Comparative constraint-based modelling of fruit development across species highlights nitrogen metabolism in the growth-defence trade-off.

Although primary metabolism is well conserved across species, it is useful to explore the specificity of its network to assess the extent to which some pathways may contribute to particular outcomes. Constraint-based metabolic modelling is an established framework for predicting metabolic fluxes and phenotypes and helps to explore how the plant metabolic network delivers specific outcomes from temporal series. After describing the main physiological traits during fruit development, we confirmed the correlations between fruit relative growth rate (RGR), protein content and time to maturity. Then a constraint-based method is applied to a panel of eight fruit species with a knowledge-based metabolic model of heterotrophic cells describing a generic metabolic network of primary metabolism. The metabolic fluxes are estimated by constraining the model using a large set of metabolites and compounds quantified throughout fruit development. Multivariate analyses showed a clear common pattern of flux distribution during fruit development with differences between fast- and slow-growing fruits. Only the latter fruits mobilise the tricarboxylic acid cycle in addition to glycolysis, leading to a higher rate of respiration. More surprisingly, to balance nitrogen, the model suggests, on the one hand, nitrogen uptake by nitrate reductase to support a high RGR at early stages of cucumber and, on the other hand, the accumulation of alkaloids during ripening of pepper and eggplant. Finally, building virtual fruits by combining 12 biomass compounds shows that the growth-defence trade-off is supported mainly by cell wall synthesis for fast-growing fruits and by total polyphenols accumulation for slow-growing fruits.

Metabolic profiles in C3, C3-C4 intermediate, C4-like, and C4 species in the genus Flaveria.

C4 photosynthesis concentrates CO2 around Rubisco in the bundle sheath, favouring carboxylation over oxygenation and decreasing photorespiration. This complex trait evolved independently in >60 angiosperm lineages. Its evolution can be investigated in genera such as Flaveria (Asteraceae) that contain species representing intermediate stages between C3 and C4 photosynthesis. Previous studies have indicated that the first major change in metabolism probably involved relocation of glycine decarboxylase and photorespiratory CO2 release to the bundle sheath and establishment of intercellular shuttles to maintain nitrogen stoichiometry. This was followed by selection for a CO2-concentrating cycle between phosphoenolpyruvate carboxylase in the mesophyll and decarboxylases in the bundle sheath, and relocation of Rubisco to the latter. We have profiled 52 metabolites in nine Flaveria species and analysed 13CO2 labelling patterns for four species. Our results point to operation of multiple shuttles, including movement of aspartate in C3-C4 intermediates and a switch towards a malate/pyruvate shuttle in C4-like species. The malate/pyruvate shuttle increases from C4-like to complete C4 species, accompanied by a rise in ancillary organic acid pools. Our findings support current models and uncover further modifications of metabolism along the evolutionary path to C4 photosynthesis in the genus Flaveria.

Ascophyllum nodosum Based Extracts Counteract Salinity Stress in Tomato by Remodeling Leaf Nitrogen Metabolism.

Biostimulants have rapidly and widely been adopted as growth enhancers and stress protectants in agriculture, however, due to the complex nature of these products, their mechanism of action is not clearly understood. By using two algal based commercial biostimulants in combination with the Solanum lycopersicum cv. MicroTom model system, we assessed how the modulation of nitrogen metabolites and potassium levels could contribute to mediate physiological mechanisms that are known to occur in response to salt/and or osmotic stress. Here we provide evidence that the reshaping of amino acid metabolism can work as a functional effector, coordinating ion homeostasis, osmotic adjustment and scavenging of reactive oxygen species under increased osmotic stress in MicroTom plant cells. The Superfifty biostimulant is responsible for a minor amino acid rich-phenotype and could represent an interesting instrument to untangle nitrogen metabolism dynamics in response to salinity and/or osmotic stress.

Biostimulation as a Means for Optimizing Fruit Phytochemical Content and Functional Quality of Tomato Landraces of the San Marzano Area.

