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Societal Impact Statement Summary Humans and honey bees have a long history of interaction to yield valued products and services. However, honey bees are under pressure from changes in vegetation, agricultural practices and climate change. We investigate if pasture legumes can be harnessed to support honey bees. We use a diverse set of species that originated from the Mediterranean and western Europe but are now developed as commercial cultivars for the southern Australian livestock industries. We find that pasture legumes show great promise as a means to support honey bees and suggest that further targeted research is warranted. Globally, humans and honey bees (Apis mellifera) have a long history of interaction and this is now under pressure. In temperate southern Australia, exotic honey bees rely on an unstable native floral resource base, yet a diverse set of exotic pasture legume species are sown as staple rotational or permanent crops. We investigated if these pasture legumes could be strategically used to support honey bee populations for the apiary and horticulture industries in this region. A literature review on temperate pasture legumes and honey bees was undertaken and integrated with data from an Australian industry case study. Comparisons were made to the widely‐grown red clover (Trifolium pratense) and white clover (T. repens). We found that many southern Australian pasture legumes could provide significant floral resources as well as enhanced temporal and spatial stability for honey bees due to phenological variability and broad adaptation. Honey bees will likely recognise and be attracted to their flowers and other characteristics are comparable, or potentially superior, to red and white clover, such as floret number per inflorescence, floret length, nectar volume and sucrose‐dominance and pollen protein. The floral resource diversity from pasture legume mixes could aid honey bee health and support pollinator services for adjacent crops. We conclude that the exotic pasture legumes of temperate Australia show promise for development as a sown floral resource for honey bees. Globally, further investigation is merited to maximise benefits from the integration of a diverse range of pasture legumes into agricultural landscapes. [ABSTRACT FROM AUTHOR]

Integrated forage–row cropping systems provide important agronomic and economic benefits to producers. However, little attention has been given to incorporating forages into row crop systems unique to the southeastern United States. This study assessed the viability of intercropping cotton (Gossypium hirsutum L.) on perennial, cool-season legumes during the summer months in the Southeast Coastal Plain over two production years. Treatments included a weedy fallow, annual ryegrass (ARG; Lolium multiflorum Lam.) monoculture, a red clover (RC; Trifolium pratense L.) and white clover (WC; Trifolium repens L.) mixture, and a three-species mixture of ARG, RC, and WC. Plots were established in fall 2020 with forage grown until May 2021 and 2022, when plots were strip-tilled and planted with cotton. Cotton was managed with minimal herbicide use to preserve perennial clovers. Results indicated WC was more persistent than RC (>40 plants m−2 vs. <40 plants m−2), and the presence of perennial clovers suppressed weeds at similar levels to the ARG monoculture (35 weeds m−2) during the second spring. Perennial clovers grew taller (4–5 cm) when mixed with ARG. The presence of clovers mixed with ARG during the second spring reduced acid detergent fiber (ADF) concentration and increased crude protein (CP) concentration (280 g ADF kg−1; 167 g CP kg−1) compared to the ARG monoculture (315 g ADF kg−1; 126 g CP kg−1). Benefits of intercropping perennial forages with cotton were maximized during the second year of growth, but future work is necessary to improve stand survival. [ABSTRACT FROM AUTHOR]

In the experimental field of the Research Institute of Mountain Stockbreeding and AgricultureTroyan during the period 2020-2022, the following meadow legume crops were studied: bird’s foot trefoil (Lotus corniculatus L.), hybrid clover (Trifolium hibridum L.), alfalfa (Medicago sativa L.), red clover (Trifolium pratense L.) and white clover (Trifolium repens L.). The purpose of the present research is to make a statistical analysis based on average data on yield, morphological features and quality, and determine dependencies between them. High correlations were found between plant height and crude fiber content (r = 0.88); crude fiber and the relative proportion of legumes in the grass stand (r = 0.77); acid detergent fibers and cellulose content (r = 0.92). These results make it possible to derive regression dependencies between them, which are represented by linear equations and coefficients of determination. [ABSTRACT FROM AUTHOR]

