Compositional, structural and functional cuticle analysis of Prunus laurocerasus L. sheds light on cuticular barrier plasticity.

Barrier properties of the hydrophobic plant cuticle depend on its physicochemical composition. The cuticular compounds vary considerably among plant species but also among organs and tissues of the same plant and throughout developmental stages. As yet, these intraspecific modifications at the cuticular wax and cutin level are only rarely examined. Attempting to further elucidate cuticle profiles, we analysed the adaxial and abaxial surfaces of the sclerophyllous leaf and three developmental stages of the drupe fruit of Prunus laurocerasus, an evergreen model plant native to temperate regions. According to gas chromatographic analyses, the cuticular waxes contained primarily pentacyclic triterpenoids dominated by ursolic acid, whereas the cutin biopolyester mainly consisted of 9/10,ω-dihydroxy hexadecanoic acid. Distinct organ- and side-specific patterns were found for cuticular lipid loads, compositions and carbon chain length distributions. Compositional variations led to different structural and functional barrier properties of the plant cuticle, which were investigated further microscopically, infrared spectroscopically and gravimetrically. The minimum water conductance was highlighted at 1 × 10-5 m s-1 for the perennial, hypostomatous P. laurocerasus leaf and at 8 × 10-5 m s-1 for the few-month-living, stomatous fruit suggesting organ-specific cuticular barrier demands.

Ripening-related cell wall modifications in olive (Olea europaea L.) fruit: A survey of nine genotypes.

The production of olive (Olea europaea L.) is very important economically in many areas of the world, and particularly in countries around the Mediterranean basin. Ripening-associated modifications in cell wall composition and structure of fruits play an important role in attributes like firmness or susceptibility to infestations, rots and mechanical damage, but limited information on these aspects is currently available for olive. In this work, cell wall metabolism was studied in fruits from nine olive cultivars ('Arbequina', 'Argudell', 'Empeltre', 'Farga', 'Manzanilla', 'Marfil', 'Morrut', 'Picual' and 'Sevillenca') picked at three maturity stages (green, turning and ripe). Yields of alcohol-insoluble residue (AIR) recovered from fruits, as well as calcium content in fruit pericarp, decreased along ripening. Cultivar-specific diversity was observed in time-course change patterns of enzyme activity, particularly for those acting on arabinosyl- and galactosyl-rich pectin side chains. Even so, fruit firmness levels were associated to higher pectin methylesterase (PME) activity and calcium contents. In turn, fruit firmness correlated inversely with ascorbate content and with α-l-arabinofuranosidase (AFase) and ß-galactosidase (ß-Gal) activities, resulting in preferential loss of neutral sugars from cell wall polymers.

Insights Into Olive Fruit Surface Functions: A Comparison of Cuticular Composition, Water Permeability, and Surface Topography in Nine Cultivars During Maturation.

Olive (Olea europaea L.) growing has outstanding economic relevance in Spain, the main olive oil producer and exporter in the world. Fruit skin properties are very relevant for fruit and oil quality, water loss, and susceptibility to mechanical damage, rots, and infestations, but limited research focus has been placed on the cuticle of intact olive fruit. In this work, fruit samples from nine olive cultivars ("Arbequina," "Argudell," "Empeltre," "Farga," "Manzanilla," "Marfil," "Morrut," "Picual," and "Sevillenca") were harvested from an experimental orchard at three different ripening stages (green, turning, and ripe), and cuticular membranes were enzymatically isolated from fruit skin. The total contents of cuticular wax and cutin significantly differed among cultivars both in absolute and in relative terms. The wax to cutin ratio generally decreased along fruit maturation, with the exception of "Marfil" and "Picual." In contrast, increased water permeance values in ripe fruit were observed uniquely for "Argudell," "Morrut," and "Marfil" fruit. The toluidine blue test revealed surface discontinuities on green samples of "Argudell," "Empeltre," "Manzanilla," "Marfil," and "Sevillenca" fruit, but not on "Arbequina," "Farga," "Morrut," or "Picual." No apparent relationship was found between water permeability and total wax coverage or the results of the toluidine blue test. The composition of cuticular waxes and cutin monomers was analyzed in detail, and sections of fruit pericarp were stained in Sudan IV for microscopy observations. Skin surface topography was also studied by means of fringe projection, showing large differences in surface roughness among the cultivars, "Farga" and "Morrut" fruits displaying the most irregular surfaces. Cultivar-related differences in cuticle and surface features of fruit are presented and discussed.

Firmness at Harvest Impacts Postharvest Fruit Softening and Internal Browning Development in Mechanically Damaged and Non-damaged Highbush Blueberries (Vaccinium corymbosum L.).

