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1.
The effect of nitrogen form (NH 4-N, NH 4-N + NO 3−, NO 3−) on nitrate reductase activity in roots and shoots of maize ( Zea mays L. cv INRA 508) seedlings was studied. Nitrate reductase activity in leaves was consistent with the well known fact that NO 3− increases, and NH 4+ and amide-N decrease, nitrate reductase activity. Nitrate reductase activity in the roots, however, could not be explained by the root content of NO 3−, NH 4-N, and amide-N. In roots, nitrate reductase activity in vitro was correlated with the rate of nitrate reduction in vivo. Inasmuch as nitrate reduction results in the production of OH − and stimulates the synthesis of organic anions, it was postulated that nitrate reductase activity of roots is stimulated by the released OH − or by the synthesized organic anions rather than by nitrate itself. Addition of HCO 3− to nutrient solution of maize seedlings resulted in a significant increase of the nitrate reductase activity in the roots. As HCO 3−, like OH −, increases pH and promotes the synthesis of organic anions, this provides circumstantial evidence that alkaline conditions and/or organic anions have a more direct impact on nitrate reductase activity than do NO 3−, NH 4-N, and amide-N. 相似文献
2.
The effect of NaCl and Na 2SO 4 salinity on NO 3− assimilation in young barley ( Hordeum vulgare L. var Numar) seedlings was studied. The induction of the NO 3− transporter was affected very little; the major effect of the salts was on its activity. Both Cl − and SO 42− salts severely inhibited uptake of NO 3−. When compared on the basis of osmolality of the uptake solutions, Cl − salts were more inhibitory (15-30%) than SO 42− salts. At equal concentrations, SO 42− salts inhibited NO 3− uptake 30 to 40% more than did Cl − salts. The absolute concentrations of each ion seemed more important as inhibitors of NO 3− uptake than did the osmolality of the uptake solutions. Both K + and Na + salts inhibited NO 3− uptake similarly; hence, the process seemed more sensitive to anionic salinity than to cationic salinity. Unlike NO3− uptake, NO3− reduction was not affected by salinity in short-term studies (12 hours). The rate of reduction of endogenous NO3− in leaves of seedlings grown on NaCl for 8 days decreased only 25%. Nitrate reductase activity in the salt-treated leaves also decreased 20% but its activity, determined either in vitro or by the `anaerobic' in vivo assay, was always greater than the actual in situ rate of NO3− reduction. When salts were added to the assay medium, the in vitro enzymic activity was severely inhibited; whereas the anaerobic in vivo nitrate reductase activity was affected only slightly. These results indicate that in situ nitrate reductase activity is protected from salt injury. The susceptibility to injury of the NO3− transporter, rather than that of the NO3− reduction system, may be a critical factor to plant survival during salt stress. 相似文献
3.
In Ankistrodesmus braunii, in the absence of CO 2, i.e. in CO 2-free air or N 2, photosynthetic nitrate uptake and nitrate reduction were inhibited, especially at low pH. Under such conditions, glucose stimulated nitrate uptake and reduction to almost the same level in the pH range between 6 and 8.5. CO 2 at 0.03% effected an intermediate pH dependence of nitrate uptake; saturating CO 2 concentration (more than 1%) eliminated the pH dependence, as did glucose, but the rates were enhanced compared with glucose. Glucose and, even more, CO 2, drastically reduced the release of nitrite and ammonia to the medium, the stoichiometry between alkalinization of the medium and nitrate uptake (OH −/NO 3−) approached 1. 相似文献
4.
Chlamydomonas reinhardii cells, growing photoautotrophically under air, excreted to the culture medium much higher amounts of NO 2− and NH 4+ under blue than under red light. Under similar conditions, but with NO 2− as the only nitrogen source, the cells consumed NO 2− and excreted NH 4+ at similar rates under blue and red light. In the presence of NO 3− and air with 2% CO 2 (v/v), no excretion of NO 2− and NH 4+ occurred and, moreover, if the bubbling air of the cells that were currently excreting NO 2− and NH 4+ was enriched with 2% CO 2 (v/v), the previously excreted reduced nitrogen ions were rapidly reassimilated. The levels of total nitrate reductase and active nitrate reductase increased several times in the blue-light-irradiated cells growing on NO 3− under air. When tungstate replaced molybdate in the medium (conditions that do not allow the formation of functional nitrate reductase), blue light activated most of the preformed inactive enzyme of the cells. Furthermore, nitrate reductase extracted from the cells in its inactive form was readily activated in vitro by blue light. It appears that under high irradiance (90 w m −2) and low CO 2 tensions, cells growing on NO 3− or NO 2− may not have sufficient carbon skeletons to incorporate all the photogenerated NH 4+. Because these cells should have high levels of reducing power, they might use NO 3− or, in its absence, NO 2− as terminal electron acceptors. The excretion of the products of NO 2− and NH 4+ to the medium may provide a mechanism to control reductant level in the cells. Blue light is suggested as an important regulatory factor of this photorespiratory consumption of NO 3− and possibly of the whole nitrogen metabolism in green algae. 相似文献
5.