Aims: To develop a methodology to study uptake and redistribution by plants of NH4+ from deep soil, applying it to investigate deep root N uptake by cultivated grassland species. Methods: A slow-release 15NH4+ label adsorbed to clinoptilolite was placed into soil (depth 42 cm) well below the densest root zone in well-established monospecific stands of five grass and two clover species. Species showing a variety of deep rooting patterns, N acquisition strategy, forage qualities, and persistence in hemiboreal conditions were chosen. The label was placed in early spring and tracked throughout one or two growing seasons in two repeated experiments. Results: After two growing seasons ~ 90% of the label was tracked in the soil and harvested herbage of grasses, less in clovers. Deep N uptake was limited in spring, increased during mid-season, and was strongest in autumn in all species, despite lower herbage yield in autumn. Species differed in ability to recover and maintain 15N in the soil–plant system. In one growing season, Lolium perenne L., Phleum pratense L., Schedonorus pratensis (Huds.) P.Beauv. and Schedonorus arundinaceus (Schreb.) Dumort herbage recovered ~ 65% of the label, Poa pratensis L. 54%, and Trifolium pratense L. and Trifolium repens L. 36–48%. Label transport to topsoil was observed, mainly attributable to plant nutrient redistribution rather than physical diffusion. Conclusions: The innovative slow-release 15N label enabled tracing species differences and seasonal changes in uptake of NH4+ from deep soil. Among the tall-growing grasses, growth vigor appeared as important for deep N uptake as expected root depth. [ABSTRACT FROM AUTHOR]

During the period 2020-2022, the following types of meadow legume crops were tested on light gray pseudopodzolic soil on the territory of the experimental field of Research Institute of Mountain Stockbreeding and Agriculture-Troyan: Lotus corniculatus L., Trifolium hibridum L., Medicago sativa L., Trifolium pretense L., Trifolium repens L. The matter yield, the biodiversity of the species in the grass stand and the impact of the factors, such as conditions of the year and the type of grass stands were studied to determine the acclimatization of meadow legume crops under mountain conditions. It was established that the tested legume forage species realized a high yield of dry matter under mountain conditions, which varied from 836.68 to 971.13 kg/da, which determined their good adaptability for cultivation under the conditions of light gray pseudopodzolic soils. The low values of their variation coefficients by year and average yield of 909.35 kg/da of all species show that they are suitable for inclusion in the structure of farms in mountain conditions. Alfalfa proved to be the most rational for cultivation, followed by red clover. The botanical composition of the grass stands depends on the rate of growth and development of the herbaceous species. The high productivity of alfalfa corresponds to its high presence in the grass stand, which also indicates its high productivity. Natural factors determine the productivity of the grass stands of the various legume species, and subsequently the economic efficiency in the production of fodder. [ABSTRACT FROM AUTHOR]

Ramularia sphaeroidea was primarily identified based on the characteristics of its conidia and several sequences. The fungus causes severe leaf spot disease on hairy vetch (Vicia villosa var. glabrescens) in Yunnan Province in China. The growth, sporulation, fungicide efficacy, and host range of the pathogen were evaluated to aid in disease management. Different types of culture media and carbon and nitrogen sources were used to evaluate the growth of R. sphaeroidea. Oatmeal, maltose, and potassium nitrate agar had a higher amount of sporulation. Difenoconazole (10%) was the most effective fungicide against the leaf disease caused by R. sphaeroidea. In addition, foliar inoculation sprays were used to assess the host range of R. sphaeroidea in six different plant species, including alfalfa (Medicago sativa L.), sainfoin (Onobrychis viciifolia Scop.), erect milkvetch (Astragalus adsurgens Pall.), common vetch (Vicia sativa L.), red clover (Trifolium pratense L.), and white clover (Trifolium repens L.). R. sphaeroidea successfully infected these plants, indicating that it has a wider host range than hairy vetches. [ABSTRACT FROM AUTHOR]