Fresh blueberries are very susceptible to mechanical damage, which limits postharvest life and firmness. Softening and susceptibility of cultivars "Duke" and "Brigitta" to developing internal browning (IB) after mechanical impact and subsequent storage was evaluated during a 2-year study (2011/2012, 2012/2013). On each season fruit were carefully hand-picked, segregated into soft (<1.60 N), medium (1.61-1.80 N), and firm (1.81-2.00 N) categories, and then either were dropped (32 cm) onto a hard plastic surface or remained non-dropped. All fruit were kept under refrigerated storage (0°C and 85-88% relative humidity) to assess firmness loss and IB after 7, 14, 21, 28, and 35 days. In general, regardless of cultivar or season, high variability in fruit firmness was observed within each commercial harvest, and significant differences in IB and softening rates were found. "Duke" exhibited high softening rates, as well as high and significant r2 between firmness and IB, but little differences for dropped vs. non-dropped fruit. "Brigitta," having lesser firmness rates, exhibited almost no relationships between firmness and IB (especially for non-dropped fruit), but marked differences between dropping treatments. Firmness loss and IB development were related to firmness at harvest, soft and firm fruit being the most and least damaged, respectively. Soft fruit were characterized by greater IB development during storage along with high soluble solids/acid ratio, which could be used together with firmness to estimate harvest date and storage potential of fruit. Results of this work suggest that the differences in fruit quality traits at harvest could be related to the time that fruit stay on the plant after turning blue, soft fruit being more advanced in maturity. Finally, the observed differences between segregated categories reinforce the importance of analyzing fruit condition for each sorted group separately.

Post-storage cell wall metabolism in two sweet cherry (Prunus avium L.) cultivars displaying different postharvest performance.

The biochemical processes underlying firmness loss of sweet cherry (Prunus avium L.) fruit are poorly understood. Studies on cell wall metabolism of sweet cherry have been generally undertaken during on-tree development or at harvest maturity, while published reports on postharvest changes are scarce and fragmentary. In this work, cell wall modifications after storage at 0 ℃ were studied in two cherry cultivars ('Celeste' and 'Somerset') displaying different postharvest potential. Firmness was largely determined by the yields of the Na2CO3- and KOH-soluble fractions, enriched in covalently-bound pectins and in matrix glycans, respectively, and correlated well with ascorbic acid contents. The yields of these two cell wall fractions were correlated inversely with pectinmethylesterase and endo-1,4-ß-d-glucanase activities, indicating a relevant role of these two enzymes in postharvest firmness changes in sweet cherry. The amount of solubilised cell wall materials was closely associated to the contents of dehydroascorbic acid, suggesting the possible involvement of oxidative mechanisms in cell wall disassembly. These data may help understanding the evolution of fruit quality during the marketing period, and give hints for the design of suitable management strategies to preserve key attributes.

Characterization of cuticle composition after cold storage of "Celeste" and "Somerset" sweet cherry fruit.

Cuticle composition and structure may be relevant factors affecting the storage potential of fruits, but very few studies have analyzed fruit cuticle composition from a postharvest perspective. In this work, the chemical composition of waxes and cutin (major cuticular components) was analyzed in cuticle samples isolated from "Celeste" and "Somerset" cherries (Prunus avium L.) after cold storage at 0 °C. Total cuticle amounts per surface unit (µg cm(-2)) increased along with cold storage. The triterpene ursolic acid, the alkane nonacosane, linoleic acid, and ß-sitosterol were the most abundant components of cuticular waxes, whereas cutin composition was dominated by C18-type monomers. In spite of being comprised of similar chemical families, cultivar-related differences were found regarding the abundance and the evolution of some compound families during cold storage. To the best of our knowledge, this is the first report on changes in cuticle composition of sweet cherry during postharvest storage.

Fruit cuticle composition of a melting and a nonmelting peach cultivar.

Although postharvest quality of fruit is greatly affected by cuticle composition, structure, and properties, very few published studies have analyzed fruit cuticle composition from a postharvest perspective. In this work, the chemical composition of waxes and cutin, major cuticular components, was analyzed in fruit cuticle samples isolated from a melting ('October Sun') and a nonmelting ('Jesca') peach (Prunus persica L. Batsch.) cultivar at harvest and after a simulated shelf-life period of 5 days at 20 °C. Cutin composition was dominated by 18-hydroxyoleic acid, whereas the triterpenoid ursolic and oleanoic acids and the alkanes n-tricosane and n-pentacosane were quantitatively prominent among the wax compounds identified. Some quantitative differences were found between both cultivars for particular compound families and in their postharvest modifications. To the best of the authors' knowledge, this is the first study characterizing the composition of the cuticle of peach fruit and describing the changes therein after harvest.

Comparison of the volatile profile and sensory analysis of 'Golden Reinders' apples after the application of a cold air period after ultralow oxygen (ULO) storage.

