Nitrogen fixation in grazed and ungrazed subterranean clover pasture in south-west Australia assessed by the 15N natural abundance technique

1995 ◽  
Vol 46 (7) ◽  
pp. 1427 ◽  
Author(s):  
P Sanford ◽  
JS Pate ◽  
MJ Unkovich ◽  
AN Thompson

The progress of N2 fixation by subterranean clover (Trifolium subterraneum L.) was followed throughout a growing season in adjacent grazed and ungrazed portions of a pasture at Mount Barker, W.A. Proportions of plant nitrogen derived from the atmosphere (%Ndfa) were determined at a sequence of sampling times using the 15N natural abundance technique with capeweed (Arctotheca calendula L.) as non-fixing reference species. Cumulative yields of fixed N by above ground biomass of clover were determined from %Ndfa values, concurrent estimates of dry matter (DM) yields, and percentage nitrogen in clover shoot DM. Seasonal DM yields of clover, capeweed and mixed grasses were in the approximate ratio 60 : 20: 20. Total herbage yields were 11.8 and 7.8 t ha-1 for the grazed and ungrazed swards respectively. Poorer performance of the latter was attributed to shading by taller grasses late in the season. Starting from a low value of 58%, Ndfa of the ungrazed sward became uniformly high (73-88%) for the rest of the season. Clover of the more productive grazed sward behaved similarly except for a significant mid winter depression to 55%Ndfa, probably caused by excessive defoliation through overgrazing. Fixed N recovered from clover shoot biomass was 103 and 188 kg N ha-1 for ungrazed and grazed pasture respectively. Mineral N under the grazed sward first consisted mostly of nitrate, and then predominantly of ammonium. Soil-derived N was utilized roughly equally by clover, grasses and capeweed and a field study of %Ndfa of subterranean clover grown in varying proportion with either the main pasture grass (Lolium rigidum Gaudin) or capeweed indicated the grass to be the more effective competitor for soil N against the clover. The data suggested that reliable estimates of seasonal accumulation of fixed N by pastures would be obtained from assessments of cumulative biomass yield of clover N with a single determination of %Ndfa at peak productivity in mid to late spring.

1994 ◽  
Vol 45 (1) ◽  
pp. 165 ◽  
Author(s):  
P Sanford ◽  
JS Pate ◽  
MJ Unkovich

In an attempt to understand why pasture production in southern Australia has declined markedly in recent years a survey of the symbiotic performance of the legume component of annual pastures on 81 farms (243 sites) was undertaken in the southern coastal region of Western Australia. The 15N natural abundance technique was used to determine the percentage of plant nitrogen derived from the atmosphere (%Ndfa) using capeweed (Arctotheca calendula) as principal non-fixing reference species. %Ndfa values were then related to edaphic and management information, e.g. soil total nitrogen, soil pH, stocking rates and cropping history of the sites. The principal legume species encountered exhibited similar mean %Ndfa values but substantial variation in symbiotic performance was evident across the sites, viz, Trifolium subterraneum 72%Ndfa (n = 184, range of values encountered 0-100%), Medicago spp. 7l%Ndfa (n = 24, range 7-l00%), Lotus spp. 8l%Ndfa (n = 15, range 1-l00%), Ornithopus compressus 76%Ndfa (n = 15, range 25-100%) and Trifolium balansae 69%Ndfa (n = 7, range 0-100%). In the case of subterranean clover, the most widely occurring species, almost one third (29%) of sites surveyed recorded %Ndfa values within the range 0-65%, suggesting that symbiotic performance might well be quite widely limiting to herbage production in the study region. Of the 24 edaphic and management factors evaluated, only one, %A1 in shoot, DM showed a significant relationship with %Ndfa, with 40% of the pastures surveyed deemed at risk in terms of acidity related aluminium toxicity. Correlations of %Ndfa with soil pH and soil total N produced examples of high values %Ndfa for sub-clover being associated with very low soil pH or high soil N, suggesting possible adaptation of symbiotic partnerships to acidity or high mineral N.


