David Page

Tomato fruit ascorbic acid content is linked with monodehydroascorbate reductase activity and tolerance to chilling stress

Quantitative trait loci (QTL) mapping is a step towards the identification of factors regulating traits such as fruit ascorbic acid content. A previously identified QTL controlling variations in tomato fruit ascorbic acid has been fine mapped and reveals that the QTL has a polygenic and epistatic architecture. A monodehydroascorbate reductase (MDHAR) allele is a candidate for a proportion of the increase in fruit ascorbic acid content. The MDHAR enzyme is active in different stages of fruit ripening, shows increased activity in the introgression lines containing the wild-type (Solanum pennellii) allele, and responds to chilling injury in tomato along with the reduced/oxidized ascorbate ratio. Low temperature storage of different tomato introgression lines with all or part of the QTL for ascorbic acid and with or without the wild MDHAR allele shows that enzyme activity explains 84% of the variation in the reduced ascorbic acid levels of tomato fruit following storage at 4 degrees C, compared with 38% at harvest under non-stress conditions. A role is indicated for MDHAR in the maintenance of ascorbate levels in fruit under stress conditions. Furthermore, an increased fruit MDHAR activity and a lower oxidation level of the fruit ascorbate pool are correlated with decreased loss of firmness because of chilling injury.

GDP‐d‐mannose 3,5‐epimerase (GME) plays a key role at the intersection of ascorbate and non‐cellulosic cell‐wall biosynthesis in tomato

The GDP-D-mannose 3,5-epimerase (GME, EC 5.1.3.18), which converts GDP-d-mannose to GDP-l-galactose, is generally considered to be a central enzyme of the major ascorbate biosynthesis pathway in higher plants, but experimental evidence for its role in planta is lacking. Using transgenic tomato lines that were RNAi-silenced for GME, we confirmed that GME does indeed play a key role in the regulation of ascorbate biosynthesis in plants. In addition, the transgenic tomato lines exhibited growth defects affecting both cell division and cell expansion. A further remarkable feature of the transgenic plants was their fragility and loss of fruit firmness. Analysis of the cell-wall composition of leaves and developing fruit revealed that the cell-wall monosaccharide content was altered in the transgenic lines, especially those directly linked to GME activity, such as mannose and galactose. In agreement with this, immunocytochemical analyses showed an increase of mannan labelling in stem and fruit walls and of rhamnogalacturonan labelling in the stem alone. The results of MALDI-TOF fingerprinting of mannanase cleavage products of the cell wall suggested synthesis of specific mannan structures with modified degrees of substitution by acetate in the transgenic lines. When considered together, these findings indicate an intimate linkage between ascorbate and non-cellulosic cell-wall polysaccharide biosynthesis in plants, a fact that helps to explain the common factors in seemingly unrelated traits such as fruit firmness and ascorbate content.

Changes in volatiles and glycosides during fruit maturation of two contrasted tomato (Solanum lycopersicum) lines.

The relationship between fruit maturation and volatile contents was investigated in two contrasted Cervil (CER) and Levovil (LEV) tomato ( Solanum lycopersicum ) lines. As fruits ripened, their volatile contents mainly increased. Although some compounds displayed contrasting patterns, overall, volatiles were clearly more abundant and conferred stronger aromas to CER than to LEV fruits. This intervarietal difference in volatile contents yielding much lower volatile contents in LEV was further investigated to determine whether it is due to a higher capacity of volatile glycosylation within LEV as compared to CER. Again, glycosides mainly increased during fruit maturation and were more abundant within CER than within LEV. Overall glycoside findings were indicative of a superior capacity to biosynthesize rather than an inferior capacity to glycosylate volatiles of CER. Eugenol and 2-methoxyphenol volatiles were exceptional compounds as they remained at higher levels in maturing LEV than in CER. 2-Methylthioacetaldehyde was for the first time identified as putatively related to differences of aroma between lines, as it was abundant in Cervil but absent in Levovil. Considering the described odor value of these three products, they should contribute differently to the particular olfactive features of LEV and CER fruits.

Kinetics of temperature increase during tomato processing modulate the bioaccessibility of lycopene

The nutritional benefit of bioactive metabolites depends on their bioavailability, i.e. the proportion that leaves the food matrix, and crosses the enteral barrier to reach their cellular target. The present study focused on lycopene, the major and bioactive tomato carotenoid, the bioavailability of which is known to be enhanced in cooked products. To better understand how processing may facilitate lycopene release, we assessed whether hot-break (HB) or cold-break (CB) treatments influence the tomato lycopene bioaccessibility. HB and CB are used in the tomato industry to modulate texture of purees through endogenous cell-wall lytic enzymes activity. HB and CB processes were mimicked through microwave heating, leading to a differentiated temperature rise in the product. The HB and CB models led to the expected differences, i.e. more viscous puree for HB with low methanol. The ability of the tomato matrix to release lycopene was measured as the extractability of lycopene to oil under standardized mixing conditions. We expected that CB treatment, by enhancing cell-wall degradation, would lead to enhanced lycopene bioaccessibility. The opposite was observed: oil contained three times less lycopene when mixed by CB (around 0.3 μgml(-1); similar to results obtained with fresh purees) than when mixed by HB (around 0.9 μgml(-1)), although HB caused more lycopene degradation. Kinetics studies indicated that the quick rise of temperature at the beginning of HB treatment was a key parameter.

