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Properties of the in vivo nitrate reductase assay in maize zea mays soybean glycine max and spinach spinacia oleracea leaves

Properties of the in vivo nitrate reductase assay in maize zea mays soybean glycine max and spinach spinacia oleracea leaves

Journal of Plant Physiology 124(1-2): 123-130

ISSN/ISBN: 0176-1617

The effect of different parameters on the in vivo nitrate reductase activity in leaves of maize, soybean, and spinach has been examined. Leaf sections infiltrated under vacuum with an assay medium containing .**GRAPHIC**. and incubated under air, excreted .**GRAPHIC**. in the dark but not in the light. Nevertheless, addition of 3-(3',4'-dichlorophenyl)-1,1-dimethylurea, a potent inhibitor of the photosynthetic electron transport, promoted an immediate excretion of .**GRAPHIC**. in the light. Under those conditions, subsequent addition of ascorbate-reduced dichlorophenolindophenol or N,N,N',N'-tetramethylphenylenediamine, which can supply electrons to photosystem I, abolished the excretion of .**GRAPHIC**. by the leaf sections. These results support the view that, in green tissues, the reducing equivalents photogenerated in the chloroplasts are more directly used for .**GRAPHIC**. reduction than for .**GRAPHIC**. reduction. Incubation of leaf discs under O2-atmosphere severely impaired .**GRAPHIC**. accumulation. Under strict anaerobic conditions, the rate of .**GRAPHIC**. production was only slightly enhanced as compared with that observed in the assay under air. Exogenous .**GRAPHIC**. stimulated the activity, more markedly in soybean than in the other two species. Concentrations of exogenous .**GRAPHIC**. between 50 and 100 mM were required for maximal activities in all the three species. The external pH optimum for in vivo .**GRAPHIC**. reduction was 7.5 for maize and spinach and 7.0 for soybean. Inclusion of 1% (v/v) n-propanol in the infiltration-incubation medium enhanced the activity about 15%. However, 7% (v/v) n-propanol caused a drastic decrease in the rate of .**GRAPHIC**. accumulation which could be greatly overcome by inclusion in the medium of NADH or reduced flavin mononucleotide.

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Accession: 006204012

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Properties of the in vivo nitrate reductase assay in maize, soybean, and spinach leaves. Journal of Plant Physiology 124(1/2): 123-130, 1986

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