Abstract
Anin situ method, derived from anin vivo method, was used to determine nitrate reductase activity (NRA) in:i) excised barley and corn shoots and excised soybean leaves during a N-depletion experiment and; ii) roots and shoots of N-depleted barley and corn seedlings during induction of nitrate, reductase (NR). Nitrate reduction, calculated from thesein situ RNA measurements, was compared with estimates of each organ's nitrate reduction in light aerobic conditions from NO −3 consumption and a15N model (Gojonet al., 1986b).
Thein situ RNA of roots strongly underestimated their15NO −3 reduction. In contrast, in barley and corn shoots and in the first trifoliolate leaves from 26-day-old, soybean, thein situ NRA assay gave a fair approximation of the true NO −3 reduction rate (relative differences ranging from −14 to +32%). In young soybean leaves (from 20-day-old plants), however, thein situ NRA strongly underestimated the actual NO −3 reduction. The physiological significance of thein situ NRA assay in shoots and roots, and its value for field studies are discussed from these results.
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Aryan A P and Wallace W 1983 Involvement of alcohol dehydrogenase in the enhancement of thein vivo nitrate reductase activity of root tissue by propanol. Plant Sci. Lett. 30, 25–32.
Aston M G and Lawlor D W 1979 The relationship between transpiration, root water uptake and leaf water potential. J. Exp. Bot. 30, 169–181.
Aslam M and Oaks A 1975 Effect of glucose on the induction of nitrate reductase in corn roots. Plant Physiol. 56, 634–639.
Crafts-Brandner S J and Harper J E 1982 Nitrate reduction by roots of soybean seedlings (Glycine max L. Merr.). Plant Physiol. 69, 1298–1303.
Dale J E 1976 Nitrate reduction in the first leaf and roots of barley seedlings grown in sand and in culture solution. Ann. Bot. 40, 1177–1184.
Dean J V and Harper J E 1988 The conversion of nitrite to nitrogen oxide(s) by the constitutive NAD(P)H-nitrate reductase enzyme from soybean. Plant Physiol. 88, 389–395.
Dry I, Wallace W and Nicholas D J D 1981 Role of ATP in nitrite reduction in roots of wheat and pea. Planta 152, 234–238.
Emes M J and Fowler M W 1983 The supply of reducing power for nitrite reduction in plastids of seedlings pea roots (Pisum sativum L.). Planta 158, 97–102.
Ezeta F N and Jackson W A 1975 Nitrate translocation by detopped corn seedlings. Plant Physiol. 56, 148–156.
Gojon A, Passama L and Robin P 1986a Root contribution to nitrate reduction in barley seedlings (Hordeum vulgare L.). Plant Soil 91, 339–342.
Gojon A, Soussana J F, Passama L and Robin P 1986b Nitrate reduction in roots and shoots of barley (Hordeum vulgare L.) and corn (Zea may L.) seedlings. I.15N study. Plant Physiol. 82, 254–260.
Granstedt R C and Huffaker R C 1982 Identification of the leaf vacuole as a major nitrate storage pool. Plant Physiol. 70, 410–413.
Gray V M and Cresswell C F 1984 Nitrite utilization by excisedZea mays roots under anaerobic conditions. Plant Sci. Lett. 33, 31–38.
Hog K, Hartvigsen M B and Rasmussen O S 1983 Critical evaluation of thein vivo nitrate reductase assay for detection of two nitrate pools in wheat leaves. Physiol. Plant 59, 141–147.
Jones W R and Sheard R W 1978 Accumulation and stability of nitrite in intact aerial leaves. Plant Sci. Lett. 44, 285–291.
Lawrence J M and Herrick H E 1982 Media forin vivo nitrate reductase assay of plant tissues. Plant Sci. Lett. 24, 17–26.
Lee R B 1980 Sources of reductant for nitrate assimilation in non-photosynthetic tissue: A review. Plant Cell Environ. 3, 65–90.
Martinoia E, Heck U H and Wiemkzn A 1981 Vacuoles as storage compartments for nitrate in barley leaves. Nature 289, 292–294.
Nelson R S, Ryan S A and Harper J E 1983 Soybean mutants lacking constitutive nitrate reductase activity. I. Selection and initial characterization. Plant Physiol. 75, 503–509.
Passama L, Gojon A, Robin P and Salsac L 1987In situ nitrate reductase activity as an indicator of nitrate availability. Plant and Soil 102, 145–148.
Reed A J and Hageman R H 1980 Relationship between nitrate uptake, flux and reduction and the accumulation of reduced nitrogen in maize (Zea mays L.). II. Effect of nutrient nitrate concentration. Plant Physiol. 66, 1184–1189.
Reed A J, Canvin D T, Sherrard J H and Hageman R H 1983 Assimilation of15N-nitrate and15N-nitrite in leaves of five plant species under light and dark conditions. Plant Physiol 71, 291–294.
Robin P, Blayac D and Salsac L 1979 Influence de l'alimentation nitrique sur la teneur en nitrate et l'activité nitrate réductase des racines et des feuilles de plantules de maís. Physiol. Vég. 17, 55–56.
Robin P, Conejero G, Passama L and Salsac L 1983a Avaluation de la fraction métabolisable du nitrate par la mesurein situ de sa réduction. Physiol. Vég. 21, 115–122.
Robin P, Conejero G, Tranchant J P, Passama L and Salsac L 1983b Mesure de la réduction du nitrate dans les feuilles intactes. Physiol. Vég. 21, 123–128.
Shaner D L and Boyer J S 1976 Nitrate reductase activity in maize (Zea mays L.) leaves. I. Regulation by nitrate flux. Plant Physiol. 58, 499–504.
Smith F W and Thompson J F 1971 Regulation of nitrate reductase in excised barley roots. Plant Physiol. 48, 219–223.
Somers D A, Kuo T M, Kleinhofs A, Warner R L and Oaks A 1983 Synthesis and degradation of barley nitrate reductase. Plant Physiol. 72, 949–952.
Srinivasan R, Prakash S and Naik M S 1982 Reevaluation ofin vivo assay of nitrate reductase activity in wheat leaves. Plant Sci. Lett. 25, 9–14.
Wakrim R 1988 Régulation de l'assimilation du nitrate dans la feuille de soja par le flux xylémique de l'anion. Doc. Thesis, Univ. Montpellier II, France, 146 p.
Wery J, Turc O and Salsac L 1986 Relationship between growth, nitrogen fixation and assimilation in a legume (Medicago sativa L.). Plant and Soil 96, 17–29.
Yoneyama T 198115N studies on thein vivo assay of nitrate reductase in leaves: Occurrence of underestimation of the activity due to dark assimilation of nitrate and nitrite. Plant Cell Physiol. 22, 1507–1520.
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Soussana, J.F., Gojon, A., Passama, L. et al. Critical evaluation of thein situ nitrate reductase assay. Plant Soil 120, 243–251 (1989). https://doi.org/10.1007/BF02377074
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DOI: https://doi.org/10.1007/BF02377074

