Next generation chemical priming: with a little help from our nanocarrier friends.

Plants are exposed to multiple threats linked to climate change which can cause critical yield losses. Therefore, designing novel crop management tools is crucial. Chemical priming has recently emerged as an effective technology for improving tolerance to stress factors. Several compounds such as phytohormones, reactive species, and synthetic chimeras have been identified as promising priming agents. Following remarkable developments in nanotechnology, several unique nanocarriers (NCs) have been engineered that can act as smart delivery systems. These provide an eco-friendly, next-generation method for chemical priming, leading to increased efficiency and reduced overall chemical usage. We review novel engineered NCs (NENCs) as vehicles for chemical agents in advanced priming strategies, and address challenges and opportunities to be met towards achieving sustainable agriculture.

Assessing the biodiversity and the impact of pollinators on carob production.

As the current climate crisis intensifies, drought resistant crops are becoming more important due to their ability to withstand the increasingly hotter and drier summers. Such crops are valuable for pollinators as they provide food resources for wild and managed species. The carob tree (Ceratonia siliqua L.) represents an example of a heat- and drought- resistant crop, able to grow in dry areas with practically no inputs. The current study assessed over two growing seasons the diversity of wild bees and other pollinators relying on carob flowers, as well as the contribution of animal pollination to carob production. Carob flowers were subjected to two treatments: Open pollination, where inflorescences were left untreated, and wind pollination, where inflorescences were bagged in a mesh during blooming. Weekly observations during blooming showed that Apis mellifera was the most frequent floral visitor followed by wild bees and wasps. Carob flowers were visited by at least 10 different wild bee species. Open-pollinated flowers produced significantly more pods, with the benefit ranging from 4 to 16 times higher production, depending on the region. Open pollination led to pods with greater weight, length and number of seeds compared to pods derived from wind pollination. The results of the current study highlight the importance of animal pollination to carob production, as well as the significance of carob trees to wild bee conservation.

The beneficial effect of leaf removal during fruit set on physiological, biochemical, and qualitative indices and volatile organic compound profile of the Cypriot reference cultivar 'Xynisteri'.

BACKGROUND: 'Xynisteri' is the reference Cypriot white cultivar that, despite its significant societal and economic impact, is poorly characterized regarding its qualitative properties, while scarce information exists regarding its aroma profile. In the current study, the effect of leaf removal during fruit set (BBCH 71) on 6-year cordon-trained, spur-pruned grapevines was assessed and an array of physiological, biochemical, and qualitative indices were monitored during successive developmental stages (BBCH 75, BBCH 85, BBCH 87, and BBCH 89). Grapes were additionally monitored for the volatile organic compounds (VOCs) profile during the advanced on-vine developmental stages (BBCH 85-BBCH 89) with the employment of gas chromatography-mass spectrometry (GC-MS), Fourier-transform near infrared (FT-NIR) spectra and electronic nose (E-nose) techniques. RESULTS: Grape berries from the vines subjected to leaf removal were characterized by higher solid soluble sugars (SSC), titratable acidity (TA), tartaric acid, and ammonium nitrogen contents, while this was not the case for assimilable amino nitrogen (primary amino nitrogen). A total of 75 compounds were identified and quantified, including aliphatic alcohols, benzenic compounds, phenols, vanillins, monoterpenes, and C13 -norisoprenoids. Leaf removal led to enhanced amounts of glycosylated aroma compounds, mainly monoterpenes, and C13 -norisoprenoids. Chemometric analysis, used through FT-NIR and E-nose, showed that the aromatic patterns detected were well associated to the grape ripening trend and differences between leaf removal-treated and control grapes were detectable during fully ripe stage. CONCLUSION: Leaf removal at fruit set resulted in an overall induction of secondary metabolism, with special reference to glycosylated aroma compounds, namely monoterpenes and C13 -norisoprenoids. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A comprehensive review on carotenoids in foods and feeds: status quo, applications, patents, and research needs.