Soybean ( Glycine max [L.] Merr.) seeds were imbibed and germinated with or without NO 3−, tungstate, and norflurazon (San 9789). Norflurazon is a herbicide which causes photobleaching of chlorophyll by inhibiting carotenoid synthesis and which impairs normal chloroplast development. After 3 days in the dark, seedlings were placed in white light to induce extractable nitrate reductase activity. The induction of maximal nitrate reductase activity in greening cotyledons did not require NO 3− and was not inhibited by tungstate. Induction of nitrate reductase activity in norflurazon-treated cotyledons had an absolute requirement for NO 3− and was completely inhibited by tungstate. Nitrate was not detected in seeds or seedlings which had not been treated with NO 3−. The optimum pH for cotyledon nitrate reductase activity from norflurazon-treated seedlings was at pH 7.5, and near that for root nitrate reductase activity, whereas the optimum pH for nitrate reductase activity from greening cotyledons was pH 6.5. Induction of root nitrate reductase activity was also inhibited by tungstate and was dependent on the presence of NO 3−, further indicating that the isoform of nitrate reductase induced in norflurazon-treated cotyledons is the same or similar to that found in roots. Nitrate reductases with and without a NO 3− requirement for light induction appear to be present in developing leaves. In vivo kinetics (light induction and dark decay rates) and in vitro kinetics (Arrhenius energies of activation and NADH:NADPH specificities) of nitrate reductases with and without a NO 3− requirement for induction were quite different. Km values for NO 3− were identical for both nitrate reductases. 相似文献
6.
Six genotypes of winter wheat ( Triticum aestivum L.) differing in grain protein concentration were grown on a nutrient solution containing low concentrations of NO 3− (2 millimolar). Total NO 3− uptake varied between genotypes but was not related to grain protein content. An in vivo nitrate reductase assay was used to determine the affinity of the enzyme for NO 3−, and large phenotypic variations were observed. In vivo estimations of the concentration and size of the metabolic pool were variable. However, the three genotypes with the higher ratios of metabolic pool size to leaf total NO 3− concentration were the high protein varieties. It is proposed that a high affinity of nitrate reductase for nitrate might be a biochemical marker for the capacity of the plant to continue assimilating NO 3− for a longer period during the last stage of growth. 相似文献
7.
Membrane associated nitrate reductase (NR) was detected in plasma membrane (PM) fractions isolated by aqueous two-phase partitioning from barley ( Hordeum vulgare L. var CM 72) roots. The PM associated NR was not removed by washing vesicles with 500 millimolar NaCl and 1 millimolar EDTA and represented up to 4% of the total root NR activity. PM associated NR was stimulated up to 20-fold by Triton X-100 whereas soluble NR was only increased 1.7-fold. The latency was a function of the solubilization of NR from the membrane. NR, solubilized from the PM fraction by Triton X-100 was inactivated by antiserum to Chlorella sorokiniana NR. Anti-NR immunoglobulin G fragments purified from the anti-NR serum inhibited NO 3− uptake by more than 90% but had no effect on NO 2− uptake. The inhibitory effect was only partially reversible; uptake recovered to 50% of the control after thorough rinsing of roots. Preimmune serum immunoglobulin G fragments inhibited NO 3− uptake 36% but the effect was completely reversible by rinsing. Intact NR antiserum had no effect on NO 3− uptake. The results present the possibility that NO 3− uptake and NO 3− reduction in the PM of barley roots may be related. 相似文献
8.
In leaves of spinach plants ( Spinacia oleracea L.) performing CO 2 and NO 3− assimilation, at the time of sudden darkening, which eliminates photosystem I-dependent nitrite reduction, only a minor temporary increase of the leaf nitrite content is observed. Because nitrate reduction does not depend on redox equivalents generated by photosystem I activity, a continuation of nitrate reduction after darkening would result in a large accumulation of nitrite in the leaves within a very short time, which is not observed. Measurements of the extractable nitrate reductase activity from spinach leaves assayed under standard conditions showed that in these leaves the nitrate reductase activity decreased during darkening to 15% of the control value with a half-time of only 2 minutes. Apparently, in these leaves nitrate reductase is very rapidly inactivated at sudden darkness avoiding an accumulation of the toxic nitrite in the cells. 相似文献
9.
