Impact of nitrogen uptake on field water balance in fertirrigated melon.

Agronomic management in Ciudad Real, a province in central Spain, is characteristic of semi-arid cropped areas whose water supplies have high nitrate (NO3?) content due to environmental degradation. This situation is aggravated by the existence of a restrictive subsurface layer of ?caliche? or hardpan at a depth of 0.60 m. Under these circumstances, fertirrigation rates, including nitrogen (N) fertilizer schedules, must be carefully calibrated to optimize melon yields while minimizing the N pollution and water supply. Such optimization was sought by fertilizing with different doses of N and irrigating at 100% of the ETc (crop evapotranspiration), adjusted for this crop and area. The N content in the four fertilizer doses used was: 0, 55, 82 and 109 kg N ha?1. Due to the NO3? content in the irrigation water, however, the actual N content was 30 kg ha?1 higher in all four treatments repeated in two different years. The results showed correlation between melon plant N uptake and drainage (Dr), which in turn affects the amount of N leached, as well as correlation between Dr and LAI (leaf area index) for each treatment. A fertilizer factor (?) was estimated through two methods, from difference in Dr and in LAI ratio with respect to the maximum N dose, to correct ETc based on N doses. The difference was found in the adjusted evapotranspiration in both years using the corresponding ? achieved 42?49 mm at vegetative period, depending on the method, and it was not significant at senescent period. Finally, a growth curve between N uptake and plant dry weight (DW) for each treatment was defined to confirm that the observed higher plant vigour, showing higher LAI and reduced Dr, was due mainly to higher N doses.

The role of cover crops in irrigated systems: water balance, nitrate leaching and soil mineral nitrogen accumulation

Soil salinity and salt leaching are a risk for sustainable agricultural production in many irrigated areas. This study was conducted over 3.5 years to determine how replacing the usual winter fallow with a cover crop (CC) affects soil salt accumulation and salt leaching in irrigated systems. Treatments studied during the period between summer crops were: barley (Hordeum vulgare L.), vetch (Vicia villosa L.) and fallow. Soil water content was monitored daily to a depth of 1.3 m and used with the numerical model WAVE to calculate drainage. Electrical conductivity (EC) was measured in soil solutions periodically, and in the soil saturated paste extracts before sowing CC and maize. Salt leaching was calculated multiplying drainage by total dissolved salts in the soil solution, and use to obtain a salt balance. Total salt leaching over the four winter fallow periods was 26 Mg ha−1, whereas less than 18 Mg ha−1 in the presence of a CC. Periods of salt gain occurred more often in the CC than in the fallow. By the end of the experiment, net salt losses occurred in all treatments, owing to occasional periods of heavy rainfall. The CC were more prone than the fallow to reduce soil salt accumulation during the early growth stages of the subsequent cash crop.

Nitrogen fertigation: An integrated agronomic and environmental study

Proper management of the N applied to crops is necessary in order to increase yield, improve water use efficiency (WUE) and reduce the pollutions risks with the least economic, environmental and health costs. A field study with melon crops was conducted during 2005, 2006 and 2007 in central Spain, using 11 different amounts of N. Some environmental indexes have been proposed, to provide an essential tool for determining the groundwater pollution risks associated with common agricultural practices. These indexes are related to variation in the nitrate concentration of drinking water (Impact Index (II)) and groundwater (Environmental Impact Index (EII)). Also, the Management Efficiency (ME) was calculated, which is related to the amount of fruit produced per gram of N leached (Nl). To determine the optimum dose of N, it was also necessary to know the N mineralisation (NM). Our results show that 160 kg ha−1 of available N (Nav) produced the maximum fruit yield (FY), enhanced WUE and gave an NM of 85 kg ha−1, while the impact indexes did not exceed the fixed maximum allowable limits and ME was adequate. The proposed indexes proved to be an effective tool for determining the risk of nitrate contamination and confirmed that the optimum dose of N corresponded to the maximum FY with minimal loss of Nl.

