Valentina Baldazzi

Profiling sugar metabolism during fruit development in a peach progeny with different fructose-to-glucose ratios

BackgroundFruit taste is largely affected by the concentration of soluble sugars and organic acids and non-negligibly by fructose concentration, which is the sweetest-tasting sugar. To date, many studies investigating the sugars in fruit have focused on a specific sugar or enzyme and often on a single variety, but only a few detailed studies addressing sugar metabolism both as a whole and dynamic system are available. In commercial peach fruit, sucrose is the main sugar, followed by fructose and glucose, which have similar levels. Interestingly, low fructose-to-glucose ratios have been observed in wild peach accessions. A cross between wild peach and commercial varieties offers an outstanding possibility to study fruit sugar metabolism.ResultsThis work provides a large dataset of sugar composition and the capacities of enzymes that are involved in sugar metabolism during peach fruit development and its genetic diversity. A large fraction of the metabolites and enzymes involved in peach sugar metabolism were assayed within a peach progeny of 106 genotypes, of which one quarter displayed a low fructose-to-glucose ratio. This profiling was performed at six stages of growth using high throughput methods. Our results permit drawing a quasi-exhaustive scheme of sugar metabolism in peach. The use of a large number of genotypes revealed a remarkable robustness of enzymatic capacities across genotypes and years, despite strong variations in sugar composition, in particular the fructose-to-glucose ratio, within the progeny. A poor correlation was also found between the enzymatic capacities and the accumulation rates of metabolites.ConclusionsThese results invalidate the hypothesis of the straightforward enzymatic control of sugar concentration in peach fruit. Alternative hypotheses concerning the regulation of fructose concentration are discussed based on experimental data. This work lays the foundation for a comprehensive study of the mechanisms involved in sugar metabolism in developing fruit.

Importance of metabolic coupling for the dynamics of gene expression following a diauxic shift in Escherichia coli

Gene regulatory networks consist of direct interactions, but also include indirect interactions mediated by metabolism. We investigate to which extent these indirect interactions arising from metabolic coupling influence the dynamics of the system. To this end, we build a qualitative model of the gene regulatory network controlling carbon assimilation in Escherichia coli, and use this model to study the changes in gene expression following a diauxic shift from glucose to acetate. In particular, we compare the relative variation in the steady-state concentrations of enzymes and transcription regulators during growth on glucose and acetate, as well as the dynamic response of gene expression to the exhaustion of glucose and the subsequent assimilation of acetate. We find significant differences between the dynamics of the system in the absence and presence of metabolic coupling. This shows that interactions arising from metabolic coupling cannot be ignored when studying the dynamics of gene regulatory networks.

Metabolic studies in plant organs: don't forget dilution by growth

The recent development of high-density technologies has enabled a dramatic extension of the exploration of living organisms. Such exploration typically consists in the parallel measurement of large numbers of compounds, at different developmental stages or following the application of contrasted environmental stimuli. Correlation analysis or more sophisticated statistical approaches are then used to identify clusters of co-varying compounds with the aim of reconstructing the underlying regulatory network governing system responses. Such approaches are now routinely used to identify predictive biomarkers (Steinfath et al., 2010; Riedelsheimer et al., 2012), including candidate genes (Carreno-Quintero et al., 2012). Originally developed for unicellular organisms, these approaches are now applied to plants for the analysis of transcripts, proteins, metabolites (Gibon et al., 2006; Stitt et al., 2010; Liberman et al., 2012) and more recently enzymes activities (Gibon et al., 2004; Saito et al., 2008; Moreno-Risueno et al., 2010). Most studies involve homogenization of specific tissues or even whole organs (e.g., fruit pericarp, leaf), without considering the subcellular localization of the measured compounds. Plant cells distinguish from other cells in possessing a large central vacuole, which size may vary dramatically between tissues, genotypes (species, cultivars) and developmental stages. Whereas young cells have small vacuoles, mature cells have large vacuoles that can encompass more than 95% of the cell volume. Here, we will demonstrate that without taking into account the volumes of the cell compartments, the analyses of the dynamics of the compounds and the subsequent compound-compound correlation analyses might be biased, especially when the functional significance of the study is bound to concentrations, as it is the case for enzymes and metabolites. Possible correction strategies are discussed, with special emphasis to their pertinence and applicability to specific questions.

