Effects of cessation of grazing on leaf‐level photosynthesis of Periploca laevigata

Abstract. The impact of grazing pressure on gas exchange of Periploca laevigata was studied in relation to its nutritional status. P. laevigata, a woody and highly palatable shrub, is an important forage species in the dry season in Cabo de Gata Natural Park (Almeria, SE Spain) where this study was carried out. Seasonal and diurnal courses of net photosynthesis, transpiration and stomatal conductance were measured in the field. In addition, soil and plant nutrient status were measured as potential factors influencing leaf photosynthetic levels. Early in the growing season, ungrazed P. laevigata had higher photosynthetic activity than grazed plants suggesting that the defoliated plants were still recovering from herbivore damage in the previous summer. Later in the growing season grazed P. laevigata had higher photosynthetic activity than ungrazed plants, suggesting higher metabolic activity and a possible investment in chemical defences. Concentrations of P and N in the soil were higher in the grazed area. The nutrient concentration of the leaves of P. laevigata was similar in grazed and ungrazed plants.     

The Role of White Clover in the Loss of Diversity in Grassland Habitat Restoration

A field experiment was designed to recreate a species‐rich mesotrophic grassland community of conservation worth. Trifolium repens (white clover) was observed to increase significantly in both frequency and abundance in sown plots grazed by cattle, but not in plots cut in June and subsequently grazed by cattle. In both these treatments permanent quadrats containing clover patches were found to be lower in species richness than were quadrats without clover. In both treatments botanical diversity was seen to decline over time. In the grazed‐only treatment the loss of diversity may be linked to the increase in clover. In the cut and grazed plots, T. repens did not become so abundant but diversity was still seen to decline, possibly due to the loss of low growing species from the taller sward. A pot experiment which varied the sowing density of a mix of seven wild flower species in full factorial combination with cutting frequency was established on soils from an arable field also sown with a single density of clover. T. repens was seen to decline from initial high cover estimates in infrequently cut and uncut treatments. In the pot experiment where a grass component to the vegetation was absent, clover was seen to have less impact on the other forbs than it did in the field. It is suggested that, being a nitrogen fixer, T. repens may have a competitive edge in ex‐arable soils low in available nitrogen. The observed reduction in botanical diversity may be a result of this increase in available nitrogen, facilitating the spread of the sown grasses and preventing the recovery of the sown forbs that were excluded by the invasion of T. repens. It is suggested that reducing the proportion of grass in the seed mixtures during grassland habitat creation on these soils may help reduce or delay this effect.     

Effects of some systemic imidazole and triazole fungicides on white clover and symbiotic nitrogen fixation by Rhizobium trifolii

Effects of prochloraz, imazalil, diclobutrazol, triadimefon and propiconazole were studied. Compared with most systemic fungicides previously tested they were all relatively toxic to R. trifolii on agar. Effects of soil residues of the compounds on the growth and nitrogen fixing capacity of white clover were measured. The lowest concentration of the fungicides used was that which would be present if all the active ingredient from a single application remained unchanged and evenly distributed in the top 5 cm of soil. This concentration of diclobutrazol decreased the size and weight of 11-wk-old clover plants. The rate of nitrogen fixation was reduced by twice this concentration and root nodulation by four times this amount. None of the other fungicides had any significant effect. The possibility of field application of diclobutrazol to cereals causing damage to clover crops is discussed. It is questionable whether harmful soil residues will ever accumulate from normal usage of the fungicide but exceeding the recommended rate of application, or application to cereals undersown with clover, might cause undesirable effects on the clover.     

Symbiotic and competitive properties of motility mutants of Rhizobium trifolii TA1

Non-motile mutants of Rhizobium trifolii defective in either flagellar synthesis or function were isolated by transposon Tn5 mutagenesis. they were indistinguishable from motile control strains in growth in both laboratory media and in the rhizosphere of clover roots. When each non-motile mutant was grown together with a motile strain in continuous culture, the numbers of motile and non-motile organisms remained in constant proportion, implying that their growth rates were essentially identical. When inoculated separately onto clover roots, the mutants and wildtype did not differ significantly in the number of nodules produced or in nitrogen fixing activity. However, when mixtures of equal numbers of mutant and wild-type cells were inoculated onto clover roots, the motile strain formed approximately five times more nodules than the flagellate or non-flagellate, non-motile mutants, suggesting that motility is a factor in competition for nodule formation.     