Meta-analysis of strategies to control nitrate leaching in irrigated agricultural systems and their effects on crop yield.

Nitrate leaching (NL) is an important N loss process in irrigated agriculture that imposes a cost on the farmer and the environment. A meta-analysis of published experimental results from agricultural irrigated systems was conducted to identify those strategies that have proven effective at reducing NL and to quantify the scale of reduction that can be achieved. Forty-four scientific articles were identified which investigated four main strategies (water and fertilizer management, use of cover crops and fertilizer technology) creating a database with 279 observations on NL and 166 on crop yield. Management practices that adjust water application to crop needs reduced NL by a mean of 80% without a reduction in crop yield. Improved fertilizer management reduced NL by 40%, and the best relationship between yield and NL was obtained when applying the recommended fertilizer rate. Replacing a fallow with a non-legume cover crop reduced NL by 50% while using a legume did not have any effect on NL. Improved fertilizer technology also decreased NL but was the least effective of the selected strategies. The risk of nitrate leaching from irrigated systems is high, but optimum management practices may mitigate this risk and maintain crop yields while enhancing environmental sustainability.

Nitrate-induced early transcriptional changes during imbibition in non-after-ripened Sisymbrium officinale seeds.

We have here demonstrated for the first time that nitrate not only accelerates testa rupture of non- AR seeds but also modifies expression pattern of the cell-wall remodeling proteins (mannanases; SoMAN6 and SoMAN7) and key genes belonging to metabolism and signaling of ABA (SoNCED6, SoNCED9, SoCYP707A2 and SoABI5) and GAs (SoGA3ox, SoGA20ox, SoGA2ox and SoRGL2). These results were obtained during Sisymbrium officinale seed imbibition in the absence of endosperm rupture. Exogenous ABA induced a notable inhibition of testa rupture in both absence and presence of nitrate being this effect sharply reversed by GA4+7. However, nitrate was capable to provoke testa rupture in absence of ABA synthesis. The expression of SoMAN6 and SoMAN7 were positively altered by nitrate. Although ABA synthesis seems apparent at the start of non-AR seed imbibition, taken together the results of SoNCED6, SoNCED9 and SoCYP707A2 expression seem to suggest that nitrate leads to a strong net ABA decrease. Likewise, nitrate positively affected the SoABI5 expression when the SoNCED9 expression was also stimulated. By contrast, at the early and final of imbibition, nitrate clearly inhibited the SoABI5 expression. The expression of SoGA2ox6 and SoGA3ox2 are strongly inhibited by nitrate whereas of SoGA20ox6 was stimulated. On the other hand, SoRGL2 transcript level decreased in the presence of nitrate. Taken together, the results presented here suggest that the nitrate signaling is already operative during the non-AR S. officinale seeds imbibition. The nitrate, in cross-talk with the AR network likely increases the favorable molecular conditions that trigger germination.

The nitrate-afterripening crosstalk is involved in the testa rupture of sisymbryum offinales seed

The loss of seed dormancy can occur by exposing the seed at low moisture storage conditions (afterripening; AR). Since a positive GA:ABA ratio play a key role in the reactivation of germination of non-dormant seeds, it seems obvious that a remarkable effect of AR is the decreasing of both ABA levels and sensitivity, as well as the increment of GA synthesis and sensitivity. ABA levels are regulated by control both of its biosynthesis thorough the 9-cis-epoxycarotenoid dioxygenase (NCED) encoding genes and its catabolism mediated mainly by ABA-8¿-hydroxylases (CYP707A). On the other hand, the last steps of the GA biosynthesis pathway should be involved to control its levels. Namely, GA20ox and GA3ox catalyzing the biosynthesis of active GA and GA2ox which catalyzes the GA inactivation. The presence of nitrate accelerates the sensu stricto germination of non-AR S. officinale seeds. Here, we demonstrate that in AR seeds nitrate also alters the expression pattern of key genes involved in ABA and GA metabolism and signalling (i.e. SoNCED6, SoNCED9, SoCYP707A2, SoABI5, SoGA3ox2, SoGA20ox6, SoGA2ox6 and SoRGL2). These results suggest that the nitrate signalling is also operative during imbibition of AR S. officinale seeds.

