The bacterial wilt,uptake of phosphate,and phosphate ester levels in the resistant and susceptible alfalfa plants

7 days or 7 weeks old alfalfa plants (Medicago sativa L.), susceptible (S) and resistant (R) to bacterial wilt, were inoculated withCorynebacterium michiganense pv.insidiosum and on day 8 and 15 after inoculation the levels of acid-soluble phosphate esters (P-esters) were determinated by means of³²P labelling in the shoots or roots. The most significant changes were recorded in the roots of the older R plants grown in full Knop nutrient solutions on day 8 after inoculation. The marked reduction of inorganic phosphate (P₁) uptake by whole R plants is accompanied by a decrease in the levels of fructose-l, 6-bisphosphate (Fru-P₂), glucose-6-phosphate (Glc-6-P), fructose-6-phosphate (Fru-6-P), adenosine mono-, and diphosphate (AMP and ADP), phosphorylcholine (P-choline) and a proportional increase in the level of P₁. In the S plants, infection affected neither P₁ uptake nor P₁ proportions. In the plants grown after inoculation in diluted Knop’s solutions (0.147 mM KH₂PO₄), infection induced a reduction of the radial transport of P₁ to the segments of R roots whereas a reduction of the levels was only recorded in some P-esters [AMP, ADP, dihydroxyacetone phosphate (DHAP), and P-choline, but no decrease of Fru-P₂, Glc-6-P and Fru-6-P]. In the S plants, P₁ transport and the levels of P-esters were increased by the infection. P₁ transport exhibited considerable metabolic dependence (DNP, DCCD). Bacterial infection probably had no influence on the activity of the plasma membrane ATPases.     

Characterization of phosphate esters in the shoots and roots of alfalfa plants susceptible and resistant to bacterial wilt

Acid-soluble phosphate esters were determined in extracts of plant material after a 24 h exposure of the roots of intact alfalfa plants to nutrient media labelled with³²P_i. Similar phosphate ester patterns were found in 2-, 3-, 8-, and 9-week-old plants with the exception of Gra-P which could be detected only in shoot extracts. However, phosphate ester levels differed in the shoots and roots. Whereas Fru-P₂, Glc-6-P, Fru-6-P, and adenine nucleotides were more abundant in the shoots, Grn-P and P-choline levels were higher in the roots. Certain differences in the levels of P-esters were also recorded between plants susceptible and resistant to bacterial wilt.     

Inhibition by trifluoperazine of ATP synthesis and hydrolysis by particulate and soluble mitochondrial F1: Competition with H2PO 4 −

The effect of trifluoperazine (TFP) on the ATPase activity of soluble and paniculate F₁ATPase and on ATP synthesis driven by succinate oxidation in submitochondrial particles from bovine heart was studied at pH 7.4 and 8.8. At the two pH. TFP inhibited ATP hydrolysis. Inorganic phosphate protected against the inhibiting action of TFP. The results on the effect of various concentrations of phosphate in the reversal of the action of TFP on hydrolysis at pH 7.4 and 8.8 showed that H₂PO ₄ ^– is the species that competes with TFP. The effect of TFP on oxidative phosphorylation was studied at concentrations that do not produce uncoupling or affect the aerobic oxidation of succinate (<15M). TFP inhibited oxidative phosphorylation to a higher extent at pH 8.8 than at pH 7.4; this was through a diminution in theV _max, and an increase in theK _m for phosphate. Data on phosphate uptake during oxidative phosphorylation at several pH showed that H₂PO ₄ ^– is the true substrate for oxidative phosphorylation. Thus, in both synthesis and hydrolysis of ATP, TFP and H₂PO ₄ ^– interact with a common site. However, there is a difference in the sensitivity to TFP of ATP synthesis and hydrolysis; this is more noticeable at pH 8.8, i.e. ATPase activity of soluble F₁ remains at about 40% of the activity of the control in a concentration range of TFP of 40–100M, whereas in oxidative phosphorylation 14M TFP produces a 60% inhibition of phosphate uptake.     