Carotenoids are isoprenoids widely distributed in foods that have been always part of the diet of humans. Unlike the other so-called food bioactives, some carotenoids can be converted into retinoids exhibiting vitamin A activity, which is essential for humans. Furthermore, they are much more versatile as they are relevant in foods not only as sources of vitamin A, but also as natural pigments, antioxidants, and health-promoting compounds. Lately, they are also attracting interest in the context of nutricosmetics, as they have been shown to provide cosmetic benefits when ingested in appropriate amounts. In this work, resulting from the collaborative work of participants of the COST Action European network to advance carotenoid research and applications in agro-food and health (EUROCAROTEN, www.eurocaroten.eu, https://www.cost.eu/actions/CA15136/#tabs|Name:overview) research on carotenoids in foods and feeds is thoroughly reviewed covering aspects such as analysis, carotenoid food sources, carotenoid databases, effect of processing and storage conditions, new trends in carotenoid extraction, daily intakes, use as human, and feed additives are addressed. Furthermore, classical and recent patents regarding the obtaining and formulation of carotenoids for several purposes are pinpointed and briefly discussed. Lastly, emerging research lines as well as research needs are highlighted.

Elaboration of novel and comprehensive protocols toward determination of textural properties and other sensorial attributes of canning peach fruit.

Peach (Prunus persica) products are destined for fresh consumption or are being consumed after processing in various forms. Despite its huge economic importance, no standardized protocols to define sensorial attributes and mechanical properties of canned peaches exist. Thus, the aim of the current study was dual and included the setting up of a list of sensorial descriptors and the elaboration of a toolkit to evaluate the textural properties of canned peaches using large deformation mechanical testing. A standardized vocabulary ("consensus language") was initially developed toward the determination and quantification of 15 sensorial attributes through a descriptive quantitative analysis (QDA) approach. Textural properties were additionally evaluated with a TA-XT Plus texture analyzer by applying three discrete large deformation tests [(a) puncture test with a flat cylindrical probe; (b) texture profile analysis (TPA) with a flat compression plunger; and (c) Kramer shear test (KST) cell with a bladed fixture]; that is, a total of nine textural properties, namely, "puncture firmness" (individual halves), "Kramer" hardness (applied in a complex mixture of peach slices), "TPA" hardness (central section of halves), fracturability, consistency, cohesiveness, springiness, chewiness, and total hardness were assessed. We hereby present novel protocols that encompass the comprehensive determination of sensorial and textural properties. The established protocols, providing complementary information, are readily applicable to the canning industry in setting up qualitative tests to determine product shelf life as well as to assist on going breeding programs for the evaluation of new candidate clingstone cultivars destined for canning purposes.

Dissection of the incidence and severity of purple spot physiological disorder in loquat fruit through a physiological and molecular approach.

Loquat (Eriobotrya japonica) fruit marketability is affected by the incidence and severity of purple spot (PS), a pre-harvest physiological disorder showing an evident skin discoloration with depressed surface. Despite its impact in limiting the cultivation and economic potential of loquat, the etiology of this disorder is still poorly understood. To this end, our study aimed to investigate and disclose possible mechanisms underlying PS development. The intensity and severity of PS in three loquat cultivars ('Morphitiki', 'Karantoki' and 'Obusa') was phenotypically monitored during successive on-tree fruit developmental stages. 'Obusa' fruits harvested at commercial maturity stage showed the highest incidence of purple spot (58.6%), while 'Morphitiki' fruits did not show any symptoms. 'Karantoki' fruits demonstrated an intermediate response, with 31.3% of the fruit being affected. Thereafter, fruits with 30-50% PS severity were selected and used for further analysis; peel tissue was removed from both symptomatic and asymptomatic tissue of the same fruit for all examined cultivars. 'Karantoki' fruit with PS were characterized by the highest accumulation of total soluble sugars, sucrose, glucose and fructose contents, while the concentration of these primary metabolites was the lowest in asymptomatic fruit of 'Obusa', exception made for the sucrose. The incidence of PS was also transcriptionally investigated by assessing the mRNA profile of important genes involved in polyphenolic (PAL1, PAL2 and PPO1) and carbohydrate (CWI2, CWI3, SPS1, SPS2, NI2, NI3, SuSy, HXK, FRK and VI) pathway. The enhanced expression levels of CWI3 and VI genes in symptomatic fruit of the highly susceptible cultivar 'Obusa' highlight a cultivar-specific type of response. Notably, SuSy registered significantly suppressed levels in symptomatic tissue of both 'Obusa' and 'Karantoki'. To what extent PPO is associated with PS incidence and whether the etiology of the disorder can be assigned to an oxidative process triggered and coordinated by its action need to be further elucidated. The aforementioned genes are suggested to be further examined as potential markers towards a more sophisticated and informed characterization of purple spot detection in loquat fruit.

