Synergistic effects of grass competition and insect herbivory on the weed Rumex obtusifolius in an inundative biocontrol approach.

Outcomes of weed biological control projects are highly variable, but a mechanistic understanding of how top-down and bottom-up factors influence the success of weed biological control is often lacking. We grew Rumex obtusifolius, the most prominent native weed in European grasslands, in the presence and absence of competition from the grass Lolium perenne and subjected it to herbivory through targeted inoculation with root-boring Pyropteron spp. To explore whether the interactive effects of competition and inundative biological control were size-dependent, R. obtusifolius was planted covering a large range of plant sizes found in managed grasslands. Overall, competition from the grass sward reduced aboveground biomass and final root mass of R. obtusifolius about 62- and 7.5-fold, respectively, and increased root decay of R. obtusifolius from 14 to 58%. Herbivory alone increased only root decay. However, grass competition significantly enhanced infestation by Pyropteron spp. and, as a consequence, enhanced the impact of herbivory on aboveground biomass and final root mass. The synergistic effect was so strong that R. obtusifolius plants grown from initially smaller roots did no longer develop. Inoculating R. obtusifolius with Pyropteron species in grasslands should be further pursued as a promising inundative biological control strategy in the weed's native range.

Can the soil seed bank of Rumex obtusifolius in productive grasslands be explained by management and soil properties?

Rumex obtusifolius is a problematic weed in temperate grasslands worldwide as it decreases yield and nutritional value of forage. Because the species can recruit from the seed bank, we determined the effect of management and soil properties on the soil seed bank of R. obtusifolius in intensively managed, permanent grasslands in Switzerland (CH), Slovenia (SI), and United Kingdom (UK). Following a paired case-control design, soil cores were taken from the topsoil of grassland with a high density of R. obtusifolius plants (cases) and from nearby parcels with very low R. obtusifolius density (controls). Data on grassland management, soil nutrients, pH, soil texture, and density of R. obtusifolius plants were also collected. Seeds in the soil were germinated under optimal conditions in a glasshouse. The number of germinated seeds of R. obtusifolius in case parcels was 866 ±152 m-2 (CH, mean ±SE), 628 ±183 m-2 (SI), and 752 ±183 m-2 (UK), with no significant difference among countries. Densities in individual case parcels ranged from 0 up to approximately 3000 seeds m-2 (each country). Control parcels had significantly fewer seeds, with a mean of 51 ±18, 75 ±52, and 98 ±52 seeds m-2 in CH, SI, and UK, respectively, and a range between 0 and up to 1000 seeds m-2. Across countries, variables explaining variation in the soil seed bank of R. obtusifolius in case parcels were soil pH (negative relation), silt content (negative), land-use intensity (negative), and aboveground R. obtusifolius plant density (positive). Because a large soil seed bank can sustain grassland infestation with R. obtusifolius, management strategies to control the species should target the reduction in the density of mature plants, prevention of the species' seed production and dispersal, as well as the regulation of the soil pH to a range optimal for forage production.

Multispecies for multifunctions: combining four complementary species enhances multifunctionality of sown grassland.

Assessing the overall performance of ecosystems requires a quantitative evaluation of multifunctionality. We investigated plant species diversity effects on individual functions and overall multifunctionality in a grassland experiment with sown monocultures and mixtures comprising four key grass and legume species. Nitrogen fertilisation rates were 50, 150, and 450 kg N ha-1 yr-1 (N50, N150, N450). Ten functions were measured representing forage production, N cycling, and forage quality, all being related to either productivity or environmental footprint. Multifunctionality was analysed by a novel approach using the mean log response ratio across functions. Over three experimental years, mixture effects benefited all forage production and N cycling functions, while sustaining high forage quality. Thus, mixture effects did not provoke any trade-off among the analysed functions. High N fertilisation rates generally diminished mixture benefits. Multifunctionality of four-species mixtures was considerably enhanced, and mixture overall performance was up to 1.9 (N50), 1.8 (N150), and 1.6 times (N450) higher than in averaged monocultures. Multifunctionality of four-species mixtures at N50 was at least as high as in grass monocultures at N450. Sown grass-legume mixtures combining few complementary species at low to moderate N fertilisation sustain high multifunctionality and are a 'ready-to-use' option for the sustainable intensification of agriculture.