Dynamic QTLs for sugars and enzyme activities provide an overview of genetic control of sugar metabolism during peach fruit development

Highlight Forty-five QTLs controlling sugars and enzyme activities related to sugar metabolism in peach fruit were identified. Dynamic QTLs revealed changing effects of alleles during fruit development.

In-silico analysis of water and carbon relations under stress conditions. A multi-scale perspective centered on fruit.

Fruit development, from its early stages, is the result of a complex network of interacting processes, on different scales. These include cell division, cell expansion but also nutrient transport from the plant, and exchanges with the environment. In the presence of nutrient limitation, in particular, the plant reacts as a whole, by modifying its architecture, metabolism, and reproductive strategy, determining the resources available for fruit development, which in turn affects the overall source-sink balance of the system. Here, we present an integrated model of tomato that explicitly accounts for early developmental changes (from cell division to harvest), and use it to investigate the impact of water deficit and carbon limitation on nutrient fluxes and fruit growth, in both dry and fresh mass. Variability in fruit response is analyzed on two different scales: among trusses at plant level, and within cell populations at fruit level. Results show that the effect of stress on individual cells strongly depends on their age, size, and uptake capabilities, and that the timing of stress application, together with the fruit position on the plant, is crucial in determining the final phenotypic outcome. Water deficit and carbon depletion impacted either source size, source activity, or sink strength with contrasted effects on fruit growth. An important prediction of the model is the major role of symplasmic transport of carbon in the early stage of fruit development, as a catalyst for cell and fruit growth.

Inter-Species Comparative Analysis of Components of Soluble Sugar Concentration in Fleshy Fruits.

The soluble sugar concentration of fleshy fruit is a key determinant of fleshy fruit quality. It affects directly the sweetness of fresh fruits and indirectly the properties of processed products (e.g., alcohol content in wine). Despite considerable divergence among species, soluble sugar accumulation in a fruit results from the complex interplay of three main processes, namely sugar import, sugar metabolism, and water dilution. Therefore, inter-species comparison would help to identify common and/or species-specific modes of regulation in sugar accumulation. For this purpose, a process-based mathematical framework was used to compare soluble sugar accumulation in three fruits: grape, tomato, and peach. Representative datasets covering the time course of sugar accumulation during fruit development were collected. They encompassed 104 combinations of species (3), genotypes (30), and growing conditions (19 years and 16 nutrient and environmental treatments). At maturity, grape showed the highest soluble sugar concentrations (16.5–26.3 g/100 g FW), followed by peach (2.2 to 20 g/100 g FW) and tomato (1.4 to 5 g/100 g FW). Main processes determining soluble sugar concentration were decomposed into sugar importation, metabolism, and water dilution with the process-based analysis. Different regulation modes of soluble sugar concentration were then identified, showing either import-based, dilution-based, or import and dilution dual-based. Firstly, the higher soluble sugar concentration in grape than in tomato is a result of higher sugar importation. Secondly, the higher soluble sugar concentration in grape than in peach is due to a lower water dilution. The third mode of regulation is more complicated than the first two, with differences both in sugar importation and water dilution (grape vs. cherry tomato; cherry tomato vs. peach; peach vs. tomato). On the other hand, carbon utilization for synthesis of non-soluble sugar compounds (namely metabolism) was conserved among the three fruit species. These distinct modes appear to be quite species-specific, but the intensity of the effect may significantly vary depending on the genotype and management practices. These results provide novel insights into the drivers of differences in soluble sugar concentration among fleshy fruits.