Primary Metabolism in Fresh Fruits During Storage.

The extension of commercial life and the reduction of postharvest losses of perishable fruits is mainly based on storage at low temperatures alone or in combination with modified atmospheres (MAs) and controlled atmospheres (CAs), directed primarily at reducing their overall metabolism thus delaying ripening and senescence. Fruits react to postharvest conditions with desirable changes if appropriate protocols are applied, but otherwise can develop negative and unacceptable traits due to the onset of physiological disorders. Extended cold storage periods and/or inappropriate temperatures can result in development of chilling injuries (CIs). The etiology, incidence, and severity of such symptoms vary even within cultivars of the same species, indicating the genotype significance. Carbohydrates and amino acids have protective/regulating roles in CI development. MA/CA storage protocols involve storage under hypoxic conditions and high carbon dioxide concentrations that can maximize quality over extended storage periods but are also affected by the cultivar, exposure time, and storage temperatures. Pyruvate metabolism is highly reactive to changes in oxygen concentration and is greatly affected by the shift from aerobic to anaerobic metabolism. Ethylene-induced changes in fruits can also have deleterious effects under cold storage and MA/CA conditions, affecting susceptibility to chilling and carbon dioxide injuries. The availability of the inhibitor of ethylene perception 1-methylcyclopropene (1-MCP) has not only resulted in development of a new technology but has also been used to increase understanding of the role of ethylene in ripening of both non-climacteric and climacteric fruits. Temperature, MA/CA, and 1-MCP alter fruit physiology and biochemistry, resulting in compositional changes in carbon- and nitrogen-related metabolisms and compounds. Successful application of these storage technologies to fruits must consider their effects on the metabolism of carbohydrates, organic acids, amino acids and lipids.

Leaf removal at veraison stage differentially affects qualitative attributes and bioactive composition of fresh and dehydrated grapes of two indigenous Cypriot cultivars.

BACKGROUND: The effect of leaf removal on postharvest performance of dehydrated grapes has been poorly analyzed. The aim of this study was to evaluate the effect of leaf removal at veraison stage on the metabolites of fresh and dehydrated grapes of two indigenous Cypriot cultivars ('Mavro' and 'Xynisteri'), which are destined for the production of 'Commandaria', a protected designation of origin (PDO) premium sweet wine. RESULTS: Leaf removal led to a reduction of soluble solids, titratable acidity, aroma potential and most of the phenolic groups in the musts of both cultivars. Dehydration led to a significant increase in all of these parameters in both cultivars, being more pronounced in cv. 'Mavro'. Interestingly, leaf removal indicated differential response in the dehydrated product of the cultivars examined; liquid chromatography (LC-DAD-qTOF-MS) data showed a significant decrease in phenolic compounds in 'Xynisteri' must (from 66.73 to 44.15 mg L-1 ), while 'Mavro' must registered similar values of phenolic compounds (from 94.78 to 96.72 mg L-1 ), but with a different distribution among phenolic groups. Intriguingly, flavonols and flavan-3-ols that present significant health-promoting properties showed higher concentrations in the must from dehydrated 'Mavro' grapes that were subjected to preharvest leaf removal. CONCLUSIONS: Leaf removal at veraison stage followed by sun drying differentially affected the chemical composition of the examined cultivars. This preharvest application followed by postharvest dehydration was beneficial for cv. 'Mavro', while this was not the case for cv. 'Xynisteri'. The effects of leaf removal in other developmental stages (i.e. pre/post bloom stage) need to be dissected with the ultimate goal of providing an end product with high bioactive content. © 2018 Society of Chemical Industry.

