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James R. Gorny and Adel A. Kader

Autocatalytic C2H4 biosynthesis in preclimacteric apple fruit (Malus domestica Borkh. `Golden Delicious') was prevented by storage in atmospheres of 20% CO2-enriched air (17% O2 + 63% N2) or 0.25% O2 (balance N2). In preclimacteric fruit, both treatments inhibited C2H2 biosynthesis by suppressing expression of ACC synthase (ACC-S) at the mRNA level. ACC oxidase (ACC-O) mRNA abundance and in vitro enzyme activity also were impaired by these treatments. However, the conversion of ACC to C2H4 never became the rate limiting step in C2H4 biosynthesis. C2H4 biosynthesis also was effectively inhibited in climacteric apple fruit kept in air + 20% CO2 or 0.25% O2. Climacteric apples also exhibited suppressed expression of ACC-S at the mRNA level, while ACC-O transcript abundance, enzyme activity, and protein abundance were reduced only slightly. ACC-S is the key regulatory enzyme of C2H4 biosynthesis and is the major site at which elevated CO2 and reduced O2 atmospheres inhibit C2H4 biosynthesis, irrespective of fruit physiological maturity. Chemical names used: 1-aminocyclopropane-1-carboxcylic acid (ACC).

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Dana F. Faubion and Adel A. Kader

Tightly stacked pallets of wrapped, place-packed `Anjou' pears cooled slower than those that were tray packed. Half-cooling times ranged from 2.0 to 15.7 days for wrapped, place-packed pears vs. 1.7 to 7.4 days for tray-packed pears. More time was required to remove heat from the middle than from the top or bottom of the pallets in both packaging systems; however, the difference in temperature between the middle and the top or bottom of the pallet was greater for a longer period of time when pears were wrapped and place packed. The large range in temperature within a pallet illustrated that a pallet is not a uniform unit. A change in how fruit are packed can dramatically change how fruit cool within a pallet. Accumulation of carbon dioxide and ethylene in slower-cooling boxes suggests that tray packing can enhance `Anjou' quality by facilitating faster product cooling.

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James R. Gorny and Adel A. Kader

Preclimacteric `Golden Delicious' apples (Malus domestica Borkh.) were stored at 0 °C in: air; air + 5% CO2; 2% O2 + 98% N2; or 2% O2 + 5% CO2 + 93% N2, and sampled monthly for 4 months to investigate the mechanism(s) by which reduced O2 and/or elevated CO2 atmospheres inhibit C2H4 biosynthesis. Ethylene biosynthesis rates and in vitro ACS activity were closely correlated in all treatments, while in vitro ACO activity significantly increased over time regardless of the treatment. Only a small amount of C2H4 biosynthesis inhibition by lowered O2 and/or elevated CO2 atmospheres could be accounted for by suppressed induction of ACO activity. Western blot analysis demonstrated that apples held for 2 months in lowered O2 and/or elevated CO2 atmospheres had significantly reduced abundance of ACO protein, compared to fruit held in air. Northern blot analysis of ACS and ACO transcript abundance revealed that reduced O2 and/or elevated CO2 atmospheres delay induction and reduce the abundance of both transcripts. Reduced O2 and/or elevated CO2 atmospheres reduce C2H4 biosynthesis by delaying and suppressing expression of ACS at the transcriptional level and by reducing the abundance of active ACO protein. Chemical names used: 1-aminocyclopropane-1-carboxylic acid (ACC), ACC synthase (ACS), ACC oxidase (ACO), ethylene (C2H4), S-adenosylmethionine (AdoMet).

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Diana L. Lange and Adel A. Kader

Preclimacteric avocado [Persea americana (Mill.) cv. Hass] fruit or fruit disks as well as fruit harvested in either June (midseason) or August (late season) and partially ripened were kept in air (21% O2 + 78% N2), 20% CO2 + 17% O2 (63% N2), or 40% CO2 + 13% O2 (47% N2) at either 10 or 20 °C. Ethylene production by preclimacteric fruit was completely inhibited during CO2 exposure, whereas there was only partial inhibition of ethylene production when partially ripened fruit were exposed. Compared to the fruit stored in air, O2 uptake of fruit stored in 20% CO2 was decreased by 20%, whereas the fruit stored in 40% CO2 showed 25% more O2 uptake than air-stored fruit. Fruit subjected to a storage regime of 40% CO2 at 10 °C followed by 2 d in air had the best visual quality. In general, climacteric fruit treated with 20% CO2 at 10 °C showed increased pyruvate dehydrogenase (PDH) activity and decreased cytochrome oxidase (CytOx) activity. Fruit stored in 40% CO2 had reduced CytOx activity compared to air-stored fruit, and PDH activity was variable depending on the harvest season of the fruit. Our results show that the effect of elevated CO2 on a given enzyme depends on concentration of CO2, duration of exposure, physiological state of the fruit, and type of tissue exposed.

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Diana L. Lange and Adel A. Kader

Partially ripened avocado [Persea americana (Mill.) cv. Hass] fruit harvested in either June or Aug. 1994 were kept at 10 °C in air (21% O2), 20% CO2 (17% O2, balance N2), or 40% CO2 (13% O2, balance N2) for 7 to 12 days and then were transferred to air at 10 °C for 2 to 3 days. Mitochondrial respiration was stimulated in response to elevated CO2 treatments at 10 °C. A shift to alternative pathway (Alt) respiration occurred on day 4 in experiments using avocados from both harvest dates, with a return to initial levels in only the 20% CO2-treated fruit (June-harvested fruit after return to air). Elevated CO2 at 20 °C decreased the in vitro O2 consumption of isolated mitochondria compared to mitochondria kept in air. The Alt pathway contributed less to the total O2 uptake of CO2-treated mitochondria compared to mitochondria kept in air. The respiratory control ratios of the CO2-treated fruit and mitochondria were higher and lower, respectively, than the air controls. Induction of 33 to 37 kD proteins (corresponding to the size of the alternative oxidase proteins) occurred in avocados after 4 days in 40% CO2. These results indicate that elevated CO2 has various effects depending on concentration, duration and temperature of exposure, and mitochondrial function of avocado fruit, such as increased and altered respiratory oxidation and up-regulation of alternative oxidase proteins.

