N2-fixation and residual N effect of four legume species and four companion grass species

Inclusion of forage legumes in low-input forage mixtures improves herbage production and soil fertility through addition of nitrogen (N) from N2-fixation. The impact of different grass-legume mixtures on the N contribution of the forage mixture has rarely been investigated under comparable soil and climatic conditions. We conducted a field experiment on a sandy soil at two nitrogen levels with seven two-species forage mixtures: alfalfa, bird’s-foot trefoil, red clover, or white clover in mixture with perennial ryegrass, and white clover in mixture with meadow fescue, timothy, or hybrid ryegrass. We found high N2-fixation of more than 300 kg N ha-1 from both red clover and alfalfa even when the two mixtures received 300 kg total-N ha-1 in cattle slurry. The addition of cattle slurry N fertilizer lowered N2-fixation for white clover and red clover as expected, but for bird’s-foot trefoil and alfalfa no changes in the proportion of N derived from N2-fixation was observed. We conclude that the competition for available soil N from perennial ryegrass in mixture was an important factor for the proportion of N in alfalfa, white clover, and bird’s-foot trefoil obtained from N2-fixation. White clover had a high proportion of N derived from atmosphere for all companion grasses despite significant differences in white clover proportion. Although the perennial ryegrass-alfalfa mixture in the grass phase yielded more than twice the N from N2-fixation compared to white clover in the perennial ryegrass mixture, this did not in the following year lead to higher residual N effects of alfalfa. Both in terms of N yield in the grass phase and N yield in the subsequent spring barley red clover contributed most to the improvement of soil N fertility

15Nitrogen uptake from shallow- versus deep-rooted plants in multi-species mixtures and monoculture grassland

Only few studies have explored the importance of functional diversity in temperate agricultural grasslands in relation to nitrogen (N) uptake. This study investigates the consequence of growing deep-rooted plants together with grass-clover mixtures in terms of N uptake efficiency from deep soil layers. The objective was to compare the N uptake of the shallow-rooted grassland species Lolium perenne and Trifolium repens; and the deep-rooted species Cichorium intybus and Medicago sativa in monocultures and mixtures. We hypothesized that growing deep-rooted plant species in mixture with shallow-rooted species increases the N uptake from deep soil layers partly through competition. A 15N tracer study was carried out with 15N enriched ammonium-sulphate placed at three different soil depths (40, 80 and 120 cm). To recover 15N, above-ground plant biomass was harvested after 10 days. We described the decline of 15N uptake with depth by using an exponential decay function. The studied plant communities showed the same relative decline in 15N uptake by increasing soil depths, but different capacities in total 15N uptake. Monoculture L. perenne foraged less 15N in all depths compared to the other four plant communities. The relative 15N uptake of individual plant species grown in mixture decreased stronger with depth than in monoculture. Thus, both findings rejected our hypothesis

15Nitrogen transfer from legumes to neighbouring plants in multi-species grassland

This study investigates the N transfer from legumes to neighbouring plants, grasses, legumes and herbs in a temperate grassland. In a field experiment white clover (Trifolium repens), red clover (Trifolium pratense) and lucerne (Medicago sativa) were leaf-labelled with 15N enriched urea. The 15N tracer was measured in above-ground plant tissue of eight neighbouring plants in two subsequent harvests in 2008. The three legumes donated 15N to all neighbouring plants, of which grasses, white and red clover were strong receivers. Results show that N transfer increases with N application and from the 1st to the 2nd cut

Forage herbs improve mineral composition of grassland herbage

Provision of an adequate mineral supply in the diets of ruminants fed mainly on grassland herbage can present a challenge if mineral concentrations are suboptimal for animal nutrition. Forage herbs may be included in grassland seed mixtures to improve herbage mineral content, although there is limited information about mineral concentrations in forage herbs. To determine whether herbs have greater macro- and micromineral concentrations than forage legumes and grasses, we conducted a 2-year experiment on a loamy-sand site in Denmark sown with a multi-species mixture comprised of three functional groups (grasses, legumes and herbs). Herb species included chicory (Cichorium intybus L.), plantain (Plantago lanceolata L.), caraway (Carum carvi L.) and salad burnet (Sanguisorba minor L.). We also investigated the effect of slurry application on the macro- and micromineral concentration of grasses, legumes and herbs. In general, herbs had greater concentrations of the macrominerals P, Mg, K and S and the microminerals Zn and B than grasses and legumes. Slurry application indirectly decreased Ca, S, Cu and B concentrations of total herbage because of an increase in the proportion of mineral-poor grasses. Our study indicates that including herbs in forage mixtures is an effective way of increasing mineral concentrations in herbage

