A comparison of nitrate transport in four different rice (Oryza sativa L.) cultivars

As rice can use both nitrate (NO₃^-) and ammonium (NH₄⁺), we have tested the hypothesis that the shift in the pattern of cultivars grown in Jiangsu Province reflects the ability of the plants to exploit NO₃^- as a nitrogen (N) source. Four rice cultivars were grown in solution culture for comparison of their growth on NO₃^- and NH₄⁺ nitrogen sources. All four types of rice, Xian You 63 (XY63), Yang Dao 6 (YD), Nong Keng 57 (NK) and Si You 917 (SY917), grew well and produced similar amounts of shoot biomass with 1 mmol/L NH₄⁺ as the only N source. However, the roots of NK were significantly smaller in comparison with the other cultivars. When supplied with 1 mmol/L NO₃^-, YD produced the greatest biomass; while NK achieved the lowest growth among the four cultivars. Electrophysiological measurements on root rhizodermal cells showed that the NO₃^--elicited changes in membrane potential (ΔE_m) of these four rice cultivars were significantly different when exposed to low external NO₃^- (<1 mmol/L); while they were very similar at high external NO₃^- (10 mmol/L). The root cell membrane potentials of YD and XY63 were more responsive to low external NO₃^- than those of NK and SY917. The ΔE_m values for YD and XY63 rhizodermal cells were almost the same at both 0.1 mmol/L and 1 mmol/L NO₃^-; while for the NK and SY917 the values became larger as the external NO₃^- increased. For YD cultivar, ΔE_m was measured over a range of NO₃^- concentrations and a Michaelis-Menten fit to the data gave a K_m value of 0.17 mmol/L. Net NO₃^- uptake depletion kinetics were also compared and for some cultivars (YD and XY63) a single-phase uptake system with first order kinetics best fitted the data; while other cultivars (ND and SY917) showed a better fit to two uptake systems. These uptake systems had two affinity ranges: one had a similar K_m in all the cultivars (0.2 mmol/L); the other much higher affinity system (0.03 mmol/L) was only present in NK and SY917. The expression pattern of twelve different NO₃^- transporter genes was tested using specific primers, but only OsNRT1.1 and OsNRT2.1 expression could be detected showing significant differences between the four rice cultivars. The results from both the physiological and molecular experiments do provide some support for the hypothesis that the more popular rice cultivars grown in Jiangsu Province may be better at using NO₃^- as an N source.     

A comparison of nitrate transport in four different rice(Oryza sativa L.) cultivars

As rice can use both nitrate (NO3-) and ammonium (NH4+), we have tested the hypothesis that the shift in the pattern of cultivars grown in Jiangsu Province reflects the ability of the plants to exploit NO3- as a nitrogen (N) source. Four rice cultivars were grown in solution culture for comparison of their growth on NO3- and NH4+ nitrogen sources. All four types of rice,Xian You 63 (XY63), Yang Dao 6 (YD), Nong Keng 57 (NK) and Si You 917 (SY917), grew well and produced similar amounts of shoot biomass with 1 mmol/L NH4+ as the only N source.However, the roots of NK were significantly smaller in comparison with the other cultivars. When supplied with 1 mmol/L NO3-, YD produced the greatest biomass; while NK achieved the lowest growth among the four cultivars. Electrophysiological measurements on root rhizodermal cells showed that the NO3--elicited changes in membrane potential (ΔEm) of these four rice cultivars were significantly different when exposed to low external NO3- (＜1 mmol/L); while they were very similar at high external NO3- (10 mmol/L). The root cell membrane potentials of YD and XY63 were more responsive to low external NO3- than those of NK and SY917. The ΔEm values for YD and XY63 rhizodermal cells were almost the same at both 0.1 mmol/L and 1 mmol/L NO3-;while for the NK and SY917 the values became larger as the external NO3- increased. For YD cultivar, ΔEm was measured over a range of NO3- concentrations and a Michaelis-Menten fit to the data gave a Km value of 0.17 mmol/L. Net NO3- uptake depletion kinetics were also compared and for some cultivars (YD and XY63) a single-phase uptake system with first order kinetics best fitted the data; while other cultivars (ND and SY917) showed a better fit to two uptake systems. These uptake systems had two affinity ranges: one had a similar Km in all the cultivars (0.2 mmol/L); the other much higher affinity system (0.03 mmol/L) was only present in NK and SY917. The expression pattern of twelve different NO3- transporter genes was tested using specific primers, but only OsNRT1.1 and OsNRT2. 1 expression could be detected showing significant differences between the four rice cultivars. The results from both the physiological and molecular experiments do provide some support for the hypothesis that the more popular rice cultivars grown in Jiangsu Province may be better at using NO3- as an N source.     

