content in both shoots and roots, but it was higher in roots. One difference emerged: line 432 had a higher nitrate content at the beginning of the starvation phase. This suggests that to overcome NOFormula-starvation, line 432 did not directly rely on the N pool composition, nor on nitrate remobilization efficiency, but on higher nitrate storage capacities prior to NOFormula-starvation. Moreover, the higher resistance of 432 corresponded to a higher nitrate uptake capacity and a 2–9-fold higher expression of AtNRT1.1, AtNRT2.1, and AtNRT2.4 genes, suggesting that the corresponding nitrate transporters may be preferentially involved under fluctuating N supply conditions.

The accumulation of lead and cadmium in maize plants, grown on liquid nutrient solutions, was studied with a two level factorial design. The accumulation of cadmium in roots was correlated with the concentration of cadmium in the solution (sol). In the shoots however, it followed a model with a negative cadmium‐lead interaction

A soybean cytosolic glutamine synthetase gene ( GS15 ) fused to a constitutive promoter (CaMV 35S), a putative nodule-specific promoter ( LBC 3 ), or a putative root-specific promoter ( rolD ) was transformed into Pisum sativum L. cv. Greenfeast. Four lines with single copies (Lines 1, 7, 8 and 9) and four lines with two copies each of GS15 (Lines 2, 4, 6 and 11) were compared to the wild-type (WT) parental line for levels of cytosolic glutamine synthetase (GS1), glutamine synthetase (GS) activity, N accumulation, N derived form the atmosphere (NDFA), and biomass of plants grown on 0.0, 0.1, 1.0 or 10.0 mM NH 4 + . Enhanced levels of GS1 were detected in leaves of one of the two lines transformed with the 35S-GS15 construct, and all three lines containing the rolD-GS15 construct. All three lines containing the LBC 3 -GS15 construct had increased levels of GS1 in nodules. Despite the increased levels of GS1 in many transformants, only the roots of lines containing the rolD-GS15 construct consistently demonstrated enhanced levels of GS activity (up to 12-fold). Positive responses in plant N content, NDFA, and biomass were rare, but increases in plant biomass and N content of up to 17% and 54%, respectively, occurred in some of the rolD-GS15 lines at certain levels of ammonium. In general, GS15 copy number did not seem to differentially affect phenotype of the transformants, and transformants respond to ammonium concentrations in similar patterns to that previously observed with nitrate. Despite the fact that the rolD-GS15 transformants consistently resulted in increased GS activity in roots and resulted in some occurrences of increases in biomass and plant N content, the lack of consistent positive growth effect across all transformants indicates that the generalized overexpression of GS1 in tissues holds little potential for positive growth responses in pea.

In plants, water and anion parameters are linked, for example through the integration of nutritional signaling and the response to diverse stress. In this work, Arabidopsis thaliana is used as a model system to dissect the genetic variation of these parameters by quantitative trait loci (QTL) mapping in the 415 recombinant inbred lines of the Bay-0 × Shahdara population. Water, nitrate, chloride, and phosphate contents were measured at the vegetative stage in the shoots of plants grown in controlled conditions. Two contrasting nitrogen (N) conditions were studied, one leading to the complete depletion of the nitrate pool in the plants. Most of the observed genetic variation was identified as QTL, with medium but also large phenotypic contributions. QTL colocalization provides a genetic basis for the correlation between water and nitrate contents in nonlimiting N conditions and water and chloride contents in limiting N conditions. The 34 new QTL described here represent at least 19 loci polymorphic between Bay-0 and Shahdara; some may correspond to known genes from water/anion transport systems, while others clearly identify new genes controlling or interacting with water/anion absorption and accumulation. Interestingly, flowering-time genes probably play a role in the regulation of water content in our conditions.

Improving plant nitrogen (N) use efficiency or controlling soil N requires a better knowledge of the regulation of plant N metabolism. This could be achieved using Arabidopsis as a model genetic system, taking advantage of the natural variation available among ecotypes. Here, we describe an extensive study of N metabolism variation in the Bay-0 × Shahdara recombinant inbred line population, using quantitative trait locus (QTL) mapping. We mapped QTL for traits such as shoot growth, total N, nitrate, and free-amino acid contents, measured in two contrasting N environments (contrasting nitrate availability in the soil), in controlled conditions. Genetic variation and transgression were observed for all traits, and most of the genetic variation was identified through QTL and QTL × QTL epistatic interactions. The 48 significant QTL represent at least 18 loci that are polymorphic between parents; some may correspond to known genes from the N metabolic pathway, but others represent new genes controlling or interacting with N physiology. The correlations between traits are dissected through QTL colocalizations: The identification of the individual factors contributing to the regulation of different traits sheds new light on the relations among these characters. We also point out that the regulation of our traits is mostly specific to the N environment (N availability). Finally, we describe four interesting loci at which positional cloning is feasible.

