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

Volatile emissions and chlorophyll fluorescence were investigated as potential signals of heat injury for apple [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.] fruit. `McIntosh', `Cortland', `Jonagold', and `Northern Spy' apples were exposed to 46 °C for 0, 4, 8, or 12 hours (heat treatments). Following treatments, fruit were kept at 20 °C and evaluated after 1, 2, 4, or 7 days. Heat treatments induced volatile production including ethanol and ethyl acetate. The 8 and 12 hours heat treatments increased ethanol and ethyl acetate production in all four cultivars by as much as 170- and 11-fold, respectively, 1 day after treatments. Heat treatments also reduced ethylene production and chlorophyll fluorescence. Heat for 12 hours caused serious flesh browning. Among the cultivars investigated, `Northern Spy' and `McIntosh' were most susceptible to heat stress based on the degree of flesh browning. Correlation coefficients of heat stress induced ethanol emission and chlorophyll fluorescence with flesh browning were 0.82 and -0.66, respectively. The nondestructive measurements of ethanol emission and chlorophyll fluorescence have potential to identify stressed fruit with reduced quality or compromised storage life.

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I. Lara and M. Vendrell

Endogenous ABA, free and conjugated ACC concentrations, ethylene-forming capacity (EFC), and presence of ACC oxidase (ACO) and ACC synthase (ACS) proteins were monitored during the preharvest maturation period of `Granny Smith' apple fruit (Malus sylvestris L. Mill. var. domestica (Borkh.) Mansf. `Granny Smith'). Total proteins from peel and pulp tissues were also extracted at different maturity stages and separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis, providing evidence of differential protein accumulation during fruit development. Endogenous ABA concentration in the peel tissue was higher than in pulp, the highest level occurring ≈2 months before commercial harvest. In the pulp tissue, concomitant increases in ACC and ABA concentrations were observed, preceded by a peak in EFC. However, no ACO or ripening-related ACS proteins were detectable throughout the period considered, suggesting that very low levels of both enzymes are present during the preclimacteric stage of `Granny Smith' apples. A hypothesis on the possible interaction between ABA and ethylene during maturation of `Granny Smith' apples is proposed. Chemical names used: abscisic acid (ABA); 1-aminocyclopropane-1-carboxylic acid (ACC).

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Zhiguo Ju and William J. Bramlage

Influences of fruit maturity, AVG and ethephon preharvest treatments, and storage conditions on cuticular phenolic concentration, α-farnesene accumulation and oxidation, and scald development of `Delicious' apples [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.] were studied. Advanced maturity and ethephon treatment increased free phenolics in fruit cuticle at harvest, while AVG treatment caused a reduction. Free cuticular phenolics increased during early storage in ethephon-treated and nontreated fruit but not in AVG-treated apples. Advanced maturity and ethephon did not alter α-farnesene accumulation overall, but reduced conjugated triene (CT281) formation and scald development. When stored in a low-ethylene room (<1 μL·L-1), AVG-treated fruit accumulated very low levels of α-farnesene and CT281 and did not develop scald after 6 months at 0 °C. When stored in a commercial room (ambient ethylene >5 μL·L-1), the AVG-treated and control fruit accumulated similar amounts of α-farnesene and CT281 and developed similar percentages of scald. In general, free phenolic concentrations in fruit cuticle were negatively correlated with CT281 formation and scald development of apples. Chemical names used: aminoethoxyvinylglycine (AVG); 2-chloroethylphosphonic acid (ethephon).

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Takashi Sato, Tsuyoshi Kudo, Tomoko Akada, Yuhya Wakasa, Minoru Niizeki, and Takeo Harada

The onset of apple [Malus sylvestris (L.) Mill. Var. domestica (Borkh.)Mansf.] fruit maturity is preceded by the production of ethylene, the ripening hormone, which induces fruit ripening. The amount of ethylene produced by the fruit correlates with the level of transcription of the ripening-specific 1-aminocyclopropane-1-carboxylate (ACC) synthase genes. We have found that an allele (MdACS1-2), which contains an inserted retroposon-like sequence at the 5'-flanking region, is transcribed at a lower level than the wild-type (MdACS1-1). MdACS1-2/2 homozygous fruit produce a lower level of ethylene at the climacteric stage than do the wild type fruit. We have also found that the preharvest drop rates of apple cultivars and strains of MdACS1-2/2 trees have less fruit drop than the MdACS1-1/1 or MdACS1-1/2 trees. Treatment of the MdACS1-1/2 trees with 1-MCP, an ethylene receptor blocker, further decreased fruit drop. Analysis of commercial apple cultivars for the presence of the MdACS1-2/2 allele may help in the early detection of apple cultivars with a low fruit drop rate.

