Spatial and temporal evolution of quantitative magnetic resonance imaging parameters of peach and apple fruit - relationship with biophysical and metabolic traits.

Fruits are complex organs that are spatially regulated during development. Limited phenotyping capacity at cell and tissue levels is one of the main obstacles to our understanding of the coordinated regulation of the processes involved in fruit growth and quality. In this study, the spatial evolution of biophysical and metabolic traits of peach and apple fruit was investigated during fruit development. In parallel, the multi-exponential relaxation times and apparent microporosity were assessed by quantitative magnetic resonance imaging (MRI). The aim was to identify the possible relationships between MRI parameters and variations in the structure and composition of fruit tissues during development so that transverse relaxation could be proposed as a biomarker for the assessment of the structural and functional evolution of fruit tissues during growth. The study provides species-specific data on developmental and spatial variations in density, cell number and size distribution, insoluble and soluble compound accumulation and osmotic and water potential in the fruit mesocarp. Magnetic resonance imaging was able to capture tissue evolution and the development of pericarp heterogeneity by accessing information on cell expansion, water status and distribution at cell level, and microporosity. Changes in vacuole-related transverse relaxation rates were mostly explained by cell/vacuole size. The impact of cell solute composition, microporosity and membrane permeability on relaxation times is also discussed. The results demonstrate the usefulness of MRI as a tool to phenotype fruits and to access important physiological data during development, including information on spatial variability.

Insights into the Intraspecific Variability of the above and Belowground Emissions of Volatile Organic Compounds in Tomato.

The in-vivo monitoring of volatile organic compound (VOC) emissions is a potential non-invasive tool in plant protection, especially in greenhouse cultivation. We studied VOC production from above and belowground organs of the eight parents of the Multi-Parent Advanced Generation Intercross population (MAGIC) tomato population, which exhibits a high genetic variability, in order to obtain more insight into the variability of constitutive VOC emissions from tomato plants under stress-free conditions. Foliage emissions were composed of terpenes, the majority of which were also stored in the leaves. Foliage emissions were very low, partly light-dependent, and differed significantly among genotypes, both in quantity and quality. Soil with roots emitted VOCs at similar, though more variable, rates than foliage. Soil emissions were characterized by terpenes, oxygenated alkanes, and alkenes and phenolic compounds, only a few of which were found in root extracts at low concentrations. Correlation analyses revealed that several VOCs emitted from foliage or soil are jointly regulated and that above and belowground sources are partially interconnected. With respect to VOC monitoring in tomato crops, our results underline that genetic variability, light-dependent de-novo synthesis, and belowground sources are factors to be considered for successful use in crop monitoring.

Fruit water content as an indication of sugar metabolism improves simulation of carbohydrate accumulation in tomato fruit.

Although fleshy fruit is mainly made up of water, little is known about the impact of its water status on sugar metabolism and its composition. In order to verify whether fruit water status is an important driver of carbohydrate composition in tomato fruit, an adaptation of the SUGAR model proposed previously by M. Génard and M. Souty was used. Two versions of the model, with or without integrating the influence of fruit water content on carbohydrate metabolism, were proposed and then assessed with the data sets from two genotypes, Levovil and Cervil, grown under different conditions. The results showed that, for both genotypes, soluble sugars and starch were better fitted by the model when the effects of water content on carbohydrate metabolism were taken into consideration. Water content might play a regulatory role in the carbon metabolism from sugars to compounds other than sugars and starch in Cervil fruit, and from sugars to starch in Levovil fruit. While water content influences tomato fruit carbohydrate concentrations by both metabolism and dilution/dehydration effects in the early developmental stage, it is mainly by dilution/dehydration effects in the late stage. The possible mechanisms underlying the effect of the fruit water content on carbohydrate metabolism are also discussed.

Coupling water fluxes with cell wall mechanics in a multicellular model of plant development.

The growth of plant organs is a complex process powered by osmosis that attracts water inside the cells; this influx induces simultaneously an elastic extension of the walls and pressure in the cells, called turgor pressure; above a threshold, the walls yield and the cells grow. Based on Lockhart's seminal work, various models of plant morphogenesis have been proposed, either for single cells, or focusing on the wall mechanical properties. However, the synergistic coupling of fluxes and wall mechanics has not yet been fully addressed in a multicellular model. This work lays the foundations of such a model, by simplifying as much as possible each process and putting emphasis on the coupling itself. Its emergent properties are rich and can help to understand plant morphogenesis. In particular, we show that the model can display a new type of lateral inhibitory mechanism that amplifies growth heterogeneities due e.g to cell wall loosening.

