What limits nitrate uptake from soil?

Abstract. An accepted view, that unless nitrate concentrations in the soil solution are very low (e.g. below 0.1–0.2 mol m^?3) the growth of high-yielding crops is not limited by the availability of nitrogen, is challenged. Conventional analyses of nutrient supply and demand, based on calculations of apparent inflow rates (uptake rates per unit total root length) are invalid. Apparent inflow rates are inversely proportional to root length. The convention of using total root length grossly overestimates the fraction of the root system active in nutrient uptake. Consequently, inflow rates based on total root lengths underestimate the true values, indicating unrealistically low nutrient concentration differentials between bulk soil and root surfaces required to drive uptake. An alternative method of analysis is suggested. This is based on total nutrient uptake rather than on inflow rate. Measurements of the former do not depend on estimates of active root length and can be made directly and reliably. The method was applied to data obtained from a pot experiment using spring wheat (Triticum aestivum L., cv. Wembley) grown in soil without nitrogen fertilizer (N₀) or with nitrogen fertilizer equivalent to 200kg N ha^?1 (N⁺). Soil nitrate concentrations calculated using the conventional method based on total root length, suggested that any increases in concentration above those measured in the N⁰ treatment should not have resulted in increased uptake and growth. However, the N⁺ plants were always bigger than those in the No treatment, refuting this suggestion. Theoretical uptakes of nitrogen (calculated initially on the basis of a fully active root system) were adjusted, by reducing the effective root length incrementally, until the theoretical uptake matched the measured net uptake of nitrogen. The mean fractions of the root systems likely to have been involved in nitrate uptake were 11% and 3.5% of the total lengths of root in the N⁰ and N⁺ treatments, respectively.     

An ex-situ and in vitro approach towards the bioremediation of carcinogenic hexavalent chromium

Abstract

Chromium, ranking the second most among toxic heavy metal pollutants in the world, causing respiratory, cardiovascular and renal problems in human beings is under study herein. We examined the biological remediation of the carcinogenic Cr (VI) polluted soils by indigenous yeast isolates. The total element analysis of the treated sample was determined by Energy Dispersion X-ray Micro Analysis (EDXMA). The sample under study was observed to have a high concentration of 458.29 mgKg^?1 Cr (VI), determined by Atomic Absorption Spectroscopy (AAS) and DPC analysis. The most tolerant isolate designated as CSR was used for in vitro and ex-situ bioremediation studies of Cr (VI). The isolate achieved significant bioremediation of 86% in vitro and 75.12% in ex-situ method. The optimal conditions for in vitro bioremediation were found to be 28?°C and a pH of 6. The ITS1, 5.8S rRNA and D1, D2 domain of LSU rRNA gene characterization of the isolate CSR illustrated that it belongs to Ustilago genera. The isolate was deposited in NCBI GenBank as Ustilago sp. CSR (KY284846). Although, Ustilago is generally a pathogenic fungus, our study opens up the scope of using Ustilago spp. for bioremediation of the carcinogenic heavy metal Chromium.     

Characteristics of inorganic nitrogen assimilation of plants in fire-prone Mediterranean-type vegetation

A range of approaches was used to investigate how species within a fire-prone Banksia woodland in South West Australia exploited inorganic soil nitrogen sources and how this changes through the development of the fire chronosequence. Nitrate and ammonium were present in soil solution throughout the chronosequence but nitrate predominated in recently burnt sites. Mean shoot nitrate reductase activities were high for all species in recently burnt sites and showed little increase when nitrate was supplied via the transpiration stream. Nitrate reductase of shoots of most species was low at sites not burnt for several years, but following transpirational induction with nitrate, developed activities similar to those at recently burnt sites. The principal amino compounds transported in the xylem were species specific, including asparagine, glutamine and citrulline-dominated species, and changed little in relative composition across the chronosequence. Species most active in leaf nitrate reduction transported the largest amounts of nitrate in their xylem sap and proportional amounts of nitrate in xylem tended to be greatest in recently burnt sites. Most of the species examined appeared to be shoot rather than root nitrate assimilators, but marked differences were recorded in potential of leafy shoots of different species to reduce nitrate. As a general rule, shallow-rooted herbaceous, non-mycorrhizal or VAM-positive species had the highest capacity to reduce nitrate, whereas woody species with ericoid mycorrhizae or combined vesicular arbuscular/ectomycorrhizal associations exhibited little capacity to reduce nitrate in roots or shoots. It seems likely that this latter group utilize ammonium or even organic forms of nitrogen rather than nitrate. Some putative nitrogen-fixing species were active in reducing and transporting nitrate, others were virtually inactive in these respects.     