On dairy farms, fertilization of grass-clover swards ensures stable grass yields but may increase the potential for nitrate leaching on light-textured soils. The aim of this study was to quantify the N use efficiency and nitrate leaching under fertilized grass-clover leys. The study was conducted over 2 years at two sites, with increasing applications of mineral fertilizer (0–480 kg available N ha−1) alone or in combination with a basic application of cattle slurry. For plots fertilized with mineral N, the N soil surface balance was independent of the application rate and in the same range as for unfertilized plots (− 11 to 51 kg N ha−1). However, when plots were fertilized with slurry N (+ mineral N), the surplus was substantially increased owing to the fraction of organic N applied in slurry (95–100 kg N ha−1) and higher biological N2 fixation inputs (55–228 kg N ha−1). The type of fertilizer had no effect on nitrate leaching across the full range of application rates. Nitrate leaching increased quadratically as a function of application rate, with a range of 3–117 kg N ha−1 (0.33–17 mg l−1 in soil solution sampled with suction cups) in the first year and less in the second year, when clover proportion was lower due to the self-regulatory nature of grass-clover mixtures. Importantly, the rate of marginal leaching increased with fertilization level: below 150 kg N ha−1 there was no additional leaching from fertilization and at 200 kg N ha−1 around 5% of additional fertilizer-N was leached. This is less than generally found for arable crops and thus even in intensive dairy systems, grass-clover leys are an environmentally favorable crop. [ABSTRACT FROM AUTHOR]

The local abiotic and environmental conditions of a grass‐based farming system may influence the agricultural benefits of mixtures in comparison to pure stands. We investigated the effects of species identities and interactions between grass, legume, and herb species on dry matter yield and sward digestibility and explored how contrasting environments may affect these relationships. We established experimental plots across 11 mixtures and 4 monocultures of perennial ryegrass (Lolium perenne L.), chicory (Cichorium intybus L.), white clover (Trifolium repens L.), and red clover (Trifolium pratense L.) established at two seeding densities in two sites Brody and Szelejewo (Poland) and managed them over a 3‐year period. The two sites were close geographically and had similar climates, but differed in soil types (Luvisols and Cambisols, respectively). We confirmed that the annual DM yields were generally higher on Cambisols soil as compared to Luvisols soil; both individual species effects and species interaction strengths differed across the two contrasting environments. The predicted annual DM yield for the 4‐species mixtures in general gave comparable or higher yields compared to monocultures across the 3 years, with the mixtures giving around 10%–30% higher yields than the weighted average of the monocultures of the constituent species. Analysing the transgressive overyielding it turned out that there was no significant difference between the 4‐species mixtures and best performing monoculture. The benefits of species diversity in our study was greater on less productive site because we found larger overyielding of the 4‐species sward on the Luvisols soil than on the Cambisols soil. Additionally, the effect of mixtures composed by 4‐species increased the yield stability compared to monocultures in the 3‐year period of our study, particularly on Cambisols soil. Sward digestibility applied to average values did not differ much between sites with mixtures performing similarly to monocultures. The reason for that could have been the dynamics of sward botanical composition during study years shifted towards increased perennial ryegrass and decreased proportions of chicory in the sward as well as the observed phenomenon that the species in mixed swards progressed to successive growth stages more slowly than in pure sowing. [ABSTRACT FROM AUTHOR]

