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PubMed2005

Great differences in antioxidant properties exist between 56 apple cultivars and vegetation seasons.

The contents of ascorbate, thiols, and phenolic compounds and antioxidative enzyme activity were measured in the apple peel of 56 genotypes after harvest in two vegetation seasons, 2003 and 2004. The main reason of great interest in these bioactive compounds is their well-established physiological role in all living systems. The biggest differences between tested genotypes were noted for ascorbate peroxidase and glutathione reductase (GR) activity, followed by total ascorbate, phenolics, and glutathione concentration; the least difference was observed in the case of catalase. A large cultivar variation was noted in the anthocyanins and flavonols contents. Distinguishing the cultivars with the lowest, highest, relatively stable or those in which antioxidant content greatly differed depending on growing seasons was attempted. The GR activity is proposed as an environmental stress marker of apple fruit.

Anthocyanins↗

PubMed2005

Rapid determination of diphenylamine residues in apples and pears with a single multicommuted fluorometric optosensor.

In this work, a single flow injection multicommuted system using solid-surface fluorescence spectroscopy has been explored for the determination of diphenylamine in apples and pears. The native fluorescence signal of diphenylamine retained on the solid support (C18 silica gel) was used for its determination (lambda(exc)/lambda(em) 291/372 nm). The sample treatment consists of a liquid-liquid extraction with acetonitrile followed by a cleanup step using primary-secondary amines. The use of a continuous flow assembly implementing multicommutation, based on a set of three-way solenoid valves controlled by appropriate software, provides the automatic control of sample, carrier, and eluting solution with remarkable advantages in relation to conventional flow injection approaches. Using an optimized sampling time, the proposed method was linear in the range 0.25-5 mg kg(-1) with a detection limit of 0.06 mg kg(-1) and RSD (percent) values better than 3%. The proposed method was successfully applied to the analysis of diphenylamine in different apple and pear samples fortified at different concentrations, and recoveries between 78 and 104% were found. The results obtained illustrate the usefulness of the proposed method for the screening and evaluation of postharvest treatment of crops possibly containing diphenylamine.

Diphenylamine↗

PubMed2006

Comparison of the HPLC method and FT-NIR analysis for quantification of glucose, fructose, and sucrose in intact apple fruits.

A rapid quantification method was developed and validated for simultaneous and nondestructive quantifying the constituent sugar concentrations of intact apples using Fourier transform near-infrared (FT-NIR) spectroscopy in diffuse reflectance mode. Multiplicative scatter correction (MSC), the second derivative of Savitsky-Golay, and mean centering were used as spectral preprocessing options. Calibration models were established by the partial least squares (PLS) regression analysis, and validation of the method was performed according to the high-performance liquid chromatography (HPLC) chromatographic method. Spectral range and the number of PLS factors were optimized for the lowest root-mean-square error of prediction (RMSEP) and correlation coefficient of determination (r). The best models showed satisfactory predictions as measured by the RMSEP and r values: glucose, 0.201 and 0.950; fructose, 0.298 and 0.968; sucrose, 0.335 and 0.969, respectively. FT-NIR analysis of constituent sugar concentrations in the intact apple form was found to be more flexible and much faster than performed with the HPLC method.

Analysis of Variance↗

PubMed1999

Effect of superficial scald suppression by diphenylamine application on volatile evolution by stored Cortland apple fruit.

Cortland apple fruit (Malus x domestica Borkh.) stored for 120-140 days in air at 0 degrees C were warmed to 22 degrees C and held for 8 days. A portion of the fruit was dipped in a solution of diphenylamine (DPA) at harvest to prevent scald development. Scald occurred only in those fruit not treated with DPA, and its development was accelerated after transfer to 22 degrees C. Ester production from apple fruit tended to increase from day 0 to day 6 of poststorage holding and declined thereafter in both treatments. However, ester production in scald-developing fruit was reduced by approximately 50%. The reduction in volatile production remained relatively constant during the rapid development of scald symptoms. Furthermore, the reduction in volatile production appeared to be independent of respiration and ethylene production. Production of esters derived from hexanol was most reduced in fruit developing scald, with hexyl 2-methylbutanoate production being reduced approximately 15-fold. Interestingly, the production of methyl butanoate was detected only in scalding fruit. alpha-Farnesene production in fruit developing scald was reduced 43% compared with DPA-treated fruit. In contrast, the primary volatile oxidation product of alpha-farnesene, 6-methyl-5-hepten-2-one (MHO), was present only in fruit developing scald. The data suggest that inhibition of ester production may occur as a result of the physiological changes associated with susceptibility to, rather than expression of, scald symptoms.

