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Jun Song and Fritz Bangerth

Changes in the profile of aroma volatiles during ripening and after ethylene treatment in apple fruit have revealed a close relationship exists between ethylene production and the timing and magnitude of volatile synthesis. Therefore, AVG (ethylene biosynthesis inhibitor) was used to study the relationship between volatile biosynthesis and metabolic processes affected by ethylene in apple fruits. AVG-treated fruit were stored either for 1 month at 2C in air or 5 months in ULO condition. During the post-storage ripening, the fruits were exposed to 50 μl·liter–1 of ethylene at least 12 h/day. Aroma production was determined at 20C. Total volatile compound production by AVG-treated fruit was much lower than that of untreated fruit. A significant increase in the production of most aroma volatile after 1 month storage in air was induced by ethylene treatment to AVG-treated fruits. I was noted that branched-chain volatile, such as 2-methylbutylacetate, which originates from branched-chain amino acids, has increased prior to butylacetate and hexylacetate, which are derived from fatty acids. Ethylene treatment was unable to stimulate the production of straight-chain volatile compounds following the 5 months of ULO storage. These results are consistent with observations suggesting apple fruit lose their sensitivity to ethylene after long ULO storage.

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Weimin Deng, Jun Song and Randolph M. Beaudry

The effect of polymers used in packaging on the aroma of the packaged product has been little explored. Using a package-in-a-jar system, we are able to simultaneously measure volatile production by plant organ (Malus domestica Borkh. cv. Golden Delicious) and the permeability of the packaging film to those volatiles. In this system, apple fruit were placed into a glass container or sealed in a low-density polyethylene(LDPE) package and subsequently placed into a glass container. Air or a modified atmosphere was slowly passed through the glass containers such that the O2 level in the package was similar to that in containers with no package. The package and jar head spaces were sampled for CO2, O2, ethylene, and aroma volatile analysis by gas chromatography/mass spectrometry. The effect of temperature, atmosphere and film presence to some major volatile compounds was determined. When storage temperature increased from 0°C to 22°C the production rate of hexylacetate and 2-methyl butylacetate increased 11.27- and 17.15-fold, respectively. At 0°C, as O2 decreased in concentration from 10% to 5% (v/v), hexylacetate and butylacetate declined significantly; however, 2-methyl butylacetate was not affected. This can be taken to indicate the production of 2-methyl butanol for 2-methyl butylacetate formation is not as O2 concentration dependent as straight-chain alcohols. At the same O2 concentration, non-packaged fruit evolved greater amounts of all volatiles than packaged fruit. The flux of α-farnesene, hexylacetate and 2-methyl butylacetate was 26.6-, 1.7-, and 1.4-fold higher, respectively, for fruit in glass container. The sorption of α-farnesene and some other volatiles into LDPE film is evidently considerable, altering the aroma profile of packaged produce relative to a flow-through system.

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Lihua Fan, Jun Song and Randolph Beaudry

Hexanal vapor is a natural, metabolizable fungicide that inhibits fungal activity and enhances the aroma biosynthesis in sliced apple fruit. Whole apple fruit were inoculated at two points per fruit with Penicillium expansum at a concentration of 0.5 × 105 spore/ml and treated with hexanal vapors. Inoculated fruit were exposed to hexanal for 48 hr and kept for another 72 hr in hexanal-free air at 22°C. Treatments included 8.2–12.3 μmol·L–1 (200–300 ppm), 14.5-18.6 μmol·L–1 (350–450 ppm), and 24.8-28.9 μmol·L–1 (600–700 ppm), each with an air control. At a concentration of 200–300 ppm hexanal, there was no fungal growth during treatment, but lesion development was evident on 100% of the treated fruit following cessation of treatment. After 72 hr holding in air, lesion diameter was significantly smaller for treated fruit. When inoculated apple fruit were exposed to 350–450 ppm and 600–700 ppm hexanal vapors, the decay rate was 44.7% and 23.9%, respectively, while the decay rate of inoculated control apple fruit was 100% and 98%, respectively, after 72 hr holding in air. The development of aroma volatiles was investigated for both treated and untreated whole apple fruit. Hexanal was actively converted to aroma volatiles by `Golden Delicious' fruit and there was no detectable hexanal emanations. The amount of hexylacetate, hexylbutanoate, hexylhexanoate, hexylpropionate, butylhexanoate, and hexyl-2-methybutanoate were about 2- to 4-fold higher in treated apple fruit than in untreated apple fruit. `Mutsu' apple fruit were treated with 350–450 ppm hexanal for 48 hr and processed into apple sauce within 4 hr. An informal sensory evaluation for processed `Mutsu' apple revealed no apparent flavor difference between treated and control fruit sauce.