Balanced nitrogen economy as a flexible strategy on yield stabilizing and quality of aquatic food crops in wetland ecosystem

In wetland ecosystem, nitrogen along with other elements and its management is most imperative for the production of so many aquatic food, non-food and beneficial medicinal plants and for the improvement of soil and water characteristics. With great significant importance of INM (integrated nutrient management) as sources, emphasizing on management on nitrogen as a key element and its divergence, a case study was undertaken on such aquatic food crops (starch and protein-rich, most popular and remunerative) in the farmers’ field of low-lying ‘Tal’ situation of New Alluvial Zone of Indian subtropics. The study was designed in factorial randomized block design, where, three important aquatic food crops (water chestnut (Trapa bispinosa Roxb.), makhana (Euryale ferox Salisb.) and water lily (Nymphaea spp.) as major factor and eleven combinations of organic and inorganic sources of nutrients as sub-factor was considered in the experiment. It revealed from the results that the production of fresh kernels or nuts of water chestnut (8.57 t ha⁻¹), matured nut yield of makhana (3.06t ha⁻¹) and flower stalks of water-lily as vegetables (6.38 t ha⁻¹) including its nutritional quality (starch, protein, sugar and minerals) was remarkably influenced with the application of both organic (neem oilcake @ 0.2 t ha⁻¹) and inorganic sources (NPK @ 30:20:20 kg ha⁻¹ along with spraying of NPK @ 0.5% each over crop canopy at 20 days interval after transplanting) than the other INM combinations applied to the crops. Among the crops, highest WCYE (water chestnut yield equivalence) exhibited in makhana due to its high price of popped-form in the country, which is being exported to other countries at now. Sole application of both (organic and inorganic sources) with lower range did not produce any significant outcome from the study and exhibited lower value for all the crops. Besides production of food crops, INM also greatly influenced the soil and water characterization and it was favourably reflected in this study. The physico-chemical characteristics of soil (textural class, pH, organic carbon, organic matter, ammoniacal nitrogen, nitrate nitrogen, available nitrogen, phosphorus and potassium) are most important and contributed a significant improvement due to cultivation of these aquatic crops. Analysis of such wet bodies represented the water characteristics (pH, BOD, COD, CO ⁼₃ , HCO ⁻₃ , NO ⁻₃ N, SO ⁻₄ S and Cl⁻) were most responsive, adaptable and quite favourable for the cultivation of these crops in this vast waste unused wetlands for the mankind without any environmental degradation.

     

Ammonium nutrition increases water absorption in rice seedlings (Oryza sativa L.) under water stress

