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The objective of this study was to quantify the farm gate nitrogen (N) offset potential of perennial ryegrass (Lolium perenne L.; PRG) white clover (Trifolium repens L.; WC) swards by comparing the herbage and milk production from dairy farmlets that were simulations of full farming systems. A study was established where 120 cows were randomly assigned to 4 farmlets of 10.9 ha (stocking rate: 2.75 cow/ha), composed of 20 paddocks each. Cows were fed 526 kg of DM of concentrate on average each year. The 4 grazing treatments were PRG-only at 150 or 250 kg of N/ha and PRG-WC at 150 or 250 kg of N/ha. Cows remained in their treatment group for an entire grazing season and were re-randomized as they calved across treatments each year. As cows calved in the spring as standard practice in Ireland, they were rotationally grazed from early February both day and night (weather permitting) to mid-November, to a target postgrazing sward height of 4.0 cm. Mean sward WC content was 18.1% and 15.4% for the 150 and 250 kg of N/ha PRG-WC treatments, respectively over the 3-yr period. When WC was included, lowering the N rate did not reduce pregrazing yield, pregrazing height, or herbage removed, but those factors decreased significantly when WC was absent. Total annual herbage DM production was 13,771, 15,242, 14,721, and 15,667 kg of DM/ha for PRG-only swards receiving 150 or 250 kg of N/ha and PRG-WC swards receiving 150 or 250 kg of N/ha, respectively. In addition, when WC was present, compressed postgrazing sward heights were lower (4.10 vs. 4.21 cm) and herbage allowance (approximately 17 kg/cow feed allocation per cow per day) higher than the high-N control (+ 0.7 kg of DM/cow per day). There was a significant increase in milk production, both per cow and per hectare, when WC was included in PRG swards. Over the 3-yr study, cows grazing PRG-WC had greater milk (+304 kg) and milk solids (+31 kg of fat + protein) yields than cows grazing PRG-only swards. This significant increase in milk production suggests that the inclusion of WC in grazing systems can be effectively used to increase milk production per cow and per hectare and help offset nitrogen use. This result shows the potential to increase farm gate N use efficiency and reduce the N surplus compared with PRG-dominant sward grazing systems receiving 250 kg of N/ha, without negatively affecting milk solids yield or herbage production, thus increasing farm profit by €478/ha. [ABSTRACT FROM AUTHOR]

Incorporating plantain with perennial ryegrass and white clover (RGWC) can improve the quality and quantity of grazing pastures; however, the sowing rate could affect the persistence of plantain, pasture yield, and nutritive value in different seasons. The objective of this study was to evaluate the effect of increasing sowing rates of plantain when established with RGWC on the dry matter (DM) yield, botanical composition, nutritive value, and bioactive compounds of the pasture over the first two years after sowing; and to determine the relationship between plantain content and nutritive characteristics in different seasons. The pasture treatments were RGWC, RGWC + low plantain rate (PLL), RGWC + medium plantain rate (PLM), and RGWC + high plantain rate (PLH). The results showed that annual DM yield was similar between treatments. The average plantain content (including leaves and reproductive stem) was 32, 44, and 48% in PLL, PLM, and PLH, respectively. The plantain composition increased in the first 15 months, then declined rapidly to about 20–30% at day 705 after sowing. Compared with RGWC, the plantain-RGWC pastures (PLL, PLM, PLH) had a higher content of organic matter digestibility (OMD), ash, starch, non-structural carbohydrates (NSC), P, S, Ca, Mg, Na, Cl, Zn, B, Co, aucubin, acteoside, and catalpol, while they contained a lower composition of DM%, acid detergent fibre (ADF), neutral detergent fibre (NDF), crude fat (CF), Fe, and Mn. These differences were linearly associated with the content of plantain leaves in the pasture and were higher in summer and autumn than in spring. In conclusion, incorporating plantain into the RGWC pasture can improve herbage nutritive quality, thus potentially increasing farm productivity and environmental benefits. However, further work is required to investigate management interventions to sustain plantain content beyond two years from sowing. [ABSTRACT FROM AUTHOR]

