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1.
Phosphoenolpyruvate carboxylase (EC 4.1.1.31; PEPCase) from Bryophyllum fedtschenkoi leaves has previously been shown to exist in two forms in vivo. During the night the enzyme is phosphorylated and relatively insensitive to feedback inhibition by malate whereas during the day the enzyme is dephosphorylated and more sensitive to inhibition by malate. These properties of PEPCase have now been investigated in leaves maintained under constant conditions of temperature and lighting. When leaves were maintained in continuous darkness and CO2-free air at 15°C, PEPCase exhibited a persistent circadian rhythm of interconversion between the two forms. There was a good correlation between periods during which the leaves were fixing respiratory CO2 and periods during which PEPCase was in the form normally observed at night. When leaves were maintained in continuous light and normal air at 15°C, starting at the end of a night or the end of a day, a circadian rhythm of net uptake of CO2 was observed. Only when these constant conditions were applied at the end of a day was a circadian rhythm of interconversions between the two forms of PEPCase observed and the rhythms of enzyme interconversion and CO2 uptake did not correlate in phase or period.Abbreviations CAM
Crassulacean acid metabolism
- FW
fresh weight
- PEPCase
phosphoenolpyruvate carboxylase
- RuBPCase
ribulose-1,5-bisphosphate carboxylase
To whom correspondence should be addressed. 相似文献
2.
This article reports marked modulation of the activity and regulatory properties of phosphoenolpyruvate carboxylase (PEPC) by temperature and light in leaf discs as well as leaves of Amaranthus hypochondriacus. The activity of PEPC increased by 1.7-fold at 45 degrees C over 25 degrees C. Warm temperature also stimulated the photoactivation of PEPC. The activation by light of PEPC was 1.9-fold at 25 degrees C and increased to 2.2-fold at 45 degrees C. The sensitivity of PEPC to its inhibitor malate was less and the activation by glucose-6-phosphate (G-6-P) or inorganic phosphate (Pi) was more at 45 degrees C than that at 25 degrees C. These effects of temperature were quite pronounced in light. Similar responses were observed when detached leaves were exposed to varying ambient temperature (dry heat). The activity of PEPC increased by 1.6-fold at 45 degrees C over 25 degrees C in the dark. The activation of PEPC by light was 2.1-fold at 25 degrees C and increased to 2.6-fold at 45 degrees C. Inhibition by malate was less and activation by G-6-P or Pi was more at 45 degrees C than that at 25 degrees C. Thus, there was a marked modulation of not only the activity but also the regulatory properties of the enzyme by temperature and light, independently as well as cooperatively with each other. Further experiments suggested that PEPC was able to memorize to a significant extent the changes induced by warm temperature and that these changes were complemented by subsequent illumination. These effects were not due to changes in PEPC protein levels. We conclude that temperature and light can modulate PEPC activity and regulatory properties not only individually but also in a significantly cooperative manner with each other. As significant increases in temperature are common during daytime in tropical or subtropical conditions, we suggest that the synergistic effects of temperature and light are quite relevant in optimizing the activity of PEPC in leaves of C(4) plants. 相似文献
3.