The effect of plant biostimulation on fruits of traditional tomato germplasm is largely unknown. We examined how a tropical plant-derived biostimulant impacts the nutritional, functional, and compositional characteristics of tomato fruits from four landraces, collected in the San Marzano (SM) tomato Protected Designation of Origin (PDO) region, by profiling primary and secondary metabolites. Biostimulation was not able to completely reshuffle the morpho-physiological and nutritional profile of the four landraces. Their distinct phytochemical profile indicated a genotype-specific tuning of the analyzed traits, which also included an improved yield and fruit quality. Biostimulation of SM1 and SM3 increased photosynthetic accumulation of carbohydrate reserves, improved mineral nutrient use efficiency and consequently, yield (+21% and 34%, respectively). Moreover, biostimulation augmented the nutraceutical properties of the SM2 landrace. Interestingly, the plant-derived product increased in all genotypes lycopene, but not polyphenol accumulation in fruits. Our results show the potential of biostimulatory applications towards optimizing the fruit quality of the acclaimed SM landraces, which is suitable to satisfy both the rising consumer demand for premium traditional tomatoes and the technological needs of the food industry.

Salinity Duration Differently Modulates Physiological Parameters and Metabolites Profile in Roots of Two Contrasting Barley Genotypes.

Hordeum maritimum With. is a wild salt tolerant cereal present in the saline depressions of the Eastern Tunisia, where it significantly contributes to the annual biomass production. In a previous study on shoot tissues it was shown that this species withstands with high salinity at the seedling stage restricting the sodium entry into shoot and modulating over time the leaf synthesis of organic osmolytes for osmotic adjustment. However, the tolerance strategy mechanisms of this plant at root level have not yet been investigated. The current research aimed at elucidating the morphological, physiological and biochemical changes occurring at root level in H. maritimum and in the salt sensitive cultivar Hordeum vulgare L. cv. Lamsi during five-weeks extended salinity (200 mM NaCl), salt removal after two weeks of salinity and non-salt control. H. maritimum since the first phases of salinity was able to compartmentalize higher amounts of sodium in the roots compared to the other cultivar, avoiding transferring it to shoot and impairing photosynthetic metabolism. This allowed the roots of wild plants to receive recent photosynthates from leaves, gaining from them energy and carbon skeletons to compartmentalize toxic ions in the vacuoles, synthesize and accumulate organic osmolytes, control ion and water homeostasis and re-establish the ability of root to grow. H. vulgare was also able to accumulate compatible osmolytes but only in the first weeks of salinity, while soon after the roots stopped up taking potassium and growing. In the last week of salinity stress, the wild species further increased the root to shoot ratio to enhance the root retention of toxic ions and consequently delaying the damages both to shoot and root. This delay of few weeks in showing the symptoms of stress may be pivotal for enabling the survival of the wild species when soil salinity is transient and not permanent.

Metabolic Profile and Performance Responses of Ranunculus asiaticus L. Hybrids as Affected by Light Quality of Photoperiodic Lighting.

Ranunculus asiaticus is a quantitative long day plant grown for cut flowers and flowering potted plants production. We evaluated the influence of light spectrum of three light sources for end-of-day photoperiodic treatments, with different phytochrome photoequilibria (PPE) induced at plant level, on the metabolic profiling of two hybrids of R. asiaticus L., MBO and MDR, in plants from vernalized tuberous roots. The following treatments were compared with natural day length (NL): white fluorescence lamp (FL, PPE 0.84), light emitting diodes (LEDs) Red:Far Red light at 3:1 ratio (R:FR 3:1, PPE 0.84), and LEDs Red:Far Red light at 1:3 ratio (R:FR 1:3, PPE 0.63). Measurements were carried out to evaluate the time course of carbohydrate, amino acid, and protein levels throughout the growing cycle in tuberous roots and leaves, in relation to the different plant stages (pre-planting, vegetative phase, and flowering). The study of metabolic profiling suggested that the differences between the tuberous root reserves of the two R. asiaticus hybrids could be responsible for the capacity of MBO to exert an early flowering. In particular, the proton-consuming synthesis during the pre-planting of two amino acids, alanine and γ-aminobutyric acid (GABA), is able to buffer the cytoplasmic acidosis and pH altered by the vernalization process, and GABA itself can efficiently scavenge reactive oxygen species. This fast response to the stress caused by vernalization allows MBO plants to accelerate the process of vegetative development and flowering. Some other changes in metabolites profile were certainly related to the different responses to day length and photoperiodic light quality in the two hybrids, such as dose exerted by low R:FR lighting in both MBO and MDR. However, most of the responses are under a strict genetic control.