Legume-based forage plant mixtures are known to increase biomass production over the mixture species grown as pure stands (overyielding), which has partly been attributed to enhanced nitrogen availability by legumes. However, the relative importance of underlying processes of these positive diversity effects and their drivers are not fully understood. Here we assessed if outcome and causes of diversity effects depend on the legume-species genetic identity. Over five years, we cultivated different white clover (Trifolium repens) populations, a grass and forb species in pure stands and clover-based mixtures and recorded biomass yield. Complementarity and selection effects of mixtures and relative yields of mixture species were calculated based on both unfertilized and nitrogen-fertilized non-leguminous pure stands. Results showed that the clover population altered the overall strength of diversity effects as well as the direction and magnitude of their temporal trends, at least for the grass component of mixtures. Differences in diversity effects between clover populations diminished when fertilized instead of unfertilized non-leguminous pure stands were considered. Hence, a part of these differences likely results from dissimilar effects of clover populations on nitrogen availability. The findings reveal the possibility to improve overyielding of legume-based forage plant mixtures by decisions on legume-species genetic identity. [ABSTRACT FROM AUTHOR]

Perennial ryegrass (Lolium perenne L.) is cultivated in many countries for its high yield and good quality forage. Its cultivars have specific functional traits, such as phenology, growth habit and rate, which affect the competitiveness and productivity of mixtures. The cultivation of new cultivars of perennial ryegrass or mixtures of cultivars in swards is one of the complementary measures to improve the condition of swards. The study was aimed to determine the productivity potential of three newer, less investigated Lithuanian perennial ryegrass (Lolium perenne L.) cultivars 'Elena DS', 'Raminta', 'Verseka', and cultivar mixtures in single-species swards and in mixtures with legumes - white and red clover. The experiment was set up in the central part of Lithuania in the Nemoral climate zone. Mineral fertilisers were not used for the mixtures with legumes, singlespecies crops were fertilised with N150 kg ha-1 yr-1. Over the two experimental years, the average dry matter yield of perennial ryegrass was 8.00 t ha-1 for the 'Elena DS', 7.4 t ha-1 for 'Verseka', 6.9 t ha-1 for 'Raminta' and 7.4 t ha-1 for the cultivar mixture. In the 1st year of sward use, the productivity of the perennial ryegrass cultivars did not differ significantly. In the 2nd year, the productivity of 'Elena DS' was significantly higher compared with that of the other cultivars cultivated in single-species crops. The influence of functional traits of the cultivars was significant in the first year of cultivation and persisted in single-species swards but not in mixtures from the second year onwards. The plants of the cultivar 'Elena DS' allowed the development of clover in the mixture, resulting in an increase in herbage productivity and crude protein content. The results of the experiment suggest that a mixture of perennial ryegrass cultivars can be used in swards, as a similar dry matter yield was recorded compared to a single-cultivar swards. However, the benefits of swards may vary from season to season, and more research is needed to assess the impact of a mixture of cultivars on grass productivity and quality with ageing of swards and accumulation in different environmental conditions. [ABSTRACT FROM AUTHOR]

Molecular characterization of genetically modified plants can provide crucial information for the development of detection and identification methods, to comply with traceability, and labeling requirements prior to commercialization. Detailed description of the genetic modification was previously a challenging step in the safety assessment, since it required the use of laborious and time-consuming techniques. In this study an accurate, simple, and fast method was developed for molecular characterization of genetically modified (GM) plants, following a user-friendly workflow for researchers with limited bioinformatic capabilities. Three GM events from a diverse array of crop species—perennial ryegrass, white clover, and canola—were used to test the approach that exploits long-read sequencing by the MinION device, from Oxford Nanopore Technologies. The method delivered a higher degree of resolution of the transgenic events within the host genome than has previously been possible with the standard Illumina short-range sequencing strategies. The flanking sequences, copy number, and presence of backbone sequences, and overall transgene insertion structure were determined for each of the plant genomes, with the additional identification of moderate-sized secondary insertions that would have previously been missed. The proposed workflow takes only about 1 week from DNA extraction to analyzed result, and the method will complement the existing approaches for molecular characterization of GM plants, since it makes the process faster, simpler, and more cost-effective. [ABSTRACT FROM AUTHOR]

