Enzymatic browning is one of the most important reactions that occur in fruits and vegetables, usually resulting in negative effects on color, taste, flavor, and nutritional value. The reaction is a consequence of phenolic compounds' oxidation by polyphenol oxidase (PPO), which triggers the generation of dark pigments. This is particularly relevant for apples, which are rich in polyphenols and highly susceptible to enzymatic browning. The objective of the present work was to quantify enzymatic browning and PPO activity and identify and quantify target polyphenols in apple [Malus ×sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.] pulp in the cultivars (cvs.) Aori27, Elstar, Fuji, and Mellow at three fruit developmental stages (FDS). The enzymatic browning was quantified by tristimulus colorimetry; PPO activity was quantified by an enzyme–substrate spectrophotometric assay; phenolic compounds were determined and quantified by reverse-phase high-performance liquid chromatography–ultraviolet/visible–mass spectrometry. Enzymatic browning showed significant difference among cvs. and FDSs and interaction between both factors. PPO activity showed significant difference among cultivars and FDSs. A significant difference was evidenced for polyphenol content among cultivars and FDSs with interaction between both factors. Chlorogenic acid was the major phenolic compound in ‘Aori27’ and ‘Mellow’. In ‘Fuji’, chlorogenic acid and (–)-epicatechin were the major phenolics and in ‘Elstar’ (–)-epicatechin and procyanidin B2 were the major phenolics at different FDSs. The enzymatic browning showed high correlation to polyphenol content in all cultivars and high correlation was observed between browning and PPO activity in ‘Aori27’ and ‘Elstar’. The magnitude of the correlation between browning and polyphenols and PPO activity is genotype-specific. At the commercial harvest, ‘Fuji’ showed the highest polyphenol content and ‘Aori27’ showed the lowest level for enzymatic browning. Chemical names used: 3-(3,4-dihydroxycinnamoyl) quinic acid (chlorogenic acid), (–)-cis-3,3′,4′,5,7-pentahydroxyflavane (epicatechin), and cis,cis″-4,8″-Bi(3,3′,4′,5,7-pentahydroxyflavane) (procyanidin B2).
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.