Natural soil inundation caused by frequent and intense precipitation affects carbon allocation in grassland biomass, ultimately leading to changes in soil carbon storage. Increasing forage diversity could provide resiliency to inundation of grassland. The objective was to evaluate forage and root biomass and C and N stocks in the soil particulate organic matter (POM) from pastures under recurring short‐term inundation. Three forage species combinations were evaluated in an inundated (typically lasting for a few days after heavy rain events) and a non‐inundated pasture: (1) predominantly tall‐fescue (Festuca arundinacea Schreb.); (2) mixture of cool‐season perennials composed of tall‐fescue, orchardgrass (Dactylis glomerata L.), bluegrass (Poa pratensis L.) and white (Trifolium repens L.) and red clover (Trifolium pratense L.); (3) and cool‐season mixture of perennials overseeded with oats (Avena strigosa Schreb.) and rye (Secale cereale L.). Roots and forage biomass were sampled during the growing seasons of 2021 and 2022. Soil POM was evaluated 2.5 years after establishment. Inundation reduced forage and root biomass mainly during periods of higher inundation frequency, leading to lower C‐POM stocks (p <.05). Inundation caused a shift in the forage botanical composition, that is, higher occurrence of weeds and less productive grass species with shallow roots. The perennial cool‐season mixture did not increase forage yield compared with tall fescue only but did increase root mass. This occurred mainly at deeper layers and, consequently, increased C‐POM stocks (p <.05). Overseeding of winter annuals reduced overall forage production, despite increasing spring biomass when inundated in the first year, but reduced C and N‐POM stocks (p <.05). Perennial cool‐season forage mixtures can increase the resilience of pastures to inundation events and contribute to increased carbon sequestration in grasslands where inundation is prevalent. [ABSTRACT FROM AUTHOR]

Context: Tagasaste (Cytisus proliferus), a fast-growing leguminous tree, has potential to supplement pasture production in steep-hill country and to increase pasture resilience. Aims: In the companion paper, we quantified tagasaste production characteristics. Here, we determine the effect of proximity of 10-year-old tagasaste trees on productivity of eight pasture species including grasses (perennial ryegrass, Lolium perenne ; cocksfoot, Dactylis glomerata ; prairie grass, Bromus willdenowii ; microlaena, Microlaena stipoides), perennial legumes (white clover, Trifolium repens ; red clover, T. pratense ; lotus, Lotus pedunculatus), and an annual legume (subterranean clover, T. subterraneum). Methods: A site was established in the East Coast region of the North Island of New Zealand on steep-hill country (>20° slope). Herbage production, nutritive value and survival of pasture species established as spaced transplants were measured over 3 years. Results: Cocksfoot had high survival, herbage production and metabolisable energy content but was negatively affected by proximity to tagasaste. Microlaena was not significantly affected by proximity to tagasaste; however, it was much less productive and had lower nutritive values than the other grasses. Only 40% of perennial ryegrass transplants survived 3 years, and survival of perennial legumes was negligible. Subterranean clover was able to set seed in the open and in shade. Conclusions: Cocksfoot was the most productive grass species, and microlaena was least affected by proximity to tagasaste. Given the poor persistence of perennial clovers, annual clovers may be better suited to a tagasaste silvopasture on steep, dry hillsides. Implications: Mixtures of cocksfoot and subterranean clover may be well suited to summer-dry hillsides, between and under trees in a tagasaste silvopasture. Microlaena may provide some forage and can maintain groundcover despite shade. Growth and survival of eight pasture species in proximity to the forage shrub tagasaste were quantified in drought-prone hill country. Cocksfoot and subterranean clover were highly productive only in lightly shaded areas, whereas microlaena, although less productive than other grasses, was not significantly affected by shade treatment, and perennial legumes showed negligible survival. Cocksfoot and subterranean clover could be established between trees in a tagasaste silvopasture, with naturalised microlaena maintained beneath the canopy to provide groundcover. [ABSTRACT FROM AUTHOR]