Genomic Sequencing of Japanese Plum (Prunus salicina Lindl.) Mutants Provides a New Model for Rosaceae Fruit Ripening Studies.

It has recently been described that the Japanese plum "Santa Rosa" bud sport series contains variations in ripening pattern: climacteric, suppressed-climacteric and non-climacteric types. This provides an interesting model to study the role of ethylene and other key mechanisms governing fruit ripening, softening and senescence. The aim of the current study was to investigate such differences at the genomic level, using this series of plum bud sports, with special reference to genes involved in ethylene biosynthesis, signal transduction, and sugar metabolism. Genomic DNA, isolated from leaf samples of six Japanese plum cultivars ("Santa Rosa", "July Santa Rosa", "Late Santa Rosa", "Sweet Miriam", "Roysum", and "Casselman"), was used to construct paired-end standard Illumina libraries. Sequences were aligned to the Prunus persica genome, and genomic variations (SNPs, INDELS, and CNV's) were investigated. Results determined 12 potential candidate genes with significant copy number variation (CNV), being associated with ethylene perception and signal transduction components. Additionally, the Maximum Likelihood (ML) phylogenetic tree showed two sorbitol dehydrogenase genes grouping into a distinct clade, indicating that this natural group is well-defined and presents high sequence identity among its members. In contrast, the ethylene group, which includes ACO1, ACS1, ACS4, ACS5, CTR1, ERF1, ERF3, and ethylene-receptor genes, was widely distributed and clustered into 10 different groups. Thus, ACS, ERF, and sorbitol dehydrogenase proteins potentially share a common ancestor for different plant genomes, while the expansion rate may be related to ancestral expansion rather than species-specific events. Based on the distribution of the clades, we suggest that gene function diversification for the ripening pathway occurred prior to family extension. We herein report all the frameshift mutations in genes involved in sugar transport and ethylene biosynthesis detected as well as the gene CNV implicated in ripening differences.

Metabolic fingerprinting of must obtained from sun-dried grapes of two indigenous Cypriot cultivars destined for the production of 'Commandaria': A protected destignation of origin product.

Grapes's sun-dried process is one of the most critical steps in the production of 'Commandaria', a dessert wine product that is exclusively produced in Cyprus and derived from must obtained from two indigenous grape cultivars, namely 'Mavro' and 'Xynisteri'. Despite its significant economic importance, no data regarding the primary and secondary metabolites of the aforementioned cultivars exist. Thus, in the current study, the effect of sun drying process on the qualitative attributes and phenolic profile of 'Mavro' and 'Xynisteri' musts was dissected. Musts were analyzed at harvest and at the end of the sun-drying process that corresponds to ca. 30-40% water loss. Results highlighted significant differences in chemical composition of the must before and after the sun-drying process. Except for the increase of soluble solids content, a significant increment in glucose, fructose, total acidity, total phenols and total flavonoids contents was monitored. Subsequently, forty-two phenolic compounds were identified by LC-DAD-qTOF-MS revealing the polyphenolic fingerprint of the two cultivars. Results also indicated that changes in the phenolic composition of the obtained must are not only correlated with the dehydration effect, but both synthesis or degradation reactions occurred. In particular, the increases in the concentration of hydroxybenzoic acids were higher than the concentration effect for both cultivars. Regarding to hydroxycinnamates, dehydration caused a six-fold increase of hydroxycinnamic acid content in both cultivars. Intriguingly, the concentration of some hydroxycinnamic acids such as caffeic acid dihexoside and fertaric acid isomer went descending. Although the degradation of the internal side of the skin facilitate improved extractability from the skins to the grape pulp and therefore to the grape must, the sun-drying process may also induced stilbene and lignans synthesis production. A significant effect of dehydration on the postharvest biosynthesis of three groups of flavonoids (flavonols, flavan-3-ols, flavanonols), was also observed. This study sheds some light in the substantial changes that occur in specific metabolites during the sun drying process; such metabolites can be considered as potential factors that may determine organoleptic characteristics and biological properties of the end-product.