Rapid modulation of nitrate reductase in pea roots 总被引:10,自引:0,他引:10
The regulatory properties of nitrate reductase (NR; EC 1.6.6.1) in root extracts from hydroponically grown pea (Pisum sativum L. cv. Kleine Rheinländerin) plants were examined and compared with known properties of NR from spinach and pea leaves. Nitrate-reductase activity (NRA) extracted from pea roots decreased slowly when plants were kept in the dark, or when illuminated plants were detopped, with a half-time of about 4 h (= slow modulation in vivo). In contrast, the half-time for the dark-inactivation of NR from pea leaves was only 10 min. However, when root tip segments were transferred from aerobic to anaerobic conditions or vice versa, changes in NRA were as rapid as in leaves (= rapid modulation in vivo). Nitrate-reductase activity was low when extracted from roots kept in solutions flushed with air or pure oxygen, and high in nitrogen. Okadaic acid, a specific inhibitor of type-1 and type-2A protein phosphatases, totally prevented the in vivo activation by anaerobiosis of NR, indicating that rapid activation of root NR involved protein dephosphorylation. Under aerobic conditions, the low NRA in roots was also rapidly increased by incubating the roots with either uncouplers or mannose. Under these conditions, and also under anaerobiosis, ATP levels in roots were much lower than in aerated control roots. Thus, whenever ATP levels in roots were artificially decreased, NRA increased rapidly. The highly active NR extracted from anaerobic roots could be partially inactivated in vitro by preincubation of desalted root extracts with MgATP (2 mM), with a half-time of about 20 min. It was reactivated by subsequently incubating the extracts with excess AMP (2 mM). Thus, pea root NR shares many of the previously described properties of NR from spinach leaves, suggesting that the root enzyme, like the leaf enzyme, can be rapidly modulated, probably by reversible protein phosphorylation/ dephosphorylation. 相似文献
4.
Purified maize leaf phosphoenolpyruvate carboxylase (EC 4.1.1.31) was completely inactivated by several thiol-modifying reagents, including, CuCl2, CdCl2 and N-ethylmaleimide. The inactivation by CuCl2 could be reversed by dithiothreitol, suggesting the involvement of vicinal dithiols in the inactivation process.Complete inactivation of phosphoenolpyruvate carboxylase was correlated with the incorporation of two mol (3H)N-ethylmaleimide per 100-kilodalton subunit. The total protection of the enzyme against N-ethylmaleimide inactivation afforded by the substrate, phosphoenolpyruvate, was correlated with the protection of one mol (3H)N-ethylmaleimide reactive residue per mol subunit.The complete inactivation of phosphoenolpyruvate carboxylase by N-ethylmaleimide and the protection afforded by phosphoenolpyruvate against modification suggest the presence of an essential cysteine residue in the catalytic site of the C4 leaf enzyme.Abbreviations PEP,
phosphoenolpyruvate
- Mops,
4-morpholinepropanesulphonic acid
(Consejo Nacional de Investigaciones Científicas y Técnicas, Fundación M. Lillo y U.N. de Rosario). 相似文献
5.
In the present work, the effect of LiCl on phosphoenolpyruvate carboxylase kinase (PEPCase-k), C4 phosphoenolpyruvate carboxylase (PEPCase: EC 4.1.1.31) and its phosphorylation process has been investigated in illuminated
leaf disks and leaves of the C4 plant Sorghum vulgare. Although this salt induced severe damages to older leaves, it did not significantly alter the physiological parameters (photosynthesis,
transpiration rate, intercellular CO2 concentration) of young leaves. An immunological approach was used to demonstrate that the PEPCase-k protein accumulated
rapidly in illuminated leaf tissues, consistent with the increase in its catalytic activity. In vivo, LiCl was shown to strongly
enhance the light effect on PEPCase-k protein content, this process being dependent on protein synthesis. In marked contrast,
the salt was found to inhibit the PEPCase-k activity in reconstituted assays and to decrease the C4 PEPCase content and phosphorylation state in LiCl treated plants. Short-term (15 min) LiCl treatment increased IP3 levels, PPCK gene expression, and PEPCase-k accumulation. Extending the treatment (1 h) markedly decreased IP3 and PPCK gene expression, while PEPCase-k activity was kept high. The cytosolic protein synthesis inhibitor cycloheximide (CHX), which
blocked the light-dependent up-regulation of the kinase in control plants, was found not to be active on this process in preilluminated,
LiCl-treated leaves. This suggested that the salt causes the kinase turnover to be altered, presumably by decreasing degradation
of the corresponding polypeptide. Taken together, these results establish PEPCase-k and PEPCase phosphorylation as lithium
targets in higher plants and that this salt can provide a means to investigate further the organization and functioning of
the cascade controlling the activity of both enzymes. 相似文献
6.
