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PubMed2004

Production of volatile compounds by Fuji apples following exposure to high CO2 or low O2.

The emission of volatile compounds by Fuji apples following short- or long-term exposure to high CO(2) was studied. The production of ethanol, methyl and ethyl esters, octanal, nonanal, and decanal was enhanced while the production of C(3)-C(6) alcohols, propyl, butyl, pentyl, and hexyl esters and butanal decreased in fruit exposed to 10 kPa O(2) + 20 kPa CO(2) at 20 degrees C for up to 12 days. The impact of high CO(2) exposure on volatile production was dependent on fruit maturity at harvest. Apples stored for 8 months in an ultralow O(2)-controlled atmosphere (CA) (0.5 kPa O(2) + 0.05 kPa CO(2)) or high CO(2) CA (1.5 kPa O(2) + 3 kPa CO(2)) at 0.5 degrees C had reduced production of most volatiles, especially butyl and hexyl esters, as compared to fruit stored in air. Two exceptions were ethanol and ethyl acetate for which the production was enhanced by both CA regimes. Treatment with the antioxidant diphenylamine prior to storage prevented most of the high CO(2)-induced and ultralow O(2)-induced changes in volatile production. The results of this study do not indicate that changes in volatile production following the exposure of Fuji apples to high CO(2) are causally related to the development of CO(2) injury.

Alcohols↗

PubMed2004

Impact of suppression of ethylene action or biosynthesis on flavor metabolites in apple (Malus domestica Borkh) fruits.

To understand the role of ethylene in regulating the overall flavor of apple fruits, ethylene production or action was reduced using transgenic apple trees suppressed for ACC-synthase or ACC-oxidase enzyme activity or by the addition of 1-methylcyclopropene (1-MCP), an ethylene action inhibitor. Flavor components were differentially regulated in response to the suppression of both ethylene biosynthesis and action. Headspace analysis of aroma production, an ethylene-associated event, showed a reduction in ester and alcohol production in the ethylene-suppressed lines and in the apples treated with 1 microL L(-1) 1-MCP for 20 h at 20 degrees C. However, no major differences were observed in the concentrations of aldehyde volatiles. Other flavor metabolites that showed an ethylene-dependent pattern were organic acids and sugars. Malic acid degradation was significantly reduced under ethylene-suppressed conditions, showing a recovery after the fruit was exposed to ethylene. Sucrose and fructose concentrations were influenced by suppression or enhancement of ethylene. Total phenolics as well as individual phenolic compounds showed an ethylene-dependent regulation only in response to the suppression of ethylene biosynthesis, but not when ethylene action was inhibited.

Amino Acid Oxidoreductases↗

PubMed2005

Interactive responses of gala apple fruit volatile production to controlled atmosphere storage and chemical inhibition of ethylene action.

Gala apples exposed to the ethylene action inhibitor 1-methylcyclopropene (1-MCP) for 12 h at 20 degrees C were stored at 1 degrees C in air or a controlled atmosphere (CA) maintained at 1 kPa O2 and 2 kPa CO2. Volatile compounds were measured after 4, 12, 20, and 28 weeks plus 1 or 7 days at 20 degrees C. Treatment with 1-MCP and then storage in air or CA or storage in CA without 1-MCP treatment reduced volatile production as compared to apples not treated with 1-MCP stored in air. The reduced production of esters, alcohols, aldehydes, acetic acid, and 1-methoxy-4-(2-propenyl)benzene was observed. Ester production by fruit stored in CA decreased throughout the storage period regardless of previous 1-MCP treatment. The production of esters, alcohols, aldehydes, acetic acid, and 1-methoxy-4-(2-propenyl)benzene by 1-MCP-treated fruit stored in air plus 7 days at 20 degrees C increased after 20 or 28 weeks of storage. Continuous exposure to 417 micromol m(-3) ethylene for 7 days at 20 degrees C after 12 or 28 weeks of storage stimulated production of many volatile compounds, primarily esters and alcohols, by fruit stored in CA or 1-MCP-treated apples stored in air. However, exposure to ethylene had no effect on the production of aldehydes or acetic acid.

Acetic Acid↗

PubMed2006

Characterization of odor-active volatiles in apples: influence of cultivars and maturity stage.