Diphenylamine↗

PubMed1999

Extending storage life of fresh-cut apples using natural products and their derivatives.

Prevention of browning of apples slices has been difficult to achieve because of the rapidity of the enzymatic oxidation of phenolic substrates even under reduced atmospheric pressure storage. Combinations of enzymatic inhibitors, reducing agents, and antimicrobial compounds containing calcium to extend storage life were tested to decrease the browning of Red Delicious apple slices stored at 5 and 10 degrees C under normal atmospheric conditions. Treatments were devised to prevent browning for up to 5 weeks at 5 degrees C with no apparent microbial growth using dipping solutions of compounds derived from natural products consisting of 4-hexylresorcinol, isoascorbic acid, a sulfur-containing amino acid (N-acetylcysteine), and calcium propionate. Analyses of organic acids and the major sugars revealed that the slices treated with the combinations of antibrowning compounds retained higher levels of malic acid and had no deterioration in sugar levels at 5 and 10 degrees C, indicating that higher quality was maintained during storage.

Biological Products↗

PubMed1999

Relationship between ethylene response manipulation and volatile production in Jonagold variety apples.

The ripening of Jonagored Jonagold apple fruit (Malus x domestica Borkh.) during development was manipulated with preharvest applications of ReTain or a combination of ReTain plus Ethrel. The fruits, harvested preclimacteric at approximately the same stages of maturity, were stored in refrigerated air (RA) for 45 days or in controlled atmosphere (CA) for 180 days at 0 degrees C. Volatile evolution, ethylene production, and respiration of stored fruit were studied during poststorage holding at 22 degrees C. ReTain reduced volatile production by 19%, but application of Ethrel to ReTain-treated fruit increased production to control levels. The inhibition of volatile production by ReTain appears to be independent of respiration but may be related to the ethylene-producing capacity of the fruit. Although ReTain reduced flavor-related volatile esters, it did not affect levels of the compound responsible for the typical spicy flavor in Jonagored Jonagold fruit, 4-methoxy-2-propenylbenzene. The CA-stored fruit had a much reduced production of volatile compounds compared to RA-stored fruit, with more discernible effects in ReTain-treated fruit. Ethrel application to ReTain-treated fruit improved the volatile production intermediate between the ReTain alone and control in CA-stored fruit. The data collectively suggest that ReTain may have some promise for better scheduling of harvest of apples with no appreciable loss in RA-stored fruit quality. Reduction in production of alpha-farnesene by ReTain may also reduce the potential for scald development in CA-stored fruit.

Chromatography, Gas↗

PubMed1999

Protective effect of ascorbic acid against the browning developed in apple fruit treated with high hydrostatic pressure.

Apple Reineta variety was used as an apple dessert. The 1-1.5-cm cubes were immersed in a sucrose solution (30% w/v) and subjected to high pressure (HP) of 400 MPa for 30 min at 5 degrees C. Different ascorbic acid concentrations were used to protect the fruit from the browning developed after the HP treatment. After 2 months of storage at 5 degrees C, no brown color was observed in the samples treated with 20 mM ascorbic acid, and they were acceptable to consumers. However, untreated samples presented fermentation, and they were not acceptable to consumers. The electric conductivity and potassium content were found to be good indicators of the metabolites released from the fruit to the solution in samples treated with high pressure. HP did not affect the peroxidase activity but eliminated the microbial population.

Ascorbic Acid↗

PubMed1999

Capillary gas chromatographic detection of invert sugar in heated, adulterated, and adulterated and heated apple juice concentrates employing the equilibrium method.

