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• W1479462911 abstract "Previous studies demonstrated that continuous 10 μL L−1 ethylene exposure induces acute watersoaking of immature beit-alpha cucumber fruit (Cucumis sativus L. cv. Manar). This phenomenon includes increases in nuclease activities and loss of total protein, supporting the idea that watersoaking is a form of programmed cell death (PCD). The present study was designed to further characterize ethylene-induced, PCD-associated nucleases and proteases in immature cucumber fruit. DNases were found to possess bifunctional (DNA/RNA) activities. In addition, four RNA-specific nucleases (17.5, 22.3, 24.4, and 26.1 kDa) were detected by in-gel assays, all of which were ethylene inducible. All nucleases were inhibited by β-mercaptoethanol, indicating a requirement for disulfide bond(s) for activity. Both the bifunctional nucleases and the 17.5 kDa RNase exhibited Ca2+ dependency. Bifunctional nuclease activities were detected above pH 7, but strongly inhibited at weakly acidic pH (6). By contrast, RNA-specific nuclease activity was optimal at acidic pH and strongly inhibited above pH 8. Ethylene induced significant protein degradation after only 2 d of exposure, with total protein levels declining by about 60% after 6 d. The decline in total protein was accompanied by significant increases in protease activity. Increased activity was also observed after 2 d, reaching levels 9.3-fold higher than initial values after 6 d. Total protease activity was greatest at pH 5 and declined sharply through pH 8. In-gel assays revealed three ethylene-responsive proteases with apparent molecular masses of 56, 106, and 180 kDa. Levels of the 106 and 180 kDa proteases increased after 2 d of ethylene exposure, reaching maxima at 4–5 d coincident with symptoms of watersoaking. A 56 kDa protease was observed only in ethylene-treated fruit and was first detected after 4 d of ethylene exposure. Together with up-regulated nuclease activities and reactive oxygen species (ROS)-generating capacity, the early appearance of proteases in response to ethylene provides additional evidence that watersoaking in immature cucumber fruit reflects the execution phase of a PCD pathway." @default.
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• W1479462911 date "2015-12-01" @default.
• W1479462911 modified "2023-10-16" @default.
• W1479462911 title "Characterization of proteases and nucleases associated with ethylene-induced programmed cell death in immature cucumber fruit" @default.
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• W1479462911 doi "https://doi.org/10.1016/j.postharvbio.2015.08.009" @default.
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