Water stress is a primary limitation on plant growth. In previous studies, it has been found that ammonium enhances the tolerance of rice plants to water stress, but how water is related to nitrogen form and water stress remains unknown. To study the effects of nitrogen form (NH ₄ ⁺ , NO ₃ ^? , and a mixture of NH ₄ ⁺ and NO ₃ ^? ) on the growth and water absorption of rice (Oryza sativa L.) seedlings, a hydroponic experiment with water stress, simulated by the addition of polyethylene glycol (PEG, 10% w/v, MW 6000), was conducted in a greenhouse. The results showed that, compared with non-water stress, under water stress, the fresh weight of rice seedlings increased by 14% with NH ₄ ⁺ nutrition, whereas it had decreased by about 20% with either NO ₃ ^? or mixed nitrogen nutrition. No significant difference was found in the transpiration rate of excised shoots or in xylem exudation of excised roots in NH ₄ ⁺ supply between the two water situations, whereas xylem flow decreased by 57% and 24% under water stress in NO ₃ ^? and mixed nutrition, and root hydraulic conductivity decreased by 29% and 54% in plants in NH ₄ ⁺ and NO ₃ ^? nutrition conditions, respectively. Although water absorption ability decreased in both NH ₄ ⁺ and NO ₃ ^? nutrition, aquaporin activity was higher in NH ₄ ⁺ than in NO ₃ ^? nutrition, regardless of water stress. We conclude that NH ₄ ⁺ nutrition can improve water handling in rice seedlings and subsequently enhance their resistance to drought.     

A bacterial view of the periodic table: genes and proteins for toxic inorganic ions

Essentially all bacteria have genes for toxic metal ion resistances and these include those for Ag⁺, AsO ⁻₂ , AsO ³⁻₄ , Cd²⁺, Co²⁺, CrO ²⁻₄ , Cu²⁺, Hg²⁺, Ni²⁺, Pb²⁺, TeO ²⁻₃ , Tl⁺ and Zn²⁺. The largest group of resistance systems functions by energy-dependent efflux of toxic ions. Fewer involve enzymatic transformations (oxidation, reduction, methylation, and demethylation) or metal-binding proteins (for example, metallothionein SmtA, chaperone CopZ and periplasmic silver binding protein SilE). Some of the efflux resistance systems are ATPases and others are chemiosmotic ion/proton exchangers. For example, Cd²⁺-efflux pumps of bacteria are either inner membrane P-type ATPases or three polypeptide RND chemiosmotic complexes consisting of an inner membrane pump, a periplasmic-bridging protein and an outer membrane channel. In addition to the best studied three-polypeptide chemiosmotic system, Czc (Cd²⁺, Zn²⁺, and Co²), others are known that efflux Ag⁺, Cu⁺, Ni²⁺, and Zn²⁺. Resistance to inorganic mercury, Hg²⁺ (and to organomercurials, such as CH₃Hg⁺ and phenylmercury) involve a series of metal-binding and membrane transport proteins as well as the enzymes mercuric reductase and organomercurial lyase, which overall convert more toxic to less toxic forms. Arsenic resistance and metabolizing systems occur in three patterns, the widely-found ars operon that is present in most bacterial genomes and many plasmids, the more recently recognized arr genes for the periplasmic arsenate reductase that functions in anaerobic respiration as a terminal electron acceptor, and the aso genes for the periplasmic arsenite oxidase that functions as an initial electron donor in aerobic resistance to arsenite.

     

Identifying the species of copper that are toxic to plant roots in alkaline nutrient solutions

Background and aims

The pH of the growth medium influences Cu speciation in solution, the negativity of plasma membrane (PM) surface potential, and hence the rhizotoxicity of Cu.

Methods

Solution culture experiments were conducted with wheat (Triticum aestivum L.) seedlings to examine the toxicity of various Cu species at pH values ranging from 4.50 to 8.25. The toxic species of Cu was identified, giving particular consideration to the electrical properties at the plant cell membrane and ion activities at the PM surface.

Results

The solution culture studies showed that at pH?<?6.60 (i.e., free Cu²⁺ >95 % of total Cu), the addition of cations (Ca²⁺ or H⁺) decreased the toxic effects of Cu by decreasing the negativity of the PM surface potential (and hence decreasing the activity of Cu²⁺ at the PM surface). For solutions with pH values from 7.50 to 8.25 (CuCO ⁰₃ >50 % of total Cu), an increase in pH significantly enhanced the toxicity of Cu, whilst the addition of Ca had negligible influence on toxicity.