In soils, ammonium (NH4 +) mainly results from the mineralisation of organic matter and represents besides nitrate (NO3 −) the quantitatively most important source of nitrogen (N) for plant nutrition. In well-aerated agricultural soils, however, average annual NH4 + concentrations are often 10 to 1000 times lower than those of NO3 −, rarely exceeding 50.µM (Marschner 1995). Despite these low concentrations in soils, NH4 + uptake by plant roots can proceed at very high rates, due to the presence of transport systems in the root plasma membrane with a particularly high substrate affinity. Indeed, NH4 + uptake is of major importance for N nutrition under numerous circumstances. On the one hand, NH4 + nutrition plays an essential role in waterlogged and acid soils, or in cold climates where nitrification is inhibited (Marschner 1995). On the other hand, under mixed N nutrition (NO3 − plus NH4 +), NH4 + is often the preferential form of N taken up by the plant (Sasakawa and Yamamoto 1978; Gojon et al. 1986; Glass and Siddiqi 1995; Gazzarrini et al. 1999). NH4 + is also probably the main form of N exported from symbiotic N2-fixing microorganisms to their host plants, thereby making a major contribution to N nutrition in several plant families (Udvardi and Day 1997). In root nodules, NH4 + is transported across the symbiosome membrane which segregates the bacteroids from the plant cytosol (see Chap. 3). In this case, NH4 + concentrations in the plant cytosol can be about 50-fold lower than in the bacteroids (Streeter 1989), requiring low-affinity and high-capacity transport systems on the plant side, to ensure an efficient import of microbially fixed N (Tyerman et al. 1995).

When NO3 - is maintained at concentrations greater than those required to sustain maximum cumulative uptake by wheat (Triticum aestivum) and nonnodulated soybean (Glycine max [L.] Merrill), the net rate of uptake is not constant but oscillates between maxima and minima. The amplitudes of both the maxima and the minima increase with increases in NO3 - concentration of the solution. To determine if the amplitudes of oscillations decline when NO3 - in solution is maintained at suboptimal concentrations, net rates of NO3 - uptake were monitored daily during 22 d of vegetative growth under controlled environmental conditions for wheat from solutions containing 0.1, 0.25, 0.4, 0.5, 0.8, 1.5, 3.0, and 5.0 mM NO3 - and for soybean from solutions containing 0.05, 0.1, 0.5, and 1.0 mM NO3 -. The amplitudes of variations in daily net rates of NO3 - uptake increased with the increases in concentration of NO3 - in the nutrient solutions. The differences in magnitude of the amplitudes in oscillation, expressed as coeff...

Les agents de solidification influent sur la croissance des pousses du noyer cultivees in vitro. La gelrite a un effet benefique sur l'elongation des explants et la production de bourgeons, tandis que l'agar inhibe la croissance et provoque la maturation des feuilles ou encore des necroses. Les 2 agents de solidification presentent des differences importantes dans leurs teneurs en elements mineraux. Ils alterent la composition minerale du milieu de culture, comme celle des explants (.)

As a part of a project aimed at elucidating the causal relationship between reserve mobilisation and the extent of shedding in Vitis vinifera L., we compared storage and fate of carbon (C) and nitrogen (N) reserves in two varieties differing in their susceptibility to fruitlet abscission. Merlot (susceptible) and Pinot Noir (P. Noir, not susceptible) vines were grown in trenches under semi-controlled conditions over a 3-y period after planting. Mobilisation of stored C and N, distribution of reserve materials within the vines and 15N uptake were followed particularly during the spring growth flush and floral development in the third year. At dormancy, starch levels in the perennial tissues (roots, trunk, canes) were higher in Merlot than in P. Noir. During the spring growth flush, starch level decreased markedly in the roots of both cultivars until early bloom. At that time, starch started to accumulate in P. Noir but not in Merlot. Similar variations were found with total N. Accordingly, 15N analysis showed that translocation of storage N to the annual tissues was nearly achieved at early bloom in P. Noir while it continued until pea berry size in Merlot. In parallel, N uptake increased during the spring growth flush, and it was higher in P. Noir than in Merlot. These results indicate that transition between heterotrophic (root) and autotrophic (leaf) mode of nutrient allocation towards the developing inflorescences occurs earlier in P. Noir. Possible consequences are discussed in relation to the susceptibility of each cultivar to shedding.

