Soluble proteins extracted from alfalfa roots of hardy and nonhardy varieties were studied in relation to cold hardiness with polyacrylamide gel electrophoresis and quantitative enzyme analysis. Soluble protein content of alfalfa roots increased during hardening in all varieties. Two new isoenzymes with peroxidase activities were found in the fully hardened samples but no large shifts in the electrophoretic pattern were detected with polyacrylamide gel electrophoresis. Peroxidase and catalase activities increased during hardening in all varieties, but only small differences among hardy and nonhardy varieties were detectable. The studies indicated that protein metabolism was altered during the hardening process.
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