The role of light quality of photoperiodic lighting on photosynthesis, flowering and metabolic profiling in Ranunculus asiaticus L.

Photoperiodic light quality affects flowering of long day plants, by influencing the phytochrome photoequilibria (PPE) at plant level; however, the most effective light spectrum to promote flowering is still unknown for most of the flower crops. We evaluated the influence of light spectrum of three light sources, with different induced PPE, on photosynthesis, metabolic profiling, plant growth and flowering in two hybrids of Ranunculus asiaticus L., MBO (early flowering) and MDR (medium earliness). Three photoperiodic treatments were compared to natural day length (NL): white fluorescent light (PPE 0.84), light emitting diodes (LEDs) with red:far red (R:FR) light at 3:1 ratio (PPE, 0.84) and LEDs with R:FR light at 1:3 ratio (PPE 0.63). Under natural light, net photosynthesis was higher in MDR than in MBO, while photochemistry was similar in the hybrids. Compared to NL, photoperiodic treatments did not affect net photosynthesis, while they promoted the quantum yield of PSII and reduced the non-photochemical quenching. Under NL, plant growth was greater in MBO, while flowering started earlier in MDR and flowers characteristics were similar in the hybrids. Despite the greater sensitivity of MDR plants in terms of metabolism, photoperiodic lighting improved plant growth and reduced the flowering time only in MBO, with a stronger effect under R:FR 3:1 light. MDR plants were characterized by higher soluble sugars, polyphenols, photosynthetic pigments and proteins, while MBO plants by higher starch and amino acid content. The morphological effects of photoperiodic light quality and the hybrid-specific response should be taken into account to optimize lighting protocols in commercial farms.

Omeprazole Treatment Enhances Nitrogen Use Efficiency Through Increased Nitrogen Uptake and Assimilation in Corn.

Omeprazole is a selective proton pump inhibitor in humans that inhibits the H+/K+-ATPase of gastric parietal cells. Omeprazole has been recently shown to act as a plant growth regulator and enhancer of salt stress tolerance. Here, we report that omeprazole treatment in hydroponically grown maize improves nitrogen uptake and assimilation. The presence of micromolar concentrations of omeprazole in the nutrient solution alleviates the chlorosis and growth inhibition induced by low nitrogen availability. Nitrate uptake and assimilation is enhanced in omeprazole treated plants through changes in nitrate reductase activity, primary metabolism, and gene expression. Omeprazole enhances nitrate assimilation through an interaction with nitrate reductase, altering its activation state and affinity for nitrate as a substrate. Omeprazole and its targets represent a novel method for enhancing nitrogen use efficiency in plants.

Sensory and functional quality characterization of protected designation of origin 'Piennolo del Vesuvio' cherry tomato landraces from Campania-Italy.

