Ectopic overexpression of ShCBF1 and SlCBF1 in tomato suggests an alternative view of fruit responses to chilling stress postharvest.

Postharvest chilling injury (PCI) is a physiological disorder that often impairs tomato fruit ripening; this reduces fruit quality and shelf-life, and even accelerates spoilage at low temperatures. The CBF gene family confers cold tolerance in Arabidopsis thaliana, and constitutive overexpression of CBF in tomato increases vegetative chilling tolerance, in part by retarding growth, but, whether CBF increases PCI tolerance in fruit is unknown. We hypothesized that CBF1 overexpression (OE) would be induced in the cold and increase resistance to PCI. We induced high levels of CBF1 in fruit undergoing postharvest chilling by cloning it from S. lycopersicum and S. habrochaites, using the stress-inducible RD29A promoter. Harvested fruit were cold-stored (2.5°C) for up to three weeks, then rewarmed at 20°C for three days. Transgene upregulation was triggered during cold storage from 8.6- to 28.6-fold in SlCBF1-OE, and between 3.1- to 8.3-fold in ShCBF1-OE fruit, but developmental abnormalities in the absence of cold induction were visible. Remarkably, transgenic fruit displayed worsening of PCI symptoms, i.e., failure to ripen after rewarming, comparatively higher susceptibility to decay relative to wild-type (WT) fruit, lower total soluble solids, and the accumulation of volatile compounds responsible for off-odors. These symptoms correlated with CBF1 overexpression levels. Transcriptomic analysis revealed that the ripening and biotic and abiotic stress responses were altered in the cold-stored transgenic fruit. Seedlings grown from 'chilled' and 'non-chilled' WT fruit, in addition to 'non-chilled' transgenic fruit were also exposed to 0°C to test their photosynthetic response to chilling injury. Chilled WT seedlings adjusted their photosynthetic rates to reduce oxidative damage; 'non-chilled' WT seedlings did not. Photosynthetic parameters between transgenic seedlings were similar at 0°C, but SlCBF1-OE showed more severe photoinhibition than ShCBF1-OE, mirroring phenotypic observations. These results suggest that 1) CBF1 overexpression accelerated fruit deterioration in response to cold storage, and 2) Chilling acclimation in fructus can increase chilling tolerance in seedling progeny of WT tomato.

Integrative analysis of the methylome and transcriptome of tomato fruit (Solanum lycopersicum L.) induced by postharvest handling.

Tomato fruit ripening is triggered by the demethylation of key genes, which alters their transcriptional levels thereby initiating and propagating a cascade of physiological events. What is unknown is how these processes are altered when fruit are ripened using postharvest practices to extend shelf-life, as these practices often reduce fruit quality. To address this, postharvest handling-induced changes in the fruit DNA methylome and transcriptome, and how they correlate with ripening speed, and ripening indicators such as ethylene, abscisic acid, and carotenoids, were assessed. This study comprehensively connected changes in physiological events with dynamic molecular changes. Ripening fruit that reached 'Turning' (T) after dark storage at 20°C, 12.5°C, or 5°C chilling (followed by 20°C rewarming) were compared to fresh-harvest fruit 'FHT'. Fruit stored at 12.5°C had the biggest epigenetic marks and alterations in gene expression, exceeding changes induced by postharvest chilling. Fruit physiological and chronological age were uncoupled at 12.5°C, as the time-to-ripening was the longest. Fruit ripening to Turning at 12.5°C was not climacteric; there was no respiratory or ethylene burst, rather, fruit were high in abscisic acid. Clear differentiation between postharvest-ripened and 'FHT' was evident in the methylome and transcriptome. Higher expression of photosynthetic genes and chlorophyll levels in 'FHT' fruit pointed to light as influencing the molecular changes in fruit ripening. Finally, correlative analyses of the -omics data putatively identified genes regulated by DNA methylation. Collectively, these data improve our interpretation of how tomato fruit ripening patterns are altered by postharvest practices, and long-term are expected to help improve fruit quality.

Influence of chitin nanofibers and gallic acid on physical-chemical and biological performances of chitosan-based films.

In this work, chitosan films loaded with gallic acid and different content of chitin nanofibers were prepared and subjected to different characterization techniques. The results showed that the inclusion of gallic acid to chitosan films caused moderate decrease in water vapor permeability (by 29 %) and increased tensile strength of films (by 169 %) in comparison to the neat chitosan films. Furthermore, it was found that the addition of chitin nanofibers up to 30 % into chitosan/gallic acid films additionally improved tensile strength (by 474 %) and reduced plasticity of films (by 171 %), when compared to the chitosan/gallic acid films. Increased concentration of chitin nanofibers in films reduced the overall water vapor permeability of films by 51 %. In addition, gallic acid and chitin nanofibers had synergic effect on high chitosan film's antioxidant and antifungal activity toward Botrytis cinerea (both above 95 %). Finally, chitosan/gallic acid/chitin nanofibers films reduced decay incidence of strawberries, increased total soluble solid content, and promoted high production of some polyphenols during cold storage, in comparison to the control chitosan films and uncoated strawberry samples. Hence, these results suggest that chitosan/gallic acid/chitin nanofibers can present eco-sustainable approach for preservation of strawberries, giving them additional nutritional value.