Cover crops effect on farm benefits and nitrate leaching: Linking economic and environmental analysis.

Introducing cover crops (CC) interspersed with intensively fertilized crops in rotation has the potential to reduce nitrate leaching. This paper evaluates various strategies involving CC between maize and compares the economic and environmental results with respect to a typical maize?fallow rotation. The comparison is performed through stochastic (Monte-Carlo) simulation models of farms? profits using probability distribution functions (pdfs) of yield and N fertilizer saving fitted with data collected from various field trials and pdfs of crop prices and the cost of fertilizer fitted from statistical sources. Stochastic dominance relationships are obtained to rank the most profitable strategies from a farm financial perspective. A two-criterion comparison scheme is proposed to rank alternative strategies based on farm profit and nitrate leaching levels, taking the baseline scenario as the maize?fallow rotation. The results show that when CC biomass is sold as forage instead of keeping it in the soil, greater profit and less leaching of nitrates are achieved than in the baseline scenario. While the fertilizer saving will be lower if CC is sold than if it is kept in the soil, the revenue obtained from the sale of the CC compensates for the reduced fertilizer savings. The results show that CC would perhaps provide a double dividend of greater profit and reduced nitrate leaching in intensive irrigated cropping systems in Mediterranean regions.

Fluctuating silicate:nitrate ratios and coastal plankton food webs

Marine diatoms require dissolved silicate to form an external shell, and their growth becomes Si-limited when the atomic ratio of silicate to dissolved inorganic nitrogen (Si:DIN) approaches 1:1, also known as the “Redfield ratio.” Fundamental changes in the diatom-to-zooplankton-to-higher trophic level food web should occur when this ratio falls below 1:1 and the proportion of diatoms in the phytoplankton community is reduced. We quantitatively substantiate these predictions by using a variety of data from the Mississippi River continental shelf, a system in which the Si:DIN loading ratio has declined from around 3:1 to 1:1 during this century because of land-use practices in the watershed. We suggest that, on this shelf, when the Si:DIN ratio in the river decreases to less than 1:1, then (i) copepod abundance changes from >75% to <30% of the total mesozooplankton, (ii) zooplankton fecal pellets become a minor component of the in situ primary production consumed, and (iii) bottom-water oxygen consumption rates become less dependent on relatively fast-sinking (diatom-rich) organic matter packaged mostly as zooplankton fecal pellets. This coastal ecosystem appears to be a pelagic food web dynamically poised to be either a food web composed of diatoms and copepods or one with potentially disruptive harmful algal blooms. The system is directed between these two ecosystem states by Mississippi River water quality, which is determined by land-use practices far inland.

The Arabidopsis CHL1 protein plays a major role in high-affinity nitrate uptake

The CHL1 (NRT1) gene of Arabidopsis encodes a nitrate-inducible nitrate transporter that is thought to be a component of the low-affinity (mechanism II) nitrate-uptake system in plants. A search was performed to find high-affinity (mechanism I) uptake mutants by using chlorate selections on plants containing Tag1 transposable elements. Chlorate-resistant mutants defective in high-affinity nitrate uptake were identified, and one had a Tag1 insertion in chl1, which was responsible for the phenotype. Further analysis showed that chl1 mutants have reduced high-affinity uptake in induced plants and are missing a saturable component of the constitutive, high-affinity uptake system in addition to reduced low-affinity uptake. The contribution of CHL1 to constitutive high-affinity uptake is higher when plants are grown at more acidic pH, conditions that increase the level of CHL1 mRNA. chl1 mutants show reduced membrane depolarization in root epidermal cells in response to low (250 μM) and high (10 mM) concentrations of nitrate. Low levels of nitrate (100 μM) induce a rapid increase in CHL1 mRNA. These results show that CHL1 is an important component of both the high-affinity and the low-affinity nitrate-uptake systems and indicate that CHL1 may be a dual-affinity nitrate transporter.

