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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.
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
3.
The effects of N source (6 m m nitrogen as NO 3− or urea) and tungstate (0, 100, 200, 300, and 400 μ m Na 2 WO 4) on nitrate metabolism, nodulation, and growth of soybean ( Glycine max [L.] Merr.) plants were evaluated. Nitrate reductase activity and, to a lesser extent, NO 3− content of leaf tissue decreased with the addition of tungstate to the nutrient growth medium. Concomitantly, nodule mass and acetylene reduction activity of NO 3−-grown plants increased with addition of tungstate to the nutrient solution. In contrast, nodule mass and acetylene reduction activity of urea-grown plants decreased with increased nutrient tungstate levels. The acetylene reduction activity of nodulated roots of NO 3−-grown plants was less than 10% of the activity of nodulated roots of urea-grown plants when no tungstate was added. At 300 and 400 μ m tungstate levels, acetylene reduction activity of nodulated roots of NO 3−-grown plants exceeded the activity of comparable urea-grown plants. 相似文献
4.
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
5.
The nature of the injury and recovery of nitrate uptake (net uptake) from NaCl stress in young barley ( Hordeum vulgare L, var CM 72) seedlings was investigated. Nitrate uptake was inhibited rapidly by NaCl, within 1 minute after exposure to 200 millimolar NaCl. The duration of exposure to saline conditions determined the time of recovery of NO 3− uptake from NaCl stress. Recovery was dependent on the presence of NO 3− and was inhibited by cycloheximide, 6-methylpurine, and cerulenin, respective inhibitors of protein, RNA, and sterol/fatty acid synthesis. These inhibitors also prevented the induction of the NO 3− uptake system in uninduced seedlings. Uninduced seedlings exhibited endogenous NO 3− transport activity that appeared to be constitutive. This constitutive activity was also inhibited by NaCl. Recovery of constitutive NO 3− uptake did not require the presence of NO 3−. 相似文献
6.
Nitrate reductase activity (NRA) was found in primary roots, but not in foliage of red spruce ( Picea rubens Sarg.) seedlings. Nitrate induced NRA:NH 4+ did not induce and slightly depressed NRA in older seedlings. Induction required 8 hours and, once induced, NRA decreased slowly in the absence of exogenous NO 3−. Seedlings were grown in perlite with a complete nutrient solution containing NH 4+ to limit NR induction. Established seedlings were stressed with nutrient solutions at pH 3, 4, or 5 supplemented with Cl − salts of Al, Cd, Pb, or Zn each at two concentrations. NRA in primary root tips was measured at 2, 14, 28, and 42 days. NRA induction was greatest at pH 3, and remained high during the period of study. NRA induction at pH 4 was lower. Metal ions suppressed NRA at pH 3 and 5, but enhanced NRA at pH 4. It is concluded that acidity and soluble metals in the root environment of red spruce are unlikely to be important factors in nitrogen transformations in red spruce roots. 相似文献
7.
Radish ( Raphanus sativus L.) seedlings pretreated with different hormones viz. kinetin, gibberellic acid and abscisic acid were subjected to different
N-forms. The seedlings were treated with different concentrations of KNO 3, NH 4Cl and NH 4NO 3 and the changes in nitrate reductase activity were seen in light and dark conditions in the cotyledons. Nitrate reductase
activity was affected differently by hormone application. Nitrate increased and ammonia decreased nitrate reductase activity;
in both light and dark-grown seedlings KNO 3 induced more in vitro nitrate reductase activity. NH
4
+
when combined with NO
3
−
, however, could level up to some extent, with KNO 3 in light, except in kinetin. A transient response of induction of NR activity was evident with decreased levels after a certain
specific ambient N-concentration, despite the presence of high N in the medium. However, the pattern of transition varied
with the hormones applied. Further, hormones are found to affect induction of different isoforms of nitrate reductase by NH
4
+
and NO
3
−
. NH
4
+
induced isoform was prominently promoted by kinetin treatment in dark. The data documents a particular kind of interaction
between controlling factors (light, N-source and phytohormones) which affect nitrate reductase levels. 相似文献
8.
Pyrobaculum aerophilum, a hyperthermophilic archaeon, can respire either with low amounts of oxygen or anaerobically with nitrate as the electron acceptor. Under anaerobic growth conditions, nitrate is reduced via the denitrification pathway to molecular nitrogen. This study demonstrates that P. aerophilum requires the metal oxyanion WO 42− for its anaerobic growth on yeast extract, peptone, and nitrate as carbon and energy sources. The addition of 1 μM MoO 42− did not replace WO 42− for the growth of P. aerophilum. However, cell growth was completely inhibited by the addition of 100 μM MoO 42− to the culture medium. At lower tungstate concentrations (0.3 μM and less), nitrite was accumulated in the culture medium. The accumulation of nitrite was abolished at higher WO 42− concentrations (<0.7 μM). High-temperature enzyme assays for the nitrate, nitrite, and nitric oxide reductases were performed. The majority of all three denitrification pathway enzyme activities was localized to the cytoplasmic membrane, suggesting their involvement in the energy metabolism of the cell. While nitrite and nitric oxide specific activities were relatively constant at different tungstate concentrations, the activity of nitrate reductase was decreased fourfold at WO 42− levels of 0.7 μM or higher. The high specific activity of the nitrate reductase enzyme observed at low WO 42− levels (0.3 μM or less) coincided with the accumulation of nitrite in the culture medium. This study documents the first example of the effect of tungstate on the denitrification process of an extremely thermophilic archaeon. We demonstrate here that nitrate reductase synthesis in P. aerophilum occurs in the presence of high concentrations of tungstate. 相似文献
9.