Leaves ofBryophyllum fedtschenkoi show a persistent circadian rhythm in CO2 assimilation when kept in continuous illumination and normal air at 15°C. The induction of phase shifts in this rhythm by
exposing the leaves for four hours at different times in the circadian cycle to 40° C, 2° C, darkness and 5% CO2 have been investigated. Exposure to high temperature has no effect on the phase at the apex of the peak but is effective
at all other times in the cycle, whereas exposure to low temperature, darkness or 5% CO2 is without effect between the peaks and induces a phase shift at all other times. The next peak of the rhythm occurs 17 h
after a 40° C treatment and 7–10 h after a 2° C, dark or 5% CO2 treatment regardless of their position in the cycle. When these treatments are given at times in the cycle when they induce
maximum phase shifts, they cause no change in the gross malate status of the leaf. The gross malate content of the leaf in
continuous light and normal air at 15% shows a heavily damped circadian oscillation which virtually disappears by the time
of the third cycle, but the CO2 assimilation rhythm persists for many days. The generation of the rhythm, and the control of its phase by environmental factors
are discussed in terms of mechanisms that involve the synthesis and metabolism of malate in specific localised pools in the
cytoplasm of the leaf cells. 相似文献
7.
The circadian rhythm of CO2 assimilation in detached leaves of Bryophyllum fedtschenkoi at 15° C in normal air and continuous illumination is inhibited both by exposure to darkness, and to an atmosphere enriched with 5% CO2. During such exposures substantial fixation of CO2 takes place, and the malate concentration in the cell sap increases from about 20 mM to a constant value of 40–50 mM after 16 h. On transferring the darkened leaves to light, and those exposed to 5% CO2 to normal air, a circadian rhythm of CO2 assimilation begins again. The phase of this rhythm is determined by the time the transfer is made since the first peak occurs about 24 h afterwards. This finding indicates that the circadian oscillator is driven to, and held at, an identical, fixed phase point in its cycle after 16 h exposure to darkness or to 5% CO2, and it is from this phase point that oscillation begins after the inhibiting condition is removed. This fixed phase point is characterised by the leaves having acquired a high malate content. The rhythm therefore begins with a period of malate decarboxylation which lasts for about 8 h, during which time the malate content of the leaf cells must be reduced to a value that allows phosphoenolpyruvate carboxylase to become active. Inhibition of the rhythm in darkness, and on exposure to 5% CO2 in continuous illumination, appears to be due to the presence of a high concentration of CO2 within the leaf inhibiting malic enzyme which leads to the accumulation of high concentrations of malate in the leaf cells. The malate then allosterically inhibits phosphoenolpyruvate carboxylase upon which the rhythm depends. The results give support to the view that malate synthesis and breakdown form an integral part of the circadian oscillator in this tissue.Abbreviations
B.
Bryophyllum
- PEPCase
phosphoenolpyruvate carboxylase 相似文献
8.
Nimmo HG 《Archives of biochemistry and biophysics》2003,414(2):189-196
Phosphoenolpyruvate (PEP) carboxylase is regulated by reversible phosphorylation in higher plants. Recently several genes encoding PEP carboxylase kinase have been cloned. The purpose of this article is to assess the contribution that information on the structure and expression of these genes is making to our understanding of the posttranslational control of PEP carboxylase activity. 相似文献
9.
Isolated intact mesophyll protoplasts from Zea mays L. were used as an enzyme source for studying properties of phosphoenolpyruvate (PEP) carboxylase (EC 4.1 1 31) just after release from cells into the reaction medium. After the injection of protoplasts into the assay mixture, an initial lag of activity was observed, mainly due to the time necessary for complete disruption of protoplasts by the osmotic shock. The final specific activity obtained was ca 18 μmol mg-1 of liberated protein min-1 , a value comparable to that usually achieved after arduous purification. Under the assay conditions employed, the chloroplasts were not disrupted and the retention of their proteins, together with the use of purified mesophyll protoplasts, were obviously the reasons for the high specific activity obtained. The activity and properties of phosphoenolpyruvate carboxylase stored in isolated protoplasts were stable for at least 24 h at 5°C. The main difference between the protoplast-derived and the routinely extracted enzyme was the sensitivity to malate inhibition, which was partially lost in the extracted phosphoenolpyruvate carboxylase; no difference was found in the Km(PEP) . The stress imposed by the protoplast isolation procedure diminished the sensitivity of the enzyme to malate inhibition, so that it can be inferred that the real malate sensitivity of pbosphocnolpyruvale carboxylase is even greater and that it is grossly underestimated with routinely extracted enzyme. 相似文献
10.