Higher species richness enhances yield stability in intensively managed grasslands with experimental disturbance.

Climate models predict increased frequency and severity of drought events. At an Irish and Swiss site, experimental summer droughts were applied over two successive years to grassland plots sown with one, two or four grassland species with contrasting functional traits. Mean yield and plot-to-plot variance of yield were measured across harvests during drought and after a subsequent post-drought recovery period. At both sites, there was a positive relationship between species richness and yield. Under rainfed control conditions, mean yields of four-species communities were 32% (Wexford, Ireland) and 51% (Zürich, Switzerland) higher than in monocultures. This positive relationship was also evident under drought, despite significant average yield reductions (-27% at Wexford; -21% at Zürich). Four-species communities had lower plot-to-plot variance of yield compared to monoculture or two-species communities under both rainfed and drought conditions, which demonstrates higher yield stability in four-species communities. At the Swiss but not the Irish site, a high degree of species asynchrony could be identified as a mechanism underlying increased temporal stability in four-species communities. These results indicate the high potential of multi-species grasslands as an adaptation strategy against drought events and help achieve sustainable intensification under both unperturbed and perturbed environmental conditions.

Grass-legume mixtures sustain strong yield advantage over monocultures under cool maritime growing conditions over a period of 5 years.

Background and Aims: Grassland-based livestock systems in cool maritime regions are commonly dominated by grass monocultures receiving relatively high levels of fertilizer. The current study investigated whether grass-legume mixtures can improve the productivity, resource efficiency and robustness of yield persistence of cultivated grassland under extreme growing conditions over a period of 5 years. Methods: Monocultures and mixtures of two grasses (Phleum pratense and Festuca pratensis) and two legumes (Trifolium pratense and Trifolium repens), one of which was fast establishing and the other temporally persistent, were sown in a field trial. Relative abundance of the four species in the mixtures was systematically varied at sowing. The plots were maintained under three N levels (20, 70 and 220 kg N ha-1 year-1) and harvested twice a year for five consecutive years. Yields of individual species and interactions between all species present were modelled to estimate the species diversity effects. Key Results: Significant positive diversity effects in all individual years and averaged across the 5 years were observed. Across years, the four-species equi-proportional mixture was 71 % (N20: 20 kg N ha-1 year-1) and 51 % (N70: 70 kg N ha-1 year-1) more productive than the average of monocultures, and the highest yielding mixture was 36 % (N20) and 39 % (N70) more productive than the highest yielding monoculture. Importantly, diversity effects were also evident at low relative abundances of either species group, grasses or legumes in the mixture. Mixtures suppressed weeds significantly better than monocultures consistently during the course of the experiment at all N levels. Conclusions: The results show that even in the less productive agricultural systems in the cool maritime regions grass-legume mixtures can contribute substantially and persistently to a more sustainable agriculture. Positive grass-legume interactions suggest that symbiotic N2 fixation is maintained even under these marginal conditions, provided that adapted species and cultivars are used.

Weed suppression greatly increased by plant diversity in intensively managed grasslands: A continental-scale experiment.

