Natural15N abundance as a method of estimating the contribution of biologically fixed nitrogen to N2-fixing systems: Potential for non-legumes

The¹⁵N abundance of plants usually closely reflects the¹⁵N abundance of their major immediate N source(s); plant-available soil N in the case of non-N₂-fixing plants and atmospheric N₂ in the case of N₂ fixing plants. The¹⁵N abundance values of these sources are usually sufficiently different from each other that a significant and systematic difference in the¹⁵N abundance between the two kinds of plants can be detected. This difference provides the basis for the natural¹⁵N abundance method of estimating the relative contribution of atmospheric N₂ to N₂-fixing plants growing in natural and agricultural settings. The natural¹⁵N abundance method has certain advantages over more conventional methods, particularly in natural ecosystems, since disturbance of the system is not required and the measurements may be made on samples dried in the field. This method has been tested mainly with legumes in agricultural settings. The tests have demonstrated the validity of this method of arriving at semi-quantitative estimates of biological N₂-fixation in these settings. More limited tests and applications have been made for legumes in natural ecosystems. An understanding of the limits and utility of this method in these systems is beginning to emerge. Examples of systematic measurements of differences in¹⁵N abundance between non-legume N₂-fixing systems and neighbouring non-fixing systems are more unusual. In principle, application of the method to estimate N₂-fixation by nodulated non-legumes, using the natural¹⁵N abundance method, is as feasible as estimating N₂-fixation by legumes. Most of the studies involving N₂-fixing non-legumes are with this type of system (e.g., Ceanothus, Chamabatia, Eleagnus, Alnus, Myrica, and so forth). Resuls of these studies are described. Applicability for associative N₂-fixation is an empirical question, the answer to which probably depends upon the degree to which fixed N goes predominantly to the plant rather than to the soil N pool. The natural¹⁵N abundance method is probably not well suited to assessing the contribution of N₂-fixation by free-living microorganisms in their natural habitat, particularly soil microorganisms.This work was supported in part by subcontracts under grants from the US National Science Foundation (DEB79-21971 and BSR821618)     

Effects of root pruning of wheat at different nitrogen levels

In two experiments, wheat plants growing in solutions of different nitrogen concentration were subjected to root pruning. In higher concentrations of nitrogen the growth rate was higher, and the proportional allocation of growth to shoot higher, but pruning did not affect the allocation of growth at either level of nitrogen. This result gives no support to Thornley's source-sink model of the control of shoot: root ratio.     

The effectiveness of some Indonesian strains of Rhizobium on four tropical legumes

Nodules were collected from 14 legume species from the Indonesian Islands of South Sulawesi, Java and Sumatra. Their rhizobia were isolated and growth characteristics, nodulation ability and nitrogen fixing effectiveness were assessed against recommended commercially available Australian strains. The test legumes wereMacroptilium atropurpureum Urb. cv. Siratro,Vigna unguiculata (L.) Walp. cv Eureka,Centrosema pubescens Benth cv. Belalto andDesmodium heterocarpon (L) DC. A significant portion of the native rhizobial isolates were of the fast growing type. Dry matter and total nitrogen production forM. atropurpureum andV. unguiculata was highest when inoculated with native strains while the commerical strains produced superior dry matter production forC. pubescens andD. heterocarpon. However the total nitrogen production of native and commercial strains was not significantly different for the latter two legumes. The study indicated that a potential exists for developing inocula from local Rhizobium strains.     

The response ofPlantago lanceolata L. andP. major L. spp.major to nitrate depletion

To study aspects of the ecology of grassland species, in a comparative experiment, plants ofP. lanceolata andP. major were grown in pots in a greenhouse, and subjected to a gradual nitrate depletion for several weeks. Control plants were weekly supplied with nitrate. Growth, leaf appearance and disappearance, concentrations of cations and inorganic anions, soluble and insoluble reduced nitrogen concentrations,in vivo nitrate reductase activity (NRA) and the concentration of non-structural carbohydrates in several parts of the plants were followed. Depletion of nitrate caused a reduction of shoot growth, both in biomass and number of leaves. Withering of leaves increased. Accumulation of root dry matter was little (P. lanceolata), or not (P. major) affected. The concentration of reduced nitrogen in all tissues also decreased, both that of the soluble and that of the insoluble fraction. As a result, nitrogen use efficiency (NUE, g dry matter produced per mmol N incorporated) increased by nitrate depletion. NRA was higher in the roots than in the leaves, and decreased with increasing nitrate depletion. In control plants, nitrate became also limiting. This resulted in decreasing nitrate concentrations in leaves and roots. In the leaves, the decrease in nitrate concentration was preceded by a decrease in NRA. The decrease of the nitrate concentration was parallelled by an increase in the concentration of soluble sugar. No major differences in the response towards nitrate depletion were observed between the two species. Grassland Species Research Group, publication no. 129     