Michael A. Nicodemus K. Francis Salifu Douglass F. Jacobs 《Trees - Structure and Function》2008,22(5):685-695
Nitrogen (N) limits plant productivity and its uptake and assimilation may be regulated by N source, N availability, and nitrate
reductase activity (NRA). Knowledge of how these factors interact to affect N uptake and assimilation processes in woody angiosperms
is limited. We fertilized 1-year-old, half-sib black walnut (Juglans nigra L.) seedlings with ammonium (NH4
+) [as (NH4)2SO4], nitrate (NO3
−) (as NaNO3), or a mixed N source (NH4NO3) at 0, 800, or 1,600 mg N plant−1 season−1. Two months following final fertilization, growth, in vivo NRA, plant N status, and xylem exudate N composition were assessed.
Specific leaf NRA was higher in NO3
−-fed and NH4NO3-fed plants compared to observed responses in NH4
+-fed seedlings. Regardless of N source, N addition increased the proportion of amino acids (AA) in xylem exudate, inferring
greater NRA in roots, which suggests higher energy cost to plants. Root total NRA was 37% higher in NO3
−-fed than in NH4
+-fed plants. Exogenous NO3
− was assimilated in roots or stored, so no difference was observed in NO3
− levels transported in xylem. Black walnut seedling growth and physiology were generally favored by the mixed N source over
NO3
− or NH4
+ alone, suggesting NH4NO3 is required to maximize productivity in black walnut. Our findings indicate that black walnut seedling responses to N source
and level contrast markedly with results noted for woody gymnosperms or herbaceous angiosperms. 相似文献
11.
Phosphoenolpyruvate (PEP) carboxylase activity in epidermal extracts of Commelina communis has been compared in the presence of malate and glucose-6-phosphate. The activity of PEP carboxylase was inhibited by increasing malate concentrations at several substrate (PEP) concentrations and changes in both the apparent K
m (PEP) and V
max values in the presence of malate suggested the occurence of mixed-type inhibiton. In the presence of glucose-6-phosphate no increase in enzyme activity was observed, although there was a slight decrease in the K
m (PEP). However, glucose-6-phosphate appeared to alleviate the inhibition caused by malate. The possible implications of these properties in the control of malate production in guard cells is discussed.Abbreviations PEP
phosphoenolpyruvate
- Glc6P
glucose-6-phosphate 相似文献
12.
The phosphorylation state and the malate sensitivity of phosphoenolpyruvate carboxylase (PEPCase, EC 4.1.1.31) in Bryophyllum fedtschenkoi Hamet et Perrier are altered by changes in the ambient temperature. These effects, in turn alter the in-vivo activity of the enzyme. Low temperature (3 °C or less), stabilizes the phosphorylated form of the enzyme, while high temperature (30 °C) promotes its dephosphorylation. The catalytic activity of the phosphorylated and dephosphorylated forms of PEPCase increases with temperature, but the apparent K
i values for malate of both forms of the enzyme decrease. Results of experiments with detached leaves maintained in darkness in normal air indicate that the changes in malate sensitivity and phosphorylation state of PEPCase with temperature are of physiological significance. When the phosphorylated form of PEPCase is stabilized by reducing the temperature of leaves 9 h after transfer to constant darkness at 15 °C, a prolonged period of CO2 fixation follows. When leaves are maintained in constant darkness at 15 °C until CO2 output reaches a low steady-state level and the PEPCase is dephosphorylated, reducing the temperature to 3 °C results in a further period of CO2 fixation even though the phosphorylation state of PEPCase does not change.Abbreviations CAM
Crassulacean acid metabolism
- PEP
phosphoenolpyruvate
- PEPCase
phosphoenolpyruvate carboxylase
We thank the Agricultural and Food Research Council for financial support for this work. 相似文献
13.