Model-Assisted Estimation of the Genetic Variability in Physiological Parameters Related to Tomato Fruit Growth under Contrasted Water Conditions

Drought stress is a major abiotic stress threatening plant and crop productivity. In case of fleshy fruits, understanding mechanisms governing water and carbon accumulations and identifying genes, QTLs and phenotypes, that will enable trade-offs between fruit growth and quality under Water Deficit (WD) condition is a crucial challenge for breeders and growers. In the present work, 117 recombinant inbred lines of a population of Solanum lycopersicum were phenotyped under control and WD conditions. Plant water status, fruit growth and composition were measured and data were used to calibrate a process-based model describing water and carbon fluxes in a growing fruit as a function of plant and environment. Eight genotype-dependent model parameters were estimated using a multiobjective evolutionary algorithm in order to minimize the prediction errors of fruit dry and fresh mass throughout fruit development. WD increased the fruit dry matter content (up to 85%) and decreased its fresh weight (up to 60%), big fruit size genotypes being the most sensitive. The mean normalized root mean squared errors of the predictions ranged between 16–18% in the population. Variability in model genotypic parameters allowed us to explore diverse genetic strategies in response to WD. An interesting group of genotypes could be discriminated in which (i) the low loss of fresh mass under WD was associated with high active uptake of sugars and low value of the maximum cell wall extensibility, and (ii) the high dry matter content in control treatment (C) was associated with a slow decrease of mass flow. Using 501 SNP markers genotyped across the genome, a QTL analysis of model parameters allowed to detect three main QTLs related to xylem and phloem conductivities, on chromosomes 2, 4, and 8. The model was then applied to design ideotypes with high dry matter content in C condition and low fresh mass loss in WD condition. The ideotypes outperformed the RILs especially for large and medium fruit-size genotypes, by combining high pedicel conductance and high active uptake of sugars. Interestingly, five small fruit-size RILs were close to the selected ideotypes, and likely bear interesting traits and alleles for adaptation to WD.

Process-Based Simulation Models Are Essential Tools for Virtual Profiling and Design of Ideotypes: Example of Fruit and Root

Process-based simulation models (PBSMs) combine, in various mathematical frameworks, many biological functional hypotheses on responses of plant processes to environmental fluctuations. Model simulated responses can be analysed in the context of adapting the current agricultural systems to the changing environment. From loads of simulations made with various cultural practices, these models allow the virtual profiling of plants and a mere analysis of how processes interact when crops are perturbed by one or several changes. They allow also describing the development of plant traits as a consequence of environmental and genetic conditions. Such knowledge is required to decipher the genotype × environment × management (G × E × M) interactions so as to build genotypes adapted to particular conditions, i.e., plant ideotypes. Two PBSMs dealing with (1) fruit quality and sensitivity to diseases and (2) root system architecture, respectively, are shortly described in this chapter. These models have been used to analyse various fruit and root properties, to deconvolute G × E × M interactions and to identify ecophysiological traits related to crop yield improvement, root foraging performance and fruit quality. PBSMs appear to be powerful tools to phenotype plants at the process level in a comprehensive and “costless” way.

Integrating Physiology and Architecture in Models of Fruit Expansion

Architectural properties of a fruit, such as its shape, vascular patterns, and skin morphology, play a significant role in determining the distributions of water, carbohydrates, and nutrients inside the fruit. Understanding the impact of these properties on fruit quality is difficult because they develop over time and are highly dependent on both genetic and environmental controls. We present a 3D functional-structural fruit model that can be used to investigate effects of the principle architectural properties on fruit quality. We use a three step modeling pipeline in the OpenAlea platform: (1) creating a 3D volumetric mesh representation of the internal and external fruit structure, (2) generating a complex network of vasculature that is embedded within this mesh, and (3) integrating aspects of the fruits function, such as water and dry matter transport, with the fruits structure. We restrict our approach to the phase where fruit growth is mostly due to cell expansion and the fruit has already differentiated into different tissue types. We show how fruit shape affects vascular patterns and, as a consequence, the distribution of sugar/water in tomato fruit. Furthermore, we show that strong interaction between tomato fruit shape and vessel density induces, independently of size, an important and contrasted gradient of water supply from the pedicel to the blossom end of the fruit. We also demonstrate how skin morphology related to microcracking distribution affects the distribution of water and sugars inside nectarine fruit. Our results show that such a generic model permits detailed studies of various, unexplored architectural features affecting fruit quality development.

Challenges in integrating genetic control in plant and crop models

Predicting genotype-to-phenotype relationships under contrasting environments is a great challenge for plant biology and breeding. Classical crop models have been developed to predict crop yield or product quality under fluctuating environments but they are usually calibrated for a single genotype, restricting the validity range of the model itself. To overcome this limitation, genetic control has to be integrated into crop models and genotype × environment interactions have to be made explicit.