1999 ◽  
Vol 50 (6) ◽  
pp. 1047 ◽  
Author(s):  
B. S. Dear ◽  
M. B. Peoples ◽  
P. S. Cocks ◽  
A. D. Swan ◽  
A. B. Smith

The proportions of biologically fixed (Pfix) plant nitrogen (N) and the total amounts of N2 fixed by subterranean clover (Trifolium subterraneum L.) growing in pure culture and in mixtures with different densities (5, 10, 20, or 40plants/m2) of newly sown phalaris (Phalaris aquatica L.) or lucerne (Medicago sativa L.) were followed over 3 years in a field study using the 15N natural abundance technique. The amount of fixed N in subterranean clover was linearly related to shoot biomass. Over the 3-year period, subterranean clover fixed 23–34 kg N/t shoot biomass compared with 17–29 kg N/t shoot biomass in lucerne. Based on above-ground biomass, pure subterranean clover fixed 314 kg N/ha over the 3 years compared with 420–510 kg N/ha by lucerne–clover mixtures and 143–177 kg N/ha by phalaris–clover mixtures. The superior N2 fixation by the lucerneŒsubterranean clover mixtures was due to the N fixed by the lucerne and the presence of a higher subterranean clover biomass relative to that occurring in the adjacent phalaris plots. In the first year, 92% of subterranean clover shoot N was derived from fixation compared with only 59% of lucerne. The reliance of clover upon fixed N2 remained high (73–95%) throughout the 3 years in all swards, except in pure subterranean clover and lucerne in August 1996 (56 and 64%, respectively). Subterranean clover usually fixed a higher proportion of its N when grown in mixtures with phalaris than with lucerne. The calculated Pfix values for lucerne (47–61% in 1995 and 39–52% in 1996) were consistently lower than in subterranean clover and tended to increase with lucerne density. Although lucerne derived a lower proportion of its N from fixation than subterranean clover, its tissue N concentration was consistently higher, indicating it was effective at scavenging soil mineral N. It was concluded that including lucerne in wheat-belt pastures will increase inputs of fixed N. Although lucerne decreased subterranean clover biomass, it maintained or raised Pfix values compared with pure subterranean clover swards. The presence of phalaris maintained a high dependence on N2 fixation by subterranean clover, but overall these swards fixed less N due to the lower clover herbage yields. Perennial and annual legumes appear compatible if sown in a mix and can contribute more N2 to the system than where the annual is sown alone or with a perennial grass. These findings suggest that increases in the amount of N2 fixed can be achieved through different legume combinations without interfering greatly with the N fixation process. Different combinations may also result in more efficient use of fixed N2 through reduced leaching. Further work looking at combinations of annuals possibly with different maturity times, different annual and perennial legume combinations, and pure combinations of perennial (e.g. lucerne) could be investigated with the aim of maximising N2 fixation and use. Grazing management to encourage clover production in mixtures with phalaris will be necessary before the potential of subterranean clover to contribute fixed N2 in these swards is fully realised.


1994 ◽  
Vol 45 (1) ◽  
pp. 119 ◽  
Author(s):  
MJ Unkovich ◽  
JS Pate ◽  
P Sanford ◽  
EL Armstrong

Precision of estimation of the proportion of legume N derived from N2 fixation (%Ndfa) was assessed in relation to subterranean clover (Trifolium subterraneum L.) pastures and crops of pea (Pisum sativum L.) and lupin (Lupinus angustifolius L.) under south-west Australian conditions. By using a standardized 10-point sampling procedure of paired sampling of legume and reference plant and reference plant 15N natural abundance (S15N) values in the range from +2.9 to +4. 0%o, %Ndfa of sample crops of lupin and field pea and a clover pasture were assessed with respective precisions of 93� O.6%, 76� 2.4% and 91�1.3% (� s.e., n = 10). Effects on S15N due to isotope discrimination during fixation and subsequent distribution of N by the three study legumes were studied using sand-cultured, fully symbiotic plant material. The resulting S15N data (B values) showed consistently more negative values for shoots than roots (all species), no significant effects of cultivar on B values (all species), a marked effect of rhizobial strain on B value (subclover) and a tendency for B values to fall with plant age (pea and lupin). The likely magnitude of errors in %Ndfa estimates due to incorrect choice of B value was indicated. By using data for reference plant S15N values from field surveys and previously assessed error factors in mass spectrometric measurement of S15N, precision of estimation of %Ndfa by using bulked material from the 10-point field sampling procedure was predicted for situations ranging from where a legume was obtaining only minimal amounts (10%) through to the bulk (90%) of its N by atmospheric fixation.