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
10.
A comparison of induction and inactivation of nitrate reductase and two of its component activities, namely FMNH 2-nitrate reductase and NO 3−-induced NADH-cytochrome c reductase, was made in roots and leaves of corn ( Zea mays L. var. W64A × 182E). The three activities were induced in parallel in both tissues when NO 3− was supplied. WO 4= suppressed the induction of NADH- and FMNH 2-nitrate reductase activities in root tips and leaves. The NO 3−-induced NADH-cytochrome c reductase activity showed a normal increase in roots treated with WO 4=. In leaves, on the other hand, there was a marked superinduction of the NO 3−-induced NADH-cytochrome c reductase in the presence of WO 4=. 相似文献
11.
Nitrate reductase utilizing NADH or reduced flavin mononucleotide (FMNH 2) as electron donor was extracted from the leaves, stems and petioles, and roots of apple seedlings. Successful extraction was made possible by the use of insoluble polyvinylpyrrolidone (Polyclar AT) which forms insoluble complexes with polyphenols and tannins. The level of nitrate reductase per gram fresh weight was highest in the leaf tissue although the nitrate content of the roots was much higher than that of the leaves. Nitrite reductase activity was detected only in leaf extracts and was 4 times higher than nitrate reductase activity. Nitrate was found in all parts of young apple trees and trace amounts were also detected in mature leaves from mature trees. Nitrate reductase was induced in young leaves of apple seedlings and in mature leaves from 3 fruit-bearing varieties. An inhibitor of polyphenoloxidase, 2-mercaptobenzothiazole was used in both the inducing medium and the extracting medium in concentrations from 10 −3 to 10 −5m with no effect upon nitrate reductase activity. 相似文献
12.
Induction of nitrate reductase activity and mRNA by nitrate and light is prevented if chloroplasts are destroyed by photooxidation in norflurazon-treated squash ( Cucurbita maxima L.) cotyledons. The enzyme activity and mRNA can be induced if norflurazon-treated squash seedlings are kept in low-intensity red light, which minimizes photodamage to the plastids. It is concluded that induction of nitrate reductase activity and nitrate reductase mRNA requires intact plastids. If squash seedlings grown in low-intensity red light are transferred to photooxidative white light, nitrate reductase activity accumulates during the first 12 hours after the shift and declines thereafter. Thus photodamage to the plastids and the disappearance of nitrate reductase activity and mRNA are events separable in time, and disappearance of the enzyme activity is a consequence of the damage to the plastids. 相似文献
13.
The role of NO 3− and NO 2− in the induction of nitrite reductase (NiR) activity in detached leaves of 8-day-old barley ( Hordeum vulgare L.) seedlings was investigated. Barley leaves contained 6 to 8 micromoles NO 2−/gram fresh weight × hour of endogenous NiR activity when grown in N-free solutions. Supply of both NO 2− and NO 3− induced the enzyme activity above the endogenous levels (5 and 10 times, respectively at 10 millimolar NO 2− and NO 3− over a 24 hour period). In NO 3−-supplied leaves, NiR induction occurred at an ambient NO 3− concentration of as low as 0.05 millimolar; however, no NiR induction was found in leaves supplied with NO 2− until the ambient NO 2− concentration was 0.5 millimolar. Nitrate accumulated in NO 2−-fed leaves. The amount of NO 3− accumulating in NO 2−-fed leaves induced similar levels of NiR as did equivalent amounts of NO 3− accumulating in NO 3−-fed leaves. Induction of NiR in NO 2−-fed leaves was not seen until NO 3− was detectable (30 nanomoles/gram fresh weight) in the leaves. The internal concentrations of NO 3−, irrespective of N source, were highly correlated with the levels of NiR induced. When the reduction of NO 3− to NO 2− was inhibited by WO 42−, the induction of NiR was inhibited only partially. The results indicate that in barley leaves NiR is induced by NO 3− directly, i.e. without being reduced to NO 2−, and that absorbed NO 2− induces the enzyme activity indirectly after being oxidized to NO 3− within the leaf. 相似文献
14.