In vivo NO 3− reduction in roots and shoots of intact barley ( Hordeum vulgare L. var Numar) seedlings was estimated in light and darkness. Seedlings were placed in darkness for 24 hours to make them carbohydrate-deficient. During darkness, the leaves lost 75% of their soluble carbohydrates, whereas the roots lost only 15%. Detached leaves from these plants reduced only 7% of the NO 3− absorbed in darkness. By contrast, detached roots from the seedlings reduced the same proportion of absorbed NO 3−, as did roots from normal light-grown plants. The rate of NO 3− reduction in the roots accounted for that found in the intact dark-treated carbohydrate-deficient seedlings. The rates of NO 3− reduction in roots of intact plants were the same for approximately 12 hours, both in light and darkness, after which the NO 3− reduction rate in roots of plants placed in darkness slowly declined. In the dark, approximately 40% of the NO 3− reduction occurred in the roots, whereas in light only 20% of the total NO 3− reduction occurred in roots. A lesser proportion was reduced in roots because the leaves reduced more nitrate in light than in darkness. 相似文献
10.
Needles from phosphorus deficient seedlings of Pinus radiata D. Don grown for 8 weeks at either 330 or 660 microliters CO 2 per liter displayed chlorophyll a fluorescence induction kinetics characteristic of structural changes within the thylakoid chloroplast membrane, i.e. constant yield fluorescence (F O) was increased and induced fluorescence ([F P-F I]/F O) was reduced. The effect was greatest in the undroughted plants grown at 660 μl CO 2 L −1. By week 22 at 330 μl CO 2 L −1 acclimation to P deficiency had occurred as shown by the similarity in the fluorescence characteristics and maximum rates of photosynthesis of the needles from the two P treatments. However, acclimation did not occur in the plants grown at 660 μl CO 2 L −1. The light saturated rate of photosynthesis of needles with adequate P was higher at 660 μl CO 2 L −1 than at 330 μl CO 2 L −1, whereas photosynthesis of P deficient plants showed no increase when grown at the higher CO 2 concentration. The average growth increase due to CO 2 enrichment was 14% in P deficient plants and 32% when P was adequate. In drought stressed plants grown at 330 μl CO 2 L −1, there was a reduction in the maximal rate of quenching of fluorescence (R Q) after the major peak. Constant yield fluorescence was unaffected but induced fluorescence was lower. These results indicate that electron flow subsequent to photosystem II was affected by drought stress. At 660 μl CO 2 L −1 this response was eliminated showing that CO 2 enrichment improved the ability of the seedlings to acclimate to drought stress. The average growth increase with CO 2 enrichment was 37% in drought stressed plants and 19% in unstressed plants. 相似文献
11.
The observation that exposure of the leaf canopy to increasing concentrations of CO 2 (100-400 μl/l) decreases the influx of nitrate to the leaf blades, but not to the roots or stalks (largely leaf sheaths), was reconfirmed using 15NO 3−. Decreases in leaf nitrate supply were associated with decreases in induction of nitrate reductase, thus supporting the view that the influx of nitrate to a tissue is a major factor in regulation of the level of nitrate reductase. The whole plant 15N distribution data show that the CO 2 effects were due to decreased influx of nitrate into the leaf blade rather than CO 2-enhanced nitrate reduction. The decreases in nitrate accumulation by the leaf blade with increases in CO 2 concentration were only partially accounted for by differences in transpiration. Because the initial malate concentration of root tissue (detopped plants) had no subsequent effect on nitrate uptake, it seems unlikely that high levels of malate induced by CO 2 were responsible for the exclusion of nitrate from the leaf blades. 相似文献
12.