Decreased sink/source ratio enhances hexose transport in the fruits of greenhouse tomatoes: integration of gene expression and biochemical analyses.

To examine the physiological role of hexose transporters in determining the sink strength of individual fruits, the regulation of hexose transporters gene expression was studied when the sink/source ratio was artificially altered under the greenhouse condition; this was done in two cultivars of tomato, i.e. Grandella and Isabella. The sink/source ratio treatments included: saving one fruit per truss (1F), two fruits per truss (2F), three fruits per truss (3F) and no fruit pruning (control). The results showed that fruit thinning could increase starch, sucrose, and hexose contents in the fruits; it could also modulate the activity of the key enzymes and the expression of tomato hexose transporter genes (LeHTs). Based on the relative transcript levels, all examined LeHTs were unregulated at the end of cell division and the cell expansion stage of fruit development, but the strongest expression level observed at the onset of ripening was related to LeHT1 and LeHT2. Given the concomitancy of cell wall invertase (EC 3.2.1.26) activity and the LeHTs relative expression cell wall, invertase activity seemed to be involved in the expression level of LeHTs. The increased trends of the LeHTs expression with the decrease of the sink/source ratio confirmed the role of hexose transporters in determining the sink strength of the tomato fruits.

Organ-wide and ploidy-dependent regulation both contribute to cell-size determination: evidence from a computational model of tomato fruit.

The development of a new organ is the result of coordinated events of cell division and expansion, in strong interaction with each other. This study presents a dynamic model of tomato fruit development that includes cell division, endoreduplication, and expansion processes. The model is used to investigate the potential interactions among these developmental processes within the context of the neo-cellular theory. In particular, different control schemes (either cell-autonomous or organ-controlled) are tested and compared to experimental data from two contrasting genotypes. The model shows that a pure cell-autonomous control fails to reproduce the observed cell-size distribution, and that an organ-wide control is required in order to get realistic cell-size variations. The model also supports the role of endoreduplication as an important determinant of the final cell size and suggests that a direct effect of endoreduplication on cell expansion is needed in order to obtain a significant correlation between size and ploidy, as observed in real data.

Are compound leaves more complex than simple ones? A multi-scale analysis.

Background and Aims: The question of which cellular mechanisms determine the variation in leaf size has been addressed mainly in plants with simple leaves. It is addressed here in tomato taking into consideration the expected complexity added by the several lateral appendages making up the compound leaf, the leaflets. Methods: Leaf and leaflet areas, epidermal cell number and areas, and endoreduplication (co-) variations were analysed in Solanum lycopersicum considering heteroblastic series in a wild type (Wva106) and an antisense mutant, the Pro35S:Slccs52AAS line, and upon drought treatments. All plants were grown in an automated phenotyping platform, PHENOPSIS, adapted to host plants grown in 7 L pots. Key Results: Leaf area, leaflet area and cell number increased with leaf rank until reaching a plateau. In contrast, cell area slightly decreased and endoreduplication did not follow any trend. In the transgenic line, leaf area, leaflet areas and cell number of basal leaves were lower than in the wild type, but higher in upper leaves. Reciprocally, cell area was higher in basal leaves and lower in upper leaves. When scaled up at the whole sympodial unit, all these traits did not differ significantly between the transgenic line and the wild type. In response to drought, leaf area was reduced, with a clear dose effect that was also reported for all size-related traits, including endoreduplication. Conclusions: These results provide evidence that all leaflets have the same cellular phenotypes as the leaf they belong to. Consistent with results reported for simple leaves, they show that cell number rather than cell size determines the final leaf areas and that endoreduplication can be uncoupled from leaf and cell sizes. Finally, they re-question a whole-plant control of cell division and expansion in leaves when the Wva106 and the Pro35S:Slccs52AAS lines are compared.

Putting primary metabolism into perspective to obtain better fruits.