Growth response of barley and tomato to nitrogen stress and its control by abscisic acid,water relations and photosynthesis

Barley (Hordeum vulgare L.) and tomato Lycopersicon esculentum Mill.) were grown hydroponically and examined 2, 5, and 10 d after being deprived of nitrogen (N) supply. Leaf elongation rate declined in both species in response to N stress before there was any reduction in rate of dryweight accumulation. Changes in water transport to the shoot could not explain reduced leaf elongation in tomato because leaf water content and water potential were unaffected by N stress at the time leaf elongation began to decline. Tomato maintained its shoot water status in N-stressed plants, despite reduced water absorption per gram root, because the decline in root hydraulic conductance with N stress was matched by a decline in stomatal conductance. In barley the decline in leaf elongation coincided with a small (8%) decline in water content per unit area of young leaves; this decline occurred because root hydraulic conductance was reduced more strongly by N stress than was stomatal conductance. Nitrogen stress caused a rapid decline in tissue NO ₃ ^- pools and in NO ₃ ^- flux to the xylem, particularly in tomato which had smaller tissue NO ₃ ^- reserves. Even in barley, tissue NO ₃ ^- reserves were too small and were mobilized too slowly (60% in 2 d) to support maximal growth for more than a few hours. Organic N mobilized from old leaves provided an additional N source to support continued growth of N-stressed plants. Abscisic acid (ABA) levels increased in leaves of both species within 2 d in response to N stress. Addition of ABA to roots caused an increase in volume of xylem exudate but had no effect upon NO ₃ ^- flux to the xylem. After leaf-elongation rate had been reduced by N stress, photosynthesis declined in both barley and tomato. This decline was associated with increased leaf ABA content, reduced stomatal conductance and a decrease in organic N content. We suggest that N stress reduces growth by several mechanisms operating on different time scales: (1) increased leaf ABA content causing reduced cell-wall extensibility and leaf elongation and (2) a more gradual decline in photosynthesis caused by ABA-induced stomatal closure and by a decrease in leaf organic N.Abbreviation and symbols ABA abscisic acid - c_i leaf internal CO₂ concentration - L_p root hydraulic conductance     

Acetylene reduction and indoleacetic acid production by Azospirillum isolates from Cactaceous plants

Azospirillum isolates were obtained from rhizosphere soil and roots of three cactaceae species growing under arid conditions. All Azospirillum isolates from rhizosphere and roots ofStenocereus pruinosus andStenocereus stellatus were identified asA. brasilense; isolates of surface-sterilized roots fromOpuntia ficus-indica were bothA. brasilense andA. lipoferum. Azospirilla per g of fresh root in the three species ranged from 70×10³ to 11×10³. The most active strains in terms of C₂H₂ reduction (25–49.6 nmol/h·ml) and indoleacetic acid (IAA) production (36.5–77 μg/ml) were those identified asA. brasilense and isolated from Stenocereus roots.A. lipoferum isolated from Opuntia roots produced low amounts of IAA (6.5–17.5 μg/ml) and low C₂H₂-reduction activity (17.8–21.2 nmol/h·ml).     