The Effect of Calcium on the Uptake and Distribution of Sodium in Excised Barley Roots

The total uptake and relative distribution of Na⁺ along the root was investigated in excised barley roots (Hordeum vulgare L. cv. Union) of 6-day-old plants cultured on 0.25 mM CaSO₄. One technique involved cutting the roots at harvest time after different uptake periods into different segments each 2 cm long. These segments were ashed and sodium was measured by flame photometry. For each experiment two treatments were done, one containing calcium in the uptake solution and one without calcium. A second technique involved the measurement of sodium transport, using labelled ²²Na⁺ solution. Although no significant differences could be found between the calcium containing and the calciumless treatments at 24 h, there were clearcut differences for the first 6 to 8 h. The effect of calcium on the first stages of the sodium uptake was a large accumulation of sodium in the part close to the apex with a translocation close to nil; whereas the sodium taken up in the absence of calcium did not accumulate in such large amounts in the apex region but was immediately transported basipetally.     

Maintenance of 32P uptake and transport in Pinus serotina seedlings under hypoxic growth conditions

In the present study, we examined the effects of long- and short-term hypoxia on net uptake and transport of phosphorus to shoots of pond pine (Pinus serotina Michx.), a moderately flood-tolerant southern pine, and the influence aerenchyma formation might have in maintenance of P uptake and transport. Seedlings were grown under aerobic (250 μM O₂) or hypoxic (≤50 μM O₂) solution conditions for 5.3 weeks in continuously flowing solution culture containing 100 μM P. Intact seedlings were then labeled with ³²P for up to 24 h to determine how short- and long-term hypoxic solution conditions affected rates of unidirectional influx and the accumulation of ³²P in roots and shoots. Seedlings in the long-term hypoxic treatment were grown for 5.3 weeks in hypoxic solution and also labeled in hypoxic uptake solution. The short-term hypoxic treatments included a 24-h hypoxic pretreatment followed by time in labeled hypoxic uptake solution for seedlings grown under aerobic or hypoxic conditions; in the latter case, diffusion of atmospheric O₂ entry into stem and root collar lenticels was blocked, thus removing any influence that aerenchyma formation might have had on enhancing O₂ concentrations of root tissue. Although unidirectional influx rates of ³²P in roots of seedlings grown under long-term hypoxic conditions were 1.4 times those of aerobically grown seedlings, accumulation of ³²P in roots was similar after 24 h in labeled uptake solution. These results suggest that ³²P efflux was also higher under hypoxic conditions. Higher shoot/root fresh weight ratios and lower shoot P concentrations in seedlings grown under hypoxic solution conditions suggest that the “shoot P demand” per unit root should be high. Yet accumulation of ³²P in shoots was reduced by 50% after 24 h in hypoxic uptake solution. Both short-term hypoxic treatments decreased accumulation of ³²P in roots by more than 50%. Short-term hypoxia decreased shoot accumulation in seedlings grown under aerobic and hypoxic conditions by 84 and 50%. respectively. Short- and long-term hypoxic conditions increased the percentage of root ³²P in the nucleic acid and chelated-P pools, resulting in a significantly smaller percentage of ³²P in the soluble inorganic phosphate (p_i) pool, the pool available for transport to the shoot. However, a reduction in pool size or in labeling of the pool available for transport cannot fully account for the large reduction in accumulation of ³²P in shoots, particularly in the short-term hypoxic treatment of aerobically grown seedlings. Our results suggest that both influx and transport of ³²P to shoots of pond pine seedlings are O₂-dependent processes, and that the transport of ³²P to shoots may be more sensitive to hypoxic solution conditions than influx at the cortical and epidermal plasmalemma, with aerenchyma formation supporting a substantial amount of both ³²P uptake and transport.     