All efforts to improve fruit quality are rewarded when consumers are satisfied after tasting the fruit. Apples are often stored under controlled atmosphere conditions to preserve them over time, but this frequently results in a loss of flavor. The aim of this work, which was based on two seasons, was to evaluate the influence of a period of short-term air storage (periods of 2 and 4 weeks) after removal from ultralow oxygen (ULO) storage (1 kPa of O(2)/1 kPa of CO(2)) with respect to increases in volatile compound emissions and the effect on standard and sensory quality in 'Golden Reinders' apples. The results showed that emissions of 26 volatile compounds increased as a result of ULO + 2 weeks or ULO + 4 weeks of storage. However, the results of tastings involving a panel of consumers and trained experts revealed that this increase was not matched by corresponding increases in either the degree of consumer preference or flavor attributes.

Preharvest calcium sprays improve volatile emission at commercial harvest of 'Fuji Kiku-8' apples.

Apple (Malus × domestica Borkh.) fruit intended for long-term storage are frequently harvested commercially before becoming fully ripe, often resulting in poor aroma development. Since postharvest calcium dips have proved effective for the enhancement of flavor-related volatile esters after cold storage of apples, this study was undertaken in order to assess whether preharvest calcium sprays (7 weekly applications at 1.6%, w/v, 81-123 days after full bloom) could also aid in improving this important attribute at harvest. This procedure significantly increased calcium content in treated fruit. The emission of aroma-related volatile esters by untreated and calcium-treated 'Fuji' apples was then monitored during maturation and ripening over two months prior to commercial harvest. Results indicate that most of the compounds contributing to overall flavor in ripe fruit were enhanced in response to preharvest calcium applications, suggesting that this procedure may be suitable for the improvement of fruit aroma at harvest. The emission of acetate esters was particularly favored, consistent with higher acetaldehyde contents in treated fruit. These effects arose apparently from increased pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) activities, possibly leading to a better supply of alcohols and acyl CoAs for ester biosynthesis.

Calcium dips enhance volatile emission of cold-stored 'Fuji Kiku-8' apples.

Despite the relevance of volatile production for overall quality of apple (Malus x domestica Borkh.) fruit, only a few studies have focused on the effects of calcium treatments on this quality attribute. In this work, 'Fuji Kiku-8' apples were harvested at commercial maturity, dipped in calcium chloride (2%, w/v), stored at 1 degrees C and 92% relative humidity for 4 or 7 months under either air or ultralow oxygen (ULO; 1 kPa of O(2)/2 kPa of CO(2)), and placed subsequently at 20 degrees C. Ethylene production, standard quality parameters, emission of volatile compounds, and the activities of some related enzymes were assessed 7 days thereafter. Calcium concentration was higher in CaCl(2)-treated than in untreated fruit, suggesting that the treatment was effective in introducing calcium into the tissues. Higher calcium contents were concomitant with higher flesh firmness and titratable acidity after storage. Furthermore, calcium treatment led to increased production of volatiles in middle-term stored apples, probably arising from enhanced supply of precursors for ester production as a consequence of increased pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) activities. After long-term storage, higher volatile emission might have arisen also from the enhancement of alcohol o-acyltransferase (AAT) activity, which was increased as a result of calcium treatment. In addition to storage period, the effects of calcium treatment were also partially dependent on storage atmosphere and more noticeable for fruit stored in air.

Lipoxygenase activity is involved in the regeneration of volatile ester-synthesizing capacity after ultra-low oxygen storage of 'Fuji' apple.

An extra period under cold air after ultra-low oxygen storage has been shown to increase the concentration of some volatile compounds emitted by stored 'Fuji' apples. The purpose of this work was to assess the role, if any, of lipoxygenase and hydroperoxide lyase activities in the regeneration of fruit capacity for volatile production after ultra-low oxygen storage. Fruits were stored at 1 °C and 92% relative humidity under ultra-low oxygen (1 kPa of O2/1 kPa of CO2); one lot was kept under hypoxia for 19 or 30 weeks, a second lot was maintained for 17 or 28 weeks under these conditions and then stored for 2 weeks in cold air, and a third lot remained for either 15 or 26 weeks under ultra-low oxygen followed by 4 weeks under cold air. Samples were placed subsequently at 20 °C, and analyses of volatile emission and enzyme activities were undertaken 1 and 7 days thereafter. Fruit stored during 4 weeks in cold air after ultra-low oxygen storage showed the highest capacity for volatile regeneration. Higher emission of volatiles by these samples was concomitant with higher levels of lipoxygenase activity. Results suggest that lipoxygenase activity, particularly in the flesh tissue, was strongly related to the regeneration of the emission of volatile compounds allowed by the extra period in cold air after ultra-low oxygen storage and, thus, appears to be a key control point for successful recovery of fruit ability for volatile compound production.

Long-term storage of Pink Lady apples modifies volatile-involved enzyme activities: consequences on production of volatile esters.