Deciphering the interplay among genotype, maturity stage and low-temperature storage on phytochemical composition and transcript levels of enzymatic antioxidants in Prunus persica fruit.

The aim of this study was to understand the antioxidant metabolic changes of peach (cvs. 'Royal Glory', 'Red Haven' and 'Sun Cloud') and nectarine fruits (cv. 'Big Top') exposed to different combinations of low-temperature storage (0, 2, 4 weeks storage at 0 °C, 90% R.H.) and additional ripening at room temperature (1, 3 and 5 d, shelf life, 20 °C) with an array of analytical, biochemical and molecular approaches. Initially, harvested fruit of the examined cultivars were segregated non-destructively at advanced and less pronounced maturity stages and qualitative traits, physiological parameters, phytochemical composition and antioxidant capacity were determined. 'Big Top' and 'Royal Glory' fruits were characterized by slower softening rate and less pronounced ripening-related alterations. The coupling of HPLC fingerprints, consisted of 7 phenolic compounds (chlorogenic, neochlorogenic acid, catechin, epicatechin, rutin, quecetin-3-O-glucoside, procyanidin B1) and spectrophotometric methods disclosed a great impact of genotype on peach bioactive composition, with 'Sun Cloud' generally displaying the highest contents. Maturity stage at harvest did not seem to affect fruit phenolic composition and no general guidelines for the impact of cold storage and shelf-life on individual phenolic compounds can be extrapolated. Subsequently, fruit of less pronounced maturity at harvest were used for further molecular analysis. 'Sun Cloud' was proven efficient in protecting plasmid pBR322 DNA against ROO attack throughout the experimental period and against HO attack after 2 and 4 weeks of cold storage. Interestingly, a general down-regulation of key genes implicated in the antioxidant apparatus with the prolongation of storage period was recorded; this was more evident for CAT, cAPX, Cu/ZnSOD2, perAPX3 and GPX8 genes. Higher antioxidant capacity of 'Sun Cloud' fruit could potentially be linked with compounds other than enzymatic antioxidants that further regulate peach fruit ripening.

Metabolic and transcriptional elucidation of the carotenoid biosynthesis pathway in peel and flesh tissue of loquat fruit during on-tree development.

BACKGROUND: Carotenoids are the main colouring substances found in orange-fleshed loquat fruits. The aim of this study was to unravel the carotenoid biosynthetic pathway of loquat fruit (cv. 'Obusa') in peel and flesh tissue during distinct on-tree developmental stages through a targeted analytical and molecular approach. RESULTS: Substantial changes regarding colour parameters, both between peel and flesh and among the different developmental stages, were monitored, concomitant with a significant increment in carotenoid content. Key genes and individual compounds that are implicated in the carotenoid biosynthetic pathway were further dissected with the employment of molecular (RT-qPCR) and advanced analytical techniques (LC-MS). Results revealed significant differences in carotenoid composition between peel and flesh. Thirty-two carotenoids were found in the peel, while only eighteen carotenoids were identified in the flesh. Trans-lutein and trans-ß-carotene were the major carotenoids in the peel; the content of the former decreased with the progress of ripening, while the latter registered a 7.2-fold increase. However, carotenoid profiling of loquat flesh indicated trans-ß-cryptoxanthin, followed by trans-ß-carotene and 5,8-epoxy-ß-carotene to be the most predominant carotenoids. High amounts of trans-ß-carotene in both tissues were supported by significant induction in a chromoplast-specific lycopene ß-cyclase (CYCB) transcript levels. PSY1, ZDS, CYCB and BCH were up-regulated and CRTISO, LCYE, ECH and VDE were down-regulated in most of the developmental stages compared with the immature stage in both peel and flesh tissue. Overall, differential regulation of expression levels with the progress of on-tree fruit development was more evident in the middle and downstream genes of carotenoid biosynthetic pathway. CONCLUSIONS: Carotenoid composition is greatly affected during on-tree loquat development with striking differences between peel and flesh tissue. A link between gene up- or down-regulation during the developmental stages of the loquat fruit, and how their expression affects carotenoid content per tissue (peel or flesh) was established.