Studies on symbiotic nitrogen fixation by a new strain of tetraploid red clover (UO36)

Summary Some characteristics of the symbiotic nitrogen fixation by tetraploid red clover (UO36) have been studied in laboratory experiments. Cross-inoculations were made on tetraploid and diploid clover plants usingRhizobium isolates from both types of clover. Nodulation activity and nitrogen fixation were determined. No sign of specific strains necessary for optimal fixation of nitrogen by theUO36-symbiosis was observed in comparison to the diploid clover strain.Rhizobium cultures effective in connection with cultivation of diploid clover will probably also be effective when cultivating tetraploid clover strains.     

Dietary preference of dairy heifers grazing ryegrass and white clover,with and without an anti-bloat treatment

The effect of an anti-bloat treatment on the dietary preference of non-pregnant Holstein×Friesian heifers for ryegrass and white clover was studied. Eight groups of two heifers grazed 0.51 ha plots containing adjacent monocultures of grass alongside clover with either 25 or 75% clover (by ground area) and either with or without a monensin intra-ruminal anti-bloat device. Preference was measured by recording the time spent grazing grass or clover between dawn and dusk by manual scan sampling at 2 min intervals over a 48 h period. The observers also recorded a visual assessment of bloat (on a scale of 0–5). The heifers selected 51.2% clover when offered 25% clover and 76.6% clover when offered 75% clover. Analysis showed an overall partial preference for clover of 63.9% (mean value across all treatments), although the relative abundance of clover did influence preference. There was a diurnal pattern to preference, with a stronger preference for clover in the morning, and the preference for grass increasing towards the evening. These results are very similar to those previously reported for sheep, suggesting that there may be a common biological basis for partial preference for grass and clover in sheep and cattle. None of the animals showed any significant signs of bloat, and the anti-bloat treatment had no effect on the proportion of clover selected. This indicates that avoidance of moderate or severe bloat is unlikely to be the mechanism for partial preference in cattle. However, it is possible that mild, sub-clinical bloat could still play a role in diet selection.     

Measurement of biological dinitrogen fixation in grassland: Comparison of the enriched 15N dilution and the natural 15N abundance methods at different nitrogen application rates and defoliation frequencies

A plant mixture of white clover (Trifolium repens L.), red clover (Trifolium pratense L.), and ryegrass (Lolium perenne L.) was established in the spring of 1991 under a cover-crop of barley. Treatments were two levels of nitrogen (400 and 20 kg N ha^-1) and two cutting intensities (3 and 6 cuts per season). Fixation of atmospheric derived nitrogen was estimated by two ¹⁵N dilution methods, one based on application of ¹⁵N to the soil, the other utilising small differences in natural abundance of ¹⁵N.Both methods showed that application of 400 kg N ha^-1 significantly reduced dinitrogen fixation, while cutting frequency had no effect. Atmospheric derived nitrogen constituted between 50 and 64% of harvested clover nitrogen in the high-N treatment, while between 73% and 96% of the harvested clover nitrogen was derived from the atmosphere in the low-N treatment. The amounts of fixed dinitrogen varied between 31–72 kg N ha^-1 and 118–161 kg N ha^-1 in the high-N and low-N treatment, respectively. The highest values for biological dinitrogen fixation were estimated by the enriched ¹⁵N dilution method.Estimates of transfer of atmospheric derived nitrogen from clover to grass obtained by the natural ¹⁵N abundance method were consistently higher than those obtained by the enriched ¹⁵N dilution method. Neither mineral nitrogen application nor defoliation frequency affected transfer of atmospheric derived nitrogen from clover to grass.Isotopic fractionation of ¹⁴N and ¹⁵N (B value) was estimated by comparing results for nitrogen fixation obtained by the enriched ¹⁵N dilution and the natural ¹⁵N abundance method, respectively. B was on average +1.20, which was in agreement with a B value determined by growing white clover in a nitrogen free media.     

Acetylene reduction assay on white clover genotypes and on grass/clover swards

Acetylene reduction assay was used to measure the nitrogenase activity of white clover genotypes in pots and of grass/clover swards in situ. Much of the variation in nitrogenase activity of single genotypes and hybrid populations was associated with plant dry weight. After adjustment for plant dry weight it was concluded that there was limited scope for selection for increased nitrogenase activity. In plant breeding this technique would seem to have greatest application in the selection for continued nitrogen fixation activity in the presence of inhibitory factors such as high levels of mineral nitrogen. The in situ studies revealed differences in nitrogenase activity of grass/clover swards based on contrasting cultivars of white clover. These differences were due to variation in clover density and also to variation in activity per unit clover dry weight. It was concluded that the variation in nitrogenase activity per unit dry weight reflected differences in growth pattern of the cultivars in the autumn when these assays were made. The in situ studies offer a means whereby the nitrogenase activity of cultivars and selected families can be monitored under varying levels of mineral nitrogen and other husbandry treatments without disruption of the sward.     