Physicochemical parameters that influence carotenoids bioaccessibility from a tomato juice

In vitro digestion models have been developed to estimate carotenoid bioavailability but most do not consider that their diffusion from fruit matrix to the lipid phase of the bolus could be a limiting step. Therefore we designed a model in which tomato juice is mixed with oil or oil/water emulsions, and the carotenoids diffusing to oil are measured by spectrometry. Temperature, pH and tomato juice/peanut oil ratio were evaluated for their influence on carotenoid diffusion. When oil/tomato ratio was between 0.11 and 1, extraction of lycopene was limited by the saturation of the oil phase. With a large excess of oil, diffusion was also limited, as only 31 ± 1% of lycopene could be extracted from the juice. Diffusion did not vary significantly with pH but doubled when temperature rose from 10°C to 37°C. When the juice was mixed in an emulsion stabilised with bovine serum albumin or phospholipids the maximum extraction decreased to 14.5 ± 0.2% and 18.5 ± 1.5% respectively, indicating that in addition to the saturation of the oil phase at low oil/tomato ratio and in addition to intrinsic properties of the tomato juice in non-saturating conditions, lycopene diffusion was limited by the structure of the interface in emulsions.

Combined Effects of Irrigation Regime, Genotype, and Harvest Stage Determine Tomato Fruit Quality and Aptitude for Processing into Puree

Industry tomatoes are produced under a range of climatic conditions and practices which significantly impact on main quality traits of harvested fruits. However, the quality of tomato intended for processing is currently addressed on delivery through color and Brix only, whereas other traits are overlooked. Very few works provided an integrated view of the management of tomato puree quality throughout the chain. To gain insights into pre- and post-harvest interactions, four genotypes, two water regimes, three maturity stages, and two processes were investigated. Field and glasshouse experiments were conducted near Avignon, France, from May to August 2016. Two irrigation regimes were applied: control plants were irrigated in order to match 100% of evapotranspiration (ETP); water deficit (WD) plants were irrigated as control plants until anthesis of the first flowers, then irrigation was reduced to 60 and 50% ETP in field, and glasshouse respectively. Fruits were collected at three stages during ripening. Their color, fresh weight, dry matter content, and metabolite contents were determined before processing. Pericarp cell size was evaluated in glasshouse only. Two laboratory-scaled processing methods were applied before structural and biochemical analyses of the purees. Results outlined interactive effects between crop and process management. WD hardly reduced yield, but increased dry matter content in the field, in contrast to the glasshouse. The puree viscosity strongly depended on the genotype and the maturity stage, but it was disconnected from fruit dry matter content or Brix. The process impact on puree viscosity strongly depended on water supply during fruit production. Moreover, the lycopene content of fresh fruit may influence puree viscosity. This work opens new perspectives for managing puree quality in the field showing that it was possible to reduce water supply without affecting yield and to improve puree quality.

Increased diffusivity of lycopene in hot break vs. cold break purees may be due to bioconversion of associated phospholipids rather than differential destruction of fruit tissues or cell structures

Lycopene bioaccessibility is enhanced by processing, as explained by the destructuration of plant tissues, making diffusion easier. However, in tomato, the relationship between grinding intensity and lycopene release from purees suffers from uncertainty. In particular, hot break puree exhibited twice as much diffusible lycopene as compared to cold break, while both were processed with the same grinding intensity. To explain the difference, we systematically studied the diffusivity of particles according to their size and integrity, and used microscopic and physical analyses to reveal structural differences. Neither particle size distribution, nor cell destruction, nor plastid transformation exhibited any correlation to the differences in diffusivity. However, Raman microspectroscopy combined with a chemometric analysis revealed significant changes in lycopene spectra and a putative linkage to phospholipid transformation. Phospholipid profiling of five pairs of contrasted purees revealed that, during the cold break, a transition from complex phospholipids to more simple phosphatidic acid molecules systematically occurred.

A Systems Biology Study in Tomato Fruit Reveals Correlations between the Ascorbate Pool and Genes Involved in Ribosome Biogenesis, Translation, and the Heat-Shock Response

Changing the balance between ascorbate, monodehydroascorbate, and dehydroascorbate in plant cells by manipulating the activity of enzymes involved in ascorbate synthesis or recycling of oxidized and reduced forms leads to multiple phenotypes. A systems biology approach including network analysis of the transcriptome, proteome and metabolites of RNAi lines for ascorbate oxidase, monodehydroascorbate reductase and galactonolactone dehydrogenase has been carried out in orange fruit pericarp of tomato (Solanum lycopersicum). The transcriptome of the RNAi ascorbate oxidase lines is inversed compared to the monodehydroascorbate reductase and galactonolactone dehydrogenase lines. Differentially expressed genes are involved in ribosome biogenesis and translation. This transcriptome inversion is also seen in response to different stresses in Arabidopsis. The transcriptome response is not well correlated with the proteome which, with the metabolites, are correlated to the activity of the ascorbate redox enzymes—ascorbate oxidase and monodehydroascorbate reductase. Differentially accumulated proteins include metacaspase, protein disulphide isomerase, chaperone DnaK and carbonic anhydrase and the metabolites chlorogenic acid, dehydroascorbate and alanine. The hub genes identified from the network analysis are involved in signaling, the heat-shock response and ribosome biogenesis. The results from this study therefore reveal one or several putative signals from the ascorbate pool which modify the transcriptional response and elements downstream.