Dual pathways for regulation of root branching by nitrate

Root development is extremely sensitive to variations in nutrient supply, but the mechanisms are poorly understood. We have investigated the processes by which nitrate (NO3−), depending on its availability and distribution, can have both positive and negative effects on the development and growth of lateral roots. When Arabidopsis roots were exposed to a locally concentrated supply of NO3− there was no increase in lateral root numbers within the NO3−-rich zone, but there was a localized 2-fold increase in the mean rate of lateral root elongation, which was attributable to a corresponding increase in the rate of cell production in the lateral root meristem. Localized applications of other N sources did not stimulate lateral root elongation, consistent with previous evidence that the NO3− ion is acting as a signal rather than a nutrient. The axr4 auxin-resistant mutant was insensitive to the stimulatory effect of NO3−, suggesting an overlap between the NO3− and auxin response pathways. High rates of NO3− supply to the roots had a systemic inhibitory effect on lateral root development that acted specifically at the stage when the laterals had just emerged from the primary root, apparently delaying final activation of the lateral root meristem. A nitrate reductase-deficient mutant showed increased sensitivity to this systemic inhibitory effect, suggesting that tissue NO3− levels may play a role in generating the inhibitory signal. We present a model in which root branching is modulated by opposing signals from the plant’s internal N status and the external supply of NO3−.

Nitrate-Ammonium Synergism in Rice. A Subcellular Flux Analysis1

Many reports have shown that plant growth and yield is superior on mixtures of NO3− and NH4+ compared with provision of either N source alone. Despite its clear practical importance, the nature of this N-source synergism at the cellular level is poorly understood. In the present study we have used the technique of compartmental analysis by efflux and the radiotracer 13N to measure cellular turnover kinetics, patterns of flux partitioning, and cytosolic pool sizes of both NO3− and NH4+ in seedling roots of rice (Oryza sativa L. cv IR72), supplied simultaneously with the two N sources. We show that plasma membrane fluxes for NH4+, cytosolic NH4+ accumulation, and NH4+ metabolism are enhanced by the presence of NO3−, whereas NO3− fluxes, accumulation, and metabolism are strongly repressed by NH4+. However, net N acquisition and N translocation to the shoot with dual N-source provision are substantially larger than when NO3− or NH4+ is provided alone at identical N concentrations.

Nitrate Transport and Not Photoinhibition Limits Growth of the Freshwater Cyanobacterium Synechococcus Species PCC 6301 at Low Temperature1

The effect of low temperature on cell growth, photosynthesis, photoinhibition, and nitrate assimilation was examined in the cyanobacterium Synechococcus sp. PCC 6301 to determine the factor that limits growth. Synechococcus sp. PCC 6301 grew exponentially between 20°C and 38°C, the growth rate decreased with decreasing temperature, and growth ceased at 15°C. The rate of photosynthetic oxygen evolution decreased more slowly with temperature than the growth rate, and more than 20% of the activity at 38°C remained at 15°C. Oxygen evolution was rapidly inactivated at high light intensity (3 mE m−2 s−1) at 15°C. Little or no loss of oxygen evolution was observed under the normal light intensity (250 μE m−2 s−1) for growth at 15°C. The decrease in the rate of nitrate consumption by cells as a function of temperature was similar to the decrease in the growth rate. Cells could not actively take up nitrate or nitrite at 15°C, although nitrate reductase and nitrite reductase were still active. These data demonstrate that growth at low temperature is not limited by a decrease in the rate of photosynthetic electron transport or by photoinhibition, but that inactivation of the nitrate/nitrite transporter limits growth at low temperature.

