Stéphane Adamowicz

DART: a software to analyse root system architecture and development from captured images

Image analysis is used in numerous studies of root system architecture (RSA). To date, fully automatic procedures have not been good enough to completely replace alternative manual methods. DART (Data Analysis of Root Tracings) is freeware based on human vision to identify roots, particularly across time-series. Each root is described by a series of ordered links encapsulating specific information and is connected to other roots. The population of links constitutes the RSA. DART creates a comprehensive dataset ready for individual or global analyses and this can display root growth sequences along time. We exemplify here individual tomato root growth response to shortfall in solar radiation and we analyse the global distribution of the inter-root branching distances. DART helps in studying RSA and in producing structured and flexible datasets of individual root growth parameters. It is written in JAVA and relies on manual procedures to minimize the risks of errors and biases in datasets.

Is the C:N ratio a reliable indicator of C allocation to primary and defence-related metabolisms in tomato?

Plant growth and defence are both fuelled by compounds synthesized from a common pool of carbon and nitrogen, implying the existence of a competition for carbon and nitrogen allocation to both metabolisms. The ratio of carbon to nitrogen (C:N) of an organ is often regarded as a convenient indicator of growth and quality. The purpose of this work was to assess whether or not it is possible to extend its use to characterize the trade-off between growth and defence processes. Therefore, we calculated C:N ratios in the pool of resources and in the total plant, and correlated them to the concentrations of diverse compounds of the primary and secondary metabolisms in young tomatoes. Plants were grown hydroponically at N availabilities either limiting (0.1 mM) or not (7 mM) for growth in two glasshouses maintained either under ambient or enriched (700 vpm) air CO(2). These conditions yielded a large array of C:N in fully developed leaves, developing leaves, stem and roots, sampled 27, 35 and 47 days after sowing. Growth parameters and tissue concentrations of primary metabolites (carbohydrates, starch), defence-related compounds (polyphenols, glycoalkaloids), lignin, nitrate, ammonium, C and N were analyzed. Net CO(2) exchange rate was also measured at the last sampling date. Low N limited plant growth more than photosynthesis. The C:N in the resource pool was far higher than the total C:N. Starch was the most responsive compound, attaining high concentration under high C:N, whereas lignin remained stable. Chlorogenic acid, rutin, kaempferol-rutinoside and tomatine concentrations correlated positively to C:N. The same patterns were observed for most organs and molecules, except soluble carbohydrates in fully developed leaves whose concentration was not influenced. Among the organs, developing leaves showed the highest concentrations of secondary compounds and were the most responsive to C:N variations. Neither the biochemical nature of the compounds (C-based or N- containing metabolites) nor the calculation mode of C:N, influenced the patterns observed. Within the range of N availabilities considered (up to N limitation but not deficiency), the C:N can be considered as a good indicator of the secondary compounds concentrations in organs, especially for those involved in the chemical defence.

Organ-specific responses of tomato growth and phenolic metabolism to nitrate limitation

Phenolic compounds are secondary metabolites involved in plant innate chemical defence against pests and diseases. Their concentration varies depending on plant tissue and also on genetic and environmental factors, e.g. availability of nutrient resources. This study examines specific effects of low (LN) and high (HN) nitrogen supply on organ (root, stem and leaf) growth and accumulation of major phenolics [chlorogenic acid (CGA); rutin; kaempferol rutinoside (KR)] in nine hydroponically grown tomato cultivars. LN limited shoot growth but did not affect root growth, and increased concentrations of each individual phenolic in all organs. The strength of the response was organ-dependent, roots being more responsive than leaves and stems. Significant differences were observed between genotypes. Nitrogen limitation did not change the phenolic content in shoots, whereas it stimulated accumulation in roots. The results show that this trade-off between growth and defence in a LN environment can be discussed within the framework of the growth-differentiation balance hypothesis (i.e. GDBH), but highlight the need to integrate all plant organs in future modelling approaches regarding the impact of nitrogen limitation on primary and secondary metabolism.