Conclusions

Root growth in solution cultures was influenced primarily by the surface activities of free Cu²⁺ and CuCO ⁰₃ . Across all experiments, the data indicate that it was CuCO ⁰₃ , rather than CuOH⁺, that contributed Cu toxicity over pH?>?7.00. Although our data do not explore the mechanism of toxicity, we propose that CuCO ⁰₃ has an important role in Cu rhizotoxicity in alkaline growth media.

     

Characterization of ammonium and nitrate uptake and assimilation in roots of tea plants

It has been pointed out that tea (Camellia sinensis (L.) O. Kuntze) prefers ammonium (NH ₄ ⁺ ) over nitrate (NO ₃ ^? ) as an inorganic nitrogen (N) source. ¹⁵N studies were conducted using hydroponically grown tea plants to clarify the characteristics of uptake and assimilation of NH ₄ ⁺ and NO ₃ ^? by tea roots. The total ¹⁵N was detected, and kinetic parameters were calculated after feeding ¹⁵NH ₄ ⁺ or ¹⁵NO ₃ ^? to tea plants. The process of N assimilation was studied by monitoring the dynamic ¹⁵N abundance in the free amino acids of tea plant roots by GC-MS. Tea plants supplied with ¹⁵NH ₄ ⁺ absorbed significantly more ¹⁵N than those supplied with ¹⁵NO ₃ ^? . The kinetics of ¹⁵NH ₄ ⁺ and ¹⁵NO ₃ ^? influx into tea plants followed a classic biphasic pattern, demonstrating the action of a high affinity transport system (HATS) and a low affinity transport system (LATS). The V _max value for NH ₄ ⁺ uptake was 54.5 nmol/(g dry wt min), which was higher than that observed for NO ₃ ^? (39.3 nmol/(g dry wt min)). K_M estimates were approximately 0.06 mM for NH ₄ ⁺ and 0.16 mM for NO ₃ ^? , indicating a higher rate of NH ₄ ⁺ absorption by tea plant roots. Tea plants fed with ¹⁵NH ₄ ⁺ accumulated larger amounts of assimilated N, especially glutamine (Gln), compared with those fed with ¹⁵NO ₃ ^? . Gln, Glu, theanine (Thea), Ser, and Asp were the main free amino acids that were labeled with ¹⁵N under both conditions. The rate of N assimilation into Thea in the roots of NO ₃ ^? -supplied tea plants was quicker than in NH ₄ ⁺ -supplied tea plants. NO ₃ ^? uptake by roots, rather than reduction or transport within the plant, seems to be the main factor limiting the growth of tea plants supplied with NO ₃ ^? as the sole N source. The NH ₄ ⁺ absorbed by tea plants directly, as well as that produced by NO ₃ ^? reduction, was assimilated through the glutamine synthetase-glutamine oxoglutarate aminotransferase pathway in tea plant roots. The ¹⁵N labeling experiments showed that there was no direct relationship between the Thea synthesis and the preference of tea plants for NH ₄ ⁺ .     

Inducibility of Rat Liver Drug-Metabolizing Enzymes as an Index of Food-Screeing for Safety Assessment

D-Lactate dehydrogenase (D-LDH) from Pediococcus pentosaceus ATCC 25745 was found to produce D-3-phenyllactic acid from phenylpyruvate. The optimum pH and temperature for enzyme activity were pH 5.5 and 45 °C. The Michaelis-Menten constant (K _m), turnover number (k _cat), and catalytic efficiency (k _cat?K _m) values for the substrate phenylpyruvate were estimated to be 1.73 mmol/L, 173 s^?1, and 100 (mmol/L)^?1 s^?1 respectively.     

