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Plant and Cell Physiology, 1991, Vol. 32, No. 5 635-641
© 1991

Article

Peroxide-Scavenging Systems during Cold Acclimation of Apple Callus in Culture

Haruyuki Kuroda1, Shonosuke Sagisaka2, Minoru Asada2 and Kazuhiko Chiba1

1The Hokkaido National Agricultural Experiment Station Sapporo, 062 Japan
2The Institute of Low Temperature Science, Hokkaido University Sapporo, 060 Japan

The relationship between peroxide-scavenging systems and cold acclimation was studied in apple callus in culture during acclimation under artificial conditions. Unacclimated callus did not survive freezing at –10°C, whereas callus acclimated at 0°C exhibited gradually increased resistance to freezing and, after acclimation for 20 days, it survived at temperatures as low as –15–C. During acclimation of callus, there was an immediate and abrupt increase in the activities of ascorbate peroxidase (EC 1.11.1.11 [EC] ), peroxidase (EC 1.11.1.7 [EC] ) and catalase (EC 1.11.1.6 [EC] ), which reached maximum values after acclimation for 10 days, at the same time as the very beginning of the increase in cold hardiness was observed. An increase in the activity of glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12 [EC] ) occurred during the first 5 days of cold treatment. The activities of glucose-6-phosphate dehydrogenase (EC 1.1.1.49 [EC] ), hexokinase (EC 2.7.1.1 [EC] ), glutathione reductase (EC 1.6.4.2 [EC] ), glutathione peroxidase (EC 1.11.1.9 [EC] ) and dehydro-ascorbate reductase (EC 1.8.5.1 [EC] ) increased gradually during the cold treatment. In contrast, the activity of glucosephosphate isomerase (EC 5.3.1.9 [EC] ) decreased gradually during acclimation. Furthermore, during acclimation, the levels of glucose-6-phosphate, fructose-6-phosphate and glucose-1-phosphate increased slowly and steadily, and the levels of GSH and ascorbate remained at consistently higher levels. In addition, acclimation caused marked cytological changes. The most striking of these changes was the microvacuolation and thickening of the cell wall. These results indicate that the enhancement of peroxide-scavenging systems at the time of cold acclimation proceeds in two stages: during the first stage, the enzymatic activities involved in the degradation of peroxides (i.e., the activities of ascorbate peroxidase, peroxidase and catalase) increase; and, in the second stage, an alternative enzymatic system develops for detoxification of peroxides, coupled with the pentose phosphate cycle.

(Received July 20, 1990; Accepted April 16, 1991)
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