Grassland diversity can support sustainable intensification of grassland production through increased yields, reduced inputs and limited weed invasion. We report the effects of diversity on weed suppression from 3 years of a 31-site continental-scale field experiment.At each site, 15 grassland communities comprising four monocultures and 11 four-species mixtures based on a wide range of species' proportions were sown at two densities and managed by cutting. Forage species were selected according to two crossed functional traits, "method of nitrogen acquisition" and "pattern of temporal development".Across sites, years and sown densities, annual weed biomass in mixtures and monocultures was 0.5 and 2.0 t  DM ha-1 (7% and 33% of total biomass respectively). Over 95% of mixtures had weed biomass lower than the average of monocultures, and in two-thirds of cases, lower than in the most suppressive monoculture (transgressive suppression). Suppression was significantly transgressive for 58% of site-years. Transgressive suppression by mixtures was maintained across years, independent of site productivity.Based on models, average weed biomass in mixture over the whole experiment was 52% less (95% confidence interval: 30%-75%) than in the most suppressive monoculture. Transgressive suppression of weed biomass was significant at each year across all mixtures and for each mixture.Weed biomass was consistently low across all mixtures and years and was in some cases significantly but not largely different from that in the equiproportional mixture. The average variability (standard deviation) of annual weed biomass within a site was much lower for mixtures (0.42) than for monocultures (1.77). Synthesis and applications. Weed invasion can be diminished through a combination of forage species selected for complementarity and persistence traits in systems designed to reduce reliance on fertiliser nitrogen. In this study, effects of diversity on weed suppression were consistently strong across mixtures varying widely in species' proportions and over time. The level of weed biomass did not vary greatly across mixtures varying widely in proportions of sown species. These diversity benefits in intensively managed grasslands are relevant for the sustainable intensification of agriculture and, importantly, are achievable through practical farm-scale actions.

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

Current 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 homegrown crops, and reduced N losses to the environment. Such challenges were addressed in a continental-scale field experiment conducted over 3 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 nonfixing 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 3 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.

Simulating evolutionary responses of an introgressed insect resistance trait for ecological effect assessment of transgene flow: a model for supporting informed decision-making in environmental risk assessment.

Predicting outcomes of transgene flow from arable crops requires a system perspective that considers ecological and evolutionary processes within a landscape context. In Europe, the arable weed Raphanus raphanistrum is a potential hybridization partner of oilseed rape, and the two species are ecologically linked through the common herbivores Meligethes spp. Observations in Switzerland show that high densities of Meligethes beetles maintained by oilseed rape crops can lead to considerable damage on R. raphanistrum. We asked how increased insect resistance in R. raphanistrum - as might be acquired through introgression from transgenic oilseed rape - would affect seed production under natural herbivore pressure. In simulation experiments, plants protected against Meligethes beetles produced about twice as many seeds as unprotected plants. All stages in the development of reproductive structures from buds to pods were negatively affected by the herbivore, with the transition from buds to flowers being the most vulnerable. We conclude that resistance to Meligethes beetles could confer a considerable selective advantage upon R. raphanistrum in regions where oilseed rape is widely grown.

Rapid and high seed germination and large soil seed bank of Senecio aquaticus in managed grassland.

Senecio aquaticus, regionally a Red List species in Europe, has become increasingly abundant in agricultural grassland of medium to high management intensity in Switzerland, Southern Germany, and Austria in recent years, where it is a threat for animal and human health due to its toxicity. In this study, we investigated the seed ecology of S. aquaticus to help protection of the species in relic populations while improving its control when abundant in managed grassland. Germination percentages of fresh ripe seeds of S. aquaticus were on average 68% in 2008, but only 45% in 2010, indicating yearly variation. Germination was generally fast: ten days after the onset of the tests, often more than 45% of all seeds had germinated. When covered with a soil layer of 5 mm, germination was only 16% compared to 63% in full light. Seeds buried in the soil for one and two years showed a germination of 78%, significantly higher than that of fresh ripe seeds, thus suggesting a stimulating effect of cold-wet stratification on germination and long seed survival in the soil. In grasslands with established populations of S. aquaticus, the number of germinable seeds of the species ranged from 361 to 1875 m⁻² in topsoil (0-10 cm) with an average of 1139 m⁻². The large seed bank and the rapid and high germination of S. aquaticus suggest that allowing seed formation is important for its preservation in relic populations. With respect to agricultural grassland, strategies to control the species should initially target hindering seed production and dispersal.

Nitrogen deposition but not ozone affects productivity and community composition of subalpine grassland after 3 yr of treatment.