We tested whether selection of ryegrass for reduced root hair length and density would reduce root competition on white clover for phosphate (P). A Rhizosheath Selection Tool was used to select two ryegrass populations with contrasting root hair traits. These were grown at five soil P levels in a glasshouse experiment and rhizosheath trait selection had no effect on growth or shoot P concentrations. The ryegrass populations were also grown with white clover in soils with Olsen P12 or 19 mg L−1. In the Olsen P12 soil, clover grown with the smaller rhizosheath ryegrass had significantly heavier shoots and contained more P than clover grown with the larger rhizosheath ryegrass. In the Olsen P19 soil differences in ryegrass rhizosheath had no effect on clover growth. The results showed that under the experimental conditions, grass-clover competition for soil P could be modified by the rhizosheath traits of ryegrass. [ABSTRACT FROM AUTHOR]

• 13C-CO 2 was pulsed labeled in an agroforestry system. • PLFA biomarker 16:1ω5c was enriched with 13C in soil under grassland. • 13C incorporation into gram positive bacteria was higher under willow compare to grassland and rape. • Gram negative bacteria utilized the highest relative amount of 13C. • Microbial utilization of 13C depends on vegetation types in an agroforestry system. The release of organic compounds from plant roots (rhizodeposits) and their subsequent utilization by soil microbial communities is a key process linking atmospheric and terrestrial carbon (C) and nutrient cycling. However, the effects of land-use types on rhizodeposit-mediated alteration of microbial community structure and its consequences for soil C cycle are still, largely, unknown. Therefore, we used 13C-PLFA-SIP to evaluate the rhizosphere C fluxes to microorganisms in a monoculture crop (oilseed rape, Brassica napus L.), a mixed grassland (dominated by Lolium perenne L. mixed with clover Trifolium repens L.), and a tree (willow, Salix schwerinii E.L. Wolf and Salix viminalis L.) within an agroforestry system. Though harboring similar total microbial biomass, less 13C was recovered in microorganisms in soil under willow (0.017 % of 13C input) after 14 days as compared to rape (0.03 %) and grassland (0.09 %). Across three land use types, gram negative bacteria incorporated most of the 13C (28–42 % of total 13C labeled PLFAs), followed by fungi and gram positive bacteria (12–18 %) and less abundance groups (e.g. actinomycetes, <5 %). The arbuscular mycorrhizal fungal biomarker 16:1ω5c was enriched with 13C in soil under grassland (8 % of total 13C labeled PLFAs). The 13C incorporation into gram positive bacteria was higher in soil under willow than under grassland and rape. By contrast, 42 % of total 13C labeled PLFAs were attributed to gram negative bacteria in soil under rape. Overall, our results demonstrated that pronounced differences in microbial community structure and subsequent microbial C utilization between land-use types could affect soil C cycles and storage. [ABSTRACT FROM AUTHOR]

Backgrounds and aims: N rhizodeposition by legumes leads to enrichment of N in soils and in companion plants. N rhizodeposition can be divided into two major components, root exudation and root senescence. Our aim was to quantify N root exudation in white clover ( Trifolium repens L.) through an estimation of short-term N rhizodeposition and to assess its impact on N transfer to companion perennial ryegrass ( Lolium perenne L.) grown in mixture with clover. Method: N provided in the root atmosphere for 3 days was used to estimate transfer of symbiotically fixed nitrogen (SFN) to the growing medium by clover grown in pure stand and to ryegrass by clover grown in mixture for 2 months. Results: The proportion of N rhizodeposited over the 3 days increased from 3.5 % of SFN in pure stand to 5.3 % in mixture. The N-enrichment of ammonium from the adhering substrate shows that a part of the rhizodeposited N was released in the form of ammonium. 4 % of the rhizodeposited N was taken up by ryegrass during the labelling period. Conclusions: This study showed a significant contribution of root N exudation to the total N rhizodeposition of legumes and in the transfer of N to grasses. [ABSTRACT FROM AUTHOR]