The equilibrium method is introduced for the detection of invert sugar addition to apple juice. The method consists of a pre-equilibration of the sample with dry pyridine at 50 degrees C for 20 min followed by the addition of trimethylsilylimidazole and heating at 75 degrees C for 40 min. The resulting derivatized carbohydrates are then analyzed by capillary gas chromatography. This method was successfully used by independent laboratories to distinguish heated pure, intentionally adulterated (with invert sugar), and intentionally adulterated and then heated apple juice concentrates. The equilibrium method was shown to give significantly lower coefficients of variation for this sample set when compared to the original capillary gas chromatographic method. In addition, these results indicate that it may also be an effective method for the detection of medium invert sugar, depending on the level of the fingerprint oligosaccharides in this sweetener.

Beverages↗

PubMed2000

Effect of freezing on the activity of catalase in apple flesh tissue.

Catalase (CAT, EC 1.11.1.6) activity was measured in flesh tissue of six apple cultivars (Malus domestica Borkh. cvs. Braeburn, Gala, Jonagold, McIntosh, Red Delicious, and Spartan). Activity of CAT was determined for fresh and frozen tissue of the same fruit. Freezing resulted in reductions of 50 to 90% in CAT activity compared with the activity measured in crude extracts from fresh tissues. The rate of freezing had an impact on the level of reduction of CAT activity, with slower freezing procedures leading to greater losses in activity. Six additives to the extraction buffer were tested to evaluate their potential to reduce the inactivation of CAT from frozen tissue, but only EDTA and Tween 20 showed any benefit. However, EDTA and Tween 20 provided only partial recovery in CAT activity. In contrast, crude extracts prepared from fresh tissue showed no appreciable loss in CAT activity after frozen storage for two weeks at -80 degrees C. Gel electrophoresis and immunological analysis indicated that the loss in CAT activity from tissue freezing could be attributed to loss of both the tetrameric CAT enzyme structure and total CAT protein. The implications of using freezing to preserve apple tissue samples prior to catalase activity analysis is discussed.

Catalase↗

PubMed2000

Application of high-temperature gas chromatography-mass spectrometry to the investigation of glycosidically bound components related to cashew apple (Anacardium occidentale L. Var. Nanum) volatiles.

Free and bound volatile components of a Brazilian cashew apple variety (Anacardium occidentale L. var. nanum) were obtained by simultaneous distillation-extraction (SDE) and XAD-2 adsorption. According to gas chromatography-mass spectrometry (GC-MS) analyses and retention indices, 62 free volatile constituents were characterized and quantified. They were esters (40%), terpenes (20%), hydrocarbons (14%), fatty acids (9%), aldehydes (8%), alcohols (3%), lactones (3%), ketones (1%), phenols (1%), and norisoprenoids (1%). The glycosidically bound volatile precursors were analyzed by high-temperature GC-MS, after room temperature silylation. Several conjugated alcohols and cinnamic acids were detected and reported as cashew apple glycosyl constituents for the first time.

Alcohols↗

PubMed2000

Cholesterol oxidation in meat products and its regulation by supplementation of sodium nitrite and apple polyphenol before processing.

The levels of cholesterol oxidation derivatives (OxChol) in eight commercial species of meat products were examined. These products contained more than 1 mg/100 g of OxChol, and 7beta-hydroxycholesterol + 5beta-epoxycholesterol (111-1092 microg/100 g), 5alpha-epoxycholesterol (80-712 microg/100 g), cholestanetriol (0-368 microg/100 g), and 7-ketocholesterol (708-1204 microg/100 g) were detected. To know the interaction of sodium nitrite supplementation against cholesterol oxidation in meat products, sausage was produced with or without varying levels of sodium nitrite and stored in the refrigerator for 15 days. As a result, cholesterol oxidation in sausage was inhibited by addition of sodium nitrite in a dose-dependent manner. This observation may be associated with inactivation of O(2)(-) radical and stabilization of polyunsaturated fatty acids (PUFAs). In fact, the levels of OxChol in sausage increased, accompanying the decrease of coexisting linoleic acid when sodium nitrite was not added to sausage meat. Thus, cholesterol oxidation in meat products seems to be considarably promoted by the oxidation of coexisting PUFAs. On the other hand, additive apple polyphenol also inhibited linoleic acid oxidation in sausage and then suppressed cholesterol oxidation through its radical scavenging effects. Therefore, apple polyphenol, having a large amount of an oligomer of catechin, may interfere with cholesterol oxidation in meat processing or storage of meat products through its antioxidative action and be useful as a new antioxitant for meat products when it is added to the original meat before processing.