1995 ◽  
Vol 46 (1) ◽  
pp. 225 ◽  
Author(s):  
JA Doughton ◽  
PG Saffigna ◽  
I Vallis ◽  
RJ Mayer

The 15N enrichment and 15N natural abundance methods for estimating N2 fixation in chickpea were compared over a range of soil NO3-N levels at crop establishment varying from 10 to 326 kg N/ha (0-120 cm depth). Barley was used as a non-N2 fixing control crop. Both methods estimated reduced N2 fixation as soil NO3-N levels at crop establishment increased. Similar estimates of % N2 fixation were obtained at high values, but at low values the enrichment method gave lower estimates, some of which were negative. The 15N natural abundance method provided realistic estimates of % N2 fixation across all soil N03-N levels at crop establishment. An asymptotic curve described a close ( R2 = 0.95) relationship between these factors. Standard errors of estimates of means for the 15N natural abundance method remained acceptable and relatively stable over the full range of measurements; however, with the 15N enrichment method they became unacceptably large at low values of % N2 fixation. These large errors may have been partly due to legume and control plants assimilating mineral N of differing 15N enrichment. High mineral N levels associated with low values of % N2 fixation were also shown to reduce reliability of N2 fixation values estimated by the 15N enrichment method. These errors caused potentially greater inaccuracy at low values of % N2 fixation than at high values. To compare N2 fixation means statistically, transformations were necessary to stabilize variance and to impart lower weightings to plots with low values of % N2 fixation.


1997 ◽  
Vol 48 (7) ◽  
pp. 1033 ◽  
Author(s):  
R. B. Thompson ◽  
I. R. P. Fillery

Nitrogen (N) mineralisation from mature subterranean clover (Trifolium subterraneum L.) shoots and roots and from sheep urine and faeces, and N uptake by wheat from the shoots, urine, and faeces, were determined with 15 N in a field study in the Western Australian wheatbelt. Treatments were applied to the soil surface of confined micro-plots in autumn and incorporated into soil immediately before wheat was sown in winter. Mature subterranean clover shoots containing 18 kg N/ha were applied to the soil surface, and root material containing 17 kg N/ha was mixed into soil. 15N-labelled urine and faeces were obtained from housed sheep fed 15N-labelled wheat straw and grain. Urine was applied at the rates of 151 and 301 kg N/ha, and faeces was added at the rate of 47 kg N/ha. There was a loss of 14% of shoot 15N in the 2 months this residue was on the soil surface, although very little mineralisation occurred. On the assumption that wind-blow caused the initial loss of 15N, 28% of shoot N mineralised in 6 months following incorporation of shoot residues into soil, and crop recovery was 11% of the 15N applied. N mineralisation from the mature roots was 26% in 6 months. NH3 volatilisation from urine, estimated by difference, was 25% for high urine (0·517 mL/cm2) and 33% for low urine (0·258 mL/cm2) application rates, the loss occurring in the first 2 weeks. Wheat uptake was 23% of the high urine 15N and 22% of the low urine 15N. Leaching losses from unplanted micro-plots were approximately 25-30% of urine 15N. In contrast, leaching losses from planted micro-plots were estimated to be approximately 10% of urine 15N. Approximately 30% of faecal N was mineralised and recovery of faeces N by wheat was 1% of applied 15N. The relative contributions of these components to N turnover in the ley pasture wheat rotation are discussed. It is concluded that assessments of the potential turnover of N in pastures to cropping phases need to consider the low rates of N mineralisation of above-ground herbage, the potential for supply of N from the total root system, the effect of grazing on NH3volatilisation, and consequent loss of N fixed by legumes.


1987 ◽  
Vol 38 (1) ◽  
pp. 15 ◽  
Author(s):  
GL Turner ◽  
RR Gault ◽  
L Morthorpe ◽  
DL Chase ◽  
FJ Bergersen