Background: One of the key goals of fruit biology is to understand the factors that influence fruit growth and quality, ultimately with a view to manipulating them for improvement of fruit traits. Scope: Primary metabolism, which is not only essential for growth but is also a major component of fruit quality, is an obvious target for improvement. However, metabolism is a moving target that undergoes marked changes throughout fruit growth and ripening. Conclusions: Agricultural practice and breeding have successfully improved fruit metabolic traits, but both face the complexity of the interplay between development, metabolism and the environment. Thus, more fundamental knowledge is needed to identify further strategies for the manipulation of fruit metabolism. Nearly two decades of post-genomics approaches involving transcriptomics, proteomics and/or metabolomics have generated a lot of information about the behaviour of fruit metabolic networks. Today, the emergence of modelling tools is providing the opportunity to turn this information into a mechanistic understanding of fruits, and ultimately to design better fruits. Since high-quality data are a key requirement in modelling, a range of must-have parameters and variables is proposed.

Reduced susceptibility of tomato stem to the necrotrophic fungus Botrytis cinerea is associated with a specific adjustment of fructose content in the host sugar pool.

Background and aims: Plant soluble sugars, as main components of primary metabolism, are thought to be implicated in defence against pathogenic fungi. However, the function of sucrose and hexoses remains unclear. This study aimed to identify robust patterns in the dynamics of soluble sugars in sink tissues of tomato plants during the course of infection by the necrotrophic fungus Botrytis cinerea . Distinct roles for glucose and fructose in defence against B. cinerea were hypothesized. Methods: We examined sugar contents and defence hormonal markers in tomato stem tissues before and after infection by B. cinerea , in a range of abiotic environments created by various nitrogen and water supplies. Key Results: Limited nitrogen or water supplies increased tomato stem susceptibility to B. cinerea . Glucose and fructose contents of tissues surrounding infection sites evolved differently after inoculation. The fructose content never decreased after inoculation with B. cinerea , while that of glucose showed either positive or negative variation, depending on the abiotic environment. An increase in the relative fructose content (defined as the proportion of fructose in the soluble sugar pool) was observed in the absence of glucose accumulation and was associated with lower susceptibility. A lower expression of the salicylic acid marker PR1a , and a lower repression of a jasmonate marker COI1 were associated with reduced susceptibility. Accordingly, COI1 expression was positively correlated with the relative fructose contents 7 d after infection. Conclusions: Small variations of fructose content among the sugar pool are unlikely to affect intrinsic pathogen growth. Our results highlight distinct use of host glucose and fructose after infection by B. cinerea and suggest strongly that adjustment of the relative fructose content is required for enhanced plant defence.

Genotype by watering regime interaction in cultivated tomato: lessons from linkage mapping and gene expression.

KEY MESSAGE: In tomato, genotype by watering interaction resulted from genotype re-ranking more than scale changes. Interactive QTLs according to watering regime were detected. Differentially expressed genes were identified in some intervals. ABSTRACT: As a result of climate change, drought will increasingly limit crop production in the future. Studying genotype by watering regime interactions is necessary to improve plant adaptation to low water availability. In cultivated tomato (Solanum lycopersicum L.), extensively grown in dry areas, well-mastered water deficits can stimulate metabolite production, increasing plant defenses and concentration of compounds involved in fruit quality, at the same time. However, few tomato Quantitative Trait Loci (QTLs) and genes involved in response to drought are identified or only in wild species. In this study, we phenotyped a population of 119 recombinant inbred lines derived from a cross between a cherry tomato and a large fruit tomato, grown in greenhouse under two watering regimes, in two locations. A large genetic variability was measured for 19 plant and fruit traits, under the two watering treatments. Highly significant genotype by watering regime interactions were detected and resulted from re-ranking more than scale changes. The population was genotyped for 679 SNP markers to develop a genetic map. In total, 56 QTLs were identified among which 11 were interactive between watering regimes. These later mainly exhibited antagonist effects according to watering treatment. Variation in gene expression in leaves of parental accessions revealed 2259 differentially expressed genes, among which candidate genes presenting sequence polymorphisms were identified under two main interactive QTLs. Our results provide knowledge about the genetic control of genotype by watering regime interactions in cultivated tomato and the possible use of deficit irrigation to improve tomato quality.