Pink Lady apples were harvested at commercial maturity and stored at 1 degrees C and 92% relative humidity under either air or controlled atmosphere conditions (2 kPa O 2:2 kPa CO 2 and 1 kPa O 2:1 kPa CO 2) for 27 weeks. Data on the emission of volatile compounds and on the activity of some related enzymes in both skin and flesh tissues were obtained during subsequent shelf life at 20 degrees C. Major effects of storage atmosphere and poststorage period were observed on the emission of volatile esters and their precursors. Changes in the production of volatile esters were partly due to alterations in the activity of alcohol o-acyltransferase, but the specific esters emitted by fruit after storage also resulted largely from modifications in the supply of the corresponding substrates. Samples stored under air were characterized by higher availability of acetaldehyde, whereas those stored under CA showed enhanced emission of the alcohol precursors ethanol and 1-hexanol (2 kPa O 2) and 1-butanol (1 kPa O 2), with accordingly higher production of ethyl, hexyl, and butyl esters. Multivariate analysis revealed that a large part of the observed differences in precursor availability arose from modifications in the activity of the enzymes considered. Higher pyruvate decarboxylase activity in air-stored fruit possibly accounted for higher acetaldehyde levels in these samples, while storage under 1 kPa O 2 led to significantly decreased lipoxygenase activity and thus to lessened production of 1-hexanol and hexyl esters. Low acetaldehyde availability together with enhanced hydroperoxide lyase and alcohol dehydrogenase levels in these fruits are suggested to have led to higher emission of 1-butanol and butyl esters.

Regeneration of volatile compounds in Fuji apples following ultra low oxygen atmosphere storage and its effect on sensory acceptability.

The aim of this work was to assess whether extra time spent under AIR conditions after storage in an ultra low oxygen (ULO) atmosphere could allow the regeneration of volatile compound emission without negatively affecting quality parameters and the consumer acceptability of Fuji apples. Fruits were stored for 19 and 30 weeks at 1 degrees C and 92% RH under ULO atmosphere conditions (1 kPa O 2:1 kPa CO 2) or under ULO conditions followed by different periods (2 and 4 weeks) in cold AIR atmosphere (ULO + 2w or ULO + 4w, respectively). Standard quality and emission of volatile compounds were analyzed after storage plus 1 and 7 days at 20 degrees C. Sensory attributes and acceptability were also determined after 7 days at 20 degrees C. The extra period of 30 weeks in an AIR atmosphere after ULO storage resulted in an increase in the concentration of the compounds that most contribute to the flavor of Fuji apples. These fruits were relatively well accepted by consumers despite a slight decline in firmness and acidity.

Volatile emission after controlled atmosphere storage of Mondial Gala apples (Malus domestica): relationship to some involved enzyme activities.

Mondial Gala apples were harvested at commercial maturity and stored at 1 degrees C under either air or controlled atmosphere (CA) conditions (2 kPa O2/2 kPa CO2 and 1 kPa O2/1 kPa CO2), where they remained for 3 or 6 months. Data on emission of selected volatile esters, alcohol precursors, and activity of some aroma-related enzymes in both peel and pulp tissues were obtained during subsequent shelf life of fruit and submitted to multivariate analysis procedures. CA storage caused a decrease in the emission of volatile esters in comparison to storage in air. Results suggest that lessened ester production was the consequence of modifications in activities of alcohol o-acyltransferase (AAT) and lipoxygenase (LOX) activities. For short-term storage, inhibition of lipoxygenase activity in CA stored fruit possibly led to a shortage of lipid-derived substrates, resulting in decreased production of volatile esters in spite of substantial ester-forming capacity that allowed for some recovery of fruit capacity for ester emission during the shelf life. For long-term storage, strong inhibition of AAT activity in CA stored fruit in combination with low LOX activities resulted in unrecoverable diminution of biosynthesis of volatile esters.

Characterization of Fuji apples from different harvest dates and storage conditions from measurements of volatiles by gas chromatography and electronic nose.

Volatile compounds in Fuji apples harvested at two different maturities were measured at harvest and after 5 and 7 months of cold storage (1 degrees C) in four different atmospheres. When the samples were characterized by both chromatographic measurements of volatiles and responses of an electronic nose, the analyses showed a clear separation between fruits from different storage conditions (a normal cold atmosphere and three controlled atmospheres). During poststorage, the apples were left to ripen for 1, 5, and 10 days at 20 degrees C before analytical measurements were done involving headspace-gas chromatography methods and electronic nose type quartz crystal microbalances. Electronic nose responses registered by seven different sensors were used to classify the apples using principal component analysis. It was possible to identify the samples from different storage periods, days of shelf life, and harvest dates, but it was not possible to differentiate the fruits corresponding to different cold storage atmospheres.