The aroma and texture of three different apple cultivars, harvested at three maturity stages, were analyzed by sensory and instrumental analysis. The emphasis was on the identification of the most potent odorant volatiles, and the challenge was to separate the few most important flavor compounds, which may be trace chemicals, from the vast number of nonodorant compounds present in apple aroma extracts. Thirty-six odorant compounds were detected, 24 of which were common to all extracts. A significant correlation coefficient was found between the aroma intensity scores and overall quantity of the odorant volatiles, which shows that the development of sensory aroma is similar to that of odorant volatiles. This study also showed that the parameters measured by penetrometry and compression were highly correlated with sensory textural attributes. The determination of the optimal maturity stage for different apple cultivars by the usual parameters, such as color, diameter, total soluble solids, and titrable acidity, may not be sufficient to determine the optimal sensory quality for consumers. Moreover, the sensory quality of fruits changes during maturation in a different way from one cultivar to another, and this should be taken into account.

Fruit↗

PubMed2006

Postharvest variation in apple (Malus x domestica Borkh.) Flavonoids following harvest, storage, and 1-MCP treatment.

The impact of 1-methylcyclopropene (1-MCP) on the synthesis and retention of flavonoid compounds during storage and ripening of red Delicious (Malus x domestica Borkh.) apples was investigated. Numerous anthocyanins, flavonols, flavan-3-ols, and a hydroxycinnamic acid from three different fruit harvest maturities were monitored after a 120 day storage and 1 week shelf life period using high-performance liquid chromatography/diode array detector analysis. The total flavonoid concentration was 5% greater in fruit treated with 1-MCP, whereas chlorogenic acid levels were 24% lower. All compounds analyzed increased in concentration during fruit harvest; however, the anthocyanins generally declined after storage, while chlorogenic acid levels increased. 1-MCP treatment resulted in the retention of anthocyanins in the latter stages of storage but did not affect the flavonols and flavan-3-ols. Chlorogenic acid biosynthesis from early and optimal fruit harvest maturities was greatly inhibited by 1-MCP during storage and the 1 week shelf life period. However, 1-MCP did not affect chlorogenic acid concentrations in late-harvested fruit. Results suggest that 1-MCP may inhibit the activity of phenylalanine ammonia-lyase and subsequent biosynthesis of flavonoid compounds. However, because very little postharvest biosynthesis of flavonoids occurs in apples, 1-MCP treatment may be useful for maintaining some of the intrinsic flavonoid levels of red Delicious apples, if applied at the proper harvest maturity.

Chlorogenic Acid↗

PubMed2006

Free-radical-scavenging and antioxidant activities of secondary metabolites from reddened cv. Annurca apple fruits.

Forty-three secondary metabolites were isolated and characterized from cv. Annurca apple fruit, an apple variety cultivated in the south of Italy. This apple cultivar undergoes a typical reddening treatment after collection. All of the compounds were characterized on the basis of their spectroscopic data. The compounds were tested for their radical-scavenging and antioxidant activities by measuring their capacity to scavenge DPPH* (2,2'-diphenyl-1-picrylhydrazyl radical), H2O2, and NO (nitric oxide) and to inhibit the formation of methyl linoleate conjugated diene hydroperoxides or TBARS (thiobarbituric acid reactive species).

Antioxidants↗

PubMed2006

Effect of thermal treatment on the quality of cloudy apple juice.

Apple juice from eight different varieties of apples was heated at high-temperature (60-90 degrees C) and short-time (20-100 s) (HTST) combinations. To determine the effect of heating conditions on enzymatic browning and cloud stability in apple juices, the activity of polyphenol oxidase and pectinesterase was analyzed and correlated with the thermal treatment conditions and the quality of the juice. Additional investigations included the measurement of pH value, soluble solid content, titratable acidity, color, and turbidity after 3 and 6 months. The results showed that HTST treatment at 80 degrees C already inactivated polyphenol oxidase, whereas pectinesterase activity was reduced to half and could even at 90 degrees C not be inactivated completely. In fact, highest residual pectinesterase activity was found at 60 degrees C. Heating at 70 degrees C caused stable pectinesterase activity and even a slight increase for 50 and 100 s heating times. Turbidity and lightness increased after HTST treatment. In particular, differences in cloud stability between the varieties were measured. HTST parameters did not correlate with the residual cloud stability after 6 months. The sensory evaluation revealed that only a few combinations were distinguishable. The best stability of cloud and color in relation to heat impact was achieved by HTST treatment between 70 degrees C/100 s and 80 degrees C/20 s.

Beverages↗

PubMed1999

Development of apple superficial scald, soft scald, core flush, and greasiness is reduced by MCP.