Water shortage and quality of fleshy fruits--making the most of the unavoidable.

Extreme climatic events, including drought, are predicted to increase in intensity, frequency, and geographic extent as a consequence of global climate change. In general, to grow crops successfully in the future, growers will need to adapt to less available water and to take better advantage of the positive effects of drought. Fortunately, there are positive effects associated with drought. Drought stimulates the secondary metabolism, thereby potentially increasing plant defences and the concentrations of compounds involved in plant quality, particularly taste and health benefits. The role of drought on the production of secondary metabolites is of paramount importance for fruit crops. However, to manage crops effectively under conditions of limited water supply, for example by applying deficit irrigation, growers must consider not only the impact of drought on productivity but also on how plants manage the primary and secondary metabolisms. This question is obviously complex because during water deficit, trade-offs among productivity, defence, and quality depend upon the intensity, duration, and repetition of events of water deficit. The stage of plant development during the period of water deficit is also crucial, as are the effects of other stressors. In addition, growers must rely on relevant indicators of water status, i.e. parameters involved in the relevant metabolic processes, including those affecting quality. Although many reports on the effects of drought on plant function and crop productivity have been published, these issues have not been reviewed thus far. Here, we provide an up-to-date review of current knowledge of the effects of different forms of drought on fruit quality relative to the primary and secondary metabolisms and their interactions. We also review conventional and less conventional indicators of water status that could be used for monitoring purposes, such as volatile compounds. We focus on fruit crops owing to the importance of secondary metabolism in fruit quality and the importance of fruits in the human diet. The issue of defence is also briefly discussed.

Variety, growing conditions and processing method act on different structural and biochemical traits to modify viscosity in tomato puree.

The texture of tomato products can be modified by choice of variety, their growing conditions and/or processing method, but no clear explanation exists of the mechanisms that transform fruit tissue, how they act on texture, or whether genetics and processing impact the same physical parameters. We therefore conducted a study that processed 4 varieties produced under low/high nitrogen supply, into puree using both hot and cold break processes. No specific rheological signature allows discrimination between cultivar-induced or process-induced textural changes, but that they can be distinguished by sensory analysis. Growth conditions impacted but was not sensory distinguished. Both caused significant variations in 7 of 11 physico-chemical parameters, but the order of importance of these traits controlling texture varied, depending on whether the cause was genetic or process-related. Analysis of alcohol insoluble solids revealed a specific signature in pectin composition and conformation that could be linked to particle aggregation in the presence of lycopene-rich particles.

Tomato genotype but not crop water deficit matters for tomato health benefits in diet-induced obesity of C57BL/6JRj male mice.

Several studies have linked the intake of lycopene and/or tomato products with improved metabolic health under obesogenic regime. The aim was to evaluate the differential impact of supplementations with several tomato genotypes differing in carotenoid content and subjected to different irrigation levels on obesity-associated disorders in mice. In this study, 80 male C57BL/6JRj mice were assigned into 8 groups to receive: control diet, high fat diet, high fat diet supplemented at 5 % w/w with 4 tomato powders originating from different tomato genotypes cultivated under control irrigation: H1311, M82, IL6-2, IL12-4. Among the 4 genotypes, 2 were also cultivated under deficit irrigation, reducing the irrigation water supply by 50 % from anthesis to fruit harvest. In controlled irrigation treatment, all genotypes significantly improved fasting glycemia and three of them significantly lowered liver lipids content after 12 weeks of supplementation. In addition, IL6-2 genotype, rich in ß-carotene, significantly limited animal adiposity, body weight gain and improved glucose homeostasis as highlighted in glucose and insulin tolerance tests. No consistent beneficial or detrimental impact of deficit irrigation to tomato promoting health benefits was found. These findings imply that the choice of tomato genotype can significantly alter the composition of fruit carotenoids and phytochemicals, thereby influencing the anti-obesogenic effects of the fruit. In contrast, deficit irrigation appears to have an overall insignificant impact on enhancing the health benefits of tomato powder in this context, particularly when compared to the genotype-related variations in carotenoid content.

Effect of long-term deficit irrigation on tomato and goji berry quality: from fruit composition to in vitro bioaccessibility of carotenoids.