A rearranging ligand enables allosteric control of catalytic activity in copper-containing nitrite reductase

In Cu-containing nitrite reductase from Alcaligenes faecalis S-6 the axial methionine ligand of the type-1 site was replaced (M150G) to make the copper ion accessible to external ligands that might affect the enzyme's catalytic activity. The type-1 site optical spectrum of M150G (A(460)/A(600)=0.71) differs significantly from that of the native nitrite reductase (A(460)/A(600)=1.3). The midpoint potential of the type-1 site of nitrite reductase M150G (E(M)=312(+/-5)mV versus hydrogen) is higher than that of the native enzyme (E(M)=213(+/-5)mV). M150G has a lower catalytic activity (k(cat)=133(+/-6)s(-1)) than the wild-type nitrite reductase (k(cat)=416(+/-10)s(-1)). The binding of external ligands to M150G restores spectral properties, midpoint potential (E(M)<225mV), and catalytic activity (k(cat)=374(+/-28)s(-1)). Also the M150H (A(460)/A(600)=7.7, E(M)=104(+/-5)mV, k(cat)=0.099(+/-0.006)s(-1)) and M150T (A(460)/A(600)=0.085, E(M)=340(+/-5)mV, k(cat)=126(+/-2)s(-1)) variants were characterized. Crystal structures show that the ligands act as allosteric effectors by displacing Met62, which moves to bind to the Cu in the position emptied by the M150G mutation. The reconstituted type-1 site has an otherwise unaltered geometry. The observation that removal of an endogenous ligand can introduce allosteric control in a redox enzyme suggests potential for structural and functional flexibility of copper-containing redox sites.     

Regulation of nitrate reductase activity in higher plants

Nitrate reductase is one of the most important enzymes in the assimilation of exogenous nitrate—the predominant form of nitrogen available to green plants growing in soil. Activity of this enzyme in plants gives a good estimate of the nitrogen status of the plant and is very often correlated with growth and yield. Although it is difficult to explain the physiological significance and the mechanism of effects of several factors on the enzyme activity, in some cases suitable postulates have been advanced. In general, the enzyme activity in a plant tissue is a balance between its relative rates of synthesis/degradation and activation/inactivation. Factors may affect the overall activity by interfering with either of these processes.     

Possible roles of calcium and ammonium in the development of bitter pit in apple

Apple trees ( Malus pumila Mill . var. domestica Fuji/ Malus prunifolia rootstock) showed a high susceptibility to bitter pit when supplyed with ammonium salt instead of nitrate (control) in the nutrient solution. When apple fruit was affected by bitter pit, a lower calcium as well as a higher nitrogen and ammonium-nitrogen contents was observed in the fruit flesh near the calyx end. The activity of the mitochondrial Ca²⁺-uptake of the fruit flesh near the calyx end was higher when the tree was grown with ammonium salt than when grown with nitrate. Both the activities of succinate: cytochrome c oxidoreductase and the mitochondrial Ca²⁺-uptake per g of tissue were higher in affected fruit than in healthy fruit. Each of chlorpromazine, N-(6-aminohexyl)-5-chloro-l-napthalenesulfonamide (W-7) and N-(6-aminohexyl)-l-naphthalenesulfonamide (W-5), calmodulin antagonists, was infiltrated into the fruit for 20 min under reduced pressure (about 1 × 10⁴ Pa). Few days later, numerous bitter pit-like spots were observed in both fruit treated with W-7 and chlorpromazine, while only a few spots were observed after the infiltration with W-5, a less potent calmodulin antagonist. A possible mechanism for the occurrence of bitter pit is discussed.     