A Conserved Acidic Motif in the N-Terminal Domain of Nitrate Reductase Is Necessary for the Inactivation of the Enzyme in the Dark by Phosphorylation and 14-3-3 Binding1

It has previously been shown that the N-terminal domain of tobacco (Nicotiana tabacum) nitrate reductase (NR) is involved in the inactivation of the enzyme by phosphorylation, which occurs in the dark (L. Nussaume, M. Vincentz, C. Meyer, J.P. Boutin, and M. Caboche [1995] Plant Cell 7: 611–621). The activity of a mutant NR protein lacking this N-terminal domain was no longer regulated by light-dark transitions. In this study smaller deletions were performed in the N-terminal domain of tobacco NR that removed protein motifs conserved among higher plant NRs. The resulting truncated NR-coding sequences were then fused to the cauliflower mosaic virus 35S RNA promoter and introduced in NR-deficient mutants of the closely related species Nicotiana plumbaginifolia. We found that the deletion of a conserved stretch of acidic residues led to an active NR protein that was more thermosensitive than the wild-type enzyme, but it was relatively insensitive to the inactivation by phosphorylation in the dark. Therefore, the removal of this acidic stretch seems to have the same effects on NR activation state as the deletion of the N-terminal domain. A hypothetical explanation for these observations is that a specific factor that impedes inactivation remains bound to the truncated enzyme. A synthetic peptide derived from this acidic protein motif was also found to be a good substrate for casein kinase II.

Phosphorylated Nitrate Reductase and 14-3-3 Proteins1 : Site of Interaction, Effects of Ions, and Evidence for an AMP-Binding Site on 14-3-3 Proteins

The inactivation of phosphorylated nitrate reductase (NR) by the binding of 14-3-3 proteins is one of a very few unambiguous biological functions for 14-3-3 proteins. We report here that serine and threonine residues at the +6 to +8 positions, relative to the known regulatory binding site involving serine-543, are important in the interaction with GF14ω, a recombinant plant 14-3-3. Also shown is that an increase in ionic strength with KCl or inorganic phosphate, known physical effectors of NR activity, directly disrupts the binding of protein and peptide ligands to 14-3-3 proteins. Increased ionic strength attributable to KCl caused a change in conformation of GF14ω, resulting in reduced surface hydrophobicity, as visualized with a fluorescent probe. Similarly, it is shown that the 5′ isomer of AMP was specifically able to disrupt the inactive phosphorylated NR:14-3-3 complex. Using the 5′-AMP fluorescent analog trinitrophenyl-AMP, we show that there is a probable AMP-binding site on GF14ω.

Chilling Delays Circadian Pattern of Sucrose Phosphate Synthase and Nitrate Reductase Activity in Tomato1

Overnight low-temperature exposure inhibits photosynthesis in chilling-sensitive species such as tomato (Lycopersicon esculentum) and cucumber by as much as 60%. In an earlier study we showed that one intriguing effect of low temperature on chilling-sensitive plants is to stall the endogenous rhythm controlling transcription of certain nuclear-encoded genes, causing the synthesis of the corresponding transcripts and proteins to be mistimed when the plant is rewarmed. Here we show that the circadian rhythm controlling the activity of sucrose phosphate synthase (SPS) and nitrate reductase (NR), key control points of carbon and nitrogen metabolism in plant cells, is delayed in tomato by chilling treatments. Using specific protein kinase and phosphatase inhibitors, we further demonstrate that the chilling-induced delay in the circadian control of SPS and NR activity is associated with the activity of critical protein phosphatases. The sensitivity of the pattern of SPS activity to specific inhibitors of transcription and translation indicates that there is a chilling-induced delay in SPS phosphorylation status that is caused by an effect of low temperature on the expression of a gene coding for a phosphoprotein phosphatase, perhaps the SPS phosphatase. In contrast, the chilling-induced delay in NR activity does not appear to arise from effects on NR phosphorylation status, but rather from direct effects on NR expression. It is likely that the mistiming in the regulation of SPS and NR, and perhaps other key metabolic enzymes under circadian regulation, underlies the chilling sensitivity of photosynthesis in these plant species.