Partitioning of previously-accumulated nitrate to translocation,reduction, and efflux in corn roots

The effect of nitrate uptake, or its absence, on the utilization of nitrate previously accumulated by dark-grown, decpitated maize (Zea mays L., cv. DeKalb XL-45) seedlings was examined. Five-d-old plants that had been pretreated with 50 mM ¹⁴NO ₃ ^? for 20 h were exposed for 8 h to nutrient solutions containing either no nitrate or 50 mM ¹⁵NO ₃ ^? , 98.7 atom % ¹⁵N. The ambient solution, xylem exudate, and plant tissue were analyzed to determine the quantities of previously-accumulated (endogenous) ¹⁴NO ₃ ^? that were translocated to the xylem, lost to the solution, or reduced within the tissue during the 8-h period. Energy was continuously available to the roots from the attached endosperm. In the absence of incoming nitrate, appreciable reduction and translocation of the endogenous ¹⁴NO ₃ ^? occurred, but efflux of ¹⁴NO ₃ ^? to the external solution was minimal. In contrast, during ¹⁵NO ₃ ^? uptake, there was considerable efflux of ¹⁴NO ₃ ^? as well as translocation of ¹⁴NO ₃ ^? to the xylem, but little ¹⁴NO ₃ ^? was reduced. Thus there appeared to be an inverse relationship between ¹⁴NO ₃ ^? efflux and reduction. The data are tentatively interpreted on the basis of a model which envisages (a) two storage locations within roots, one of which primarily supplies nitrate for translocation and the other of which primarily supplies nitrate for outward passage through plasmalemma, and (b) the majority of nitrate reduction as occurring during or immediately following influx across the plasmalemma, with endogenous ¹⁴NO ₃ ^? initially moving outward being recycled inward and thereby being reduced.     

Denitrification in soil amended with thermophile-fermented compost suppresses nitrate accumulation in plants

NO ₃ ^? is a major nitrogen source for plant nutrition, and plant cells store NO ₃ ^? in their vacuoles. Here, we report that a unique compost made from marine animal resources by thermophiles represses NO ₃ ^? accumulation in plants. A decrease in the leaf NO ₃ ^? content occurred in parallel with a decrease in the soil NO ₃ ^? level, and the degree of the soil NO ₃ ^? decrease was proportional to the compost concentration in the soil. The compost-induced reduction of the soil NO ₃ ^? level was blocked by incubation with chloramphenicol, indicating that the soil NO ₃ ^? was reduced by chloramphenicol-sensitive microbes. The compost-induced denitrification activity was assessed by the acetylene block method. To eliminate denitrification by the soil bacterial habitants, soil was sterilized with γ irradiation and then compost was amended. After the 24-h incubation, the N₂O level in the compost soil with presence of acetylene was approximately fourfold higher than that in the compost soil with absence of acetylene. These results indicate that the low NO ₃ ^? levels that are often found in the leaves of organic vegetables can be explained by compost-mediated denitrification in the soil.     

Dominance of biologically produced nitrate in upland waters of Great Britain indicated by stable isotopes

Atmospheric deposition of nitrogen (N) compounds is the major source of anthropogenic N to most upland ecosystems, where leaching of nitrate (NO ₃ ^? ) into surface waters contributes to eutrophication and acidification as well as indicating an excess of N in the terrestrial catchment ecosystems. Natural abundance stable isotopes ratios, ¹⁵N/¹⁴N and ¹⁸O/¹⁶O (the “dual isotope” technique) have previously been used in biogeochemical studies of alpine and forested ecosystems to demonstrate that most of the NO ₃ ^? in upland surface waters has been microbially produced. Here we present an application of the technique to four moorland catchments in the British uplands including a comparison of lakes and their stream inflows at two sites. The NO ₃ ^? concentrations of bulk deposition and surface waters at three sites are very similar. While noting the constraints imposed by uncertainty in the precise δ¹⁸O value for microbial NO ₃ ^? , however, we estimate that 79–98% of the annual mean NO ₃ ^? has been microbially produced. Direct leaching of atmospheric NO ₃ ^? is a minor component of catchment NO ₃ ^? export, although greater than in many similar studies in forested watersheds. A greater proportion of atmospheric NO ₃ ^? is seen in the two lake sites relative to their inflow streams, demonstrating the importance of direct NO ₃ ^? deposition to lake surfaces in catchments where terrestrial ecosystems intercept a large proportion of deposited N. The dominance of microbial sources of NO ₃ ^? in upland waters suggests that reduced and oxidised N deposition may have similar implications in terms of contributing to NO ₃ ^? leaching.     

