635 PB 334 HISTOCHEMICAL LOCALIZATION OF LIPOXYGENASE, PEROXIDASE AND POLYPHENOL OXIDASE IN POTATO TUBER TISSUE

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  • 1 USDA, Agricultural Research Service, National Center for Agricultural Utilization Research, Peoria, IL 61604

Localization of enzymes in specific plant tissues is crucial to understanding their role in processes such as differentiation and disease resistance. The oxidative enzymes lipoxygenase (LOX; EC 1.13.11.12), peroxidase (PER, EC 1.11.1.7) and polyphenol oxidase (PPO; EC 1.10.3.1) have all been implicated as playing critical roles in plant disease resistance. The histochemical localization of all three enzymes in potato tuber slices was accomplished either directly on the tissue slices (for LOX) or by blotting of the tissue onto nitrocellulose membranes (for PER and PPO). LOX was visualized in specific tissues by the oxidation of KI to I2 via lipid peroxides and the subsequent reaction of I2 and endogenous starch to form a colored, insoluble complex. PER and PPO activities were visualized with 4-methoxy-α-naphthol and 3,4-dihydroxy-phenylalanine, respectively. Fractionation of the slices and determination of enzyme activities in the fractions confirmed the reliability of these techniques.

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