Transformations and fate of sheep urine-N applied to an upland U.K. pasture at different times during the growing season

The fate of sheep urine-N applied to an upland grass sward at four dates representing widely differing environmental conditions, was followed in soil (0–20 cm) and in herbage. Urine was poured onto 1-m² plots to simulate a single urination in August 1984 (warm and dry), May (cool), July and August 1985 (cool and wet) at rates equivalent to 40–52 g N m⁻². The transformation of urine-N (61–69% urea-N) in soil over a 6–7 week period followed the same general pattern when applied at different times during the season; rapid hydrolysis of urea, the appearance of large amounts of urine-N as ammonium in soil extracts, and the appearance of nitrate about 14 days after application. The magnitude of “apparent” nitrification however differed markedly with environmental conditions, being greatest in May 1985 when a maximum of 76% of the inorganic soil N was in the form of nitrate. At all other application dates nitrate levels were relatively low. With the August 1984 application soil inorganic N returned to control levels (given water only) after 31 days but considerable amounts remained in soil for 60–90 days with the other applications. Weekly cuts to 3-cm indicated that increases in herbage dry matter and N yields in response to urine application were greatest in absolute terms after the May 1985 application and continued for at least 70 days with all applications. Relative to control plots the May application resulted in a 3-fold increase in herbage DM compared with corresponding values of 6-, 5-, and 7-fold increases with the August 1984, July and August 1985 applications. Recovery of urine-N in herbage was poor averaging only 17% of that applied at different dates, while recovery in soil extracts was incomplete. The exact routes of loss (volatilisation, leaching, denitrification or immobilisation) were not quantified but it is evident that substantial amounts of urine-N can be lost from the soil-plant system under upland conditions.     

Effect of Rhizobium andAzospirillum lipoferum inoculation on the nodulation,yield and nitrogen uptake of peanut cultivars

Peanut (Arachis hypogaea Linn.) Cvs. Robut 33-1 and JL 24 were inoculated with Rhizobium strain NC 92 and a strain ofAzospirillum lipoferum singly and as mixed inoculum. Seed inoculation with these bacteria enhanced nodulation, N content and yield of these cultivars under field conditions. While a mix inoculation of these two diazotrophic cultures had an adverse effect on these parameters as compare to single inoculation.     

Effects of CaSO4 and NaCl on growth and nitrogen fixation ofLeucaena leucocephala

Effects and interactions of varying CaSO₄ and NaCl levels on growth and nitrogen fixation ofLeucaena leucocephala K8 were examined. Leucaena was grown in nutrient solution at four levels of CaSO₄ (0.5, 1.0, 2.5 and 5.0 mM) and NaCl (1, 25, 50 and 100 mM) in randomized blocks with five replications. While NaCl significantly reduced plant growth, additions of CaSO₄ increased plant height, leaf number, and biomass of salt treated plants. For the nonsaline treatments, high CaSO₄ levels slightly depressed growth, which contradicts suggestions that Leucaena has a high calcium requirement. A significant calcium/sodium interaction was not seen for nodule number or weight. Nodule number was significantly depressed by 100 mM NaCl and nodule weight of the salt stressed plants significantly increased as CaSO₄ concentration increased from 0.5 to 2.5 mM. Effects of NaCl and CaSO₄ on nitrogen content of plant parts were inconclusive. The promotion of Leucaena salinity tolerance by addition of CaSO₄ may be attributed to the effect of calcium in maintaing the selective permeability of membranes.     

Survival ofSclerotinia minor Jagger sclerotia in solarized soil

Calcium, magnesium and potassium dynamics in decomposing litter of three tree species were measured over a two-year period. The speices studied were flowering dogwood (Cornus florida), red maple (Acer rubrum) and chestnut oak (Quercus prinus). The order of decomposition was:C. florida>A. rubrum>Q. prinus.Calcium concentrations increased following any initial leaching losses. However, there were net releases of Ca from all three litter types since mass loss exceeded the increases in concentration. Net release of Ca by the end of two years from all three species combined was 42% of initial inputs in litterfall. Magnesium concentrations increased in the second year, following decreases due to leaching during the first year inC. florida andA. rubrum litter. Net release of Mg by the end of two years was 58% of initial inputs. Potassium concentrations decreased rapidly and continued to decline throughout the study. Net release of K by the end of two years was 91% of initial inputs.These data on cation dynamics, and similar data on N, S and P dynamics from a previous study, were combined with annual litterfall data to estimate the release of selected nutrients from foliar litter of these tree species at the end of one and two years of decomposition. The relative mobility of all six elements examined in relation to mass loss after two years was; K>Mg>mass>Ca>S>P>N.