Pea seedlings (Pisum sativum L. cv ‘Kleine Rheinlän-derin’) were grown hydroponically in solutions containing either nitrate (3 or 14 mol m−3) or ammonium (3 mol m−3) as the nitrogen source. Ammonium nutrition as such had no negative effect on plant biomass production, but drastically increased the sensitivity to moderate salinity (50 mol m−3 NaCl). The reasons for this effect are investigated here and in a subsequent paper. The appearance of visible symptoms of salt damage (wilting of marginal leaf areas followed by progressive necrosis) was paralleled by the development of several characteristic modifications in the solute and metabolite contents. Major changes were: (i) high salt (NaCl) accumulation in leaves; (ii) accumulation of ammonium (up to 20 mol m−3) and amino acids (up to 110 mol m−3) in leaves, but at decreased ammonium uptake rates; and (iii) decreased protein content. In a comparison paper we report on the subcellular distribution of salts, ammonium and metabolites under the above conditions. 相似文献
14.
Barley leaf protoplasts were incubated in light or darkness in the presence of various inhibitors, metabolites or weak acids/bases. Nitrate reductase (NR) and phosphoenolpyruvate carboxylase (PEPCase) were rapidly extracted from the protoplasts and assayed under sub-optimal conditions, i.e. in the presence of Mg2+ and malate, respectively. Under these conditions changes in activities are thought to reflect changes in the phosphorylation states of the enzymes. The NR was activated by illumination to 90% of its maximal activity within 10 min. Photosynthetic electron transport appeared necessary for light activation of NR since activation was inhibited by the photosynthetic electron-transport inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), and, additionally, an electron acceptor (HCO
3
-
) was required. The PEPCase was also activated by light. However, this activation was not prevented by DCMU or lack of HCO
3
-
. Loading of protoplasts in the dark with a weak acid resulted in activation of both NR and PEPCase. For NR, full activation was completed within 5 min, whereas for PEPCase a slower, modest activation continued for at least 40 min. Incubation of protoplasts with a weak base also gave activation of PEPCase, but not of NR. On the contrary, base loading counteracted light activation of NR. Since several treatments tested resulted in the modulation of either NR or PEPCase activity, but not both, signal transduction cascades leading to changes in activities appear to be very different for the two enzymes.Abbreviations DCMU
3-(3,4-dichlorophenyl)-1,1-dimethylurea (diuron)
- DMO
5,5-dimethyl-2,4 oxazolidinedione
- NR
nitrate reductase
- PEPCase
Phosphoenolpyruvate carboxylase
This work was supported by the Norwegian Research Council by a Grant to C.L: L.H.S. was supported by the Biotechnology and Biological Sciences Research Council. 相似文献
15.
Xiao Ying Dong Ren Fang Shen Rong Fu Chen Zhao Liang Zhu Jian Feng Ma 《Plant and Soil》2008,306(1-2):139-147
Lespedeza bicolor (Lespedeza bicolor Turcz. cv. Jiangxi) is a leguminous shrub that is well adapted to acid infertile soils. However, the mechanisms of aluminum
resistance in this species have not been established. This study aimed to assess the possible resistance mechanisms of this
plant to Al. An Al-sensitive species of Lespedeza, sericea lespedeza [Lespedeza cuneata (Dum.-Cours.) G. Don cv. Zhejiang], was used as a reference. The roots of L. bicolor secreted both malate and citrate after exposure to Al, but roots of L. cuneata did not. The secretion of organic acids from L. bicolor was specific to Al; neither 15-day P starvation nor 50 μM lanthanum induced the secretion of these organic acid anions. Secretion
of organic acid anions in L. bicolor was detected after 3–6 h exposure to Al, and the amount increased significantly after 6 h exposure, suggesting that this
plant shows a pattern II-type organic acid secretion. This is supported by the finding that the secretion was significantly
inhibited by a protein-synthesis inhibitor, cycloheximide. Two kinds of anion-channel inhibitors had different effects on
Al-induced secretion of organic acids: 9-anthracene carboxylic acid completely inhibited secretion, phenylglyoxal had no effect.