Cholesterol↗

PubMed2000

Stripped corn oil controls scald and maintains volatile production potential in golden supreme and delicious apples.

Effects of stripped (alpha-tocopherol < 5 mg L(-)(1)) corn oil on flesh firmness, skin color, acidity, soluble solids content (SSC), scald, and fruit volatiles during 6 months at 0 degrees C were studied using Golden Supreme and Delicious apples. Treatment with 10% oil emulsion reduced production of ethylene, alpha-farnesene, and major volatile esters in the first 3 months of storage, but this trend reversed after 5 months. After 6 months at 0 degrees C plus 7 days at 20 degrees C, oil-treated fruit were firmer and greener and had higher levels of titratable acidity than the controls. In addition, control fruit developed 27% and 42% scald in Golden Supreme and Delicious apples, respectively, whereas oil-treated fruit were free from scald. Soluble solids content and ethanol production were unaffected by oil treatment.

Corn Oil↗

PubMed1998

Apple messenger RNAs related to bacterial lignostilbene dioxygenase and plant SAUR genes are preferentially expressed in flowers.

In an attempt to use a differential display procedure to identify organ-specific genes in apple, cDNA fragments of two transcripts preferentially expressed in flowers were isolated and corresponding full-length cDNA inserts were subsequently obtained. One of these clones, Md-FS1, belongs to the SAUR gene family, originally identified as a set of auxin-inducible genes in soybean. The second one, Md-FS2, encodes a polypeptide with sequence similarities to bacterial lignostilbene-alpha,beta-dioxygenase isozymes, which are thought to be involved in lignin biodegradation. Northern blot analysis confirmed that both genes are preferentially expressed in floral organs at full bloom, while being expressed at lower or undetectable levels in vegetative organs (leaves, shoots or roots) as well as in immature, green and unopened blossoms. Furthermore, Md-FS1 transcripts also appeared to accumulate in vegetative tissues after auxin treatment of micropropagated apple shoots.

Amino Acid Sequence↗

PubMed2000

Pollen availability for predaceous mites on apple: spatial and temporal heterogeneity.

It has been suggested that an abundance of alternate food early in the spring may be critical to the ability of generalist predaceous mites to suppress spider mite pests. One alternate food that is typically very abundant in spring is wind-dispersed pollen. Here we investigate, at several spatial scales, the heterogeneity in the availability of pollen to predaceous mites on apple. We found pollen to be abundant on apple leaves very soon after they opened (> 100 grains/cm2), and that the dominant pollen types at this time were wind-dispersed tree pollens (Betulaceae and Pinaceae). We found that most of the spatial variation in pollen abundance occurred at either small spatial scales (within trees) or very large spatial scales (among orchard blocks). Variability among orchards was clearly influenced by the surrounding vegetation, and probably also by the management regime (frequency of mowing). Spatial heterogeneity in pollen availability may affect the build-up of predatory mite populations in the spring, as we found early season abundances of Typhlodromus pyri (Phytoseiidae) and Zetzellia mali (Stigmaeidae) to be better correlated with early season pollen density than with abundance of mite prey (Aculus schlechtendali).

Animals↗

PubMed1999

Effect of gamma irradiation on storability of apples (Malus domestica L.).

The aim of this study was to investigate the effect of gamma irradiation on storability of the two main apple varieties, Golden Delicious and Starking, in Syria. The experiments were performed in 1995 and 1996. Fruits were irradiated with 0, 0.5, 1.0 and 1.5 KGy. Irradiated and unirradiated fruits were stored at 1 to 2 degrees C and under a relative humidity of 80 to 90%. Weight loss and spoilage due to physiological disorders and fungal diseases were evaluated throughout the different storage periods. Firmness, coloration and pH values were estimated immediately after irradiation. The results showed that, in both varieties, gamma irradiation increased the weight loss after 45 days of storage in apples gathered in 1995 but not in the 1996 season. After 180 days of storage, gamma irradiation had different effects on weight loss depending on the growing year and variety, and increased fungal spoilage. Application of gamma irradiation prevented the growth of Aspergillus niger and the formation of skin scald in 'Golden Delicious' fruits. Immediately after treatment, gamma irradiation increased the softening of fruits, changed their color from green to yellow and decreased the pH value of the juice.