Interaction between NO 3 − and abscisic acid in the regulation of lateral root elongation in Zea mays L.

Lateral root (LR) elongation rate of 7–8-day maize seedlings depends on the availability of NO ₃ ^? , NO ₂ ^? , and abscisic acid (ABA) in an environment. Four-hour exposure to 0.01–1.5 mM NO ₂ ^? increases the relative LR elongation rate; in the case of NO ₂ ^? , the stimulation occurs only at an NO ₂ ^? concentration equal to 0.01 mM. Exogenous ABA (10^?6 M) inhibits the LR elongation process. In the case of a combined influence of NO ₃ ^? and ABA or NO ₂ ^? and ABA, the character of the response elongation reaction is different. The NO role in the regulation of LR elongation is discussed.     

Influence of different mineral nitrogen sources (NO 3 − -N vs. NH 4 + -N) on arbuscular mycorrhiza development and N transfer in a Glomus intraradices–cowpea symbiosis

Labeled nitrogen (¹⁵?N) was applied to a soil-based substrate in order to study the uptake of N by Glomus intraradices extraradical mycelium (ERM) from different mineral N (NO ₃ ^? vs. NH ₄ ⁺ ) sources and the subsequent transfer to cowpea plants. Fungal compartments (FCs) were placed within the plant growth substrate to simulate soil patches containing root-inaccessible, but mycorrhiza-accessible, N. The fungus was able to take up both N-forms, NO ₃ ^? and NH ₄ ⁺ . However, the amount of N transferred from the FC to the plant was higher when NO ₃ ^? was applied to the FC. In contrast, analysis of ERM harvested from the FC showed a higher ¹⁵?N enrichment when the FC was supplied with ¹⁵NH ₄ ⁺ compared with ¹⁵NO ₃ ^? . The ¹⁵?N shoot/root ratio of plants supplied with ¹⁵NO ₃ ^? was much higher than that of plants supplied with ¹⁵NH ₄ ⁺ , indicative of a faster transfer of ¹⁵NO ₃ ^? from the root to the shoot and a higher accumulation of ¹⁵NH ₄ ⁺ in the root and/or intraradical mycelium. It is concluded that hyphae of the arbuscular mycorrhizal fungus may absorb NH ₄ ⁺ preferentially over NO ₃ ^? but that export of N from the hyphae to the root and shoot may be greater following NO ₃ ^? uptake. The need for NH ₄ ⁺ to be assimilated into organically bound N prior to transport into the plant is discussed.     

Alteration of nitrogen metabolism in rice variety 'Nipponbare' induced by alkali stress

Aims

Alkali stress (AS) is an important agricultural contaminant and has complex effects on plant metabolism, specifically root physiology. The aim of this study was to test the role of nitrogen metabolism regulation in alkali tolerance of rice variety 'Nipponbare'.

Methods

In this study, the rice seedlings were subjected to salinity stress (SS) or AS. Growth, the contents of inorganic ions, NH ₄ ⁺ -nitrogen (free amino acids), and NO ₃ ^? -nitrogen in the stressed seedlings were then measured. The expression of some critical genes involved in nitrogen metabolism were also assayed to test their roles in the regulation of nitrogen metabolism during adaptation of rice variety 'Nipponbare' to AS.

Results

AS showed a stronger inhibiting effect on rice variety 'Nipponbare' growth than SS. AS may have more complex effects on nitrogen metabolism than SS.