Compositional characterization was performed on seven 'Pomodorino del Piennolo del Vesuvio' (PPV) tomato landraces, a signature product of Campania (Italy) threatened by genetic erosion. Characterization encompassed determinations of macro-minerals, soluble carbohydrates, starch, acidity, lycopene, polyphenols, anthocyanins, protein and free amino acids. Exceptionally high dry matter (13.0 ±â€¯0.2%) and sugar content (101.3 ±â€¯3.8 µmol g-1 fw) and very low (0.007-0.009) Na/K ratio were invariably obtained across landraces, contrasted by significant variation in acidity (28.5-3.9 g kg-1 dw). Giagiù, Acampora and Riccia San Vito differentiated by high polyphenols content (131.8 ±â€¯2.5 mg 100 g-1) while Acampora, Cozzolino and Fofò by high lycopene content (13.3 ±â€¯10.6 mg 100 g-1 fw). Glutamate, GABA and glutamine represented 65% of the 22 detected amino acids mean total content. Glutamate, linked to umami taste, was highest (19.2 µmol g-1 fw) in Fofò. Our results will contribute towards the systematic documentation of sensory and functional quality profiles of an important collection of tomato landraces.

Spatial and Temporal Profile of Glycine Betaine Accumulation in Plants Under Abiotic Stresses.

Several halophytes and a few crop plants, including Poaceae, synthesize and accumulate glycine betaine (GB) in response to environmental constraints. GB plays an important role in osmoregulation, in fact, it is one of the main nitrogen-containing compatible osmolytes found in Poaceae. It can interplay with molecules and structures, preserving the activity of macromolecules, maintaining the integrity of membranes against stresses and scavenging ROS. Exogenous GB applications have been proven to induce the expression of genes involved in oxidative stress responses, with a restriction of ROS accumulation and lipid peroxidation in cultured tobacco cells under drought and salinity, and even stabilizing photosynthetic structures under stress. In the plant kingdom, GB is synthesized from choline by a two-step oxidation reaction. The first oxidation is catalyzed by choline monooxygenase (CMO) and the second oxidation is catalyzed by NAD+-dependent betaine aldehyde dehydrogenase. Moreover, in plants, the cytosolic enzyme, named N-methyltransferase, catalyzes the conversion of phosphoethanolamine to phosphocholine. However, changes in CMO expression genes under abiotic stresses have been observed. GB accumulation is ontogenetically controlled since it happens in young tissues during prolonged stress, while its degradation is generally not significant in plants. This ability of plants to accumulate high levels of GB in young tissues under abiotic stress, is independent of nitrogen (N) availability and supports the view that plant N allocation is dictated primarily to supply and protect the growing tissues, even under N limitation. Indeed, the contribution of GB to osmotic adjustment and ionic and oxidative stress defense in young tissues, is much higher than that in older ones. In this review, the biosynthesis and accumulation of GB in plants, under several abiotic stresses, were analyzed focusing on all possible roles this metabolite can play, particularly in young tissues.

Hordeum vulgare and Hordeum maritimum respond to extended salinity stress displaying different temporal accumulation pattern of metabolites.

Hordeum maritimum With. (= H. marinum Huds. subsp. marinum, 2n=14) is a wild cereal present in the saline depressions of the Soliman and Kelbia Sebkhas, which contributes significantly to annual biomass production in Tunisia. This species is able to tolerate high NaCl concentrations at the seedling stage without showing symptoms of toxicity; however, the tolerance strategy mechanisms of this plant have not yet been unravelled. Our metabolite analysis, performed on leaves of H. maritimum during extended stress in comparison with Hordeum vulgare L. cv. Lamsi, has revealed an adaptive response of the wild species based on a different temporal accumulation pattern of ions and compatible metabolites. Further, wild and cultivated genotypes with contrasting salt-tolerant behaviour display different pattern of metabolites when salt stress is prolonged over 2 weeks. In particular, when exposed to up to 3 weeks of 200mM NaCl salt stress, H. maritimum is able to maintain lower leaf concentrations of sodium and chloride, and higher concentrations of potassium compared with H. vulgare. This likely restricts sodium entry into plants at the root level, and uses the toxic ions, glycine betaine and low levels of proline for osmotic adjustment. Under prolonged stress, the accumulation of proline increases, reaching the highest levels in concomitance with the decrease of potassium to sodium ratio, the increase of hydrogen peroxide and decrease of chlorophylls. The modulation of proline accumulation over time can be interpreted as an adaptive response to long-term salinity. Moreover, once synthetised glycine betaine is transported but not metabolised, it can contribute together with proline to osmotically balance H. maritimum leaves and protect them from oxidative stress. The 2-3 week delay of H. maritimum in showing the symptoms of stress and damages compared with H. vulgare could be important in the survival of plants when soil salinity is not a permanent condition, but just a transient state of stress.