A field experiment was established at 2000 m above sea level (asl) in the central Swiss Alps with the aim of investigating the effects of elevated ozone (O(3)) and nitrogen deposition (N), and of their combination, on above-ground productivity and species composition of subalpine grassland. One hundred and eighty monoliths were extracted from a species-rich Geo-Montani-Nardetum pasture and exposed in a free-air O(3)-fumigation system to one of three concentrations of O(3) (ambient, 1.2 x ambient, 1.6 x ambient) and five concentrations of additional N. Above-ground biomass, proportion of functional groups and normalized difference vegetation index (NDVI) were measured annually. After 3 yr of treatment, the vegetation responded to the N input with an increase in above-ground productivity and altered species composition, but without changes resulting from elevated O(3). N input > 10 kg N ha(-1) yr(-1) was sufficient to affect the composition of functional groups, with sedges benefiting over-proportionally. No interaction of O(3) x N was observed, except for NDVI; positive effects of N addition on canopy greenness were counteracted by accelerated leaf senescence in the highest O(3) treatment. The results suggest that effects of elevated O(3) on the productivity and floristic composition of subalpine grassland may develop slowly, regardless of the sensitive response to increasing N.

Convergence patterns and multiple species interactions in a designed plant mixture of five species.

It is known that convergence and divergence can occur in complex plant communities, but the relative importance of biotic and abiotic factors driving these processes is less clear. We addressed this issue in an experiment using a range of mixed stands of five species that are common in Swiss fens (Carex elata, C. flava, Lycopus europaeus, Lysimachia vulgaris and Mentha aquatica) and two levels of water and nutrients. One hundred and seventy-six experimental mixtures were maintained in large pots (75 l) for two consecutive growing seasons in an experimental garden. The stands varied systematically in the initial relative abundance of each of the five species and in overall initial stand abundance. The changes in biomass over 2 years were modelled as linear functions of treatments and the initial biomass of each species. The dynamics of the system were mainly driven by differences in the identity of species and by a negative feedback mechanism but also by different abiotic conditions. In all mixtures, C. elata became more dominant over time, which caused an overall convergence of community composition. In addition, the rate of change of each species' biomass was negatively related to its own initial abundance. Thus, a negative feedback further contributed to the convergence of communities. Species responded differently to water level and nutrient supply, causing community dynamics to differ among treatments. However, the different abiotic conditions only slightly modified the overall convergence pattern. Competitive interactions between more than two species were weaker than the negative feedback but still significantly influenced the species' final relative abundance. The negative feedback suggests that there is niche partitioning between the species, which permits their coexistence.

Asymptomatic carotid atherosclerosis is associated with circulating chlamydia pneumoniae DNA in younger normotensive subjects in a general population survey.

OBJECTIVE: Chlamydia pneumoniae infection has been associated with atherosclerosis, but serodiagnosis is unreliable in predicting vascular infection. Direct detection of circulating chlamydial DNA in peripheral blood mononuclear cells (PBMCs) was thus evaluated as a marker for cardiovascular risk in a general population survey using the common carotid intima-media thickness (IMT) as surrogate marker of asymptomatic atherosclerosis. METHODS AND RESULTS: C pneumoniae DNA in PBMCs was determined by nested polymerase chain reaction and associated with IMT for 1032 healthy participants of a general population survey who were within the highest or lowest IMT distribution quartile. C pneumoniae DNA was more prevalent in those with increased IMT (13.4% versus 10.7%), but this was not significant in univariate and of borderline significance in multivariate analysis. Testing for potential effect modifications by known strong determinants of an increased IMT in group interaction analysis revealed an independent association between C pneumoniae DNA and IMT in normotensive subjects (odds ratio [OR], 2.06; 95% CI, 1.05 to 4.03; P=0.04) and in those <70 years old (OR, 1.84; 95% CI, 1.06 to 3.19; P=0.03). CONCLUSIONS: Asymptomatic atherosclerosis is associated with circulating C pneumoniae DNA independently of classical cardiovascular risk factors in normotensive subjects and those <70 years old. C pneumoniae has been implicated in atherogenesis. We determined the association of chlamydial DNA in peripheral blood mononuclear cells with the carotid intima-media thickness from 1032 healthy subjects from a general population survey. A stratified group interaction analysis revealed an independent association in normotensive subjects and those <70 years old.