Antioxidant Phytochemicals in Fresh Produce: Exploitation of Genotype Variation and Advancements in Analytical Protocols.

Horticultural commodities (fruit and vegetables) are the major dietary source of several bioactive compounds of high nutraceutical value for humans, including polyphenols, carotenoids and vitamins. The aim of the current review was dual. Firstly, toward the eventual enhancement of horticultural crops with bio-functional compounds, the natural genetic variation in antioxidants found in different species and cultivars/genotypes is underlined. Notably, some landraces and/or traditional cultivars have been characterized by substantially higher phytochemical content, i.e., small tomato of Santorini island (cv. "Tomataki Santorinis") possesses appreciably high amounts of ascorbic acid (AsA). The systematic screening of key bioactive compounds in a wide range of germplasm for the identification of promising genotypes and the restoration of key gene fractions from wild species and landraces may help in reducing the loss of agro-biodiversity, creating a healthier "gene pool" as the basis of future adaptation. Toward this direction, large scale comparative studies in different cultivars/genotypes of a given species provide useful insights about the ones of higher nutritional value. Secondly, the advancements in the employment of analytical techniques to determine the antioxidant potential through a convenient, easy and fast way are outlined. Such analytical techniques include electron paramagnetic resonance (EPR) and infrared (IR) spectroscopy, electrochemical, and chemometric methods, flow injection analysis (FIA), optical sensors, and high resolution screening (HRS). Taking into consideration that fruits and vegetables are complex mixtures of water- and lipid-soluble antioxidants, the exploitation of chemometrics to develop "omics" platforms (i.e., metabolomics, foodomics) is a promising tool for researchers to decode and/or predict antioxidant activity of fresh produce. For industry, the use of optical sensors and IR spectroscopy is recommended to estimate the antioxidant activity rapidly and at low cost, although legislation does not allow its correlation with health claims.

Regulation of On-Tree Vitamin E Biosynthesis in Olive Fruit during Successive Growing Years: The Impact of Fruit Development and Environmental Cues.

The term vitamin E refers to a group of eight lipophilic compounds known as tocochromanols. The tocochromanols are divided into two groups, that is, tocopherols and tocotrienols, with four forms each, namely α-, ß-, γ-, and δ-. In order to explore the temporal biosynthesis of tocochromanols in olive (Olea europaea cv. 'Koroneiki') fruit during on-tree development and ripening over successive growing years, a combined array of analytical, molecular, bioinformatic, immunoblotting, and antioxidant techniques were employed. Fruits were harvested at eight successive developmental stages [10-30 weeks after flowering (WAF)], over three consecutive years. Intriguingly, climatic conditions affected relative transcription levels of vitamin E biosynthetic enzymes; a general suppression to induction pattern (excluding VTE5) was monitored moving from the 1st to the 3rd growing year, probably correlated to decreasing rainfall levels and higher temperature, particularly at the fruit ripening stage. A gradual diminution of VTE5 protein content was detected during the fruit development of each year, with a marked decrease occurring after 16 WAF. Alpha-tocopherol was the most abundant metabolite with an average percentage of 96.82 ± 0.23%, 91.13 ± 0.95%, and 88.53 ± 0.96% (during the 1st, 2nd, and 3rd year, respectively) of total vitamin E content in 10-30 WAF. The concentrations of α-tocopherol revealed a generally declining pattern, both during the on-tree ripening of the olive fruit and across the 3 years, accompanied by a parallel decline of the total antioxidant capacity of the drupe. Contrarily, all other tocochromanols demonstrated an inverse pattern with lowest levels being recorded during the 1st year. It is likely that, in a defense attempt against water deficit conditions and increased air temperature, transcription of genes involved in vitamin E biosynthesis (excluding VTE5) is up-regulated in olive fruit, probably leading to the blocking/deactivating of the pathway through a negative feedback regulatory mechanism.