The secret life of wild animals revealed by accelerometer data: how landscape diversity and seasonality influence the behavioural types of European hares

Context Landscape composition and configuration, as well as seasonal landscape dynamics shape the behaviour, movement and energy expenditure of animals, i.e. foraging, hiding or fleeing, and ultimately survival. Especially in highly modified agricultural systems, it is crucial to understand how animal behaviour is influenced by landscape context to develop sustainable land management concepts. Objectives We show how landscape composition and configuration, together with seasonal dynamics affect animal behavioural types, accounting for the different life-history events in both sexes. Methods We investigated 34 European hares in two contrasting agricultural landscapes (a simple and a complex landscape) by using tri-axial accelerometer data to classify the animals’ behaviour into five categories: resting, foraging, moving, grooming and standing upright (i.e. vigilance behaviour). We tested whether the amount of behaviours per category changed with landscape composition and configuration, season and sex. Results During peak breeding, hares in areas of high habitat diversity rested more, moved less and spent less time searching for resources. During winter, hares moved more and rested less. Females rested less and foraged more in areas with large agricultural fields. Conclusions A complex landscape is particularly important during the breeding season, allowing animals to allocate enough energy into reproduction. In winter, hares in areas of low habitat diversity may not find enough thermal and anti-predator shelter to move as much as they would need to meet their requirements. Hence, high habitat diversity and small field sizes guarantee species persistence in human-altered agricultural areas throughout the year

Circadian rhythms regulate the environmental responses of net CO2 exchange in bean and cotton canopies

Studies on the dependence of the rates of ecosystem gas exchange on environmental parameters often rely on the up-scaling of leaf-level response curves ('bottom-up' approach), and/or the down-scaling of ecosystem fluxes ('top-down' approach), where one takes advantage of the natural diurnal covariation between the parameter of interest and photosynthesis rates. Partly independent from environmental variation, molecular circadian clocks drive ∼24 h oscillations in leaf-level photosynthesis, stomatal conductance and other physiological processes in plants under controlled laboratory conditions. If present and of sufficient magnitude at ecosystem scales, circadian regulation could lead to different results when using the bottom-up approach (where circadian regulation exerts a negligible influence over fluxes because the environment is modified rapidly) relative to the top-down approach (where circadian regulation could affect fluxes as it requires the passage of a few hours). Here we dissected the drivers of diurnal net CO2 exchange in canopies of an annual herb (bean) and of a perennial shrub (cotton) through a set of experimental manipulations to test for the importance of circadian regulation of net canopy CO2 exchange, relative to that of temperature and vapor pressure deficit, and to understand whether circadian regulation could affect the derivation of environmental flux dependencies. Contrary to conventional wisdom, we observed how circadian regulation exerted controls over net CO2 exchange that were of similar magnitude to the controls exerted by direct physiological responses to temperature and vapor pressure deficit. Diurnal patterns of net CO2 exchange could only be explained by considering effects of environmental responses combined with circadian effects. Consequently, we observed significantly different results when inferring the dependence of photosynthesis over temperature and vapor pressure deficit when using the top-down and the bottom up approaches.We remain indebted to E. Gerardeau, D. Dessauw, J. Jean, P. Prudent (Aïda CIRAD), J.-J. Drevon, C. Pernot (Eco&Sol INRA), B. Buatois, A. Rocheteau (CEFE CNRS), A. Pra, A. Mokhtar and the full Ecotron team, in particular C. Escape, for outstanding technical assistance during experiment set-up, plant cultivation and measurements. Earlier versions of the manuscript benefitted from comments by M. Dietze, B. Medlyn, R. Duursma and Y.-S. Lin. This study benefited from the CNRS human and technical resources allocated to the ECOTRONS Research Infrastructures as well as from the state allocation ‘Investissement d'Avenir’ ANR-11-INBS-0001, ExpeER Transnational Access program, Ramón y Cajal fellowships (RYC-2012-10970 to VRD and RYC-2008-02050 to JPF), the Erasmus Mundus Master Course Mediterranean Forestry and Natural Resources Management (MEDfOR) and internal grants from UWS-HIE to VRD and ZALF to AG. We thank the Associate Editor T. Vesala and two anonymous reviewers for their help to improve this manuscript