ABSTRACT: This experiment investigated the variation in enteric methane production and associated gas exchange parameters, nutrient digestibility, rumen fermentation, and rumen microbiome when a range of silages based on different forage types (grass or clover), and different species within the 2 types, were fed as the sole feed to heifers. Three grass species (perennial ryegrass, festulolium, and tall fescue) and 2 clover species (red clover and white clover) were included. Perennial ryegrass was harvested at 2 maturity stages in the primary growth, white clover was harvested once in the primary growth, and 4 cuts of festulolium and tall fescue and 3 cuts of red clover were harvested during the growing season, giving 14 different silage batches in total. Sixteen Holstein heifers 16 to 21 mo old and 2 to 5 mo in pregnancy were fed the silages ad libitum as the sole feed in an incomplete crossover design. Each silage was fed to 4 heifers, except for the 2 perennial ryegrass silages, which were fed to 8 heifers; in total 64 observations. The CH4 production was measured for 3 d in respiration chambers. Heifers fed clover silages had higher dry matter intake (DMI) compared with heifers fed grass silages, and heifers fed tall fescue silages had the numerically the lowest DMI. Compared with grass silages, feeding clover silages led to higher crude protein digestibility but lower neutral detergent fiber (NDF) digestibility. Rumen pH was higher in heifers fed clover silages compared with those fed grass silages. Based on composition analysis, the rumen microbiota of the heifers clustered clearly according to forage type and species. More specifically, 7 of the 34 dominating rumen bacterial genus-level groups showed higher relative abundances for the clover silages, whereas 7 genus-level groups showed higher abundances for the grass silages. Methane yield was higher for heifers fed grass silages than for those fed clover silages when methane production was related to dry matter and digestible organic matter intake, whereas the opposite was seen when related to NDF digestion. The gross energy lost as methane (CH4 conversion factor, %) reduced from 7.5% to 6.7%, equivalent to an 11% reduction. The present study gives the outlines for choosing the optimal forage type and forage species with respect to nutrient digestibility and enteric methane emission in ruminants.

In addition to improving soil quality, the fertilisation of grassland with cattle slurry is often associated with seed dispersal. Most studies focus on the effects of cattle slurry on the germination and early development of weed species, but less is known about how slurry affects the germination process of grasses and forage legumes. The aim of Experiment I of our study was therefore to investigate the influence of soaking time in cattle slurry of different concentrations on Lolium multiflorum, Dactylis glomerata, Trifolium pratense and Trifolium repens. Seeds were soaked in undiluted (100%) and diluted cattle slurries (50% and 25%) for 14, 28, 42, 56, 70 and 84 days. Experiment II was conducted to study the initial growth of studied plants from seeds soaked in cattle slurry of different concentrations for 14 days. After the germination test, which was carried out under controlled conditions, the germination index (GI) was calculated. The results (Experiment I) show that a short soaking in cattle slurry (14 days) has no negative effect on the germination process for all species. However, a longer soaking resulted in significantly reduced and delayed germination, especially in undiluted slurry for grasses and diluted slurries for clovers. The slurry concentration (Experiment II) only influenced the root growth of L. multiflorum. Seedlings grown from seeds soaked in undiluted slurry had a 17% higher relative root length than the control and developed significantly longer root systems than the other two slurry concentrations. [ABSTRACT FROM AUTHOR]

Multispecies swards, comprised of different plant functional groups have comparable production potential to high N input L. perenne swards at reduced N when legumes are included. However, information on the appropriate management of multispecies swards is limited. The effect of differing management practices on the herbage dry‐matter (DM) production and botanical composition of different sward types was investigated using a 3 × 2 × 2 factorial design plot experiment. The experiment consisted of three sward types: L. perenne‐only receiving (LP; 250 kg N ha−1 a−1); L. perenne‐Trifolium repens (LP‐TR); multispecies sward containing L. perenne, Phleum pratense, Trifolium repens, Trifolium pratense, Plantago lanceolata and Cichorium intybus (MSS). LP‐TR and MSS received 90 kg N ha−1 a−1. Harvesting of plot herbage took place every 21 or 28 days to a defoliation height of 4 or 6 cm, over two growing seasons (March to November 2020 and 2021 inclusive). Annual herbage produced by both LP and LP‐TR was not significantly affected by defoliation frequency. However, MSS produced 1923 kg DM ha−1 a−1 more herbage when harvested less frequently. Unsown species contributed significantly less to the herbage DM of MSS compared to LP and LP‐TR and remained similar in 2020 and 2021, whereas the contribution of unsown species to herbage DM increased in LP and LP‐TR from 2020 to 2021, with noxious species more associated with LP and LP‐TR than MSS. Results demonstrate the role of multispecies swards in improving the sustainability of grass‐based agricultural systems in an environment of reduced fertilizer inputs. [ABSTRACT FROM AUTHOR]