The natural abundances (S15N with reference to atmospheric N2) of the stable isotope of nitrogen (15N) in the total nitrogen and in KCl-extractable mineral nitrogen (typically 96% NO-3-N and 4% NH+4-N) were measured in the surface 10 cm of a transitional red earth at Yanco, N.S.W., and of a grey soil of heavy texture at Trangie, N.S.W. Measurements were made in Autumn (May), prior to planting crops of winter oats, at the time of harvest (October) and in December, using both cropped and continuously fallowed soils. At Trangie, additional measurements were made in September, near the beginning of rapid growth in spring. Despite differences in soil type, pH .and location, both sites showed: (i) S15N in extractable mineral nitrogen varied with time (decreasing from 18.7 to 6.0% in fallowed soil at Yanco, and increasing from 5.8 to 12.0%~ under oats at Trangie), and in cropped versus fallowed treatments (12.0 and 5.3% respectively in December at Trangie), and values were different from those of the total soil nitrogen, in which S15N remained virtually unchanged (over all times and sites, S15N = 8.2 � 0.2 at Trangie); (ii) after removal of the crop, S15N in increments of extractable mineral nitrogen were higher than in the total nitrogen of previously cropped soils, whilst in the continuously fallowed soils, S15N of extractable mineral nitrogen was lower than in the total nitrogen. In addition, at Trangie, S15N in the extractable mineral nitrogen was highest late in growth of the oat crop, and this was reflected in the values for S15N of nitrogen assimilated in the crop. Values of the S15N of plant nitrogen agreed well with the S15N of extractable mineral N when the former were determined in increments of plant N during fixed periods of growth and plotted appropriately (the mid-point between sampling times) in relation to the time courses of changes in the mineral N. These results are discussed in relation to the use of 15N natural abundance techniques for estimating nitrogen fixation by nodulated legumes and in the study of other aspects of soil nitrogen cycle processes.


1994 ◽  
Vol 45 (1) ◽  
pp. 133 ◽  
Author(s):  
JS Pate ◽  
MJ Unkovich ◽  
EL Armstrong ◽  
P Sanford

The 15N natural abundance (S15N) of the shoot total N of a range of non-N2 fixing potential reference species was compared with that of nodulated field pea (Pisum sativum L.), narrow leafed lupin (Lupinus angustijolius L.) or subterranean clover (Trijolium subterraneum L.) across a range of field sites, to which N fertilizers had not been applied in the season of study. Shoot S15N values of reference species lay mostly within the range from +3 to +5%o and there was some evidence of lower S15N values in gramineaceous than dicotyledonous non-legume species. Continuous sampling within crops of each legume showed S15N values to differ consistently between and within potential reference species through the season. The S15N values of seedlings of four non legume species in a lupin crop were closer to that of soil N03-N (S15N = 4.2%o) than soil NH4-N (S15N = 7.9%0). Shoot S15N values of non-nodulated pea, lupin and subterranean clover, and a range of possible reference species all sand-cultured on a defined nitrate source (S15N = 7.5%), suggested little or no discrimination during utilization of nitrate. However, when four candidate reference species were sand cultured with nodulated actively fixing subterranean clover on the same nitrate source, the S15N of the ryegrass was lowered significantly below that of the NO3. Field plot comparisons of nine potential reference species with nodulated field pea showed certain species to resemble field pea more closely than others in terms of the S15N value of their shoots. Reference plants sampled within or well outside the rooting zone of an actively fixing legume (subterranean clover, field pea or lupin) showed significantly lower shoot S15N of mixed grass components when harvested in root contact with, as opposed to well distant from, subterranean clover. A similar effect was observed for barley within v. outside the rooting zone of pea, but no such effects were observed between capeweed and subterranean clover, pea and radish, or for any of five reference plants matched with lupin. The data are utilized to select appropriate reference plants for field assessments of N2 fixation under south-west Australian conditions.


1998 ◽  
Vol 49 (3) ◽  
pp. 459 ◽  
Author(s):  
M. B. Peoples ◽  
R. R. Gault ◽  
G. J. Scammell ◽  
B. S. Dear ◽  
J. Virgona ◽  
...  