Conclusions

Effects of AS on the nitrogen metabolism of rice variety 'Nipponbare' mainly comprised two mechanisms. Firstly, in roots, AS caused the reduction of NO ₃ ^? content, which caused two harmful consequences, the large downregulation of OsNR1 expression and the subsequent reduction of NH ₄ ⁺ production in roots. On the other hand, under AS (pH, 9.11), almost all the NH ₄ ⁺ was changed to NH₃, which caused a severe deficiency of NH ₄ ⁺ surrounding the roots. Both events might cause a severe deficiency of NH ₄ ⁺ in roots. Under AS, the increased expression of several OsAMT family members in roots might be an adaptative response to the reduction of NH ₄ ⁺ content in roots or the NH ₄ ⁺ deficiency in rhizosphere. Also, the down-regulation of OsNADH-GOGAT and OsGS1;2 in roots might be due to NH ₄ ⁺ deficiency in roots. Secondly, in shoots, AS caused a larger acuumulatiuon of Na⁺, which possibly affected photorespiration and led to a continuous decrease of NH ₄ ⁺ production in shoots, and inhibited the expression of OsFd-GOGAT and OsGS2 in chloroplasts.     

Dark induction of nitrate reductase in the halophilic alga Dunaliella salina

The effect of nitrogen starvation on the NO₃-dependent induction of nitrate reductase (NR) and nitrite reductases (NIR) has been investigated in the halophilic alga Dunaliella salina. When D. salina cells previously grown in a medium with NH ₄ ⁺ as the only nitrogen source (NH ₄ ⁺ -cells) were transferred into NO ₃ ^? medium, NR was induced in the light. In contrast, when cells previously grown in N-free medium were transferred into a medium containing NO ₃ ^? , NR was induced in light or in darkness. Nitrate-dependent NR induction, in darkness, in D. salina cells previously grown at a photon flux density of 500 umol · m^?2 s^?1 was observed after 4 h preculture in N-free medium, whilst in cells grown at 100 umol · m^?2 s^?1 NR induction was observed after 7–8 h. An inhibitor of mRNA synthesis (6-methylpurine) did not inhibit NO ₃ ^? -induced NR synthesis when the cells, previously grown in NH ₄ ⁺ medium, were transferred into NO ₃ ^? medium (at time 0 h) after 4-h-N starvation. However, when 6-methylpurine was added simultaneously with the transfer of the cells from NH ₄ ⁺ to NO ₃ ^? medium (at time 0 h), NO ₃ ^? induced NR synthesis was completely inhibited. The activity of NIR decreased in N-starved cells and the addition of NO ₃ ^? to those cells greatly stimulated NIR activity in the light. The ability to induce NR in darkness was observed when glutamine synthetase activity reached its maximal level during N starvation. Although cells grown in NO ₃ ^? medium exhibited high NR activity, only 0.33% of the total NR was found in intact chloroplasts. We suggest that the ability, to induce NR in darkness is dependent on the level of N starvation, and that NR in D. salina is located in the cytosol. Light seems to play an indirect regulatory role on NO ₃ ^? uptake and NR induction due to the expression of NR and NO ₃ ^? -transporter mRNAs.     

Molten Earth and the origin of prebiological molecules

Evidence for the molten Earth at its accretion time has been accumulated through the geochemical investigations and the observations of the surfaces of planets by space probes such as Venera 8, Mariner 9, Surveyor, Luna, and Apollo. The primitive terrestrial atmosphere might have been derived from the volcanic gases, as suggested by Rubey, but of a higher temperature than so far assumed. A thermochemical calculation of the composition of the volcanic gas suggests the following possibilities:

1. Large amounts of H₂ and CO were present in the primitive atmosphere. This gives a theoretical basis for the HCN-production experiment by Abelson.
2. HCHO and NH₃ existed in the primitive oceans, of the amount comparable with the weight of the present biosphere.
3. Plenty of NO ₃ ⁻ , SO ₄ ⁻⁻ , and PO ₄ ⁻⁻⁻ were expected in the primitive oceans. The NO ₃ ⁻ ions might have been useful for the nitrate respiration advocated by Egami.

In an appendix, it is argued, on the basis of the observational evidence of the exospheric temperatures of planets by space probes, that a highly reducing atmosphere would (if it existed on the primitive Earth) have disappeared very quickly due to the thermal escape of hydrogen from its exosphere.     