Aspergillus niger↗

PubMed2001

Emission of volatile organic compounds by apple trees under spider mite attack and attraction of predatory mites.

Emission rates of volatile organic compounds (VOCs) from Pirus malus L. subsp. mitis (Wallr.) var. Golden Delicious and var. Starking attacked by the phytophagous mite Panonychus ulmi Koch, and their attractiveness to the predatory mites Amblyseius andersoni Chant and Amblyseius californicus McGregor, were studied during three years. A large variability was found in the emission of individual VOCs depending on the infestation, the apple tree variety and the date. There were larger total VOC emission rates and larger total VOC leaf concentrations in apple trees attacked by phytophagous mites, especially in the var. Starking. In infested trees of this variety, there were also more predatory mites. An olfactometer assay showed that predatory mites preferentially chose branches infested by Panonychus ulmi (85% went to infested branches vs 15% to uninfested control branches) indicating that volatiles may be used as cues to find their prey.

Animals↗

PubMed2001

Synergistic activity of endochitinase and exochitinase from Trichoderma atroviride (T. harzianum) against the pathogenic fungus (Venturia inaequalis) in transgenic apple plants.

Genes from the biocontrol fungus Trichoderma atroviride encoding the antifungal proteins endochitinase or exochitinase (N-acetyl-beta-D-hexosaminidase) were inserted into 'Marshall McIntosh' apple singly and in combination. The genes were driven by a modified CaMV35S promoter. The resulting plants were screened for resistance to Venturia inaequalis, the causal agent of apple scab, and for effects of enzyme expression on growth. Disease resistance was correlated with the level of expression of either enzyme when expressed alone but exochitinase was less effective than endochitinase. The level of expression of endochitinase was negatively correlated with plant growth while exochitinase had no consistent effect on this character. Plants expressing both enzymes simultaneously were more resistant than plants expressing either single enzyme at the same level; analyses indicated that the two enzymes acted synergistically to reduce disease. Selected lines, especially one expressing low levels of endochitinase activity and moderate levels of exochitinase activity, were highly resistant in growth chamber trials and had negligible reduction in vigor relative to control plants. We believe that this is the first report of resistance in plants induced by expression of an N-acetylhexosaminidase and is the first report of in planta synergy between an exochitinase and an endochitinase.

Blotting, Western↗

PubMed2003

Transgene expression in the vegetative tissues of apple driven by the vascular-specific rolC and CoYMV promoters.

The ability of the heterologous promoters, rolCP and CoYMVP, to drive expression of the gusA reporter gene in the vegetative tissues of apple (Malus pumila Mill.) has been studied using transgenic plants produced by Agrobacterium-mediated transformation. Replicate plants of each transgenic clone were propagated in soil to a uniform size and samples of leaf, petiole, stem, and root were taken for the measurement of beta-glucuronidase (GUS) activity by fluorometric assay. The levels of expression were compared with those in tissues of a representative clone containing the CaMV 35S promoter. These quantitative GUS data were related to the copy number of transgene loci assessed by Southern blotting. The CoYMV promoter was slightly more active than the rolC promoter, although both expressed gusA at a lower level than the CaMV 35S promoter. In clones containing the rolC promoter with multiple transgene loci, expression values were generally among the highest or lowest in the range. The precise location of GUS activity in each tissue was identified by staining of whole leaves and tissue sections with a chromogenic substrate. This analysis demonstrated that with both the rolC and CoYMV promoters the reporter gene activity was primarily localised to vascular tissues, particularly the phloem. Our results indicate that both promoters would be suitable to drive the expression of transgenes to combat pests and diseases of apple that are dependent on interaction with the phloem.

Fruit↗