The effects of different management regimes on N2 fixation by subterranean clover (Trifolium subterraneum) in annual pastures and lucerne (Medicago sativa) in perennial-based pastures were examined in 5 experiments and 55 commercial paddocks, in which the pastures were grown in phased rotation with crops. The objectives were to quantify the inputs of fixed N2 and to determine ways of increasing nitrogen (N) inputs into ley-farming systems of southern New South Wales and north-eastern Victoria. Estimates of annual amounts of N2 fixed, based on above-ground herbage production in grazed pastures, ranged from 5 to 238 kg N/ha for subterranean clover and from 47 to 167 kg N/ha for lucerne. Legume reliance upon N2 fixation for growth (Pfix) was high (>65%) in most annual and perennial pastures examined. The levels of Pfix were generally unaffected by management treatments. As a consequence the amounts of N2 fixed were predominantly regulated by the legume content and herbage yield of pastures rather than by any marked differences in the ability of the legume to fix N. When all experimental results were combined with on-farm measurements of N2 fixation, the data indicated that lucerne and subterranean clover fixed 22-25 kg N for every tonne of legume dry matter produced. Management inputs to annual pastures which improved the productivity of subterranean clover and the amounts of N2 fixed included applications of superphosphate and the removal of grass species with herbicide, although the response to these treatments was not consistent across all sites in all years. Potential inputs from N2 fixation were high in annual pastures, and improved management during a good clover season enhanced the levels of mineral N detected in the soil profile (0-200 cm) the following autumn by 100-200 kg N/ha. However, year-to-year variability in annual pasture productivity and clover content resulted in large fluctuations in amounts of N2 fixed. Perennial pastures containing lucerne provided consistently greater annual herbage production, had more stable legume contents, and fixed on average 90-150% more N2 than neighbouring subterranean clover-based pastures. Even during the 1994 drought when annual pastures failed, lucerne still managed to fix >70 kg N/ha. It is proposed that lucerne-based pastures could represent a more reliable means of improving soil fertility for subsequent crops than annual pastures.


1984 ◽  
Vol 35 (4) ◽  
pp. 539 ◽  
Author(s):  
JH Silsbury ◽  
D Zuill ◽  
PH Brown

Effects of constant temperatures of 10, 15, 20, 25 and 30�C on the germination, emergence and early vegetative growth of Trifolium subterraneum L. cv. Mt Barker grown as swards were examined in temperature-controlled glasshouses and in a growth cabinet. Seedlings were established at a density of about 2000 plants m-2 and grown for up to 70 days. Plants were either inoculated and grown without mineral nitrogen (-N), or supplied with 7.5 mM NO-3 (+ N). Percentage germination and emergence were hardly affected by temperatures of 10-20�C, but at 25�C were reduced to 50%, and at 30�C to about 10%. The rates of germination and emergence were slowest at 10�C, but showed little change with temperature over the range 15-30�C. Time to closed canopy (leaf area index 3) and time to a dry weight of 133 g m-2 were shorter where plants were supplied with NO; than where mineral nitrogen was withheld and a symbiotic system established. Rates of N2-fixation, as measured by acetylene reduction assay, were not markedly affected by temperature over the range 10-25�C. Relative efficiency ranged from about 0.55 at 10, 15, and 20�C to about 0.66 at 25�C. At 30�C nodulation still occurred, but nitrogenase activity was very slight. It is concluded that, where swards of subterranean clover are grown in the absence of any mineral N, a period of N-starvation limits growth during the time taken for symbiotic N2-fixation to become established. Such retardation of growth is small at about 20�C, but becomes more marked at lower and higher temperatures. The establishment of subterranean clover swards in soils of low N status are likely to be retarded following an early (March) or a late (July) start in the growing season. In such cases a 'starter' application of mineral nitrogen may promote the early growth of the legume.


2004 ◽  
Vol 44 (9) ◽  
pp. 883 ◽  
Author(s):  
J. Braschkat ◽  
P. J. Randall

Excess cation concentrations (total cations – total inorganic anions) are reported for roots and shoots of 16 plant species of importance in pastures in south-eastern Australia. This information is required for the calculation of acidification in grazed pasture systems. The excess cation concentrations for shoots at flowering were [cmol(+)/kg]: perennial grasses — Lolium perenne (perennial ryegrass) 50, Phalaris aquatic (phalaris) 51, Danthonia richardsonii (wallaby grass) 30, Dactylus glomerata (cocksfoot) 62, Holcus lanatus (Fog grass) 60; annual grasses — Lolium rigidum 29, Vulpia bromoides (vulpia) 40, Hordeum leporinum (barley grass) 46, Bromus mollis (soft brome) 59; perennial legumes — Medicago sativa (lucerne) 115, Trifolium repens (white clover) 147; annual legumes — Trifolium subterraneum (subterranean clover) 142, Medicago truncatula (barrel medic) 114, Ornithopus sativus (serradella) 137; weeds — Arctotheca calendula (cape weed) 165, Echium plantagineum (Paterson’s curse) 169. Values for roots were in the same order as shoots in vulpia and wallaby grass but lower for the other species, varying between 26 and 62% of the shoot value in grasses and 29 and 49% in legumes. For a subset of 4 legumes and 3 grasses, the excess cation concentrations in shoots were measured over the main production period in spring. Excess cation concentrations generally declined during the season, with the change being relatively larger in grasses than legumes.


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