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Measuring Nitrite and Nitrate, Metabolites in the Nitric Oxide Pathway, in Biological Materials using the Chemiluminescence Method
Published on: December 25, 2016
UPTAKE OF NITRATE AND NITRITE BY DITYLUM BRIGHTWELLII-KINETICS AND MECHANISMS(1) (2)
1Institute of Marine Resources, University of California, San Diego La Jolla, California 92038.
Abstract:
Ditylum brightwellii grown on NO2 - as a nitrogen source took up and assimilated NO2 - only in the light, apparently via a photosynthetic nitrite reductase. Assimilation was inhibited by dichlorophenyldimethylurea (DCMU), KCN, partially by 2,4 dinitrophenol, and by NO3 -. Kinetics of inhibition of NO2 - assimilation by NO3 - appeared to be "competitive." D. brightwellii cells grown on NO2 - took up NO3 - in both light and dark and in both cases the uptake was inhibited by p-chloromercuribenzoate, but not by DCMU, KCN, or by NO2 -. Most of the NO3 - taken up in the dark was recovered unchanged from the cells. However only 40% of NO3 - taken up in light was recovered from the cells and no NO2 - was found. This suggests that a photosynthetic nitrate reduction mechanism was active in these cells. DCMU inhibited the light-induced NO3 - reduction. This mechanism of NO3 - reduction is distinct from that involving NADH nitrate reductase in D. brightwellii since the concentration of the latter enzyme is very low in cells grown on NO2 -. Saturation kinetics were observed for NO2 - and NO3 - uptake. Half-saturation concentrations (Ks values) were 4 and 2 μM, respectively. These values are compared with those obtained for NO2 - and NO3 - assimilation by other unicellular algae. The comparisons show lower Ks values in oceanic species compared with tide-pool or freshwater algae and they support the idea that Ks values for NO3 - assimilation may provide a key to understanding species succession when this is due to declining: nitrate concentrations in the sea.
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