Nitrate reductase activity in excised embryos of Agrostemma githago increases in response to both NO 3− and cytokinins. We asked the question whether cytokinins affected nitrate reductase activity directly or through NO 3−, either by amplifying the effect of low endogenous NO 3− levels, or by making NO 3− available for induction from a metabolically inactive compartment. Nitrate reductase activity was enhanced on the average by 50% after 1 hour of benzyladenine treatment. In some experiments, the cytokinin response was detectable as early as 30 minutes after addition of benzyladenine. Nitrate reductase activity increased linearly for 4 hours and began to decay 13 hours after start of the hormone treatment. When embryos were incubated in solutions containing mixtures of NO 3− and benzyladenine, additive responses were obtained. The effects of NO 3− and benzyladenine were counteracted by abscisic acid. The increase in nitrate reductase activity was inhibited at lower abscisic acid concentrations in embryos which were induced with NO 3−, as compared to embryos treated with benzyladenine. Casein hydrolysate inhibited the development of nitrate reductase activity. The response to NO 3− was more susceptible to inhibition by casein hydrolysate than the response to the hormone. When NO 3− and benzyladenine were withdrawn from the medium after maximal enhancement of nitrate reductase activity, the level of the enzyme decreased rapidly. Nitrate reductase activity increasd again as a result of a second treatment with benzyladenine but not with NO 3−. At the time of the second exposure to benzyladenine, no NO 3− was detectable in extracts of Agrostemma embryos. This is taken as evidence that cytokinins enhance nitrate reductase activity directly and not through induction by NO 3−. 相似文献
14.
The effect of water stress on patterns of nitrate reductase activity in the leaves and nodules and on nitrogen fixation were investigated in Medicago sativa L. plants watered 1 week before drought with or without NO 3−. Nitrogen fixation was decreased by water stress and also inhibited strongly by the presence of NO 3−. During drought, leaf nitrate reductase activity (NRA) decreased significantly particularly in plants watered with NO 3−, while with rewatering, leaf NRA recovery was quite important especially in the NO 3−-watered plants. As water stress progressed, the nodular NRA increased both in plants watered with NO 3− and in those without NO 3− contrary to the behavior of the leaves. Beyond −15.10 5 pascal, nodular NRA began to decrease in plants watered with NO 3−. This phenomenon was not observed in nodules of plants given water only. 相似文献
15.
Ricinus communis L. plants were grown in nutrient solutions in which N was supplied as NO 3− or NH 4+, the solutions being maintained at pH 5.5. In NO 3−-fed plants excess nutrient anion over cation uptake was equivalent to net OH − efflux, and the total charge from NO 3− and SO 42− reduction equated to the sum of organic anion accumulation plus net OH − efflux. In NH 4+-fed plants a large H + efflux was recorded in close agreement with excess cation over anion uptake. This H + efflux equated to the sum of net cation (NH 4+ minus SO 42−) assimilation plus organic anion accumulation. In vivo nitrate reductase assays revealed that the roots may have the capacity to reduce just under half of the total NO 3− that is taken up and reduced in NO 3−-fed plants. Organic anion concentration in these plants was much higher in the shoots than in the roots. In NH 4+-fed plants absorbed NH 4+ was almost exclusively assimilated in the roots. These plants were considerably lower in organic anions than NO 3−-fed plants, but had equal concentrations in shoots and roots. Xylem and phloem saps were collected from plants exposed to both N sources and analyzed for all major contributing ionic and nitrogenous compounds. The results obtained were used to assist in interpreting the ion uptake, assimilation, and accumulation data in terms of shoot/root pH regulation and cycling of nutrients. 相似文献
16.
The photoreversible nature of the regulation of nitrate reductase is one of the most interesting features of this enzyme. As well as other chemicals, NH 2OH reversibly inactivates the reduced form of nitrate reductase from Ankistrodesmus braunii. From the partial activities of the enzyme, only terminal nitrate reductase is affected by NH 2OH. To demonstrate that the terminal activity was readily inactivted by NH 2OH, the necessary reductants of the terminal part of the enzyme had to be cleared of dithionite since this compound reacts chemically with NH 2OH. Photoreduced flavins and electrochemically reduced methyl viologen sustain very effective inactivation of terminal nitrate reductase activity, even if the enzyme was previously deprived of its NADH-dehydrogenase activity. The early inhibition of nitrate reductase by NH 2OH appears to be competitive versus NO 3−. Since NO 3−, as well as cyanate, carbamyl phosphate and azide (competitive inhibitors of nitrate reductase versus NO 3−), protect the enzyme from NH 2OH inactivation, it is suggested that NH 2OH binds to the nitrate active site. The NH 2OH-inactivated enzyme was photoreactivated in the presence of flavins, although slower than when the enzyme was previously inactivated with CN −. NH 2OH and NADH concentrations required for full inactivation of nitrate reductase appear to be low enough to potentially consider this inactivation process of physiological significance. 相似文献
17.