Cover crops can be used to reduce leaching and erosion, introduce variability into crop rotation and fix nitrogen (N) for use by the main crops, less is however known about effects on weeds. The effects on weed seed bank, weed growth and grain yield of 4 years of annual undersown clover and ryegrass alone and in combination, and one of the 4 years with clover or clover + grass as green manure, were studied in oat and spring wheat at two experimental sites in south-eastern Norway. These treatments were compared with no undersown crop (control) and with weed harrowing. In contrast to many results in the literature, the undersown clover in this study did not suppress annual weeds, but fertilized the weeds as well as the cereals. Undersown clover resulted in a statistically significant increase of grain yield at the two sites to 116% and 121% of control. During the 4-year period relative seed bank and density of emerged weed (dominated by Spergula arvensis) increased significantly about 4.5 and 10 times respectively in the undersown clover plots at Apelsvoll. At Kise both ryegrass alone and ryegrass mixed with clover significantly suppressed the weed biomass to 70% and 74% of control respectively. It is concluded that fertilization effects of undersown clover may have dominated and overriden the competitive effects. One whole-season clover green manure did not increase the mean yield, but resulted in a significant drop in seed bank size the following year, because of limited weed establishment in an established ley. Only a slight increase in average weed biomass was observed at one of the two experimental sites. The weed seed bank and the weed biomass were essentially kept at steady state during the experimental period in harrowed plots, but harrowing decreased grain yield significantly at both sites. [ABSTRACT FROM PUBLISHER]

The aim of the present investigation was to study the effect of white clover (var. Milka and Donna), red clover (var. Fanny) and ryegrass (var. Tove) undersown in winter wheat on a succeeding oat crop. Under the climatic conditions prevailing in Sweden, growing a catch crop after winter cereals is of particular interest because the latter are usually followed by a spring sown crop, leaving the ground bare during autumn and winter. Field trials were carried out during three growing seasons in an integrated farming system and for one year in an organic farming system. Competition from the dense wheat crop in the integrated farming system had a negative effect on the undersown species, and at harvest of the wheat they showed quite poor growth in all three years, with nitrogen contents under 5 kg ha-1. No significant yield increase was measured without added nitrogen when the averages for all three years were calculated. On average, the grain yields were improved by 750 kg ha-1 (14% moisture content) for the treatments with undersown clover for all three years when 90 kg N ha-1 were added. The experiment within the organic system showed a different pattern with a better development of the undersown clover species, with nitrogen contents approximately 25 kg ha-1 and an improvement in oat grain yield, from around 2000 kg ha-1 for the control to almost 3500 kg ha -1 with clover undersown the previous year. [ABSTRACT FROM AUTHOR]

Pure stands of Trifolium repens L. N.Z. Huia and Lolium perenne L. S32 and mixtures of various densities and frequencies were treated with sulphuric acid solutions of pH 2.5, 3.5 and 5.6. The species yield was recorded over live sequential harvests. Multifactorial response equations were used to describe the behavior of the species yield as a function of acidity and the density of both species. Predictions, made through these models, showed that the response of yield of these species to simulated acid rain treatments varied greatly depending on the planting density, the time of harvesting and on whether the plants were growing in monoculture or in mixture. This suggests that it is not sufficient to consider the effects of experimentally applied acid rain on single individuals or low density stands, since the results obtained will not reflect the response in a close canopy sward. [ABSTRACT FROM AUTHOR]

In six experiments Lolium perenne L. and Plantago lanceolata L. were grown together in pots. Two experiments also included L. perenne alone, P. lanceolata alone, or L. perenne + Trifolium repens L. In five of the experiments prior sterilization of the soil followed by reinoculation was used to obtain two treatments, one having plants infected with vesicular-arbuscular mycorrhiza, the other non-mycorrhizal. The shoots of some plants in each pot ('donors') were injected with 32P. After a week these shoots were removed, and after a further week the remaining plants ('receivers') were harvested and their 32P content determined. In all experiments and species-combinations there was some 32P transfer to the shoots of both mycorrhizal and non-mycorrhizal receiver plants. The amount of transfer in mycorrhizal treatments was 2 to 8-fold greater than in non-mycorrhizal treatments, except in one experiment where mycorrhizal infection was very low. There was no clear difference between the different species combinations in the amount by which phosphorus transfer was enhanced by mycorrhizas. These results show that mycorrhizas can increase phosphorus transport between plants, but do not show whether there is direct transport from one root to another via interconnecting hyphae or whether the phosphorus must leave the donor root before being taken up by mycorrhizal hyphae. [ABSTRACT FROM AUTHOR]