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Fritz K. Bangerth, Jun Song and Josef Streif

After a brief description of the “history of research” of aroma volatiles of apple and strawberry fruit, possible reasons for the reduced production of these important quality attributes by particular pre- and postharvest procedures are given. Among the possible physiological factors in association with reduced aroma volatile production, a reduced ethylene sensitivity, a decline in the rate of respiration as well as the content of adenine nucleotides and limited free fatty acids as precursors for aroma volatiles biosynthesis are proposed. A hypothesis about how this sequence of events leads to reduced volatile production is given and finally some suggestions of how to improve volatile synthesis are discussed.

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Randolph M. Beaudry, Jun Song and Weimin Deng

Apple scald (peel browning) is hypothesized to involve a chilling disorder. Numerous studies have linked chloroplast fluorescence changes with chilling injury before symptom development. Therefore, chloroplast fluorescence was used for the prediction of scald in apples. `Red Delicious' apple fruit were harvested at three maturities and stored at 1 to 2C. They were removed from storage weekly and placed at ambient temperature (22C). Chloroplast fluorescence was measured at 0, 3, and 7 days after removal. A significant decline in quantum yield response (Fv/Fm), which indicates a reduction of chloroplast function, was recorded after 30 days in first-harvest fruit and 40 to 50 days in the second- and third-harvest fruit. The decline in Fv/Fm preceded scald development by ≈30 days in first-harvest fruit and 20 to 30 days in second- and third-harvest fruit. The data suggest that fluorescence changes and scald development may be related physiologically. Fruit firmness and other fruit ripening phenomena were also measured and their relationship to the fluorescence and scald development were investigated. The results indicated that the chloroplast fluorescence may be used as a predictive tool for scald development in stored apple fruit.

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Charles F. Forney*, Jun Song and Michael A. Jordan

Apple fruit are treatmented with diphenylamine (DPA) in the form of a postharvest dip to prevent the development of storage scald. However, DPA residues have been detected on apples not treated with DPA, which is problematic in markets where DPA residues are not acceptable. The objective of this study was to identify sources of DPA contamination and evaluate the effectiveness of ozone to reduce contamination. Concentrations of DPA in the atmosphere of commercial storage rooms was monitored during the storage season and the adsorption of DPA onto wood and plastic bin material, plastic bin liners, foam insulation, and apple fruit was assessed. DPA was sampled from headspace with solid phase micro extraction using 65 μm polyacrylate micro fibers and analyzed using GC-MS. The effectiveness of gaseous treatments of 300 and 800 ppb ozone to reduce DPA contamination on apple fruit and bin material was also determined. DPA was found to volatilize from treated apples and bins into the storage room air, where it was adsorbed onto storage room walls, bins, bin liners and other fruit. DPA was found in the atmosphere of storage rooms containing apples that were not treated with DPA. Wood and plastic bin material, bin liners, and foam insulation all had a high affinity for DPA and were determined to be potential sources of contamination. Ozone reacted with DPA and following gaseous ozone treatments, off-gassing of DPA from wood and plastic bin material and bin liners was reduced. However, ozone was not effective in removing all DPA in contaminated materials and was ineffective in removing DPA from contaminated apples. Due to the pervasive and persistent nature of DPA, fruit should be handled and stored in facilities where DPA is not used to prevent contamination of fruit.

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Sastry Jayanty, Mauricio Canoles, Alejandra Ferenczi, Jun Song and Randolph Beaudry*

Volatile aroma compounds produced by apple, banana, and tomato are produced throughout development, however, those associated with ripening and edible quality are dependent upon ethylene action. In apple and banana, characteristic aroma is, in large part, dependent upon the formation of volatile esters. In tomato, many of the characteristic aromas are dependent upon tissue disruption and result from aldehydes and alcohols following lipid degradation. For apple and banana, the enzyme alcohol acyl-CoA transferase (AAT, EC 2.3.1.84) is the enzyme responsible for the final reaction in the pathway for ester formation and catalyzes the union of an alcohol and the CoA derivative of fatty acids. In both tissues, AAT gene expression was detected prior to the onset of ester production. In apple, AAT expression was found to be closely tied with the onset of autocatalytic ethylene synthesis. In banana, ethylene synthesis peaked and began to decline well before ester synthesis began. However, the expression of AAT increased as ester production increased for both tissues. Tomato fruit, like apple and banana, produced characteristic aromas following the onset of the ethylene climacteric, suggesting changes in the activity of various components of the lipoxygenase pathway. In all three tissue types, there are continuous, significant shifts in the aroma profile as fruit ripen age, suggesting shifts in specific metabolic pathways associated with precursor synthesis or degradation.