1-Methylcyclopropene (MCP) was used to evaluate the role of ethylene in development of apple (Malus x domestica Borkh.) physiological disorders during storage. Granny Smith, Red Chief Delicious, and Fuji apple fruit were treated with MCP at a concentration of 1 microL L(-)(1) for 12 h at 20 degrees C. For all varieties stored at 0 degrees C, ethylene production and respiration rates were reduced for several months following MCP treatment, and firmness and titratable acidity of treated fruit were higher compared to controls. Apples treated with MCP did not develop superficial scald or peel greasiness through 6 months storage plus ripening at 20 degrees C for 7 days. Core flush was not observed in MCP-treated fruit until 6 months after treatment when the incidence was still lower compared to control fruit. MCP delayed the rise in production of alpha-farnesene and reduced accumulation of its oxidation products.

Cyclopropanes↗

PubMed1999

Analysis of patulin in apple juice by diphasic dialysis extraction with in situ acylation and mass spectrometric determination.

A procedure combining diphasic dialysis extraction with in situ acylation and gas chromatography/mass spectrometry (GC/MS) determination was developed for detection and quantification of the mycotoxin patulin in apple juice. Apple juice samples spiked with 4-N,N-dimethylaminopyridine were dialyzed using methane chloride and acetic anhydride inside dialysis tubing. Patulin was derivatized into its acetate and collected in the tubing after diphasic dialysis and was directly determined using GC/MS with the selective ion monitoring mode without further concentration and cleanup steps. Quantification was carried out by a calibration curve with an internal standard of correlation. The appropriate parameters of both dialysis and derivatization were examined. The linear range of the calibration curve was found to be 10-250 microg/L for patulin, and the limit of quantification was 10 microg/L. Levels of patulin ranging from 0 to 107.2 microg/L with 77-109% recovery were found in 10 apple samples. The technique combining diphasic dialysis extraction and acylation was demonstrated and showed potential for other applications.

Acylation↗

PubMed1999

Relationships between volatile production, fruit quality, and sensory evaluation in Granny Smith apples stored in different controlled-atmosphere treatments by means of multivariate analysis.

Aroma compounds, quality parameters, and sensory evaluation of Granny Smith apples were analyzed after 3, 5, and 7 months of cold storage in three controlled-atmosphere (CA) treatments, in which oxygen and carbon dioxide were held at 1, 2, and 3%. During poststorage ripening, the apples were kept at 20 degrees C for 1, 5, and 10 days before analytical measurements were made. The highest volatile emission was obtained after 5 months of storage in all CA treatments, reaching its highest value when a low-oxygen CA (LO) was used. Ultralow-oxygen CA (ULO) showed the highest ability to maintain apple firmness. The correlation among analytical and sensory parameters suggests that ethyl 2-methylbutyrate, 1-butanol, pentyl acetate, and tert-butyl propionate are the aroma compounds with the highest influence in the sensorial score. Concerning CA treatments, LO and ULO appear to be very valuable technologies for maintaining the sensorial quality even after 7 months of storage.

Alcohols↗

PubMed1999

Characterization of Royal Gala apple aroma using electronic nose technology-potential maturity indicator.

Changes in aroma of apple harvested at four different maturities were measured at harvest and after short-term storage using electronic aroma sensors ("electronic nose") and classical headspace/gas chromatography methods. Stored fruits were also evaluated by a trained sensory panel. Compared with headspace/gas chromatography, the electronic nose was found to be more sensitive ( approximately 40 times) in terms of sample size. The sampling procedure for the electronic nose was much less complex. Using discriminant function analysis, both methods classified the apples tested into groups according to harvest date. After storage, the groupings were more diffuse. Results from sensory testing showed partial separation along the first linear discriminant but did not classify the apple into distinct groups. Important differences between treatments were found for "overall flavor", "acid flavor" intensity, "crispness", "cider/fermented aroma", "vegetative aroma", and "canned pear aroma".

Electronics↗

PubMed1999

Polyphenol profiles of French cider apple varieties (Malus domestica sp.).

The cortex of 14 French apple varieties (12 cider and 2 juice varieties), one English cider variety, and one dessert apple (i.e., Golden Delicious) were studied for their polyphenol composition. Total polyphenols were assayed by the Folin-Ciocalteu method, and the precise polyphenolic composition (monomeric catechins, proanthocyanidins, hydroxycinnamic acids, and dihydrochalcones) was obtained by HPLC following thiolysis. ESI-MS and ESI-MS/MS analyses showed that chlorogenic acid and p-coumaroylquinic acid were methylated under the conditions of thiolysis. Depending on the variety, the global polyphenol concentration varied from 1 to 7 g per kilogram of fresh cortex. Cider varieties globally showed a higher polyphenol concentration than the dessert apple Golden Delicious, bitter varieties being the more concentrated. The proportion of the polyphenol classes varied greatly from one cultivar to another. For all varieties, procyanidins were always the predominant class. They were mainly constituted of (-)-epicatechin units with a small proportion of (+)-catechin as a terminal unit. The average degree of polymerization ranged between 4.2 and 7.5 depending upon the variety with an exception for the sharp varieties Guillevic and Avrolles which showed significant concentrations of procyanidins with DPn of 40 and 50, respectively.