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Richard P. Marini

For 4 years, `Redchief Delicious' apple [Malus ×sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.] trees were treated with ethephon and/or Accel (6-BA + Gibberellins 4+7) when average fruit diameter was 5.0 to 11.4 mm. Effective thinning was obtained with ethephon at concentrations >500 mg·L-1 and with Accel at concentrations of 100 to 125 mg·L-1. In only 1 year out of 4 did combinations of Accel plus ethephon reduce fruit set more than either material applied alone. Repeat applications of either material alone or in combination reduced fruit set no more than single applications. Fruit weight was negatively related to the number of fruit per tree. After adjusting fruit weight for number of fruit per tree at harvest with analysis of covariance, ethephon did not improve fruit weight at harvest, but Accel improved fruit weight in two of three experiments. The effect of combinations of Accel and ethephon on fruit weight was inconsistent. Chemical names used: 2-chloroethyl phosphonic acid (ethephon); polyothyethylleneploypropanol dihydroxy-propane 2, butoxyethanol (Regulaid); N-(phenylmethyl)-1H-purine-6-amine plus giberrellins A4 + A7 (Accel).

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Minou Hemmat, Norman F. Weeden, and Susan K. Brown

We mapped DNA polymorphisms generated by 41 sets of Simple Sequence Repeat (SSR) primers, developed independently in four laboratories. All primer sets gave polymorphisms that could be located on our `White Angel' x `Rome Beauty' map for apple [Malus sylvestris (L.) Mill. Var. domestica (Borkh.) Mansf.]. The SSR primers were used to identify homologous linkage groups in `Wijcik McIntosh', NY 75441-58, `Golden Delicious', and `Liberty' cultivars for which relatively complete linkage maps have been constructed from isozyme and Random Amplified Polymorphic DNA (RAPD) markers. In several instances, two or more SSRs were syntenic, and except for an apparent translocation involving linkage group (LG) 6, these linkages were conserved throughout the six maps. Twenty-four SSR primers were consistently polymorphic, and these are recommended as standard anchor markers for apple maps. Experiments on a pear (Pyrus communis L.) population indicated that many of the apple SSRs would be useful for mapping in pear. However some of the primers produced fragments in pear significantly different in size than those in apple.

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Satoru Kondo, Kentaro Hiraoka, Shozo Kobayashi, Chikako Honda, and Norihiko Terahara

Cyanidin 3-galactoside was the primary anthocyanin in red `Tsugaru' apples [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.]. The concentration of cyanidin 3-galactoside in the skin decreased from 20 to 62 days after full bloom (DAFB), then increased rapidly after 104 DAFB. Small amounts of cyanidin 3-arabinoside and cyanidin 3-glucoside were detected at 122 and 133 DAFB (harvest). The expression of five anthocyanin biosynthetic genes of chalcone synthase (MdCHS), flavanone 3-hydroxylase (MdF3H), dihydroflavonol 4-reductase (pDFR), anthocyanidin synthase (MdANS), and UDP glucose-flavonoid 3-O-glucosyltransferase (pUFGluT) was examined in the skin of red and nonred apples. In general, the expression of anthocyanin biosynthetic genes in red apples was strong in juvenile and ripening stages. The expression of MdCHS, MdF3H, pDFR, and MdANS was observed before ripening stage when anthocyanin was not detected. In contrast, the expression of pUFGluT was detected in the development stage only when anthocyanin was detected. However, the expression of all five genes was observed at 20 DAFB in fruit bagged after fertilization, and anthocyanin was not detected. The expression of MdCHS, MdF3H, pDFR, and MdANS, excluding pUFGluT, was detected at 98 DAFB in fruit bagged after 30 DAFB, and anthocyanin was not detected. These results suggest that pUFGluT may be closely related to the anthocyanin expression in apple skin at the ripening stage.