The comparative induction of nitrate reductase (NR) by ambient NO 3− and NO 2− as a function of influx, reduction (as NR was induced) and accumulation in detached leaves of 8-day-old barley ( Hordeum valgare L.) seedlings was determined. The dynamic interaction of NO 3− influx, reduction and accumulation on NR induction was shown. The activity of NR, as it was induced, influenced its further induction by affecting the internal concentration of NO 3−. As the ambient concentration of NO 3− increased, the relative influences imposed by influx and reduction on NO 3− accumulation changed with influx becoming a more predominant regulant. Significant levels of NO 3− accumulated in NO 2−-fed leaves. When the leaves were supplied cycloheximide or tungstate along with NO 2−, about 60% more NO 3− accumulated in the leaves than in the absence of the inhibitors. In NO 3−-supplied leaves NR induction was observed at an ambient concentration of as low as 0.02 m m. No NR induction occurred in leaves supplied with NO 2− until the ambient NO 2− concentration was 0.5 m m. In fact, NR induction from NO 2− solutions was not seen until NO 3− was detected in the leaves. The amount of NO 3− accumulating in NO 2−-fed leaves induced similar levels of NR as did equivalent amounts of NO 3− accumulating from NO 3−-fed leaves. In all cases the internal concentration of NO 3−, but not NO 2−, was highly correlated with the amount of NR induced. The evidence indicated that NO 3− was a more likely inducer of NR than was NO 2−. 相似文献
18.
The mass transfer rate of 14C-sucrose translocation from sugar beet ( Beta vulgaris, L.) leaves was measured over a range of net photosynthesis rates from 0 to 60 milligrams of CO 2 decimeters −2 hour −1 under varying conditions of light intensity, CO 2 concentration, and O 2 concentration. The resulting rate of translocation of labeled photosynthate into total sink tissue was a linear function (slope = 0.18) of the net photosynthesis rate of the source leaf regardless of light intensity (2000, 3700, or 7200 foot-candles), O 2 concentration (21% or 1% O 2), or CO 2 concentration (900 microliters/liter of CO 2 to compensation concentration). These data support the theory that the mass transfer rate of translocation under conditions of sufficient sink demand is limited by the net photosynthesis rate or more specifically by sucrose synthesis and this limitation is independent of light intensity per se. The rate of translocation was not saturated even at net photosynthesis rates four times greater than the rate occurring at 300 microliters/liter of CO 2, 21% O 2, and saturating light intensity. 相似文献
19.
Nitrate reduction was studied as a function of carbohydrate concentration in detached primary leaves of barley ( Hordeum vulgare L. cv Numar) seedlings under aerobic conditions in light and darkness. Seedlings were grown either in continuous light for 8 days or under a regimen of 16-hour light and 8-hour dark for 8 to 15 days. Leaves of 8-day-old seedlings grown in continuous light accumulated 4 times more carbohydrates than leaves of plants grown under a light and dark regimen. When detached leaves from these seedlings were supplied with NO 3− in darkness, those with the higher levels of carbohydrates reduced a greater proportion of the NO 3− that was taken up. In darkness, added glucose increased the percentage of NO 3− reduced up to 2.6-fold depending on the endogenous carbohydrate status of the leaves. Both NO 3− reduction and carbohydrate content of the leaves increased with age. Fructose and sucrose also increased NO 3− reduction in darkness to the same extent as glucose. Krebs cycle intermediates, citrate and succinate, did not increase NO 3− reduction, whereas malate slightly stimulated it in darkness. In light, 73 to 90% of the NO3− taken up was reduced by the detached leaves; therefore, an exogenous supply of glucose had little additional effect on NO3− reduction. The results indicate that in darkness the rate of NO3− reduction in primary leaves of barley depends upon the availability of carbohydrates. 相似文献
20.
Evidence is presented that chlorate is an extremely good analog for nitrate during nitrate uptake by intact barley ( Hordeum vulgare cv. Fergus) roots. The depletion of ClO 3− or NO 3− from uptake media over 2 to 6 hours by seedlings was found to be dependent on combined NO 3− plus ClO 3− concentrations, and total anion uptake was equivalent at different NO 3−/ClO 3− ratios. After loading barley seedlings with 36ClO 3− for 6 hours, kinetic parameters were derived from the analysis of efflux of [ 36Cl] chlorate into unlabeled solution. On the basis of this analysis, the half times for exchange for the cytoplasmic and vacuolar phases were 17 minutes and 20 hours, respectively. 相似文献
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