Root elongation in L. bicolor was more severely inhibited by Al in the presence of 9-anthracene carboxylic acid. All these results indicated that the secretion
of malate and citrate is a specialized response to Al stress in L. bicolor roots, which might be one of the Al-resistance mechanisms in this species. 相似文献
16.
We have identified a differential response of stomatal conductance to sulfur dioxide in two cultivars of pea ( Pisum sativum L. cvs P715 and Nugget). The response to sulfite exposure of PEPC activities present in epidermal peels obtained from the two cultivars was qualitatively in agreement with the results obtained for stomatal conductance. With epidermal tissue isolated from the more sensitive cultivar, we have investigated the effect of light and sulfite on guard cell phosphoenolpyruvate carboxylase (E.E. 4.1.1.31.) and NADP-dependent malate dehydrogenase (E.C. 1.1.1.82), two enzymes of the malate biosynthetic pathway. No difference was found between the substrate-saturated activity of phosphoenolpyruvate carboxylase in epidermal tissue incubated in the light or in the dark under the same conditions. Substratesaturated NADP-dependent malate dehydrogenase activity increased nearly 3-fold during a 60 min incubation in the light. Incubations of epidermal tissue in the light in the presence of sulfite resulted in a decrease in the activity of both enzymes. Our results suggest that the inhibition of these two enzymes of the malate biosynthetic pathway may be one cause of sulfur dioxide-mediated stomatal closure. 相似文献
17.
《Bioorganic & medicinal chemistry》2016,24(2):220-225
With the continuous increase of atmospheric CO2 in the last decades, efficient methods for carbon capture, sequestration, and utilization are urgently required. The possibility of converting CO2 into useful chemicals could be a good strategy to both decreasing the CO2 concentration and for achieving an efficient exploitation of this cheap carbon source. Recently, several single- and multi-enzyme systems for the catalytic conversion of CO2 mainly to bicarbonate have been implemented. In order to design and construct a catalytic system for the conversion of CO2 to organic molecules, we implemented an in vitro multienzyme system using mesophilic and thermophilic enzymes. The system, in fact, was constituted by a recombinant phosphoenolpyruvate carboxylase (PEPC) from the thermophilic cyanobacterium Thermosynechococcus elongatus, in combination with mesophilic/thermophilic bacterial carbonic anhydrases (CAs), for converting CO2 into oxaloacetate, a compound of potential utility in industrial processes. The catalytic procedure is in two steps: the conversion of CO2 into bicarbonate by CA, followed by the carboxylation of phosphoenolpyruvate with bicarbonate, catalyzed by PEPC, with formation of oxaloacetate (OAA). All tested CAs, belonging to α-, β-, and γ-CA classes, were able to increase OAA production compared to procedures when only PEPC was used. Interestingly, the efficiency of the CAs tested in OAA production was in good agreement with the kinetic parameters for the CO2 hydration reaction of these enzymes. This PEPC also revealed to be thermoactive and thermostable, and when coupled with the extremely thermostable CA from Sulphurhydrogenibium azorense (SazCA) the production of OAA was achieved even if the two enzymes were exposed to temperatures up to 60 °C, suggesting a possible role of the two coupled enzymes in biotechnological processes. 相似文献
18.
