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Effect of freezing and thawing on in situ nitrate reductase activity in spinach spinacia oleracea and maize zea mays leaves

Effect of freezing and thawing on in situ nitrate reductase activity in spinach spinacia oleracea and maize zea mays leaves

Journal of Plant Physiology 120(5): 409-418

ISSN/ISBN: 0176-1617

Spinach leaf discs subjected to freezing and thawing, and subsequently incubated with a buffer containing nitrate and NADH, excreted nitrite into the medium linearly for at least 90 min. Chemically-reduced FMN was also an efficient added electron donor for in situ nitrate reduction. The freeze-thaw nitrate reductase assay did not require anaerobic conditions. Nitrite production occurred equally in the light and in the dark; consequently, DCMU did not inhibit the assay in light. The freeze-thaw treatment completely abolished the photosynthetic oxygen evolution by leaf discs. In maize leaf sections, freezing and thawing caused irreversible inactivation of nitrate reductase. This inactivation was greatly prevented when both freeze-thaw treatment and further incubation were carried out under anaerobic conditions. Alternatively, the freeze-thaw assay in maize leaves could be improved by performing both the freeze-thaw treatment and the incubation in the presence of compounds which protect the enzyme against the inactivity effects of phenolics.

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Accession: 005240165

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