Determination of nitrate and nitrite by high-pressure liquid chromatography: Comparison with other methods for nitrate determination

Thayer, James R.; Huffaker, R. C.

doi:10.1016/0003-2697(80)90325-5

Cited by 190 publications

(94 citation statements)

References 16 publications

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“…Leaf NO3-was determined by first extracting N03-from dried tissue in 50°( water for 5 h. Nitrate in the extracts was determined with a Gilson (Middleton, WI) HPLC, using the procedure of Thayer and Huffaker (25). The difference between the total N as determined by the Kjeldahl procedure and leaf NO3 gave the leaf organic nitrogen content.…”

Section: Methodsmentioning

confidence: 99%

The Nitrogen Use Efficiency of C₃ and C₄ Plants

Sage¹,

Pearcy²

1987

Plant Physiol.

188

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ABSTRACIThe effect of applied nitrogen (N) on the growth, laf expansion rate, biomass padtioning and laf N levels of Cheaopodium album (C3) and Amaranthus rtroftexus (C4) were investigted. At a given applied N leveL C. album had 50% greater leaf N per unit area (N.) than A. retrojkxw. Nitrate accumulated at lower N. in A. retrojiexus thn C. album. A. rctroJiexus was more productive than C. album at high N, but C. album was more productive at low N. At high appled N, nitrogen use efficiency (NUE) expressed either as net assimilation rate (NAR) per unit N or relative growth rate per unit N, was greater in A. retroflexus than C. album. However, at low applied N, C. album had a greater NUE on both an NAR and growth basis than A. retroflexus. The leaf area partitioning coefficient was similar in the species at high N, but was greater in A. retrofjexus than C. album at low N. At low N, greater leaf area partitioning apparently lowered leaf N in A. retroflexus to levels at which necrosis occurred. In C. album by contrast, leaf area partitioning declined to a greater degree with declining N than it did in A. retroflxus, so that leaf N did not decline as much. Consequently, low N C. album plants did not lose leaf area to necrosis and had a greater NAR and NUE at low applied N than A. retrojfxus.Photosynthetic rate per unit leaf nitrogen, which has been termed photosynthetic nitrogen use efficiency (PNUE2), has been shown to be greater in C4 plants than C3 plants (3, 4, 18, 24, 26, 28). However, the effect of PNUE on the growth and reproductive yield of C4 relative to C3 plants is unclear. Under limiting N, a $reater PNUE may enable C4 plants to have greater photosynthetic rates than C3 species. Alternatively, the higher PNUE of C4 species may allow them to allocate a greater fraction of available N to new leaf, root, or stem production. Increasing N availability increases the leafexpansion rate (8,14,26,27), which is well correlated with growth rate (21). Moreover, reductions in yield following N deficiency are often due more to reduced leaf expansion rates than to a decline in photosynthesis rate per unit area (8,10,27 To assess the importance of PNUE differences between C3 and C4 species, we have examined the effect of applied N on the growth, leaf expansion rate and allocation of N to leaves in the ecologically similar C3 dicot, Chenopodium album, and C4 dicot Amaranthus retroflexus. Both are important agricultural weeds with similar growth forms, periods of activity, and microsite requirements. In addition, both have high photosynthetic capacities when grown at high N (20). Photosynthetic characteristics as a function of leaf N were also determined and are described in an accompanying report (23). MATERIALS AND METHODSLeaf Nitrogen Content. C. album and A. retroflexus plants were grown in a growth chamber at a PFD of 600 ,mol m-2 s-', 27/230C day/night temperatures and a 16 h photopenod. Plants were grown in equal volume mixtures of sand, perlite, and vermiculite. Treatments consisted of daily applications of either...

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Section: Methodsmentioning

confidence: 99%

The Nitrogen Use Efficiency of C₃ and C₄ Plants

Sage¹,

Pearcy²

1987

Plant Physiol.

188

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“…Samples of culture (1 ml each) were filtered through 0n45-µm-pore cellulose acetate membrane filters (Sartorius, Go$ ttingen, Germany) and the clear medium was automatically injected into an HPLC system (Spectra Series4, Thermo Separation Products, Darmstadt, Germany) with a silica-based column (Partisil sax 10 micron, Alltech, Deerfield, IL, USA) and NO $ − was determined in the eluate at 210 nm (Thayer & Huffaker, 1980). To determine the intracellular concentrations of NO $ − , 200 ml of culture were filtered, the cells rapidly washed with 0n1 M KP i , pH 7n65, and broken as described previously after small aliquots had been removed to determine the cell number.…”

Section: Nitrate Protein and Chlorophyll Determinationsmentioning

confidence: 99%

Soluble and membrane‐associated nitrate reductases in the dinoflagellate Peridinium gatunense