Effect of four plant species on soil 15N-access and herbage yield in temporary agricultural grasslands

Positive plant diversity-productivity relationships have been reported for experimental semi-natural grasslands (Cardinale et al. 2006; Hector et al. 1999; Tilman et al. 1996) as well as temporary agricultural grasslands (Frankow-Lindberg et al. 2009; Kirwan et al. 2007; Nyfeler et al. 2009; Picasso et al. 2008). Generally, these relationships are explained, on the one hand, by niche differentiation and facilitation (Hector et al. 2002; Tilman et al. 2002) and, on the other hand, by greater probability of including a highly productive plant species in high diversity plots (Huston 1997). Both explanations accept that diversity is significant because species differ in characteristics, such as root architecture, nutrient acquisition and water use efficiency, to name a few, resulting in composition and diversity being important for improved productivity and resource use (Naeem et al. 1994; Tilman et al. 2002). Plant diversity is generally low in temporary agricultural grasslands grown for ruminant fodder production. Grass in pure stands is common, but requires high nitrogen (N) inputs. In terms of N input, two-species grass-legume mixtures are more sustainable than grass in pure stands and consequently dominate low N input grasslands (Crews and Peoples 2004; Nyfeler et al. 2009; Nyfeler et al. 2011). In temperate grasslands, N is often the limiting factor for productivity (Whitehead 1995). Plant available soil N is generally concentrated in the upper soil layers, but may leach to deeper layers, especially in grasslands that include legumes (Scherer-Lorenzen et al. 2003) and under conditions with surplus precipitation (Thorup-Kristensen 2006). To improve soil N use efficiency in temporary grasslands, we propose the addition of deep-rooting plant species to a mixture of perennial ryegrass and white clover, which are the most widespread forage plant species in temporary grasslands in a temperate climate (Moore 2003). Perennial ryegrass and white clover possess relatively shallow root systems (Kutschera and Lichtenegger 1982; Kutschera and Lichtenegger 1992) with effective rooting depths of <0.7 m on a silt loamy site (Pollock and Mead 2008). Grassland species, such as lucerne and chicory, grow their tap-roots into deep soil layers and exploit soil nutrients and water in soil layers that the commonly grown shallow-rooting grassland species cannot reach (Braun et al. 2010; Skinner 2008). Chicory grown as a catch crop after barley reduced the inorganic soil N down to 2.5 m depth during the growing season, while perennial ryegrass affected the inorganic soil N only down to 1 m depth (Thorup-Kristensen 2006). Further, on a Wakanui silt loam in New Zealand chicory extracted water down to 1.9 m and lucerne down to 2.3 m soil depth, which resulted in greater herbage yields compared with a perennial ryegrass-white clover mixture, especially for dryland plots (Brown et al. 2005). There is little information on both the ability of deep- and shallow-rooting grassland species to access soil N from different vertical soil layers and the relation of soil N-access and herbage yield in temporary agricultural grasslands. Therefore, the objective of the present work was to test the hypotheses 1) that a mixture comprising both shallow- and deep-rooting plant species has greater herbage yields than a shallow-rooting binary mixture and pure stands, 2) that deep-rooting plant species (chicory and lucerne) are superior in accessing soil N from 1.2 m soil depth compared with shallow-rooting plant species, 3) that shallow-rooting plant species (perennial ryegrass and white clover) are superior in accessing soil N from 0.4 m soil depth compared with deep-rooting plant species, 4) that a mixture of deep- and shallow-rooting plant species has greater access to soil N from three soil layers compared with a shallow-rooting two-species mixture and that 5) the leguminous grassland plants, lucerne and white clover, have a strong impact on grassland N acquisition, because of their ability to derive N from the soil and the atmosphere