Phosphate uptake and polyphosphate metabolism of mycorrhizal and nonmycorrhizal roots of pine and of Suillus bovinus at varying external pH measured by in vivo 31P-NMR

 Comparative in vivo ³¹P-NMR analyses of mycorrhizal and nonmycorrhizal roots of Pinus sylvestris and the fungus of Suillus bovinus in pure culture were used to investigate alterations in phosphate metabolism due to changes in external pH in the range 3.5–8.5. All control samples maintained a constant pH in both cytoplasm and vacuole. Mycorrhizal roots and pure fungus, but not nonmycorrhizal roots, transformed accumulated inorganic phosphate into mobile polyphosphate with a medium chain length. Phosphate uptake rates and polyphosphate accumulation responded differently to external pH. In all cases, maximal phosphate uptake occurred at an external pH close to 5.5. At an external pH of 8.5, both roots and fungus showed a distinct lag in phosphate uptake, which was abolished when the external pH was lowered to 7.5. An irreversible effect on phosphate uptake as a consequence of variation in external pH was also observed. The central role of the fungus in regulating mycorrhizal phosphate metabolism is discussed. Accepted: 15 April 1997     

Triterpeneglycosides as Inhibitors of Fungal Growth and Metabolism 3. Effect on Uptake of Potassium, Magnesium and Inorganic Phosphate

A venacin, the resistance factor in oat roots against Ophiobolus graminis var. graminis, and a related triterpeneglycoside, aescin, inhibited the uptake of K⁺ and Mg²⁺ in the fungal mycelium both in phosphate and succinate buffers. The uptake of the cations in Neurospora crassa was similarly inhibited when the inhibitors were dissolved in phosphate or acetatebuffer, while no decrease in the uptake of K⁺ and Mg²⁺ was observed when the inhibitors were dissolved in succinate buffer. The uptake of cations in Aspergillus niger and Pythium irregulare was more or less unaffected by aescin. The uptake of inorganic phosphate was in no case inhibited, but some decrease of the accumulation of inorganic phosphate in Ophiobolus graminis and Ncurospora crassa due to inhibitor treatment in phosphate buffer was observed. No accumulation of Ca²⁺ was observed in any of the tested fungi.     

Mechanisms of phosphate uptake into brush-border membrane vesicles from goat jejunum

This study concerns the uptake of inorganic phosphate into brush-border membrane vesicles prepared from jejunal tissues of either control or Ca-and/or P-depleted goats. The brush-border membrane vesicles showed a time-dependent accumulation of inorganic phosphate with a typical overshoot phenomenon in the presence of an inwardly directed Na⁺ gradient. The Na⁺-dependent inorganic phosphate uptake was completely inhibited by application of 5 mmol·l^-1 sodium arsenate. Half-maximal stimulation of inorganic phosphate uptake into brush-border membrane vesicles was found with Na⁺ concentrations in the order of 5 mmol·l^-1. Inorganic phosphate accumulation was not affected by a K⁺ diffusion potential (inside negative), suggesting an electroneutral transport process. Stoichiometry suggested an interaction of two or more Na ions with one inorganic phosphate ion at pH 7.4. Na⁺-dependent inorganic phosphate uptake into jejunal brush-border membrane vesicles from normal goats as a function of inorganic phosphate concentration showed typical Michaelis-Menten kinetic with V _max=0.42±0.08 nmol·mg^-1 protein per 15 s^-1 and K _m=0.03±0.01 mmol·l^-1 (n=4, x ±SEM). Long-term P depletion had no effect on these kinetic parameters. Increased plasma calcitriol concentrations in Ca-depleted goats, however, were associated with significant increases of V _max by 35–80%, irrespective of the level of P intake. In the presence of an inwardly directed Na⁺ gradient inorganic phosphate uptake was significantly stimulated by almost 60% when the external pH was decreased to 5.4 (pH_out/pH_in=5.4/7.4). The proton gradient had no effect on inorganic phosphate uptake in absence of Na⁺. In summary, in goats Na⁺ and calcitriol-dependent mechanisms are involved in inorganic phosphate transport into jejunal brush-border membrane vesicles which can be stimulated by protons.Abbreviations AP activity of alkaline phosphatase - BBMV brush-border membrane vesicles - EGTA ethyleneglycol-triacetic acid - n _app apparent Hill coefficient - P _i inorganic phosphate - PTH parathyroid hormone     