NO 3 − transport across the plasma membrane of Arabidopsis thaliana root hairs: Kinetic control by pH and membrane voltage

High-affinity nitrate transport was examined in intact root hair cells of Arabidopsis thaliana using electrophysiological recordings to characterise the response of the plasma membrane to NO ₃ ^? challenge and to quantify transport activity. The NO ₃ ^? -associated membrane current was determined using a three-electrode voltage clamp to bring membrane voltage under experimental control and to compensate for current dissipation along the longitudinal cell axis. Nitrate transport was evident in the roots of seedlings grown in the absence of a nitrogen source, but only 4–6 days postgermination. In 6-day-old seedlings, additions of 5–100 μm NO ₃ ^? to the bathing medium resulted in membrane depolarizations of 8–43 mV, and membrane voltage (V _m) recovered on washing NO ₃ ^? from the bath. Voltage clamp measurements carried out immediately before and following NO ₃ ^? additions showed that the NO ₃ ^? -evoked depolarizations were the consequence of an inward-directed current that appeared across the entire range of accessible voltages (?300 to +50 mV). Both membrane depolarizations and NO ₃ ^? -evoked currents recorded at the free-running voltage displayed quasi-Michaelian kinetics, with apparent values for K_m of 23 ± 6 and 44 ± 11 μm, respectively and, for the current, a maximum of 5.1 ± 0.9 μA cm^?2. The NO ₃ ^? current showed a pronounced voltage sensitivity within the normal physiological range between ?250 and ?100 mV, as could be demonstrated under voltage clamp, and increasing the bathing pH from 6.1 to 7.4–8.0 reduced the current and the associated membrane depolarizations 3- to 8-fold. Analyses showed a well-defined interaction between the kinetic variables of membrane voltage, pH_o and [NO ₃ ^? ]_o. At a constant pH_o of 6.1, depolarization from ?250 to ?150 mV resulted in an approximate 3-fold reduction in the maximum current but a 10% rise in the apparent affinity for NO ₃ ^? . By contrast, the same depolarization effected an approximate 20% fall in the K_m for transport as a function in [H⁺]_o. These, and additional characteristics of the transport current implicate a carrier cycle in which NO ₃ ^? binding is kinetically isolated from the rate-limiting step of membrane charge transit, and they indicate a charge-coupling stoichiometry of 2(H⁺) per NO ₃ ^? anion transported across the membrane. The results concur with previous studies showing a high-affinity NO ₃ ^? transport system in Arabidopsis that is inducible following a period of nitrogen-limiting growth, but they underline the importance of voltage as a kinetic factor controlling NO ₃ ^? transport at the plant plasma membrane.     

Ultrasonic and Thermal Inactivation of Catalases from Bovine Liver,the Methylotrophic Yeast Pichia pastoris,and the Fungus Penicillium piceum

The kinetics of inactivation of catalases from bovine liver (CAT), the fungus Penicillium piceum (CAT1), and the methylotrophic yeast Pichia pastoris (CAT2) was studied in phosphate buffer (pH 5.5 or 7.4) at 45 and 50°C or under the conditions of exposure to low-frequency ultrasound (LFUS; 27 kHz, 60 W/cm²). The processes were characterized by effective first-order rate constants (s^?1): k _in (total inactivation), k ^*_in (thermal inactivation), and k ^*_in (us) (ultrasonic inactivation). The values of k _in and k ^*_in increased in the following order: CAT1 < CAT < CAT2. Circular dichroic spectra of the enzyme solutions were recorded in the course of inactivation by high temperatures (45 and 50°C ) and LFUS, and the contents of secondary structures were calculated. Processes of thermal and ultrasonic inactivation of catalases were associated with a decrease in the content of α helices and an increase in that of antiparallel β structures and irregular regions (CAT1 < CAT < CAT2). We conclude that the enzymes exhibit the following rank order of resistance: CAT1 > CAT > CAT2. Judging from the characteristics of CAT1, it appears to be an optimum component for antioxidant enzyme complexes.