Céline Richard-Molard

Plant response to nitrate starvation is determined by N storage capacity matched by nitrate uptake capacity in two Arabidopsis genotypes

In a low-input agricultural context, plants facing temporal nutrient deficiencies need to be efficient. By comparing the effects of NO(3)(-)-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 NO(3)(-)-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 NO(3)(-). It was demonstrated that line 432 coped better with NO(3)(-)-starvation, producing higher shoot and root biomass and sustaining maximal growth for a longer time. However, both lines exhibited similar features under NO(3)(-)-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 NO(3)(-)-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 NO(3)(-)-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.

Ara-rhizotron : An effective culture system to study simultaneously root and shoot development of Arabidopsis

Studying Arabidopsis thaliana (L.) Heynh. root development in situ at the whole plant level without affecting shoot development has always been a challenge. Such studies are usually carried out on individual plants, neglecting competition of a plant population, using hydroponic systems or Agar-filled Petri dishes. Those both systems, however, present some limitations, such as difficulty to study precisely root morphogenesis or time-limited culture period, respectively. In this paper, we present a method of Arabidopsis thaliana (L.) Heynh. cultivation in soil medium, named “Ara-rhizotron”. It allows the non-destructive study of shoot and root development simultaneously during the entire period of vegetative growth. In this system, roots are grown in 2D conditions, comparable to other soil cultures. Moreover, grouping several Ara-rhizotrons in a box enables the establishment of 3D shoot competition as for plants grown in a population. In comparison to a control culture grown in pots in the same environmental conditions, the Ara-rhizotron resulted in comparable shoot development in terms of dry mass, leaf area, number of leaves and nitrogen content. We used this new culture system to study the effect of irrigation modalities on plant development. We found that irrigation frequency only affected root partitioning in the soil and shoot nitrogen content, but not shoot or root growth. These effects appeared at the end of the vegetative growth period. This experiment highlights the opportunity offered by the Ara-rhizotron to point out tardy effects, affecting simultaneously shoot development and root architecture of plants grown in a population. We discuss its advantages in relation to root development and physiology, as well as its possible applications.

Nitrate (15NO3) limitation affects nitrogen partitioning between metabolic and storage sinks and nitrogen reserve accumulation in chicory (Cichorium intybus L.)

Abstract. In chicory, we examined how NO3− supply affected NO3− uptake, N partitioning between shoot and root and N accumulation in the tuberized root throughout the vegetative period. Plants were grown at two NO3− concentrations: 0.6 and 3 mM. We used 15N-labelling/chase experiments for the quantification of N fluxes between shoot and root and for determining whether N stored in the tuberized root originates from N remobilized from the shoot or from recently absorbed NO3−. The rate of 15NO3− uptake was decreased by low NO3− availability at all stages of growth. In young plants (10–55 days after sowing; DAS), in both NO3− treatments the leaves were the strongest sink for 15N. In mature (tuberizing) plants, (55–115 DAS), the rate of 15NO3− uptake increased as well as the amount of exogenous N allocated to the root. In N-limited plants, N allocation to the tuberized root relied essentially on recent N absorption, while in N-replete plants, N remobilized from the shoot contributed more to N-reserve accumulation in the root. In senescing plants (115–170 DAS) the rate of 15NO3− uptake decreased mainly in N-replete plants whereas it remained almost unchanged in N-limited plants. In both NO3− treatments the tuberized root was the strongest sink for recently absorbed N. Remobilization of previously absorbed N from shoot to tuberized root increased greatly in N-limited plants, whereas it increased slightly in N-replete plants. As a consequence, accumulation of the N-storage compounds vegetative storage protein (VSP) and arginine was delayed until later in the vegetative period in N-limited plants. Our results show that although the dynamics of N storage was affected by NO3− supply, the final content of total N, VSP and arginine in roots was almost the same in N-limited and N-replete plants. This indicates that chicory is able to build up a store of available N-reserves, even when plants are grown on low N. We also suggest that in tuberized roots there is a maximal capacity for N accumulation, which was reached earlier (soon after 100 DAS) in N-replete plants. This hypothesis is supported by the fact that in N-replete plants despite NO3− availability, N accumulation ceased and significant amounts of N were lost due to N efflux.