(1)H NMR Metabolic Fingerprinting to Probe Temporal Postharvest Changes on Qualitative Attributes and Phytochemical Profile of Sweet Cherry Fruit.

Sweet cherry fruits (Prunus avium cvs. 'Canada Giant', 'Ferrovia') were harvested at commercial maturity stage and analyzed at harvest and after maintenance at room temperature (storage at ∼20°C, shelf life) for 1, 2, 4, 6, and 8 days, respectively. Fruit were initially analyzed for respiration rate, qualitative attributes and textural properties: 'Canada Giant' fruit were characterized by higher weight losses and stem browning index, being more intense over the late stages of shelf life period; meanwhile 'Ferrovia' possessed appreciably better performance even after extended shelf life period. A gradual decrease of respiration rate was monitored in both cultivars, culminated after 8 days at 20°C. The sweet cherry fruit nutraceutical profile was monitored using an array of instrumental techniques (spectrophotometric assays, HPLC, (1)H-NMR). Fruit antioxidant capacity was enhanced with the progress of shelf life period, concomitant with the increased levels of total anthocyanin and of phenolic compounds. 'Ferrovia' fruit presented higher contents of neochlorogenic acid and p-coumaroylquinic acid throughout the shelf life period. We further developed an (1)H-NMR method that allows the study of primary and secondary metabolites in a single running, without previous separation and isolation procedures. Diagnostic peaks were located in the aliphatic region for sugars and organic acids, in the aromatic region for phenolic compounds and at 8.2-8.6 ppm for anthocyanins. This NMR-based methodology provides a unifying tool for quantitative and qualitative characterization of metabolite changes of sweet cherry fruits; it is also expected to be further exploited for monitoring temporal changes in other fleshy fruits.

Temporal analysis reveals a key role for VTE5 in vitamin E biosynthesis in olive fruit during on-tree development.

The aim of this work was to generate a high resolution temporal mapping of the biosynthetic pathway of vitamin E in olive fruit (Olea europaea cv. "Koroneiki") during 17 successive on-tree developmental stages. Fruit material was collected from the middle of June until the end of January, corresponding to 6-38 weeks after flowering (WAF). Results revealed a variable gene regulation pattern among 6-38 WAF studied and more pronounced levels of differential regulation of gene expression for the first and intermediate genes in the biosynthetic pathway (VTE5, geranylgeranyl reductase, HPPD, VTE2, HGGT and VTE3) compared with the downstream components of the pathway (VTE1 and VTE4). Notably, expression of HGGT and VTE2 genes were significantly suppressed throughout the developmental stages examined. Metabolite analysis indicated that the first and intermediate stages of development (6-22 WAF) have higher concentrations of tocochromanols compared with the last on-tree stages (starting from 24 WAF onwards). The concentration of α-tocopherol (16.15 ± 0.60-32.45 ± 0.54 mg/100 g F.W.) were substantially greater (up to 100-fold) than those of ß-, γ-, and δ-tocopherols (0.13 ± 0.01-0.25 ± 0.03 mg/100 g F.W., 0.13 ± 0.01-0.33 ± 0.04 mg/100 g F.W., 0.14 ± 0.01-0.28 ± 0.01 mg/100 g F.W., respectively). In regard with tocotrienol content, only γ-tocotrienol was detected. Overall, olive fruits (cv. "Koroneiki") exhibited higher concentrations of vitamin E until 22 WAF as compared with later WAF, concomitant with the expression profile of phytol kinase (VTE5), which could be used as a marker gene due to its importance in the biosynthesis of vitamin E. To the best of our knowledge, this is the first study that explores the complete biosynthetic pathway of vitamin E in a fruit tree crop of great horticultural importance such as olive, linking molecular gene expression analysis with tocochromanol content.