Phosphoenolpyruvate carboxykinase activity decreases when Euglena gracilis Z and ZR undergo light-induced chloroplast development in batch resting medium lacking utilizable organic carbon and CO2. This enzyme is present in heterotrophically grown cells (Briand et al. 1981) and assures gluconeogenesis. It was consistently more active in strain ZR. Decreased carboxykinase activities were accompanied by parallel increases in the activities of ribulose bisphosphate carboxylase and phosphoenolpyruvate carboxylase. The rates of O2 evolution in light were much lower than those of CO2 fixed simultaneously. The incorporation of 14CO2 into early C-4 dicarboxylic acids was higher in green cells than in etiolated cells, and it was even higher in green cells assayed in light in the presence of (DCMU). A hypothesis has been proposed, according to which there is a possible cooperation of phosphoenolpyruvate carboxylase in photosynthetic CO2 fixation, especially under conditions of limiting CO2.High temperatures (34° C) depress carboxylation enzyme activities to a greater extent than that of the carboxykinase without a great effect on cellular chlorophyll content. In the presence of 25 m DCMU, however, chlorophyll accumulation is reduced without any detectable changes in enzyme activities in the Z strain. The ZR strain displayed its characteristic resistance to DCMU.Abbreviations PEP
phosphoenolpyruvate
- RuBP
ribulose bisphosphate
- DCMU
3-(3,4-dichlorophenyl)-1,1-dimethylurea
To whom all correspondence and reprint request should be addressed 相似文献
19.
The chlorophyll a specific absorption coefficient aph* and absorption ratio (aph ratio) of Alexandrium tamarense at five concentrations of nitrate, ammonium and urea over a range from 6 to 100 μM were examined. The experimental results were compared to two coastal diatom species and a prymnesiophyte to identify differences in absorption ratios. Cells exposed to increasing nitrate concentrations were characterized by an increase in aph* at 443, 490, 510, 555 and 675 nm. However, ammonium and urea induced low aph* values at their lowest and highest concentrations. The aph relative to 510 or 555 nm was constant regardless of the concentration of the N source, but dependent on the N source. Oxidized N induced a lower aph ratio than the reduced form. Comparisons of the aph ratio among taxonomic groups revealed significant differences. The aph ratio of A. tamarense was 20–30 and >50% lower than those of two diatoms and a prymnesiophyte, respectively. The aph ratio of the present study could assist in increasing the capability for detecting harmful species such as A. tamarense. 相似文献
20.
Maize (Zea mays L.) grown on low (0.8 mM) NO
3
-
, as well as untransformed and transformed Nicotiana plumbaginifolia constitutively expressing nitrate reductase (NR), was used to study the effects of NO
3
-
on the NR activation state. The NR activation state was determined from the relationship of total activity extracted in the presence of ethylenediaminetetracetic acid to that extracted in the presence of Mg2+. Light activation was observed in both maize and tobacco leaves. In the tobacco lines, NO
3
-
did not influence the NR activation state. In excised maize leaves, no correlation was found between the foliar NO
3
-
content and the NR activation state. Similarly, the NR activation state did not respond to NO
3
-
. Since the NR activation state determined from the degree of Mg2+-induced inhibition of NR activity is considered to reflect the phosphorylation state of the NR protein, the protein phosphatase inhibitor microcystin LR was used to test the importance of protein phosphorylation in the NO
3
-
-induced changes in NR activity. In-vivo inhibition of endogenous protein phosphatase activity by microcystin-LR decreased the level of NR activation in the light. This occurred to the same extent in the presence or absence of exogenous NO
3
-
. We conclude that NO
3
-
does not effect the NR activation state, as modulated by protein phosphorylation in either tobacco (a C3 species) or maize (a C4 species). The short-term regulation of NR therefore differs from the NO
3
-
-mediated responses observed for phosphoenolpyruvate carboxylase and sucrose phosphate synthase.Abbreviations Chl
chlorophyll
- MC
microcystin-LR
- PEP-Case
phosphoenolpyruvate carboxylase
- SPS
sucrose-phosphate synthase
We are indebted to Madeleine Provot and Nathalie Hayes for excellent technical assistance. This work was funded by EEC Biotechnology Contract No. BI02 CT93 0400, project of technical priority, Network D — Nitrogen Utilisation and Efficiency. 相似文献