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I.J. Warrington, T.A. Fulton, E.A. Halligan, and H.N. de Silva

Container-grown `Delicious', `Golden Delicious', `Braeburn', `Fuji' and `Royal Gala' apple [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.] trees, on Malling 9 (M.9) rootstock, were subjected to a range of different maximum/minimum air temperature regimes for up to 80 days after full bloom (DAFB) in controlled environments to investigate the effects of temperature on fruit expansion, final fruit weight, and fruit maturation. Fruit expansion rates were highly responsive to temperature with those at a mean of 20 °C being ≈10 times greater than those at a mean of 6 °C. All cultivars exhibited the same general response although `Braeburn' consistently showed higher expansion rates at all temperatures compared with lowest rates for `Golden Delicious' and intermediate rates for both `Delicious' and `Fuji'. The duration of cell division, assessed indirectly by measuring expansion rate, appeared to be inversely related to mean temperature (i.e., prolonged under cooler conditions). Subsequently, fruit on trees from the coolest controlled temperature treatment showed greater expansion rates when transferred to the field and smaller differences in fruit size at harvest than would have been expected from the measured expansion rates under the cool treatment. Nonetheless, mean fruit weight from warm postbloom treatments was up to four times greater at harvest maturity than that from cool temperature treatments. Postbloom temperature also markedly affected fruit maturation. Fruit from warm postbloom temperature conditions had a higher soluble solids concentration, more yellow background color, lower flesh firmness, and greater starch hydrolysis than fruit from cooler temperatures.

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Manfredo J. Seufferheld, Cecil Stushnoff, Philip L. Forsline, and Gerardo H. Terrazas Gonzalez

Unlike cold-hardy apple germplasm, dormant vegetative buds from cold-tender accessions require stabilization of meristematic tissue to protect against injury during desiccation and cryopreservation. Dormant buds of six apple cultivars [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf. `Cox's Orange Pippin', `Einshemer', `Golden Delicious', `Jonagold', `K-14', and `Mutsu'] collected at specific intervals in 1993, 1994, and 1995 at Geneva, N.Y., were stabilized by encapsulation in 5% alginate, treated with step-wise imbibition of 0.5 to 1.0 m sucrose and 0.2 m raffinose solution, and desiccated with forced air at 0 °C. Sugar-alginate stabilization reduced injury during desiccation, increased cold-hardiness of the six cold-tender cultivars frozen to -30 °C, and improved recovery following cryopreservation of buds collected before optimal cold acclimation was attained. Sucrose tissue levels did not increase following stabilization treatment, but levels of glucose and fructose, and of an unknown disaccharide increased. This procedure used nontoxic cryoprotectants, and has potential to expand the scope of dormant bud cryopreservation to include cold-tender apple germplasm.

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Anne Plotto, Mina R. McDaniel, and James P. Mattheis

Aroma and flavor characters of `Gala' apples [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf. `Gala'] were identified by 10 trained panelists. A vocabulary of 13 aroma descriptors and 16 flavor descriptors were used to characterize changes in controlled atmosphere (CA) and air, or regular atmosphere (RA) storage over 20 weeks. When compared with RA storage, the intensity of fruity (pear, banana, and strawberry) and floral descriptors decreased after 10 weeks in CA for whole and cut fruit aroma and flavor. During the entire storage period under CA, aroma of cut apples retained high vegetative and citrus characters but had a less intense anise aroma. Sourness and astringency were significantly higher for CA-stored apples, and sweetness was significantly lower. A musty note was perceived in whole apples stored in CA for 20 weeks. Aroma of whole fruit stored for 16 weeks in CA followed by 4 weeks in RA was higher in fruitiness, banana, floral, and anise characters when compared with apples stored 20 weeks in CA. There was no difference between fruit stored in CA followed by RA versus CA stored apples for flavor and aroma of cut fruit. Changes in descriptor ratings during storage are discussed in relation to gas chromatography and olfactometry data obtained with the Osme method.