Ozone-induced kiwifruit ripening delay is mediated by ethylene biosynthesis inhibition and cell wall dismantling regulation.

Ozone treatments are used to preserve quality during cold storage of commercially important fruits due to its ethylene oxidizing capacity and its antimicrobial attributes. To address whether or not ozone also modulates ripening by directly affecting fruit physiology, kiwifruit (Actinidia deliciosa cv. 'Hayward') were stored in very low ethylene atmosphere at 0°C (95% RH) in air (control) or in the presence of ozone (0.3µLL(-1)) for 2 or 4 months and subsequently ripened at 20°C (90% RH) for up to 8d. Ozone-treated kiwifruit showed a significant delay of ripening during maintenance at 20°C, accompanied by a marked decrease in ethylene biosynthesis due to inhibited AdACS1 and AdACO1 expression and reduced ACC synthase (ACS) and ACC oxidase (ACO) enzyme activity. Furthermore, ozone-treated fruit exhibited a marked reduction in flesh softening and cell wall disassembly. This effect was associated with reduced cell wall swelling and pectin and neutral sugar solubilization and was correlated with the inhibition of cell wall degrading enzymes activity, such as polygalacturonase (PG) and endo-1,4-ß-glucanase/1,4-ß-glucosidase (EGase/glu). Conclusively, the present study indicated that ozone may exert major residual effects in fruit ripening physiology and suggested that ethylene biosynthesis and cell walls turnover are specifically targeted by ozone.

Sodium hydrosulfide induces systemic thermotolerance to strawberry plants through transcriptional regulation of heat shock proteins and aquaporin.

BACKGROUND: Temperature extremes represent an important limiting factor to plant growth and productivity. The present study evaluated the effect of hydroponic pretreatment of strawberry (Fragaria x ananassa cv. 'Camarosa') roots with an H2S donor, sodium hydrosulfide (NaHS; 100 µM for 48 h), on the response of plants to acute heat shock treatment (42°C, 8 h). RESULTS: Heat stress-induced phenotypic damage was ameliorated in NaHS-pretreated plants, which managed to preserve higher maximum photochemical PSII quantum yields than stressed plants. Apparent mitigating effects of H2S pretreatment were registered regarding oxidative and nitrosative secondary stress, since malondialdehyde (MDA), H2O2 and nitric oxide (NO) were quantified in lower amounts than in heat-stressed plants. In addition, NaHS pretreatment preserved ascorbate/glutathione homeostasis, as evidenced by lower ASC and GSH pool redox disturbances and enhanced transcription of ASC (GDH) and GSH biosynthetic enzymes (GS, GCS), 8 h after heat stress imposition. Furthermore, NaHS root pretreatment resulted in induction of gene expression levels of an array of protective molecules, such as enzymatic antioxidants (cAPX, CAT, MnSOD, GR), heat shock proteins (HSP70, HSP80, HSP90) and aquaporins (PIP). CONCLUSION: Overall, we propose that H2S root pretreatment activates a coordinated network of heat shock defense-related pathways at a transcriptional level and systemically protects strawberry plants from heat shock-induced damage.

Berry antioxidants: small fruits providing large benefits.

Small berry fruits are consumed because of their attractive colour and special taste, and are considered one of the richest sources of natural antioxidants. Their consumption has been linked to the prevention of some chronic and degenerative diseases. The term 'berry fruits' encompasses the so-called 'soft fruits', primarily strawberry, currants, gooseberry, blackberry, raspberry, blueberry and cranberry. The objective of this review is to highlight the nutraceutical value of berries and to summarize the factors affecting berry fruit antioxidants. Particular attention is given to postharvest and processing operation factors that may affect fruit phytochemical content. The structure-antioxidant relationships for phenolic compounds - the main group of antioxidants in this fruit group - are presented and major areas for future research are identified.