Drought is a persistent challenge for horticulture, affecting various aspects of fruit development and ultimately fruit quality, but the effect on nutritional value has been under-investigated. Here, fruit quality was studied on six tomato genotypes and one goji cultivar under deficit irrigation (DI), from fruit composition to in vitro bioaccessibility of carotenoids. For both species, DI concentrated most health-related metabolites in fresh fruit. On a dry mass basis, DI increased total phenolic and sugar concentration, but had a negative or insignificant impact on fruit ascorbic acid, organic acid, and alcohol-insoluble matter contents. DI also reduced total carotenoids content in tomato (-18.7% on average), especially ß-carotene (-32%), but not in goji berry DW (+15.5% and +19.6%, respectively). DI reduced the overall in vitro bioaccessibility of carotenoids to varying degrees depending on the compound and plant species. Consequently, mixed micelles produced by digestion of fruits subjected to DI contained either the same or lesser quantities of carotenoids, even though fresh fruits could contain similar or higher quantities. Thus, DI effects on fruit composition were species and genotype dependent, but an increase in the metabolite concentration did not necessarily translate into greater bioaccessibility potentially due to interactions with the fruit matrix.

Resource Translocation Modelling Highlights Density-Dependence Effects in Fruit Production at Various Levels of Organisation.

The size of fruit cells, seeds and fruits depends on their number. Could this density-dependence effect result from sugar resource sharing and, if so, does it involve phloem sugar flow or the intensity of sugar unloading to the sink? A density-dependence model (DDM) describing these processes was designed and parameterised for six species at five levels of organisation: cells and seeds within fruits, fruits within clusters, fruits within plants and plants within plots. Sugar flow was driven by phloem conductance, determined by parameters α, governing the shape of its relationship to population size, and κ, its value for a population size of one. Sugar unloading followed Michaelis-Menten kinetics with parameters Vm (maximal unloading rate) and Km (Michaelis constant). The DDM effectively reproduced the observed individual mass dynamics, the undercompensating density dependence observed in most species at all sub-plant levels and the undercompensating, exact and overcompensating density dependence observed at the plant level. Conductance (κ) was a scaling factor varying with the level of organisation. Vm was positively correlated with density dependence, and α was negatively correlated with density dependence only if the plant-within-plot level was not considered. Analysis of the model's behaviour indicates that density dependence of fruit growth could be a result of sugar sharing, and that both phloem sugar flow and sugar unloading contribute to these effects.

Pedicel anatomy and histology in tomato vary according to genotype and water-deficit environment, affecting fruit mass.

The growth and composition of fleshy fruits depend on resource acquisition and distribution in the plant. In tomato, the pedicel serves as the final connection between plant and fruit. However, very few quantitative data are available for the conducting tissues of the pedicel, nor is their genetic variability known. In the present study, a histological approach was combined with process-based modeling to evaluate the potential contribution made by the anatomy and histology of the pedicel to variations in fruit mass. Eleven genotypes were characterized and the impact of water deficit was studied for a single genotype using stress intensity and stage of application as variables. The results highlighted extensive variations in the relative proportions of the different pedicel tissues and in the absolute areas of xylem and phloem between genotypes. The model suggests that the variations in the area of the pedicel's vascular tissues induced by differences in genotype and water-deficit environments partly contributed to fruit mass variability. They therefore warrant phenotyping for use in the development of plant strains adapted to future environmental constraints. The results also demonstrated the need to develop non-invasive in vivo measurement methods to establish the number and size of active vessels and the flow rates in these vessels to improve prediction of water fluxes in plant architecture.

Virtual profiling: a new way to analyse phenotypes.

Simulation models can be used to perform virtual profiling in order to analyse eco-physiological processes controlling plant phenotype. To illustrate this, an eco-physiological model has been used to compare and contrast the status of a virtual fruit system under two situations of carbon supply. The model simulates fruit growth, accumulation of sugar, citric acid and water, transpiration, respiration and ethylene emission, and was successfully tested on peach (Prunus persica L. Batsch) for two leaf-to-fruit ratios (6 and 18 leaves per fruit). The development stage and the variation in leaf number had large effects of the fruit model variables dealing with growth, metabolism and fruit quality. A sensitivity analysis showed that changing a single parameter value, which could correspond to a genotypic change induced by a mutation, either strongly affects most of the processes, or affects a specific process or none. Correlation analysis showed that, in a complex system such as fruit, the intensity of many physiological processes and quality traits co-varies. It also showed unexpected co-variations resulting from emergent properties of the system. This virtual profiling approach opens a new route to explore the impact of mutations, or naturally occurring genetic variations, under differing environmental conditions.

