`Granny Smith' apples [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.] were harvested in two seasons and stored at 0 °C air storage with no pretreatment (control), after heating for 4 d at 38 °C, or after treating for 16 hours at 20 °C with 1 μL·L-1 1-methylcyclopropene (1-MCP). The effects of the two treatments on superficial scald development were consistent over both seasons. Scald began to appear after 8 weeks in control fruit, after 16 weeks in heated fruit but not on 1-MCP treated fruit. α-Farnesene accumulation and oxidation were slower in the skin of heated than in control fruit, and almost entirely absent in 1-MCP treated fruit. The activities of five antioxidant enzymes, ascorbate peroxidase, catalase, glutathione reductase, peroxidase and superoxide dismutate, were measured at two-week intervals in the apple peel, quantitatively as total activity and qualitatively by isozyme analysis. Enzyme activities either increased or remained stable during 16 weeks of storage, except for superoxide dismutase activity, which decreased. Ascorbate oxidase activity was higher in heated than control apples and there was an additional peroxidase isozyme present in activity gels. The activities of antioxidant enzymes were lower in 1-MCP treated fruit except for catalase during the first month of storage. Lipid soluble antioxidant activity was higher in 1-MCP treated fruit than the fruit of the other treatments, and water soluble antioxidant activity was higher in both treatments than in control fruit during the time that scald was developing in control apples. Both free and total phenol contents in the peel fluctuated during storage but no consistent trend was detected. The differences in enzyme activity and antioxidant content of the peel of 1-MCP and heated apples may play a role in preventing or delaying the appearance of superficial scald.
Zohar Shaham, Amnon Lers, and Susan Lurie
Hong-Wei Zhou, Lilian Sonego, Andrai Khalchitski, Ruth Ben-Arie, Amnon Lers, and Susan Lurie
Most `Flavortop' nectarines [Prunus persica (L.) Batsch (Nectarine Group)] that were placed directly into 0 °C storage developed chilling injury after removal, while preconditioning fruit for 2 days at 20 °C (delayed storage) reduced chilling injury substantially. Chilling injury was expressed as the development of a dry, woolly flesh texture during ripening. Delayed-storage fruit were as firm as control fruit when placed in storage, but softened more during storage. Analysis of cell wall components showed that in woolly fruit a higher percentage of pectin was retained in the sodium carbonate fraction, although during ripening polymers in this fraction decreased in molecular mass (Mr). In the guanidine thiocyanate hemicellulose fraction of woolly fruit, the associated pectin and hemicellulose remained as large polymers, while in delayed-storage fruit they decreased in Mr during ripening. Endo-polygalacturonase (PG), pectin esterase (PE), and endo-glucanase (EGase) activities of delayed-storage fruit were the same as control fruit at the beginning of storage, although exo-PG was higher. However, differences were observed at the end of storage. Endo-PG activity was lower in control than delayed-storage fruit at the end of storage while PE activity was higher, and exo-PG and EGase activities were similar. These differences in activity were not reflected in the mRNA abundance of the respective enzymes. Endo-PG and PE message was similar in all fruit at the end of storage and increased during ripening, while EGase message was low at all times except in control fruit after storage and development of woolliness. Prevention of chilling injury by delayed storage appears to be due to the ability of the fruit to continue a progressive, slow cell wall degradation in storage which allows normal ripening to proceed when the fruit are rewarmed. Regulation of the softening process did not appear to be by enzyme synthesis, since mRNA levels of the enzymes did not correspond with enzyme activity.
Susan Lurie, Amnon Lers, Zohar Shacham, Lilian Sonego, Shaul Burd, and Bruce Whitaker
Untreated control, 1-methylcyclopropene (1-MCP)-treated, and heated fruit of the superficial scald-susceptible `Granny Smith' cultivar of apple [Malus sylvestris (L.) Mill. var. domestica (Borkh.) Mansf.] were compared with respect to scald incidence, internal ethylene concentration (IEC), α-farnesene metabolism, expression of the genes AFS1, which encodes α-farnesene synthase, the final, rate-limiting enzyme in the α-farnesene biosynthetic pathway, and HMG2 and HMG3, which encode isozymes of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, the proposed rate-limiting enzyme in the mevalonate pathway of isoprenoid synthesis. The incidence of scald in untreated `Granny Smith' apples after 16 weeks at 0 °C plus 1 week at 20 °C was 100%; 1-MCP treatment prevented scald development, whereas heat treatment delayed and reduced scald development. 1-MCP also inhibited both α-farnesene and IEC, suggesting that ethylene induces transcription of key genes involved in α-farnesene biosynthesis. Heat treatment reduced levels of α-farnesene and and its oxidation products, conjugated trienols (CTols), but not to the extent of 1-MCP. Internal ethylene concentrations in heated apples did not differ from those in the controls. In both control and heated fruit, a sharp increase in AFS1 mRNA during the first 4 weeks of storage preceded an increase in α-farnesene and a subsequent increase in CTols. AFS1 transcript was absent from 1-MCP-treated apples for the first 10 weeks of storage, and even at 16 weeks was lower than in heated and untreated control fruit. Levels of the HMG2 and HMG3 transcripts varied during storage and among treatments, and were not correlated with the incidence of scald. HMG2 mRNA transcript accumulation was low at harvest and increased in abundance during storage in all treatments, with the greatest increase occurring in 1-MCP-treated fruit. In contrast, HMG3 transcript was constitutively present at all storage times, although it too was slightly more abundant in 1-MCP-treated fruit.