Beverages↗

PubMed2000

Temperature-dependent autoxidation of conjugated trienols from apple peel yields 6-methyl-5-hepten-2-one, a volatile implicated in induction of scald.

Conjugated triene (CT) oxidation products of alpha-farnesene have long been thought to be involved in development of superficial scald in apple fruit. Early studies found that CT hydroperoxides and the volatile 6-methyl-5-hepten-2-one (MHO) are major in vitro autoxidation products of alpha-farnesene. However, it was recently shown that > or =99% of the oxidation products of alpha-farnesene that accumulate in apple peel are conjugated trienols (CTols), isomers of 2,6,10-trimethyldodeca-2,7,9,11-tetraene-6-ol. HPLC-purified CTols from fruit of two scald-susceptible cultivars, Granny Smith (GS) and Red Delicious (RD), were used to study autoxidation of these compounds in vitro. Incubation of CTols in sealed glass vials under air resulted in accumulation of MHO. Oxygen enrichment did not increase the amount of MHO produced. Regardless of which cultivar CTols were derived from, at 0 degrees C autoxidation yielding MHO was quite slow and linear, whereas at 20 degrees C MHO production was much more rapid, and after several hours the rate increased abruptly. However, CTols isolated from GS and RD fruit differed in the duration of the initial lag phase and the overall level of MHO generated at 20 degrees C. The sharp increase in MHO production occurred after 3 h with GS CTols and at about 12 h with RD CTols. Also, the yield of MHO from GS CTols after 6 h at 20 degrees C was nearly 6-fold greater than that from RD CTols after 20 h at 20 degrees C. The antioxidants butylated hydroxytoluene and diphenyamine reduced the yield of MHO by about 97%. Recent work has shown that MHO can induce scald-like symptoms in apple peel and that tissue sensitivity increases with time in storage. This may explain the correlation between high CTol levels and scald development, and why symptoms rapidly intensify when fruits are removed from cold storage.

Fatty Acids↗

PubMed2000

Acceleration of pH variation in cloudy apple juice using electrodialysis with bipolar membranes.

The purpose of this study was to accelerate pH variation in cloudy apple juice using electrodialysis (ED). The testing was conducted using two ED configurations. The bipolar and cationic membrane configuration showed that reducing the spacing from 8 to 0.75 mm had little effect on treatment time, whereas stacking eight bipolar membranes reduced acidification time by 30%, although the treatment still took too long (21 min). Furthermore, it was not possible to acidify apple juice to a pH of 2.0 to completely inhibit enzymatic browning. The bipolar and anionic membrane configuration helped to accelerate the acidification step by a factor of 3, increasing the yield from 3.3 to 10 L of juice/m(2) membrane/min. Moreover, treatment time was inversely proportional to the size of the membrane stack. The speed at which the pH of acidified juice returned to its initial value was, however, 4 times slower than the speed of acidification, giving a yield of 2.5 L of juice/m(2) membrane/min. By accelerating the acidification step, ED treatment with bipolar and anionic membranes results in more effective polyphenol oxidase activity and more rapid control of juice browning at pH 2.0. Also, the treatment has very little effect on the chemical composition and organoleptic quality of apple juice.

Beverages↗

PubMed1998

Investigation of Agrobacterium-mediated transformation of apple using green fluorescent protein: high transient expression and low stable transformation suggest that factors other than T-DNA transfer are rate-limiting.

To investigate early events of Agrobacterium-mediated transformation of apple cultivars, a synthetic green fluorescent protein gene (SGFP) was used as a highly sensitive, vital reporter gene. Leaf explants from four apple cultivars ('Delicious', 'Golden Delicious', 'Royal Gala' and 'Greensleeves') were infected with Agrobacterium EHA101 harboring plasmid pDM96.0501. Fluorescence microscopy indicated that SGFP expression was first detected 48 h after infection and quantitative analysis revealed a high T-DNA transfer rate. Plant cells with stably incorporated T-DNA exhibited cell division and developed transgenic calli, followed by formation of transgenic shoots at low frequencies. The detection of SGFP expression with an epifluorescence stereomicroscope confirmed the effectiveness of SGFP as a reporter gene for detection of very early transformation events and for screening of putative transformants. The efficiency of the transformation and regeneration process decreased ca. 10,000-fold from Agrobacterium infection to transgenic shoot regeneration, suggesting that factors other than Agrobacterium interaction and T-DNA transfer are rate-limiting steps in Agrobacterium-mediated transformation of apple.