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Diana L. Lange and Adel A. Kader

Changes in cytosolic and vacuolar pH, ATP, ADP, and the ATP : ADP ratio were measured in whole fruit or mesocarp disks of avocado [Persea americana (Mill.) cv. Hass] during brief exposures to elevated CO2. Intact climacteric fruit exposed to air (21% O2), 20% CO2 (17% O2, balance N2), or 40% CO2 (13% O2, balance N2) had cytosolic pH values of 7.0, 6.6, and 6.4, respectively, while mesocarp disks had cytosolic pH values of 6.9, 6.7, and 6.4, respectively. The ß-ATP levels of intact climacteric fruit exposed to 20% CO2 or 40% CO2 for 2 h were reduced by 25% or 43%, respectively, relative to air-exposed fruit. HPLC analysis of nucleotide phosphates from preclimacteric avocados revealed that ATP levels and the ATP : ADP ratio increased in 40% compared to the air-stored fruit. However, 1 day after transfer to air, the effects of elevated CO2 had dissipated. These modifications in cellular state could alter the activity of respiratory enzymes in fruit exposed to elevated CO2 atmospheres.

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Dangyang Ke, Lili Zhou, and Adel A. Kader

`Chandler' strawberries (Fragaria ananassa Duck.) were kept in air, 0.25% O2, 21% O2 + 50% CO2, or 0.25 O2 + 50% CO2 (balance N2) at 5C for 1 to 7 days to study the effects of controlled atmospheres (CAs) on volatiles and fermentation enzymes. Concentrations of acetaldehyde, ethanol, ethyl acetate, and ethyl butyrate were greatly increased, while concentrations of isopropyl acetate, propyl acetate, and butyl acetate were reduced by the three CA treatments compared to those of air-control fruit. The CA treatments enhanced activities of pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH) but slightly decreased activity of alcohol acetyltransferase (AAT). The results indicate that the enhanced PDC and ADH activities by CA treatments cause ethanol accumulation, which in turn drives the biosynthesis of ethyl esters. The increased ethanol concentration also competes with other alcohols for carboxyl groups for esterification reactions. The reduced AAT activity and limited availability of carboxyl groups due to ethanol competition decrease production of other acetate esters.

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Marius Huysamer, John M. Labavitch, and Adel A. Kader

Commercially grown Granny Smith apples were stored at 0°C in air or 1% O2, and 2 sets of samples were taken every 4 weeks over a 28 week period. One set was immediately analysed for weight loss, firmness, color, soluble solids, pH and titratable acidity. Alcohol-insoluble substances were analysed for starch, water-soluble uronides, water-insoluble uronides, cellulose and neutral sugars. The second set of samples was kept in air at 20°C for an additional week, during which respiration and ethylene production rates were measured, prior to the above analyses. Storage in 1% O2 led to the improved maintenance of firmness, reduced respiration and ethylene production rates in ambient air, and a reduced content of water-soluble uronides, suggesting a reduced degree of hydrolysis. The correlation between firmness and water-soluble uronide content was not very strong. The predominant neutral sugars present in the wall were arabinose and galactose, and activities of putative hydrolyses that may be involved in the metabolism of polymers containing these sugars will be discussed.

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Marius Huysamer, John M. Labavitch, and Adel A. Kader

Commercially grown Granny Smith apples were stored at 0°C in air or 1% O2, and 2 sets of samples were taken every 4 weeks over a 28 week period. One set was immediately analysed for weight loss, firmness, color, soluble solids, pH and titratable acidity. Alcohol-insoluble substances were analysed for starch, water-soluble uronides, water-insoluble uronides, cellulose and neutral sugars. The second set of samples was kept in air at 20°C for an additional week, during which respiration and ethylene production rates were measured, prior to the above analyses. Storage in 1% O2 led to the improved maintenance of firmness, reduced respiration and ethylene production rates in ambient air, and a reduced content of water-soluble uronides, suggesting a reduced degree of hydrolysis. The correlation between firmness and water-soluble uronide content was not very strong. The predominant neutral sugars present in the wall were arabinose and galactose, and activities of putative hydrolyses that may be involved in the metabolism of polymers containing these sugars will be discussed.

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Deirdre M. Holcroft, Maria I. Gil, and Adel A. Kader

The influence of CO2 on color and anthocyanin concentration in the arils of `Wonderful' pomegranate (Punica granatum L.) was investigated. Pomegranates were placed in jars ventilated continuously with air or air enriched with 10% or 20% CO2 at 10°C for 6 weeks. Samples were taken initially, and after 1, 2, 4, and 6 weeks and anthocyanin concentration was measured by HPLC. The arils of the pomegranates stored in air were deeper red than those stored in CO2-enriched atmospheres. This increase in red color resulted from an increase in anthocyanin concentration. Arils from fruit stored in air+10% CO2 had a lower anthocyanin concentration than air-stored fruit, and atmospheres enriched with 20% CO2 suppressed anthocyanin biosynthesis. Anthocyanin concentration was well-correlated to the activity of phenylalanine ammonia lyase (PAL), but not to glucosyltransferase (GT) activity. Moderate CO2 atmospheres (10%) prolong the storage life and maintain the quality of pomegranates, including an adequate red color of the arils.