1999

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An improved method of cell fractionation allowed the extraction of soluble (sNR) and membrane-associated (mNR) forms of nitrate reductase (NR) from a dinoflagellate, even though in previous studies only mNR had been found in these algae. Both activities were assayed in cell-free extracts of Peridinium gatunense from Lake Kinneret, Israel, after disruption of the cells and differential centrifugation. In the cultures used, sNR showed much higher NO $ − -reducing activity. Only a low proportion, 2n5-3% of NR activity, was found to be associated with mNR. Moreover, mNR comprised two forms as indicated by protein solubilization : a tightly membrane-bound and a more weakly attached NR. Ascorbate inhibited all NR activities, but that of mNR recovered after its removal. Polyvinyl pyrrolidone (PVP) and DTT also diminished sNR and mNR activities. For both enzymes, pH optima (7n65) and temperature optima (13-25mC) were similar, and agreed with those for optimum growth of P. gatunense both in culture and in the lake. The most efficient electron donor was NADH, though NADPH sustained low NR activities. Carboxylic anions such as succinate and malate did not support any reduction of NO $ − , nor did they cause any stimulation of sNR or mNR activities. Both forms of NR showed a high affinity for their substrates : K m was c. 10 µM for NO $ − and c. 5 µM for NADH. The high efficiency of NO $ − assimilation by Peridinium seems to be limited mainly by energy under otherwise optimal nutritional conditions and, at low nitrate concentrations, the low K m may be one of the main reasons for the high competitivity of this alga in Lake Kinneret.

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“…A peristaltic pump (Micro Perpex Pump 2132, LKB, Broma, Sweden) sampled aliquots (1·14 cm 3 ) from the nutrient solutions for rinsing the tubes and valves and filling the sample loop of an electric switching injector (Valco EC6 W, VICI Inc., Houston, TX, USA). The HPLC system for nitrate analysis corresponded to that of Thayer & Huffaker (1980). Separation was achieved with a strong anion exchanger (Partisil SAX 10, Whatmann, Maidstone, Kent, UK) in a Vertex column (120 × 4 mm, Knauer, Bad Homburg, Germany).…”

Section: Hplc Systemmentioning

confidence: 99%

The effects of light on induction, time courses, and kinetic patterns of net nitrate uptake in barley

Peuke

Jeschke

1998

Plant Cell & Environment

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Barley seedlings (Hordeum vulgare L.) were grown hydroponically with (induced) or without (uninduced) nitrate in a light/dark cycle with high photon flux density to determine the effects of light on time courses, induction and kinetics of net nitrate uptake. Nitrate uptake was induced by external nitrate in both light and dark and was prevented by 1 mol m -3 p-fluorophenylalanine. In high light, nitrate uptake was about 2-fold higher than in low light. During time course experiments the uptake rates oscillated due to daily light-dark changes. Rates of nitrate uptake also increased at about 2200 h during continuous darkness. This increase coincided approximately with the time at which the dark period started during the previous culture of the plants, indicating that it was due to a mechanism associated with an endogenous diurnal rhythm. When calculating the kinetics of nitrate uptake, a model with two saturable systems, including a high-affinity system (HATS) and a low-affinity system (LATS), gave the best fit to data in all treatments. The apparent affinity of the HATS ranged from 7·7 to 12·2 mmol m -3 in induced plants in all light conditions. The effect of light on the HATS was mainly an increase of apparent V max in the step from low to high light. In uninduced plants the HATS operated at a very low activity which was strongly enhanced during induction. Interpretation of the calculated kinetics of the LATS was much more difficult on the basis of net uptake data. The apparent affinity of the LATS increased from 24·3 mol m -3 in low light up to 0·17 mol m -3 after acceleration in high light. These extreme changes in apparent affinity of the LATS could not be explained satisfactorily, and the nature of this system is also discussed with respect to the method used.

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Determination of nitrate and nitrite by high-pressure liquid chromatography: Comparison with other methods for nitrate determination

Cited by 190 publications

References 16 publications

The Nitrogen Use Efficiency of C₃ and C₄ Plants

The Nitrogen Use Efficiency of C₃ and C₄ Plants

Soluble and membrane‐associated nitrate reductases in the dinoflagellate Peridinium gatunense

The effects of light on induction, time courses, and kinetic patterns of net nitrate uptake in barley

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Determination of nitrate and nitrite by high-pressure liquid chromatography: Comparison with other methods for nitrate determination

Cited by 190 publications

References 16 publications

The Nitrogen Use Efficiency of C3 and C4 Plants

The Nitrogen Use Efficiency of C3 and C4 Plants

Soluble and membrane‐associated nitrate reductases in the dinoflagellate Peridinium gatunense

The effects of light on induction, time courses, and kinetic patterns of net nitrate uptake in barley

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Product

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The Nitrogen Use Efficiency of C₃ and C₄ Plants

The Nitrogen Use Efficiency of C₃ and C₄ Plants