Diel changes in nitrogen and carbon resource status and use for growth in young plants of tomato (Solanum lycopersicum)

BACKGROUND AND AIMS Modellers often define growth as the development of plant structures from endogenous resources, thus making a distinction between structural (W(S)) and total (W) dry biomass, the latter being the sum of W(S) and the weight of storage compounds. In this study, short-term C and N reserves were characterized experimentally (forms, organ distribution, time changes) in relation to light and nutrition signals, and organ structural growth in response to reserve levels was evaluated. METHODS Tomato plants (Solanum lycopersicum) were grown hydroponically in a growth room with a 12-h photoperiod and an adequate supply of NO(3)(-) (3 mol m(-3)). Three experiments were carried out 18 d after sowing: [NO(3)(-)] was either maintained at 3 mol m(-3), changed to 0.02 mol m(-3) or to 0 mol m(-3). Plants were sampled periodically throughout the light/dark cycles over 24-48 h. Organ W(S) was calculated from W together with the amount of different compounds that act as C and N resources, i.e. non-structural carbohydrates and carboxylates, nitrate and free amino acids. KEY RESULTS With adequate nutrition, carbohydrates accumulated in leaves during light periods, when photosynthesis exceeded growth needs, but decreased at night when these sugars are the main source of C for growth. At the end of the night, carbohydrates were still high enough to fuel full-rate growth, as W(S) increased at a near constant rate throughout the light/dark cycle. When nitrate levels were restricted, C reserves increased, but [NO(3)(-)] decreased progressively in stems, which contain most of the plant N reserves, and rapidly in leaves and roots. This resulted in a rapid restriction of structural growth. CONCLUSIONS Periodic darkness did not restrict growth because sufficient carbohydrate reserves accumulated during the light period. Structural growth, however, was very responsive to NO(3)(-) nutrition, because N reserves were mostly located in stems, which have limited nitrate reduction capacity.

Phenolic characterization and variability in leaves, stems and roots of Micro-Tom and patio tomatoes, in response to nitrogen limitation

Phenolics are implicated in the defence strategies of many plant species rendering their concentration increase of putative practical interest in the field of crop protection. Little attention has been given to the nature, concentration and distribution of phenolics within vegetative organs of tomato (Solanum lycopersicum. L) as compared to fruits. In this study, we extensively characterized the phenolics in leaves, stems and roots of nine tomato cultivars using high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-MS(n)) and assessed the impact of low nitrogen (LN) availability on their accumulation. Thirty-one phenolics from the four sub-classes, hydroxycinnamoyl esters, flavonoids, anthocyanins and phenolamides were identified, five of which had not previously been reported in these tomato organs. A higher diversity and concentration of phenolics was found in leaves than in stems and roots. The qualitative distribution of these compounds between plant organs was similar for the nine cultivars with the exception of Micro-Tom because of its significantly higher phenolic concentrations in leaves and stems as compared to roots. With few exceptions, the influence of the LN treatment on the three organs of all cultivars was to increase the concentrations of hydroxycinnamoyl esters, flavonoids and anthocyanins and to decrease those of phenolamides. This impact of LN was greater in roots than in leaves and stems. Nitrogen nutrition thus appears as a means of modulating the concentration and composition of organ phenolics and their distribution within the whole plant.

Influence of nitrogen availability on shoot development in young peach trees [Prunus persica (L.) Batsch]

Abstract. It is commonly stated that nitrogen (N) influences biomass accumulation in plants. For trees, however, a precise characterisation of shoot response to N and its impact on architecture is lacking. We attempted to study on the phytomer scale the effects of N limitation on shoot growth components, i.e. leaf emergence rate, final internode length and branching on the main and secondary axes of 1-year-old peach trees [Prunus persica (L.) Batsch]. Trees were grown on recycled nutrient solutions in which N concentration was restored once a week. We used two hydroponic set-ups in which weekly N availability, i.e. amount of N per tree, differed being either low (N1) or high (N2). Net N availability was defined weekly as the relative amount of N remaining in each set-up before solution replenishment. It declined with time and three periods of contrasting net N availability were identified. During these periods, leaf emergence rate and final internode length were similar on the main axis of N1 and N2 trees, so too was the distribution of secondary axes along the main axis. Secondary axes responded to N limitation by decreasing their growth components according to their position along the main axis. Differences were most important for basal secondary axes. Leaf emergence rate and final internode length responded similarly to N availability depending on axis order and position in the tree. It was concluded that N availability, particularly during the period of maximum growth of axes, influenced the shoot growth components and thus tree architecture.