Rain Cover and Netting Materials Differentially Affect Fruit Yield and Quality Traits in Two Highbush Blueberry Cultivars via Changes in Sunlight and Temperature Conditions.

The use of covers to protect blueberry orchards from adverse weather events has increased due to the variability in climate patterns, but the effects of rain covers and netting materials on yield and fruit quality have not been studied yet. This research evaluated the simultaneous effect of an LDPE plastic cover, a woven cover, and netting material on environmental components (UV light, PAR, NIR, and growing degree days (GDDs)), plant performance (light interception, leaf area index, LAI, yield, and flower development), and fruit quality traits (firmness, total soluble solids, and acidity) in two blueberry cultivars. On average, UV transmission under the netting was 11% and 43% higher compared to that under woven and LDPE plastic covers, while NIR transmission was 8-13% higher with both types of rain covers, with an increase in fruit air temperature and GDDs. Yield was 27% higher under the woven cover with respect to netting, but fruit firmness values under the netting were 12% higher than those of the LDPE plastic cover. Light interception, LAI, and flower development explained 64% (p = 0.0052) of the yield variation due to the cover material's effect. The obtained results suggest that the type of cover differentially affects yield and fruit quality in blueberries due to the specific light and temperature conditions generated under these materials.

Dissecting postharvest chilling injury through biotechnology.

Paradoxically, refrigerating many fruits and vegetables destroys their quality, and may even accelerate their spoilage. This phenomenon, known as postharvest chilling injury (PCI), affects produce from tropical and subtropical regions and leads to economic and postharvest loss and waste. Low temperatures are used to pause the physiological processes associated with senescence, but upon rewarming, these processes may resume at an accelerated rate. Chilling-injured produce may be discarded for not meeting consumer expectations or may prematurely deteriorate. In this review, we describe progress made in identifying the cellular and molecular processes underlying PCI, and point to advances in biotechnological approaches for ameliorating symptoms. Further, we identify the gaps in knowledge that must be bridged to develop effective solutions to PCI.

Can gene editing reduce postharvest waste and loss of fruit, vegetables, and ornamentals?

Postharvest waste and loss of horticultural crops exacerbates the agricultural problems facing humankind and will continue to do so in the next decade. Fruits and vegetables provide us with a vast spectrum of healthful nutrients, and along with ornamentals, enrich our lives with a wide array of pleasant sensory experiences. These commodities are, however, highly perishable. Approximately 33% of the produce that is harvested is never consumed since these products naturally have a short shelf-life, which leads to postharvest loss and waste. This loss, however, could be reduced by breeding new crops that retain desirable traits and accrue less damage over the course of long supply chains. New gene-editing tools promise the rapid and inexpensive production of new varieties of crops with enhanced traits more easily than was previously possible. Our aim in this review is to critically evaluate gene editing as a tool to modify the biological pathways that determine fruit, vegetable, and ornamental quality, especially after storage. We provide brief and accessible overviews of both the CRISPR-Cas9 method and the produce supply chain. Next, we survey the literature of the last 30 years, to catalog genes that control or regulate quality or senescence traits that are "ripe" for gene editing. Finally, we discuss barriers to implementing gene editing for postharvest, from the limitations of experimental methods to international policy. We conclude that in spite of the hurdles that remain, gene editing of produce and ornamentals will likely have a measurable impact on reducing postharvest loss and waste in the next 5-10 years.

Integrative analysis of postharvest chilling injury in cherry tomato fruit reveals contrapuntal spatio-temporal responses to ripening and cold stress.

Postharvest chilling injury (PCI) reduces fruit quality and shelf-life in tomato (Solanum lycopersicum L.). PCI has been traditionally studied in the pericarp, however its development is likely heterogeneous in different fruit tissues. To gain insight into PCI's spatio-temporal development, we used postharvest biomarkers e.g. respiration and ethylene rates, ion leakage etc., to confirm the occurrence of PCI, and compared these data with molecular (gene expression), biophysical (MRI data) and biochemical parameters (Malondialdehyde (MDA) and starch content) from the pericarp or columella. Tissues were stored at control (12.5 °C) or PCI-inducing temperatures (2.5 or 5 °C) followed by rewarming at 20 °C. MRI and ion leakage revealed that cold irreversibly impairs ripening-associated membrane liquefaction; MRI also showed that the internal and external fruit tissues responded differently to cold. MDA and especially starch contents, were affected by chilling in a tissue-specific manner. The expression of the six genes studied: ACO1 and ACS2 (ripening), CBF1 (cold response), DHN, AOX1a and LoxB (stress-related) showed non-overlapping temporal and spatially-specific responses. Overall, the data highlighted the interconnectedness of fruit cold response and ripening, and showed how cold stress reconfigures the latter. They further underscored that multidimensional spatial and temporal biological studies are needed to develop effective solutions to PCI.