Effect of exogenous nitrogen (15NO3) on utilization of vegetative storage proteins (VSP) during regrowth in chicory (Cichorium intybus)

Summary In the present work we evaluate the accumulation and further remobilization of vegetative storage proteins (VSP) in chicory. A protein with molecular weight of 17kDa, corresponding to 7 isoforms with pi ranging between 5 and 7, accumulated dramatically over the vegetative phase from spring to autumn and was extensively depleted during the flowering period in the following summer, a pattern typical for a VSP. When mature tuberized roots of chicory are harvested in autumn and forced in darkness, an etiolated bud (chicon) grows: this is the salad known as Belgian endive. In our experiments plants were fed, during the forcing process, nutrient solutions containing 1.5 or 18 mmol/L 15 NO 3 (1.79 % atom excess 15 N) or with demineralized water (control). We determined the cycling of endogenous nitrogen ( 14 N), protein (VSP) and amino acids, and the movement of concurrently absorbed nitrogen ( 15 N). Soluble proteins were remobilized at the onset of forcing as a primary response of nitrogen cycling in chicory root. Amino acid remobilization took place only when the chicon growth began with arginine remobilized first. Although 14 N flux into the chicon was similar in all three treatments, indicating that NO 3 supply did not effect endogenous N remobilization, VSP use was effected by NO 3 supply. SDS-PAGE and 2-D gel electrophoresis analyses showed an extensive depletion of VSP (especially five isoforms) only in the control. We suggested that extensive and specific depletion of VSP was delayed by NO 3 supply; with higher NO 3 availability, there was lower VSP remobilization. Furthermore, neo-synthesis of VSP could occur during the forcing process. The finding that 15 N was incorporated into the protein pool during this period supports this hypothesis. The chicon constituted a very strong sink for absorbed nitrogen. Either in high or low NO 3 supply, 15 N was translocated to the chicon almost without mixing with the bulk nitrogen of the root.

Nitrogen-induced changes in morphological development and bacterial susceptibility of Belgian endive (Cichorium intybus L.) are genotype-dependent

Abstract. Nitrogen is known to modulate plant development and resistance to pathogens. Four selected lines (Alg, NS1, NR1 and NR2) of chicory (Cichorium intybus L.) were grown on low (0.6 mM) and high (3 mM) NO−3 nutrition in order to study the effect of N on the expression of three traits, namely, shoot/root ratio, chicon morphology and resistance to soft rot caused by Erwinia sp. For all genotypes, increasing N supply led to a higher shoot/root ratio, resulting from an increased shoot biomass but with no effect on root growth. In contrast, the effect of N on chicon morphology and resistance to bacteria was genotype-dependent and we distinguished two groups of lines according to their phenotypic characteristics. In the group consisting of NR1 and NR2, increasing NO−3 supply during the vegetative phase made the chicon morphology switch from an opened to a closed type while resistance to bacteria was not affected by N supply. In the NS1 and Alg group, the effect of N on chicon morphology was the opposite to that observed in the NR1-NR2 group while NS1 and Alg exhibited a partial resistance to Erwinia sp., only expressing soft-rot disease when the N supply reached 3 mM. Characterization by DNA amplification fingerprinting (DAF) allowed the generation of 110 polymorphic bands and confirmed that the lines NR1 and NR2, on the one hand, and NS1 and Alg, on the other hand, belong to two distinct genetic groups. The DAF results indicate that chicon morphology and partial resistance to Erwinia sp. are complex traits which would be amenable to quantitative trait loci analysis. The split growth phase of chicory means that any changes in chicon related to N supply during vegetative growth were mediated by a root-originating signal. No variation in root carbon content among genotypes and NO−3 treatments was observed. In contrast, differences in root N content revealed the same grouping of the chicory lines, NR1 and NR2 being systematically richer in amino acids and NO−3 than NS1 and Alg. However, no correlation existed between N compounds and chicon morphology or pathology if all genotypes were considered together. Thus, the effect of N on plant development and pathology as well as putative identified signals might be specific for a genotype. Our study indicates that it is necessary to consider the genetic variability within a species in any signalling-pathway research.

Use of a structure-function plant model to assess the morphogenetic plasticity. How does variation in phyllochron modify plant growth and development of Brassica napus in the GreenLab model?

A functional-structural model of winter oilseed rape (WOSR) has been developed to study plant morphogenetic plasticity, i.e. how processes of morphogenesis are adapted in response to environmental constraints. The phyllochron (time between emergence of two successive leaves) is one of the variables sensitive to environment. The aim of this article is to use model sensitivity analysis to quantify the impact of an increase or a reduction in phyllochron on plant growth and source/sink functioning.