The effects of CO 2-limited photosynthesis on 15NO 3− uptake and reduction by maize ( Zea mays, DeKalb XL-45) seedlings were examined in relation to concurrent effects of CO 2 stress on carbohydrate levels and in vitro nitrate reductase activities. During a 10-hour period in CO 2-depleted air (30 microliters of CO 2/ per liter), cumulative 15NO 3− uptake and reduction were restricted 22 and 82%, respectively, relative to control seedlings exposed to ambient air containing 450 microliters of CO 2 per liter. The comparable values for roots of decapitated maize seedlings, the shoots of which had previously been subjected to CO 2 stress, were 30 and 42%. The results demonstrate that reduction of entering nitrate by roots as well as shoots was regulated by concurrent photosynthesis. Although in vitro nitrate reductase activity of both tissues declined by 60% during a 10-hour period of CO 2 stress, the remaining activity was greatly in excess of that required to catalyze the measured rate of 15NO 3− reduction. Root respiration and soluble carbohydrate levels in root tissue were also decreased by CO 2 stress. Collectively, the results indicate that nitrate uptake and reduction were regulated by the supply of energy and carbon skeletons required to support these processes, rather than by the potential enzymatic capacity to catalyze nitrate reduction, as measured by in vitro nitrate reductase activity. 相似文献
15.
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
16.
Corn seedlings ( Zea mays cv W64A × W182E) were grown hydroponically, in the presence or absence of NO 3−, with or without light and with NH 4Cl as the only N source. In agreement with earlier results nitrate reductase (NR) activity was found only in plants treated with both light and NO 3−. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by transfer of the proteins to nitrocellulose paper and reaction with antibodies prepared against a pure NR showed that crude extracts prepared from light-grown plants had a polypeptide of approximately 116 kilodaltons (the subunit size for NR) when NO 3− was present in the growth medium. Crude extracts from plants grown in the dark did not have the 116 kilodalton polypeptide, although smaller polypeptides, which reacted with NR-immunoglobulin G, were sometimes found at the gel front. When seedlings were grown on Kimpack paper or well washed sand, NR activity was again found only when the seedlings were exposed to light and NO 3−. Under these conditions, however, a protein of about 116 kilodaltons, which reacted with the NR antibody was present in light-grown plants whether NO 3− was added to the system or not. The NR antibody cross-reacting protein was also seen in hydroponically grown plants when NH 4Cl − was the only added form of nitrogen. These results indicate that the induction of an inactive NR-protein precursor in corn is mediated either by extremely low levels of NO 3− or by some other unidentified factor, and that higher levels of NO 3− are necessary for converting the inactive NR cross-reacting protein to a form of the enzyme capable of reducing NO 3− to NO 2−. 相似文献
17.
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
18.
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
19.
Nitrate and NO 2− transport by roots of 8-day-old uninduced and induced intact barley ( Hordeum vulgare L. var CM 72) seedlings were compared to kinetic patterns, reciprocal inhibition of the transport systems, and the effect of the inhibitor, p-hydroxymercuribenzoate. Net uptake of NO 3− and NO 2− was measured by following the depletion of the ions from the uptake solutions. The roots of uninduced seedlings possessed a low concentration, saturable, low Km, possibly a constitutive uptake system, and a linear system for both NO 3− and NO 2−. The low Km system followed Michaelis-Menten kinetics and approached saturation between 40 and 100 micromolar, whereas the linear system was detected between 100 and 500 micromolar. In roots of induced seedlings, rates for both NO 3− and NO 2− uptake followed Michaelis-Menten kinetics and approached saturation at about 200 micromolar. In induced roots, two kinetically identifiable transport systems were resolved for each anion. At the lower substrate concentrations, less than 10 micromolar, the apparent low Kms of NO 3− and NO 2− uptake were 7 and 9 micromolar, respectively, and were similar to those of the low Km system in uninduced roots. At substrate concentrations between 10 and 200 micromolar, the apparent high Km values of NO 3− uptake ranged from 34 to 36 micromolar and of NO 2− uptake ranged from 41 to 49 micromolar. A linear system was also found in induced seedlings at concentrations above 500 micromolar. Double reciprocal plots indicated that NO 3− and NO 2− inhibited the uptake of each other competitively in both uninduced and induced seedlings; however, Ki values showed that NO 3− was a more effective inhibitor than NO 2−. Nitrate and NO 2− transport by both the low and high Km systems were greatly inhibited by p-hydroxymercuribenzoate, whereas the linear system was only slightly inhibited. 相似文献
20.
Effects of NO 2−, ClO 3−, and ClO 2− on the induction of nitrate transport and nitrate reductase activity (NRA) as well as their effects on NO 3− influx into roots of intact barley ( Hordeum vulgare cv Klondike) seedlings were investigated. A 24-h pretreatment with 0.1 mol m −3 NO 2− fully induced NO 3− transport but failed to induce NRA. Similar pretreatments with ClO 3− and ClO 2− induced neither NO 3− transport nor NRA. Net ClO 3− uptake was induced by NO 3− but not by ClO 3− itself, indicating that NO 3− and ClO 3− transport occur via the NO 3− carrier. At the uptake step, NO 2− and ClO 2− strongly inhibited NO 3− influx; the former exhibited classical competitive kinetics, whereas the latter exhibited complex mixed-type kinetics. ClO 3− proved to be a weak inhibitor of NO 3− influx ( Ki = 16 mol m −3) in a noncompetitive manner. The implications of these findings are discussed in the context of the suitability of these NO 3− analogs as screening agents for the isolation of mutants defective in NO 3− transport. 相似文献
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