Altering young tomato plant growth by nitrate and CO2 preserves the proportionate relation linking long‐term organic‐nitrogen accumulation to intercepted radiation

* A previously published model of crop nitrogen (N) status based on intercepted photosynthetically active radiation (R(i), mol per plant) suggested that plant organic N accumulation is related to R(i) by a constant ratio, defined hereafter as the radiation use efficiency for N (NRUE). The aim of this paper was to compare the effects of N nutrition and CO2 enrichment on NRUE and RUE (radiation use efficiency for biomass accumulation). * In three unrelated glasshouse experiments, tomato plants (Solanum lycopersicum) grown in hydroponics were fed for 28 d (exponential growth) with full solutions containing constant NO3(-) concentrations ([NO3(-)]) ranging from 0.05 to 15 mol m(-3), both under ambient or CO2-enriched (1000 microl l(-1)) air. * Each experiment comprised five harvests. Low [NO3(-)] (< 0.3 mol m(-3)) limited growth via leaf area (LA) restriction and decreased light interception. CO2 enrichment enhanced dry weight and LA. RUE was not affected by [NO3(-)], but increased under CO2-enriched air. By contrast, NRUE was not affected by [NO3(-)] or CO2 enrichment. * It is suggested that the radiation efficiency for organic N acquisition (NRUE) did not depend on C or N nutrition for young plants grown under unstressed conditions.

Totomatix: a novel automatic set-up to control diurnal, diel and long-term plant nitrate nutrition.

BACKGROUND Stand-alone nutritional set-ups are useful tools to grow plants at defined nutrient availabilities and to measure nutrient uptake rates continuously, in particular that for nitrate. Their use is essential when the measurements are meant to cover long time periods. These complex systems have, however, important drawbacks, including poor long-term reliability and low precision at high nitrate concentration. This explains why the information dealing with diel dynamics of nitrate uptake rate is scarce and concerns mainly young plants grown at low nitrate concentration. SCOPE The novel system detailed in this paper has been developed to allow versatile use in growth rooms, greenhouses or open fields at nitrate concentrations ranging from a few micro- to several millimoles per litres. The system controls, at set frequencies, the solution nitrate concentration, pH and volumes. Nitrate concentration is measured by spectral deconvolution of UV spectra. The main advantages of the set-up are its low maintenance (weekly basis), an ability to diagnose interference or erroneous analyses and high precision of nitrate concentration measurements (0·025 % at 3 mm). The paper details the precision of diurnal nitrate uptake rate measurements, which reveals sensitivity to solution volume at low nitrate concentration, whereas at high concentration, it is mostly sensitive to the precision of volume estimates. CONCLUSIONS This novel set-up allows us to measure and characterize the dynamics of plant nitrate nutrition at high temporal resolution (minutes to hours) over long-term experiments (up to 1 year). It is reliable and also offers a novel method to regulate up to seven N treatments by adjusting the daily uptake of test plants relative to controls, in variable environments such as open fields and glasshouses.

Nitrogen Nutrition and Use in Horticultural Crops

Summary The fertilization practice lays mainly on two concepts, maintenance of the rooting medium fertility and balancing of the crop nutrient demand. Both benefited from recent scientific advances which serve the purposes of diagnosis, crop modelling and optimization of fertilizer use. These would be most helpful to horticulture which makes the most intensive use of N amendments, but they seem challenged by its uttermost diversity of cultural techniques and cultivated plant species. For instance, fertility must be defined for plain soil as well as for hydroponically grown crops, while N demand may concern a very large range of time scales, from a few hours in the case of fertigation on inert media to weeks or months in the open field. This paper confronts the concepts of fertility and crop nutrient demand, mostly developed for field crops, to the needs of horticulture with particular reference to the constraints imposed by genetic and time scale diversities.

Analysis of bias in the calculation and measurement of plant mineral uptake rates

AimsCalculation of net ion uptake rate (F) from hydroponic solutions relies on balanced equations where F is equal to the initial minus the final ion content, plus fertilization. Knowledge is thus required of both volume (V), concentration (C) and of their temporal variations. The literature, however, proposes simplified equations that disregard variations in V and are thus strictly inaccurate. This paper studies the bias arising from such simplified formulae and also from deviations in V and C measurements.MethodsWe used our experimental data and simulation to analyse the impact of different bias sources on F calculation, and to compare setups where C is regulated, or left to drift in order to study F = f(C).ResultsThis paper reports two major findings, the first being that simplified equations distort F diurnal dynamics and ion uptake isotherms, yielding underestimated Michaelis-Menten parameters. The second shows the advantage of using C-regulated over unregulated systems to determine F when biased V and C measurements cannot be avoided.ConclusionsRegulated systems are able to minimize the biases on F, but the measurement of water uptake rate is compulsory. Therefore, simplified formulae should not be used.