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Jun Song, Lihua Fan, Paul D. Hildebrand and Charles F. Forney

The biological effect of corona discharge on onions (Allium cepa L.) in a commercial storage was investigated. Surface discoloration and mold were modestly but significantly reduced by the corona discharge when onions were stored for 2 or 4 weeks with or without an additional 2 weeks of shelf life under high humidity. Corona discharge treatment also reduced airborne mold spores in the storage room. No significant changes in internal decay, firmness, sprouting, or rooting, in treated onions were found.

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Lihua Fan, Jun Song, Charles F. Forney and Michael A. Jordan

Ethanol concentration and chlorophyll fluorescence (CF) were measured as signs of heat stress in apple fruit [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.]. `McIntosh', `Cortland', `Jonagold', and `Northern Spy' apples were placed in trays and exposed to 46 °C for 0, 4, 8, or 12 hours. Following treatments, fruit were stored in air at 0 °C and evaluated after 0, 1, 2, or 3 months. Ethanol and ethylene production, CF, peel and flesh browning, firmness, skin color, soluble solids, and titratable acidity were measured. Increases in ethanol were apparent immediately following 12-hour heat treatments as well as after 3 months. After 3 months, ethanol concentrations were 16-, 52-, 6-, and 60-fold higher in `McIntosh', `Cortland', `Jonagold', and `Northern Spy' apples than in controls, respectively. The concentrations of ethanol accumulated reflected the degree of heat-induced fruit injury. Heat treatments reduced ethylene production relative to control values. After 3 months of storage ethylene production of fruit exposed to 46 °C for 12 h was <0.48 μmol·kg-1·h-1 compared to >4.3 μmol·kg-1·h-1 for controls. Heat treatments also reduced CF which was expressed as Fv/Fm, where Fv is the difference between the maximal and the minimal fluorescence (Fm - Fo), and Fm is the maximal fluorescence. After 3 months storage at 0 °C, Fv/Fm was ≈0.2 in fruit held at 46 °C for 12 hours compared with 0.5-0.6 for control fruit. Exposure to 46 °C for 12 hours caused severe peel and flesh browning in all cultivars. Severity of peel and flesh browning increased with increasing duration of heat treatment and subsequent storage at 0 °C. `Northern Spy' apple fruit were most susceptible to heat stress based on the degree of flesh browning. Heat treatments of 8 and 12 hours reduced firmness of `McIntosh', `Cortland', and `Northern Spy', but not `Jonagold' apples. Hue angle of the green side of fruit was also reduced in `Cortland', Jonagold' and `Northern Spy' apples receiving the 8- and 12-hour treatments. Heat treatments caused a decrease in fruit tiratable acidity, but had no effect on soluble solids content. The increase in ethanol production and decrease in CF correlated with heat-induced injury, and were apparent before browning was visually apparent. Ethanol and CF have the potential to be used to nondestructively predict the severity of injury that develops during storage.

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Jun Song, Lihua Fan, Charles F. Forney and Michael A. Jordan

Ethanol production and chlorophyll fluorescence were measured as signals of freezing and heat stress in apple fruit. `Cortland' and `Jonagold' apples were held at –8.5 °C for 0, 6, 12 or 24 h (freezing treatments), or at 46 °C for 0, 4, 8 or 12 h (heat treatments). Following treatments, fruit were stored at 0 °C and evaluated after 0, 1, 2, or 3 months. Following storage, fruit samples were kept for 12 h at 20 °C and then analyzed for ethanol production, chlorophyll fluorescence, and visible injury. Severity of flesh browning increased with increasing treatment time for both freezing and heat treatments. Freezing for 24 h and heating for 12 h caused severe flesh browning in both cultivars. Severity of heat-induced browning increased during storage. Increases in ethanol production were apparent 12 h following treatments and reflected the degree of stress-induced fruit injury. After 2 months of storage, ethanol concentrations peaked and were as much as 400-fold greater than that of controls. These stress treatments also reduced ethylene production and chlorophyll fluorescence. The degree of increase in stress-induced ethanol production and decrease in chlorophyll fluorescence correlated with stress-induced injury and could be used to predict the severity of injury that develops during storage. Other volatile production and their relationship to fruit stress will also be discussed.