Bacteriophage in relation to nitrogen fixation by red clover

Summary Cultures of Rhizobium trifolii resistant to the action of a given bacteriophage, may vary appreciably in their nitrogen fixing ability in association with the proper host plant. Likewise, cultures of Rh. trifolii, sensitive to bacteriophage may or may not benefit the host plant as judged by nitrogen fixation.Since sensitive and resistant cultures may be comparable in their nitrogen fixing capacity, it appears that the behavior of a culture of Rh. trifolii toward the lytic action of a specific bacteriophage in vitro cannot be correlated with its nitrogen fixing ability in association with the host plant.If a bacteriophage is added to a sensitive strain of Rh. trifolii used for inoculation of red clover, the cultures recovered from the nudules are often only of the resistant type. When this occurs the nitrogen fixed through association of plant and bacteria is decreased in the case of a good strain, but is unaffected if the strain is of the poor type. The addition of phage to a resistant strain of Rh. trifolii used for inoculation of red clover plants does not change the resistant type of culture recovered from the nodules, nor is the fixation of nitrogen by plant and bacteria affected.     

Uptake of 32P labelled phosphate by clover and ryegrass growing in mixed swards with different nitrogen treatments

Lolium perenne cv. S.23 and Trifolium repens cv. Olwen were sown together in 1975, fertilised then and in 1976, and finally given nitrogen doses of either 50, 100, 200 or 400 kg/ha (as N) combined with 0.64 times as much potassium (as K₂O) in 1977. As nitrogen increased, grass yield increased, but clover decreased. Grass roots absorbed more ³²P than clover roots, and nitrogen increased this difference. Grass roots bore more mycorrhiza than clover roots. The difference in ³²P uptake between grass and clover was less in June and July than in August. Clover roots took up most phosphate from the upper layers of soil, while grass absorbed ³²P rather uniformly down to 25 cm. It was concluded that optimum fertiliser placement for clover growth was a surface dressing in the early season.     

Functional plasticity of Trifolium repens L. in response to sulphur and nitrogen availability

Recent control of atmospheric SO₂ pollution is leading to important soil sulphur impoverishment. Plasticity could be a mechanism allowing species to adapt to this rapid global change. Trifolium repens L. is a key grassland species whose performances in community are strongly linked to nitrogen availability. Plasticity of three white clover lines contrasting in their ability to use atmospheric N₂ or soil N was assessed by analysing a set of functional traits along a gradient of nitrogen and sulphur fertilisation applied on a poor soil. White clover traits showed high morphological and physiological plasticity. Nitrogen appeared to be the most limiting factor for the VLF (Very Low Fixation) line. S was the element that modulated the most traits for the nitrogen fixing lines NNU (Normal Nitrate Uptake) and LNU (Low Nitrate Uptake). As expected, N fertilisation inhibited white clover fixation, but we also observed that N₂ fixation was enhanced when S was added. S fertilisation increased nodule length as well as the proportion of nodules containing leghaemoglobin. S fertilisation, with a direct effect and an indirect effect through N₂ fixation, increases white clover performances particularly with regards to photosynthesis and potential vegetative reproduction. The important plasticity in response to S availability should allow it to adapt to a large range of abiotic conditions, but its sensitivity to S nutrition would be a disadvantage for competition in a situation of soil sulphur impoverishment. In contrast, S fertilisation could help maintain this species when nitrogen status is against it.     

How season of grazing and herbivore selectivity influence monsoon tall-grass communities of northern Australia

Abstract. The hypothesis that season of defoliation and herbivore selectivity may be as important as level of use in determining plant community response to grazing was tested in a monsoon grassland in northern Australia. Plots, dominated by the tussock grasses Themeda triandra and Chrysopogon fallax, were grazed by cattle at low, medium and high rates of utilization in either the early wet, late wet or dry seasons. Effects of grazing on species composition were greatest in the early wet season when high rates of utilization significantly reduced the proportion and occurrence of Themeda and increased the proportion of forbs. Grazing in the dry season had no significant effect on composition. At medium and high levels of utilization in the early wet season, the pasture responded negatively to defoliation, only partially compensating for plant tissue lost to herbivory. The negative response to defoliation carried over to the next wet season when these same medium and high-grazing treatments produced only 80 % and 60 % growth, respectively, of that in treatments grazed at low levels of utilization or those grazed during the dry season. The frequency of Themeda was still lower, and that of annual grasses and non-leguminous forbs higher, in plots that had been grazed at a high rate of utilization for just eight weeks in the early wet season two years previously. Species richness and diversity were also significantly affected by this grazing disturbance. If species composition is to be maintained in these grasslands then stocking rates must be set at low levels to cope with the combined effect of undercompensation in response to defoliation in the wet season and strong dietary preferences for grazing sensitive species.     