Oxidative phosphorylation in pea cotyledon submitochondrial particles

Mitochondria and submitochondrial particles (SMP) from pea cotyledons were shown to catalyze oxidative phosphorylation as measured by ³²Pi uptake into phosphate esters. ATP synthesis was sensitive to the electron transport inhibitor KCN, the uncoupler carbonyl cyanide m-chlorophenylhydrazone, and the coupling factor inhibitor oligomycin. Experiments with the adenine nucleotide translocator inhibitor atractyloside indicated the SMP were inside-out. Mersalyl completely inhibited ATP synthesis by SMP, and a separate experiment indicated that mersalyl has a direct effect on the ATPase complex. The kinetics of ATP synthesis indicated a high affinity for phosphate (K_m = 0.18 millimolar). ADP kinetics gave a biphasic curve with K_m values of about 4.8 and 160 micromolar. O₂ uptake and ATP synthesis had a pH maximum of 7.6 while the ratio of micromoles phosphate esterified to microatoms O₂ taken up was highest at pH 7.2. Sodium chloride inhibited both ATP synthesis and O₂ uptake but stimulated the ATPase reaction. The SMP also catalyzed a slow ATP-phosphate exchange reaction.     

Phosphate and calcium uptake in beech (Fagus sylvatica) in the presence of aluminium and natural fulvic acids

In the present study we examine the effects of Al on the uptake of Ca²⁺ and H₂PO^-₄ in beech (Fagus sylvatica L.) grown in inorganic nutrient solutions and nutrient solutions supplied with natural fulvic acids (FA). All the solutions used were chemically well characterized. The uptake of Al by roots of intact plants exposed to solutions containing 0, 0.15 or 0.3 mM AlCl₃ for 24 h, was significantly less if FA (300 mg l⁻¹) were also present in the solutions. The Ca²⁺(⁴⁵Ca²⁺) uptake was less affected by Al in solutions supplied with FA than in solutions without FA. There was a strong negative correlation between the Al and Ca²⁺ uptake (r²=0.98). When the Al and Ca²⁺ (⁴⁵Ca²⁺) uptake were plotted as a function of the Al³⁺ activity (or concentration of inorganic mononuclear Al), almost the same response curves were obtained for the -FA and +FA treatments. We conclude that FA-complexed Al was not available for root uptake and therefore could not affect the Ca²⁺ uptake. The competitive effect of Al on the Ca²⁺ uptake was also shown in a 5-week cultivation experiment, where the Ca concentration in shoots decreased at an AlCl₃ concentration of 0.3 mM. The effect of Al on H₂PO⁻₄ uptake was more complex. The P content in roots and shoots was not significantly affected, compared with the control, by cultivation for 5 weeks in a solution supplied with 0.3 mM AlCl₃, despite a reduction of the H₂PO⁻₄ concentration in the nutrient solution to about one-tenth. At this concentration Al obviously had a positive effect on H₂PO⁻₄ uptake. The presence of FA decreased ³²P-phosphate uptake by more than 60% during 24 h, and the addition of 0.15 or 0.3 mM AlCl₃ to these solutions did not alter the uptake of ³²P-phosphate.     