Blotting, Southern↗

PubMed1998

Apple ACC-oxidase and polygalacturonase: ripening-specific gene expression and promoter analysis in transgenic tomato.

Levels of 1-aminocyclopropane-1-carboxylate (ACC) oxidase and polygalacturonase (PG) mRNAs were characterized during ripening of Royal Gala, Braeburn and Granny Smith apples. Both ACC-oxidase and PG mRNAs were up-regulated in ripening fruit of all three cultivars. Expression in Royal Gala was detected earlier than in Braeburn and Granny Smith, relative to internal ethylene concentration. Genomic clones corresponding to the ACC-oxidase and PG mRNAs expressed in ripe apple fruit were isolated and ca. 2 kb of each promoter was sequenced. The start point of transcription in each gene was mapped by primer extension, and sequences homologous to elements in other ethylene-responsive or PG promoters were identified. The fruit specificity of the apple ACC-oxidase and PG promoters was investigated in transgenic tomato plants using a nested set of promoter fragments fused to the beta-glucuronidase (gusA) reporter gene. For the ACC-oxidase gene, 450 bp of 5' promoter sequence was sufficient to drive GUS expression, although this expression was not specific to ripening fruit. Larger fragments of 1966 and 1159 bp showed both fruit and ripening specificity. For the PG gene, promoter fragments of 1460 and 532 bp conferred ripening-specific expression in transgenic tomato fruit. However GUS expression was down-regulated by 2356 bp of promoter, suggesting the presence of a negative regulatory element between positions -1460 and -2356.

Amino Acid Oxidoreductases↗

PubMed2000

MDH1: an apple homeobox gene belonging to the BEL1 family.

Differential display was used to isolate genes differentially expressed early in fruit development of apple (Malus domestica Borkh.). This approach resulted in the isolation of MDH1, a homeobox gene with a homeodomain similar to that of BELL1 (BEL1), which is involved in regulation of ovule development in Arabidopsis. However, outside the homeodomain MDH1 is quite different from BEL1. In apple, MDH1 mRNA was predominantly found in flowers, expanding leaves and expanding fruit. In pre-anthesis flowers, in situ hybridization showed that MDH1 mRNA accumulated in ovules. To further investigate the function of this new homeobox gene, MDH1 was transformed into Arabidopsis thaliana under the control of the cauliflower mosaic virus 35S promoter. The transgenic Arabidopsis plants showed dwarfing, reduced fertility and changes in carpel and fruit (silique) shape. The size and shape of the cells in the transgenic fruit was irregular. Both the transgenic phenotypes in Arabidopsis and the expression pattern of this gene in apple are consistent with the idea that MDH1 is likely to play an important role in control of plant fertility.

Amino Acid Sequence↗

PubMed1999

Calcium chloride sprays decrease physiological disorders following long-term cold storage of apple.

Field experiments were carried out at the Horticultural Research Institute, and in five commercial apple orchards, located in southwest Finland including the Aland Islands during 1994 and 1995. The experimental cultivars were Melba, Raike, Red Atlas, Akerö, Aroma, and Lobo. The treatments were untreated control and preharvest calcium chloride (CaCl2) sprays at Ca 2.0 g/l. Fruit samples were stored for two to six months at 2-4 degrees C and 85-95% RH. The percentage of the incidence of physiological disorders of stored apples was scored. Fruit macronutrients, firmness, diameter and juice titratable acidity and percentage of soluble solids were determined. Only a few nutrient effects in the flesh of the apple cultivars were a result of CaCl2 sprayings. Preharvest CaCl2 sprays increased fruit firmness and the titratable acidity but decreased soluble solids, soluble solids/titratable acidity ratio, and the incidence of physiological storage disorders of some cultivars. When looking at the mean of all cultivars, CaCl2 sprayings increased titratable acidity and decreased soluble solids/titratable acidity ratio after four months of storage during 1995 and reduced the incidence of physiological disorders after three months of storage during 1994 and after four months of storage in 1995.

Calcium Chloride↗