Consequences of including adapted white clover in northern European grassland: transfer and deposition of nitrogen

The interspecific transfer of nitrogen (N) between white clover (Trifolium repens) and smooth meadow grass (Poa pratensis) in legume-based grasslands was assessed under North European field conditions using ¹⁵N individual plant leaf labelling. On average 50% of N in the grass was transferred from the white clover and about 6% of N in white clover was transferred from the grass. This corresponds to 2.5 and 0.3 g N m⁻² being transferred over the growing season between the two species, respectively, and demonstrates that a significant part of the total N of the grass is coming through interspecific transfer. The majority of the ¹⁵N transferred was within a period of 20 days at relatively low soil temperatures. This implies that there is a need for a new focus on direct transfer pathways or exudation and transfer of organic N sources. Rhizodeposition in the top 10 cm of the soil was found to be 2.98 g N m⁻² on average over the growing season for the grass and white clover mixture. Inclusion of adapted white clover varieties in the low-input grassland systems of northern Europe will lead to a substantial contribution of N. Responsible Editor: Euan K. James.     

Growth responses of arctic graminoids following grazing by captive lesser snow geese

The effects of grazing by captive goslings of the Lesser Snow Goose on coastal vegetation at La Pérouse By. Manitoba were investigated. Swards of Carex subspathacea, Festuca rubra and Calamagrostis deschampsioides were grazed once for different periods (0–180 min) and regrowth of vegetation determined, based on measurements of standing crop, net above-ground primary production (NAPP) and forage quality (leaf nitrogen content). The amounts of foliage removed from swards of Carex subspathacea increased with the length of the grazing period, but after 44 days of regrowth there were no significant differences in above-ground biomass between control and grazed plots. While the amount of foliage removed by goslings from swards of Festuca rubra increased with the length of the grazing period (except after 150 min of grazing), the increase in biomass following defoliation was similar among treatments. Goslings removed little biomass from swards of Calamagrostis deschampsioides, even when the opportunity for grazing was 180 min. No significant differences in standing-crop or NAPP between grazed and ungrazed plots were detected by the end of summer. Grazing had no significant effect on amounts of nitrogen in leaf tissue of all species, suggesting that faecal nitrogen was not rapidly incorporated into plant biomass within the growing season. Patterns of regrowth of these species are compared to that of Puccinellia phryganodes. An increase in goose numbers in recent years has led to birds foraging on less preferred species, such as Calamagrostis deschampsiodes and Festuca rubra. Their poor nutritional quality and a lack of a rapid growth response following defoliation may explain, in part, the decline in the weight of wild goslings recorded over the last decade.     

Soil characteristics of Rocky Mountain National Park grasslands invaded by Melilotus officinalis and M. alba

Aim Invasion of nitrogen‐fixing non‐native plant species may alter soil resources and impact native plant communities. Altered soils may be the driving mechanism that provides a suitable environment to facilitate future invasions and decrease native biodiversity. We hypothesized that Melilotus invasion would increase nitrogen availability and produce soil microclimate and biochemical changes, which could in turn alter plant species composition in a montane grassland community. Location Our research addressed the effects of white and yellow sweet clover (Melilotus officinalis and M. alba) invasion on soil characteristics and nitrogen processes in the montane grasslands in Rocky Mountain National Park. Methods We sampled soil in replicate sites of Melilotus‐invaded and control (non‐invaded) patches within disturbed areas in montane grassland habitats. Soil composites were analysed for available nitrogen, net nitrogen mineralization, moisture, carbon/nitrogen (C : N ratio), texture, organic matter and pH. Data were recorded at three sample dates during the growing seasons of 1998 and 1999. Results Contrary to our expectations, we observed lower nitrogen availability and mineralization in invaded patches, and differences in soil moisture content and soil C : N. Soil C : N ratios were higher in invaded plots, in spite of the fact that Melilotus had the lowest C : N ratios of other plant tissue analysed in this study. Main conclusions These findings provide land managers of natural areas with a better perspective on the possibilities of nitrogen‐fixing species impact on soil nutrient levels.     