Globalisation as a challenge or opportunity for organic farming

During one intensive week in October 2005, the authors were gathered to discuss the impact that globalisation has on the Organic Food Systems and the opportunities that globalisation opens up for developing these systems. The meeting took place as a Ph.D. course under the auspices of the Research School of Organic Farming and Food Systems (SOAR; www.soar.dk). All participants research within Organic Agriculture and Food Production in one way or another

Herbage dry-matter production and forage quality of three legumes and four non-leguminous forbs grown in single-species stands

Agronomic data on most broad-leaved species of grasslands are scarce. The aim of this study was to obtain novel information on herbage DM yield and forage quality for several forb species, and on species differences and seasonal patterns across harvests and in successive years. Four non-leguminous forbs [salad burnet (Sanguisorba minor), caraway (Carum carvi), chicory (Cichorium intybus) and ribwort plantain (Plantago lanceolata)] and three leguminous forbs [yellow sweet clover (Melilotus officinalis), lucerne (Medicago sativa) and birdsfoot trefoil (Lotus corniculatus)] and a perennial ryegrass–white clover mixture were investigated in a small-plot cutting trial in Denmark during 2009 and 2010. Plots were harvested four times per year. On average, annual herbage yield was highest for lucerne (15.4 DM ha_1) and grass–white clover (15.4 t DM ha_1), and lowest for salad burnet (4.6 t DM ha_1) and yellow sweet clover (3.9 t DM ha_1). Ribwort plantain and lucerne had the highest concentrations of acid detergent fibre (339 and 321 g kg_1 DM respectively) and lignin (78 and 67 g kg_1 DM respectively); contents in other species were similar to grass–white clover (275 and 49 g kg_1 DM respectively). No common feature was found within the functional groups of non-leguminous forbs and leguminous forbs, other than higher crude protein contents (198–206 g kg_1 DM) in the legumes. DM yield and fibre content were lowest in October. Digestibility declined with higher temperature and increasing fibre content. Results are discussed in terms of the potential of forbs to contribute to forage resources in farming practice

Nitrogen yield advantage from grass-legume mixtures is robust over a wide range of legume proportions and environmental conditions

Coordination of this project was supported by the EU Commission through COST Action 852 ‘Quality legume-based forage systems for contrasting environments‘. A636 contribution to the research leading to these results has been conducted as part of the Animal Change project which received funding from the European Union’s Seventh Framework Programme (FP7/2007-20 13) under the grant agreement no. 266018.peer-reviewedCurrent challenges to global food security require sustainable intensification of agriculture through initiatives that include more efficient use of nitrogen (N), increased protein self-sufficiency through home-grown crops, and reduced N losses to the environment. Such challenges were addressed in a continental-scale field experiment conducted over three years, in which the amount of total nitrogen yield (Ntot) and the gain of N yield in mixtures as compared to grass monocultures (Ngainmix) was quantified from four-species grass-legume stands with greatly varying legume proportions. Stands consisted of monocultures and mixtures of two N2 fixing legumes and two non-fixing grasses.The amount of Ntot of mixtures was significantly greater (P ≤ 0.05) than that of grass monocultures at the majority of evaluated sites in all three years. Ntot and thus Ngainmix increased with increasing legume proportion up to one third of legumes. With higher legume percentages, Ntot and Ngainmix did not continue to increase. Thus, across sites and years, mixtures with one third proportion of legumes attained ~95% of the maximum Ntot acquired by any stand and had 57% higher Ntot than grass monocultures.Realized legume proportion in stands and the relative N gain in mixture (Ngainmix/Ntot in mixture) were most severely impaired by minimum site temperature (R = 0.70, P = 0.003 for legume proportion; R = 0.64, P = 0.010 for Ngainmix/Ntot in mixture). Nevertheless, the relative N gain in mixture was not correlated to site productivity (P = 0.500), suggesting that, within climatic restrictions, balanced grass-legume mixtures can benefit from comparable relative gains in N yield across largely differing productivity levels.We conclude that the use of grass-legume mixtures can substantially contribute to resource-efficient agricultural grassland systems over a wide range of productivity levels, implying important savings in N fertilizers and thus greenhouse gas emissions and a considerable potential for climate change mitigation.European Unio