Transport and metabolism of pyridoxine in rat liver

Evidence, obtained with in situ perfused rat liver, indicated that pyridoxine is taken up from the perfusate by a non-concentrative process, followed by metabolic trapping. These conclusions were reached on the basis of the fact that at low concentrations (0.125 μM), the ³H of [³H]pyridoxine accumulated against a concentration gradient, but high concentrations (333 μM) of pyridoxine or 4-deoxypyridoxine prevented this apparent concentrative uptake. Under no conditions did the tissue water : perfusate concentration ratio of [³H]pyridoxine exceed unity.The perfused liver rapidly converted the labeled pyridoxine to pyridoxine phosphate, pyridoxal phosphate and pyridoxamine phosphate and released a substantial amount of pyridoxal and some pyridoxal phosphate into the perfusate. Since muscle and erythrocytes failed to oxidize pyridoxine phosphate to pyridoxal phosphate, it is suggested that the liver plays a major role in oxidizing dietary pyridoxine and pyridoxamine as their phosphate esters to supply pyridoxal phosphate which then reaches to other organs chiefly as circulating pyridoxal.     

Effect of 4,6-Dinitro-o-sec-butylphenol on Phosphorus Accumulation and Incorporation in Tomato Leaf Disks

The accumulation and incorporation of externally applied P³² into ATP and the effect of 4,6-dinitro-o-sec-butylphenol (DNBP) on these processes was studied, using tomato (Lycopersicon esculentum Mill.) leaf disks.

P³² was, in most part, actively accumulated into leaf disks with time and was incorporated into ATP and other organic phosphates. DNBP inhibited both P³² accumulation and ATP generation. The amount of inhibition increased with time of incubation.

It is concluded that P³² accumulation is related to ATP generation. Even though DNBP greatly inhibits phosphorus accumulation, there is little or no effect on its retention.

DNBP has the ability to uncouple oxidative phosphorylation. Therefore, it is assumed that its inhibitory effect on phosphate accumulation and generation of high-energy phosphorus esters is related to its inhibition of oxidative phosphorylation.

A method is described which appears to be satisfactory to determine the relative amounts of ATP and ADP in leaf disks labeled with P³².

     

Metabolic regulation in the senescing tobacco leaf: I. Changes in pattern of p incorporation into leaf disc metabolites

Changes in phosphate metabolism were explored in discs from tobacco (Nicotiana tabacum) leaves of three contrasting types: green leaves which were fully expanded and attached to the plant, leaves which had yellowed following excision and dark starvation, and leaves which had yellowed while attached to the plant. 2,4-Dinitrophenol at 10⁻⁵m stimulated the respiration rate of discs from green and yellow-detached leaves only slightly, but markedly stimulated that of discs from yellow-attached leaves. Following a 10-minute uptake period the incorporation of ³²P-orthophosphate into phosphate esters and lipids of discs from yellow-detached leaves was resistant to 2,4-dinitrophenol, whereas in discs from green and yellow-attached leaves it was inhibited by 2,4-dinitrophenol. Incorporation into a salt-soluble fraction containing unidentified nucleotide material showed converse behavior in that it was stimulated by 2,4-dinitrophenol in discs from green and yellow-attached leaves; in discs from yellow-detached leaves it was resistant to 2,4-dinitrophenol. In discs from yellow-detached and yellow-attached leaves there was a shift in the labeling pattern of phosphate esters toward increased label in hexose phosphates at the expense of adenine nucleotides, 3-phosphoglycerate, and phosphoenolpyruvate. It is concluded that incorporation into phosphate esters in discs from yellow-detached leaves is by substrate level phosphorylation coupled to enhanced aerobic glycolysis. In discs from yellow-attached leaves, on the other hand, incorporation depends on oxidation phosphorylation, and it is suggested that the shift in labeling pattern is caused by senescence-induced changes in activity of glycolytic enzymes.     