Modelling the size and composition of fruit, grain and seed by process-based simulation models.

Understanding what determines the size and composition of fruit, grain and seed in response to the environment and genotype is challenging, as these traits result from several linked processes controlled at different levels of organization, from the subcellular to the crop level, with subtle interactions occurring at or between the levels of organization. Process-based simulation models (PBSMs) implement algorithms to simulate metabolic and biophysical aspects of cell, tissue and organ behaviour. In this review, fruit, grain and seed PBSMs describing the main phases of growth, development and storage metabolism are discussed. From this concurrent work, it is possible to identify generic storage organ processes which can be modelled similarly for fruit, grain and seed. Spatial heterogeneity at the tissue and whole-plant level is found to be a key consideration in modelling the effects of the environment and genotype on fruit, grain and seed end-use value. In the future, PBSMs may well become the main link between studies at the molecular and whole-plant levels. To bridge this phenotype-to-genotype gap, future models need to remain plastic without becoming overparameterized.

Combining ecophysiological modelling and quantitative trait locus analysis to identify key elementary processes underlying tomato fruit sugar concentration.

A mechanistic model predicting the accumulation of tomato fruit sugars was developed in order (i) to dissect the relative influence of three underlying processes: assimilate supply (S), metabolic transformation of sugars into other compounds (M), and dilution by water uptake (D); and (ii) to estimate the genetic variability of S, M, and D. The latter was estimated in a population of 20 introgression lines derived from the introgression of a wild tomato species (Solanum chmielewskii) into S. lycopersicum, grown under two contrasted fruit load conditions. Low load systematically decreased D in the whole population, while S and M were targets of genotype × fruit load interactions. The sugar concentration positively correlated to S and D when the variation was due to genetic introgressions, while it positively correlated to S and M when the variation was due to changes in fruit load. Co-localizations between quantitative trait loci (QTLs) for sugar concentration and QTLs for S, M, and D allowed hypotheses to be proposed on the processes putatively involved at the QTLs. Among the five QTLs for sugar concentration, four co-localized with QTLs for S, M, and D with similar allele effects. Moreover, the processes underlying QTLs for sugar accumulation changed according to the fruit load condition. Finally, for some genotypes, the processes underlying sugar concentration compensated in such a way that they did not modify the sugar concentration. By uncoupling genetic from physiological relationships between processes, these results provide new insights into further understanding of tomato fruit sugar accumulation.

Effects of water deficit on leaves and fruit quality during the development period in tomato plant.

In nature, plants are often exposed to a multitude of environmental constraints that severely limit crop productivity. Water deficit is one of the factors that most affects agricultural production. The aim of this work is to evaluate the effect of water deficit on morphology, development, nutritional behavior, as well as chlorophyll fluorescence and certain important metabolic parameters (soluble sugars, organic acids, starch, carotenoid, and vitamin C) of the cultivated tomato (Solanum lycopersicum cv Plovdiv). In this study, the water supply was reduced by 60% compared to control conditions. The conditions of water deficit showed that the size of the different organs (leaves, fruits) was reduced. A reduction in the number, width, and length of the leaves, respectively, 9%, 36%, and 37%, then the leaf surface was also observed. Reduction of fluorescence (Fo, Fm, and Fv) and total index performance were among the other symptoms of plants with water deficiency. For fruit, we observed a significant decrease in diameter, fresh weight, and moisture content during the cell division period, the cell expansion period, and the fruit ripening period. In contrast, the composition of the Plovdiv fruit changed only during cell division and expansion phase. On the other hand, the water deficit induces an increase in the total carotenoid and vitamin C content of the fruits.. Besides, water deficit induced a reduction of fruit size, moisture content, and production dry matter during different phases of development. Decrease levels of soluble sugars and organic acid but increase in vitamin C and carotenoid content.