Influence of the culture medium constituents and inoculum size on the accumulation of blue pigment and cell growth of Lavandula spica

The effects of the concentration of PO₄ ^3-, NO₃ ^–, Fe ²⁺, sucrose and inoculum size on the accumulation of blue pigment and growth of suspension cultures of Lavandula spica D.C. ( L. latifolia Vill.), were studied. The combination of 2.5 mM PO₄ ^3-, 14.1 mM NO₃ ^–, 1 mM Fe ²⁺, 30 g l^–1 sucrose and 10 g l^–1fresh weight of inoculum, promoted a 7-fold enhancement on the productivity of blue pigment in comparison to the control medium. Among all the culture medium constituents tested, phosphate exerted the highest beneficial effect and Fe²⁺ showed to be essential for the accumulation of the pigment. High levels of sucrose (60 or 90 g l^–1) did not stimulate the accumulation of blue pigment. In these conditions, cell growth and cell viability were drastically affected.     

Effect of Cycloheximide on Development of Mitochondria in Germinating Pea Cotyledons

Polyethylene glycol (PEG) mediated transfection of Lactobacillus casei ATCC 27092 protoplasts by phage PL-1 DNA was done. The protoplasts were obtained by treatment with purified PL-1 phage N-acetylmuramidase in the presence of citrate. Optimum conditions for transfection were 50% PEG 4,000, 15 µg protamine sulfate/ml, 0.15 m sucrose, and 10 m m MgSO₄ in MR medium (pH 6.0). The extent of transfection was proportional to the amounts of DNA added, and the greatest efficiency of transfection after a 10-min incubation was about 3.3 × 10⁵ PFU/µg DNA. The eclipse period of growth of progeny phages in the transfectants was 3 hr and the average burst size was 200.     

Biological Control of Root-Knot Nematode <i>Meloidogyne incognita</i> in <i>Psoralea corylifolia</i> Plant by Enhancing the Biocontrol Efficacy of <i>Trichoderma harzianum</i> Using Press Mud

Meloidogyne incognita is a plant pathogen causing root-knot disease and loss of crop yield. The present study aimed to use Trichoderma harzianum as a biocontrol agent against plant-parasitic nematodes and used press mud, which is a solid waste by-product of sugarcane, as a biocontrol agent and biofertilizer. Therefore, the combined application of T. harzianum and press mud may enhance nematode control and plant growth. Elemental analysis of press mud using scanning electron microscopy (SEM) integrated with an Energy Dispersive X-ray (EDX) analyzer revealed the presence of different elements such as C, O, Mg, Si, P, K, Ca, Cu and Zn. In addition, a greenhouse study was conducted to investigate the combined effects of press mud and T. harzianum on M. incognita reproduction and growth and the biochemical features of Psoralea corylifolia. The results showed that plant length, dry biomass, leaf area, the number of seeds per plant, chlorophyll a, chl b, carotenoid content, nitrate reductase, carbonic anhydrase, and nitrogen content were significantly increased (P ≤ 0.05) in the T2 plants (plants were treated with 100 g of press mud + 50 mL T. harzianum before one week of M. incognita inoculation), over inoculated plants (IC). Antioxidant enzyme activity of ascorbate peroxidase (APX), catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) in the foliage of P. corylifolia was significantly increased when plants were treated with press mud + T. harzianum. A significant reduction in the number of egg masses, nematode population, and root-knot index (RKI) was found in plants with T2 plants. These results suggest that the combined application of T. harzianum and press mud has the potential to control the M. incognita infection and can be used as an environmentally safe alternative to chemical nematicides and also help in the removal of sugarcane waste that causes environmental pollution.     

Modelling the interactions between water and nutrient uptake and root growth

A model of three-dimensional root growth has been developed to simulate the interactions between root systems, water and nitrate in the rooting environment. This interactive behaviour was achieved by using an external-supply/internal-demand regulation system for the allocation of endogenous plant resources. Data from pot experiments on lupins heterogeneously supplied with nitrate were used to test and parameterise the model for future simulation work. The model reproduced the experimental results well (R ² = 0.98), simulating both the root proliferation and enhanced nitrate uptake responses of the lupins to differential nitrate supply. These results support the use of the supply/demand regulation system for modelling nitrate uptake by lupins. Further simulation work investigated the local uptake response of lupins when nitrate was supplied to a decreasing fraction of the root system. The model predicted that the nitrate uptake activity of lupin roots will increase as the fraction of root system with access to nitrate decreases, but is limited to an increase of around twice that of a uniformly supplied control. This work is the first example of a modelled root system responding plastically to external nutrient supply. This model will have a broad range of applications in the study of the interactions between root systems and their spatially and temporally heterogeneous environment.