International audience

Aujourd’hui, la recherche dans le domaine de l’agriculture se concentre sur l’amélioration des plantes dans le but de conserver ou d’augmenter leur productivité dans un environnement en perpétuels changements. L’efficacité d’utilisation de l’azote (NuUE) est de fait un sujet de recherche privilégié en agriculture. Le sujet de ce travail de thèse est ainsi de comprendre les mécanismes naturels de la NuUE chez Arabidopsis thaliana. La variabilité naturelle d’Arabidopsis a constitué le lien commun entre les différentes approches employées dans ce travail, avec le but particulier de mieux comprendre la complexité de la réponse des plantes à une large gamme de stress azotés, mais aussi de comprendre l’interaction des stress soufrés et azotés et enfin de déterminer la connexion existant entre la disponibilité en azote et la sénescence foliaire. Une carence en azote amène à une diminution de la biomasse de la plante et affecte certains traits morphologiques. Le même phénomène est observé avec l’utilisation de concentrations élevées en azote, avec un effet probablement toxique. Selon la concentration en azote présente dans l’environnement, les pools de métabolites des plantes sont modifiés, montrant ainsi que la capacité à utiliser avantageusement des concentrations élevées en azote est une caractéristique dépendante du génotype. Cela ouvre ainsi des perspectives d’amélioration pour la sélection de plantes avec une meilleure NuUE. Dans ce travail de thèse, pour la première fois, un modèle génétique et physiologique décrivant la réponse aux perturbations azotées a été présenté chez Arabidopsis. Il a été déterminé que les métabolites du système racinaire ont un impact majeur sur la biomasse des feuilles, davantage encore que les caractères métaboliques intrinsèques aux feuilles. Cela souligne l’importance de la sélection de plantes avec un système racinaire développé et efficace pour l’assimilation de nutriments. De plus, il a été montré qu’une limitation ou une carence en soufre provoque une modification dans le pool des métabolites soufrés et que la concentration en azote influence directement le métabolisme soufré. Enfin, grâce à la production de plusieurs lignées qui surexpriment des gènes d’un QTL (Quantitative Trait Locus) de sénescence liée à l’azote, il semblerait qu’un phénomène de développement de racines secondaires en réponse à la carence azotée soit à la base du phénotype de sénescence précoce étudié.

The field of the nanotechnology exponentially increased during the last ten years. Intensive production of nanomaterials would lead to their release in the environment. Plants are one of the most sensitive organisms since they are at the interface between soil, water and air. Moreover, they represent a point of entry into the food chain. This study deals with the impact of TiO2 nanoparticle and carbon nanotube on plants, specifically on wheat and rapeseed. The physico-chemistry of nanomaterials has been deeply characterized.We prove that TiO2 nanoparticles and carbon nanotubes are internalized in roots and transferred to shoot of both wheat and rapeseed upon hydroponics exposure. TiO2 nanoparticles can also be internalized upon exposure in soil and even after an aerial contamination. TiO2 nanoparticles are accumulated in plants up to few hundred mg Ti/kg DM, their crystal phase is not modified and they do not dissolve during their transfer inside plants. CNT are less accumulated, up to few hundred µg NTC/kg DM, and seem to be altered by their transfer to and though plants.The nanomaterials we studied have very limited impact on plant development and physiology. One test out of eight has shown modulation after nanomaterial exposure: root elongation was induced when exposed to very small of both nanomaterials. Finally, we show that the nominal diameter of nanomaterials is negatively correlated with their internalization, transfer to shoot and induction of root elongation: the finest the nanomaterial, the highest their accumulation and impact on plant development.; Dans le contexte du développement exponentiel des nanotechnologies, les nanomatériaux sont susceptibles de se disséminer dans l'environnement. Les végétaux sont un élément sensible des écosystèmes car ils constituent une interface entre l'eau, le sol et l'air et se situent à la base de la chaine trophique. Cette étude avait pour but d'évaluer l'impact des nanoparticules de TiO2 et des nanotubes de carbone sur les végétaux, et plus particulièrement sur le blé et le colza. L'accent a également était mis sur la caractérisation des nanomatériaux employés. Nous avons mis en évidence l'accumulation racinaire et le transfert vers les feuilles des nanoparticules de TiO2 et des NTC dans le blé et le colza après une exposition en hydroponie. Les nanoparticules de TiO2 sont également accumulées dans les plantes lors d'une exposition racinaire sur sol ou encore lors d'une exposition par voie aérienne. Les nanoparticules de TiO2 s'accumulent dans les végétaux à hauteur de quelques dizaines voire centaines de mg Ti/kg MS, leur phase cristalline n'est pas modifiée et ces nanoparticules ne se dissolvent pas lors de leur transfert dans le végétal. Les NTC s'accumulent en plus faible quantité (centaines de µg NTC/kg MS) et il semble qu'ils soient altérés par le contact avec le végétal. Ces nanomatériaux affectent peu le développement des végétaux. Sur huit tests réalisés, seule l'élongation racinaire s'est révélée être un critère sensible, avec une induction de l'élongation pour les NPs et les NTC présentant le plus faible diamètre nominal. Enfin, une nette influence du diamètre nominal des nanomatériaux a pu être mise en évidence, contrôlant ainsi leur internalisation, leur transfert et leurs effets biologiques sur les végétaux.