Simple Summary: The present study simulated the effects of different dairy cow diets based on local feeding strategies on enteric methane (CH4) emissions and surpluses of crude protein (CP) in small-scale dairy systems (SSDS). Our study evaluated five scenarios of supplementation (S): without supplementation (control diet), meaning no supplements were provided, only pasture (S1); pasture supplemented with 4.5 kg dry matter (DM)/cow/day of commercial concentrate (CC) (S2); supplemented with 200 g DM/kg per milk produced of CC (S3); supplemented with ground maize grains and wet distiller brewery grains (S4); and S4 plus maize silage (S5). In addition, two pasture managements (cut-and-carry versus grazing) and two varieties of legumes (red clover vs. white clover) were considered. The results suggest that methane emissions and large nitrogen surpluses in the diet are affected by the type of supplementation given to cows, in addition to the management and chemical composition of the pastures offered. In SSDS, it is possible to formulate diets with local inputs to reduce excess nutrients and dependence on external inputs, increasing feed efficiency and reducing costs (excess of CP in the diet) and CH4 emissions. In cattle, greenhouse gas (GHG) emissions and nutrient balance are influenced by factors such as diet composition, intake, and digestibility. This study evaluated CH4 emissions and surpluses of crude protein, using five simulated scenarios of supplementation in small-scale dairy systems (SSDS). In addition, two pasture managements (cut-and-carry versus grazing) and two varieties of legumes (red clover vs. white clover) were considered. The diets were tested considering similar milk yield and chemical composition; CH4 emission was estimated using Tier-2 methodology from the Intergovernmental Panel on Climate Change (IPCC), and the data were analyzed in a completely randomized 5 × 2 × 2 factorial design. Differences (p < 0.05) were found in predicted CH4 emissions per kg of milk produced (g kg−1 FCM 3.5%). The lowest predicted CH4 emissions were found for S3 and S4 as well as for pastures containing white clover. Lower dietary surpluses of CP (p < 0.05) were observed for the control diet (1320 g CP/d), followed by S5 (1793 g CP/d), compared with S2 (2175 g CP/d), as well as in cut-and-carry management with red clover. A significant correlation (p < 0.001) was observed between dry matter intake and CH4 emissions (g−1 and per kg of milk produced). It is concluded that the environmental impact of formulating diets from local inputs (S3 and S4) can be reduced by making them more efficient in terms of methane kg−1 of milk in SSDS. [ABSTRACT FROM AUTHOR]

In artificial ecosystems, mixed planting of gramineous and leguminous plants can have obvious advantages and is very common. Due to their improved growth performances and stress tolerance, endophyte-infected grasses are considered to be ideal plant species for grasslands. However, endophytic fungi can inhibit the growth of neighboring nonhost leguminous plants. In this study, we chose endophyte-infected and endophyte-free tall fescue (Lolium arundinaceum Darbyshire ex. Schreb.) and clover (Trifolium repens) as the experimental materials to explore whether arbuscular mycorrhizal fungi and rhizobium can alleviate the inhibitory effect of endophyte infection on clover. The results showed that endophytic fungi significantly reduced clover biomass. Arbuscular mycorrhizal fungi inoculation significantly increased the biomass of clover in both endophyte-infected tall fescue/clover and endophyte-free tall fescue/clover systems but the beneficial contribution of arbuscular mycorrhizal fungi was more obvious in the endophyte-infected tall fescue/clover system. Rhizobia inoculation could alleviate the detrimental effect of tall fescue on the growth of clover but did not alleviate the detrimental effect of endophyte infection on the growth of clover. [ABSTRACT FROM AUTHOR]