Local measurements of nitrate and potassium fluxes along roots of maritime pine. Effects of ectomycorrhizal symbiosis

In order to assess the actual role of ectomycorrhizae in ion uptake by the ectomycorrhizal root system, we used a microelectrode ion flux estimation methodology that provided access to local values of net fluxes. This made it possible to investigate the heterogeneity of ion fluxes along the different types of roots of Pinus pinaster associated or not with ectomycorrhizal species. We compared two fungi able to grow with nitrate in pure culture, Rhizopogon roseolus and Hebeloma cylindrosporum, the former having a positive effect on host tree shoot growth (c. +30%) and the latter a negative effect (c.? 30%). In non‐mycorrhizal plants (control), NO₃^– was taken up at higher rates by the short roots than by the long ones, whereas K⁺ uptake occurred mainly in growing apices of long roots. In mycorrhizal plants, H. cylindrosporum did not modify K⁺ uptake and even decreased NO₃^– uptake at the level of ectomycorrhizal short roots, whereas R. roseolus strongly increased K⁺ and NO₃^– fluxes at the level of ectomycorrhizal short roots without any modification of the fluxes measured along the fungus‐free long roots. The measurement of ion influxes at the surface of the ectomycorrhizal roots can provide a way to reveal actual effects of mycorrhizal association on ion transport in relation to mycorrhizal efficiency in natural conditions.     

The turnover of thiamine and its phosphate esters in rat organs

Thiamine-deficient rats were injected with [³⁵S]thiamine and the turnover of the injected radioactive thiamine and its phosphate esters was measured in brain, heart, and liver after returning the rats to a normal diet. The radioactivity per gram wet tissue, as well as the specific activity, fell to half its initial value in less than 40 hr, showing that thiamine compounds were turning over rapidly in the body under normal conditions. Furthermore, there was no difference in the rate of turnover of the individual phosphate esters of thiamine. The amount of the various phosphate esters as a percentage of the total thiamine remained the same throughout the experiment.     

An easy method for preparing radioactive methyl esters of eicosanoids suitable as ligands in radioimmunoassays

A rapid and convenient method is described for methylating prostanoids and other arachidonic acid metabolites. With ³H-methyl iodide of high specific activity, tracers for radioimmunoassay can be produced at a cost which is only a fraction of that of labeled compounds currently available. In radioimmunoassays for PGE₂, TXB₂ and 6-keto -PGF_1α, labeled methyl esters gave results which were comparable to those obtained with the use of tritiated free acids as radioligands.     

Zur Wirkung von Sauerstoff auf die 32P-Markierung von Polyphosphaten und organischen Phosphaten bei Ankistrodesmus braunii im Licht

Summary Short time incorporation of ³²P was carried out with synchronised algae (young cells) depleted of phosphate. For the separation and determination of the acid-insoluble phosphate fractions of the cells an improved fractionation procedure was applied. In order to exclude competition by carbon dioxide all experiments were done in the absence of CO₂.Compared with nitrogen, CO₂-free air produces an increase in the labelling of phosphorylated compounds in the light. In strong white light, at high pH, air effects a remarkable increase of ³²P in the acid-insoluble phosphate (P _u), mainly in inorganic polyphosphates (P _ul), whereas the total phosphate uptake remains almost unchanged. The increase in labelling of acid-insoluble phosphate is, therefore, accompanied by a substantial decrease in the labelling of acid-soluble compounds (P _l). In weak white light or in far-red light, at low pH even in strong white or red light, an increase of phosphate uptake and an increased labelling of the acid-stable organic acid-soluble fraction (P _os) is observed instead. The effect of oxygen increases somewhat with increasing light intensity up to light saturation, and it increases markedly with increasing oxygen concentration.An essential contribution by oxidative phosphorylation to this oxygen effect can be ruled out on account of its much higher sensitivity to oxygen. Pseudocyclic photophosphorylation is also not regarded as the main force because of its higher oxygen affinity. Occurrence of photorespiration has not been clearly established so far in related algae (Chlorella), and its use for phosphorylation is unknown. A better, although not complete explanation is given by comparing the oxygen effect with the well-known inhibition of photosynthesis by oxygen (Warburg effect), which leads to an increase in glycolate formation and a simultaneous decrease in the pool sizes of carbon reduction cycle intermediates, even in the absence of CO₂. Since the photophosphorylation process, as well as the photosynthetic electron flow, seem unaffected by high oxygen concentrations whereas the formation of organic phosphate compounds is partially inhibited, excess ATP may be available for polyphosphate synthesis. This explanation would be consistent with the assumption that polyphosphate-ADP kinase mediates an equilibrium between ATP and polyphosphates, mainly at higher pH. At low pH and in other cases the excess ATP might be available for an increased phosphate uptake and for phosphorylation of endogenous carbohydrates.