Free amino acid, ammonium and nitrate concentrations in soil solutions of a grazed coastal marsh in relation to plant growth

Soluble free amino acids, ammonium and nitrate ions as sources of nitrogen for plant growth were measured in soils of a coastal marsh grazed by snow geese in Manitoba, Canada. Amounts of nitrogen, primarily ammonium ions, increased in the latter half of the growing season and over winter, but fell to low values early in the growing season. Free amino acid concentrations relative to ammonium concentrations were highest during the period of rapid plant growth in early summer, especially in soils in the intertidal zone, where the median ratio of amino acid nitrogen to ammonium nitrogen was 0·36 and amino acid concentrations exceeded those of ammonium ions in 24% of samples. Amino acid profiles, which were dominated by alanine, proline and glutamic acid, were similar to goose faecal profiles. In a continuous flow hydroponic experiment conducted in the field, growth of the salt‐marsh grass, Puccinellia phryganodes, on glycine was similar to growth on ammonium ions at an equivalent concentration of nitrogen. When supplies of soil inorganic nitrogen are low, amino acids represent a potentially important source of nitrogen for the re‐growth of plants grazed by geese and amino acid uptake may be as high as 57% that of ammonium ions.     

Investigations into nitrogen sources and supply in reclaimed colliery spoil

Summary Two trials were established to investigate the supply of nitrogen from ammonium and nitrate fertilizers, slow release nitrogen fertilizers, an organic nitrogen fertilizer and a legume, at two phosphate levels, to eight grass cultivars on colliery spoil. Spoil nitrogen supply and chemical characteristics and herbage dry matter and nitrogen yields were monitored for up to seven years. pH and conductivity fell at both sites. pH trends appeared to be independent of nitrogen treatment. Nitrogen in the ammonium form gave better yields than in the nitrate form when 125 kg N ha^–1 was supplied in a season but there was no difference when 62.5 kg N ha^–1 was applied. Slow release forms of nitrogen gave better yields more evenly distributed over the season than one application of ammonium sulphate per season. Once established white clover (Trifolium repens) plots had a more consistent nitrogen supply, more evenly distributed yield and better quality herbage than nitrogen fertilizer plots.Lolium perenne yielded poorly at low fertility.Festuca rubra andAgrostis castellana, although establishing slowly, yielded well under high and low fertility.     

Regulation of symbiotic nitrogen fixation in root nodules of alfalfa (Medicago sativa) infected with Rhizobium meliloti

Symbiotic nitrogen fixation of Rhizobium meliloti bacteroids in Medicago sativa root nodules was suppressed by several inorganic nitrogen sources. Amino acids like glutamine, glutamic acid and aspartic acid, which can serve as sole nitrogen sources for the unnodulated plant did not influence nitrogenase activity of effective nodules, even at high concentrations.Ammonia and nitrate suppressed symbiotic nitrogen fixation in vivo only at concentrations much higher than those needed for suppression of nitrogenase activity in free living nitrogen fixing bacteria. The kinetics of suppression were slow compared with that of free living nitrogen fixing bacteria. On the other hand, nitrite, which acts as a direct inhibitor of nitrogenase, suppressed very quickly and at low concentrations. Glutamic acid and glutamine enhanced the effect of ammonia dramatically, while the suppression by nitrate was enhanced only slightly.     

Effects of copper deficiency on root nodules of subterranean clover

Summary Effects of copper deficiency on the development of subterranean clover root nodules were studied. These included a depression of the bacteroid content, an accumulation of starch in the invaded cells and a reduction in the rate of development of the nodule apex. A similar pattern of increasing severity of metabolic effects in the youngest tissues was also found in leaves, but at a later stage of plant development than in nodules. At low copper supply the copper content of tissues and the activity of known copper-containing enzymes was depressed. The relevance of the observations to the disturbance of metabolism within the low copper nodules was discussed. It was suggested that low activity of copper containing enzymes (e.g. cytochrome c oxidase) might be important for maintenance of low oxygen tension within the nodule cells essential for nitrogen fixing activity. Most of the data in this paper are taken from the thesis submitted by B. Cartwright for the degree of Ph.D. of the University of Nottingham.     

Competition between soil- and seed-borneRhizobium trifolii in nodulation of introducedTrifolium subterraneum

Summary Native rhizobia associated withTrifolium albopurpureum, T. bifidum, T. ciliolatum, T. depauperatum, T. dichotomum, T. flavulum, T. melanthum, T. microcephalum, T. microdon, T. oliganthum andT. tridentatum were found in Northern California range soils. These rhizobia nodulate subterranean clover but are ineffective in nitrogen fixation with this host. Native rhizobia compet with those in commercial inoculants to form nodules. To ensure effective nodulation by nitrogen fixing rhizobia, commercial inoculants should be applied at rates greater than those recommended by the manufacturerse Effective nodulation was achieved by an application of 7.5×10⁴ rhizobia per seed, four times the recommended rate.