Effect of Thermal Stress on Tissue Ultrastructure and Metabolite Profiles During Initiation of Radiata Pine Somatic Embryogenesis.

Climate change will inevitably lead to environmental variations, thus plant drought tolerance will be a determinant factor in the success of plantations and natural forestry recovery. Some metabolites, such as soluble carbohydrates and amino acids, have been described as being the key to both embryogenesis efficiency and abiotic stress response, contributing to phenotypic plasticity and the adaptive capacity of plants. For this reason, our main objectives were to evaluate if the temperature during embryonal mass initiation in radiata pine was critical to the success of somatic embryogenesis, to alter the morphological and ultrastructural organization of embryonal masses at cellular level and to modify the carbohydrate, protein, or amino acid contents. The first SE initiation experiments were carried out at moderate and high temperatures for periods of different durations prior to transfer to the control temperature of 23°C. Cultures initiated at moderate temperatures (30°C, 4 weeks and 40°C, 4 days) showed significantly lower initiation and proliferation rates than those at the control temperature or pulse treatment at high temperatures (50°C, 5 min). No significant differences were observed either for the percentage of embryogenic cell lines that produced somatic embryos, or for the number of somatic embryos per gram of embryonal mass. Based on the results from the first experiments, initiation was carried out at 40°C 4 h; 50°C, 30 min; and a pulse treatment of 60°C, 5 min. No significant differences were found for the initiation or number of established lines or for the maturation of somatic embryos. However, large morphological differences were observed in the mature somatic embryos. At the same time, changes observed at cellular level suggested that strong heat shock treatments may trigger the programmed cell death of embryogenic cells, leading to an early loss of embryogenic potential, and the formation of supernumerary suspensor cells. Finally, among all the differences observed in the metabolic profile, it is worth highlighting the accumulation of tyrosine and isoleucine, both amino acids involved in the synthesis of abiotic stress response-related secondary metabolites.

Durum wheat seedling responses to simultaneous high light and salinity involve a fine reconfiguration of amino acids and carbohydrate metabolism.

Durum wheat plants are extremely sensitive to drought and salinity during seedling and early development stages. Their responses to stresses have been extensively studied to provide new metabolic targets and improving the tolerance to adverse environments. Most of these studies have been performed in growth chambers under low light [300-350 µmol m-2 s-1 photosynthetically active radiation (PAR), LL]. However, in nature plants have to face frequent fluctuations of light intensities that often exceed their photosynthetic capacity (900-2000 µmol m-2 s-1 ). In this study we investigated the physiological and metabolic changes potentially involved in osmotic adjustment and antioxidant defense in durum wheat seedlings under high light (HL) and salinity. The combined application of the two stresses decreased the water potential and stomatal conductance without reducing the photosynthetic efficiency of the plants. Glycine betaine (GB) synthesis was inhibited, proline and glutamate content decreased, while γ-aminobutyric acid (GABA), amides and minor amino acids increased. The expression level and enzymatic activities of Δ1-pyrroline-5-carboxylate synthetase, asparagine synthetase and glutamate decarboxylase, as well as other enzymatic activities of nitrogen and carbon metabolism, were analyzed. Antioxidant enzymes and metabolites were also considered. The results showed that the complex interplay seen in durum wheat plants under salinity at LL was simplified: GB and antioxidants did not play a main role. On the contrary, the fine tuning of few specific primary metabolites (GABA, amides, minor amino acids and hexoses) remodeled metabolism and defense processes, playing a key role in the response to simultaneous stresses.