---

---

Herrn Prof. Dr. W. Simonis zum 60. Geburtstag gewidmet.     

Role of sugars in nitrate utilization by roots of dwarf bean

Nitrate uptake and in vivo, nitrate reductase activity (NRA) in roots of Phaseolus vulgaris, L. cv. Witte Krombek were measured in nitrogen-depleted plants of varying sugar status, Variation in sugar status was achieved at the start of nitrate nutrition by excision, ringing, darkness or administration of sugars to the root medium. The shape of the apparent induction pattern of nitrate uptake was not influenced by the sugar status of the absorbing tissue. When measured after 6 h of nitrate nutrition (0.1 mol m^?3), steady state nitrate uptake and root NRA were in the order intact>dark>ringed>excised. Exogenous sucrose restored NRA in excised roots to the level of intact plants. The nitrate uptake rate of excised roots, however, was not fully restored by sucrose (0.03–300 mol m^?3). When plants were decapitated after an 18 h NO₃^? pretreatment, the net uptake rate declined gradually to become negative after three hours. This decline was slowed down by exogenous fructose, whilst glucose rapidly (sometimes within 5 min) stimulated NG^?₃ uptake. Presumably due to a difference in NO₃^? due to a difference in NO₃^? uptake, the NRA of excised roots was also higher in the presence of glucose than in the presence of fructose after 6 h of nitrate nutrition. The sugar-stimulation of, oxygen consumption as well as the release of ¹⁴CO₂ from freshly absorbed (U-¹⁴C) sugar was the same for glucose and fructose. Therefore, we propose a glucose-specific effect on NO₃^? uptake that is due to the presence of glucose rather than to its utilization in root respiration. A differential glucose-fructose effect on nitrate reductase activity independent of the effect on NO₃^? uptake was not indicated. A constant level of NRA occurred in roots of NO₃^? induced plants. Removal of nutrient nitrate from these plants caused an exponential NRA decay with an approximate half-life of 12 h in intact plants and 5.5 h in excised roots. The latter value was also found in roots that were excised in the presence of nitrate, indicating that the sugar status primarily determines the apparent rate of nitrate reductase decay in excised roots.     

Nitrate uptake by bean (Phaseolus vulgaris L.) roots under phosphate deficiency

Bean (Phaseolus vulgaris L.) plants were cultured for 19 d on complete or on phosphate deficient culture media. Low inorganic phosphate concentration in the roots decreased ATP level and nitrate uptake rate. The mechanisms which may control nitrate uptake rate during phosphate deficiency were examined. Plasma membrane enriched fractions from phosphate sufficient and phosphate deficient plants were isolated and compared. The decrease in total phospholipid content was observed in plasma membranes from phosphate deficient roots, but phospholipid composition was similar. No changes in ATPase and proton pumping activities measured in isolated plasma membrane of phosphate sufficient and phosphate deficient bean roots were noted. The electron microscope observations carried out on cortical meristematic cells of the roots showed that active ATPases were found in plasma membrane of both phosphate sufficient and phosphate deficient plants. The decrease in inorganic phosphate concentration in roots led to increased nitrate accumulation in roots, accompanied by a corresponding alterations in NO₃ distribution between shoots and roots. Nitrate reductase activity in roots of phosphate deficient plants estimated in vivo and in vitro was reduced to 50–60% of the control. The increased NO₃ concentration in root tissue may be explained by decreased NR activity and lower transport of nitrate from roots to shoots. Therefore, the reduction of nitrate uptake during phosphate starvation is mainly a consequence of nitrate accumulation in the roots.