Perennial cool-season grasses are the foundation of horse pastures in the North Central U.S. A majority of U.S. horses graze pastures as a way to encourage exercise and socialization, and to reduce feeding costs. However, other forages must be evaluated to help extend the grazing season and respond to emerging issues (e.g., floods and winter kill). This abstract summarizes the results of six individual grazing trials that aimed to evaluate forage yield, horse preference, forage nutritive value, and horse health parameters of perennial cool season grasses and alternative forages. Over 10 yr, multiple adult horses in good health grazed monocultures of perennial cool season grasses, annual cool season grasses, legumes, annual warm season grasses, and cover crops in replicated small plots designed in a randomized complete block. Yield was mechanically harvested, samples for forage nutritive value were hand harvested, and preference was visually assessed. Horses were allowed to rotationally graze forages using best management practices throughout multiple Minnesota growing seasons (May through October). Blood glucose and insulin concentrations were collected to determine the impact of certain forage species on horse health parameters. Data were analyzed using various procedures in SAS with P values ≤ 0.05. Forage yields varied greatly between forage types (P ≤ 0.05) and ranged from 13 to 2 Mgּ ha1 . Many forages that were higher yielding, were among the least preferred by horses, including orchardgrass, oat, alfalfa, sudangrass, radish, and turnip. The most preferred forages included Kentucky bluegrass, timothy, meadow fescue, annual ryegrass, winter wheat, red clover, white clover, teff, and berseem clover (P ≤ 0.05). Almost all forages had greater forage nutritive values that met or exceeded the nutritional needs of horses in maintenance. Many warm season grasses had inverted calcium to phosphorus ratios and increased nitrate-nitrogen content. While no consistent relationship was found, horse preference tended to be positively associated with crude protein and nonstructural carbohydrates. When evaluating health parameters, horses grazing teff, an annual warm season grass, had reduced average glucose, average insulin, and peak insulin compared with horses grazing perennial cool season grasses in the late fall (P ≤ 0.05). Collectively, these results suggest that strategic use of pasture forages can be used to help manage common horse health concerns (e.g., obesity and Equine Metabolic Syndrome). While perennial cool-season grasses will remain the foundation of horse pastures in the North Central U.S., this research identified other suitable forage alternatives. Placing a priority on the balance of forage yield and horse preference, annual ryegrass, alfalfa, teff, and berseem clover can be used as horse pasture forage alternatives to extend the grazing season, maximize yield, increase pasture nutritive value, help manage common horse health concerns, and when responding to emerging issues. [ABSTRACT FROM AUTHOR]

Red clover (Trifolium pratense L.), a key perennial pastoral species used globally, can strengthen pastural mixes to withstand increasingly disruptive weather patterns from climate change. Breeding selections can be refined for this purpose by obtaining an in-depth understanding of key functional traits. A replicated randomized complete block glasshouse pot trial was used to observe trait responses critical to plant performance under control (15% VMC), water deficit (5% VMC) and waterlogged conditions (50% VMC) in seven red clover populations and compared against white clover. Twelve morphological and physiological traits were identified as key contributors to the different plant coping mechanisms displayed. Under water deficit, the levels of all aboveground morphological traits decreased, highlighted by a 41% decrease in total dry matter and 50% decreases in both leaf number and leaf thickness compared to the control treatment. An increase in root to shoot ratio indicated a shift to prioritizing root maintenance by sacrificing shoot growth, a trait attributed to plantwater deficit tolerance. Under waterlogging, a reduction in photosynthetic activity among red clover populations reduced several morphological traits including a 30% decrease in root dry mass and total dry matter, and a 34% decrease in leaf number. The importance of root morphology for waterlogging was highlightedwith low performance of red clover: there was an 83% decrease in root dry mass compared to white clover which was able to maintain root dry mass and therefore plant performance. This study highlights the importance of germplasm evaluation across water stress extremes to identify traits for future breeding programs. [ABSTRACT FROM AUTHOR]

The EFSA Panel on Plant Health performed a pest categorisation of Hoplolaimus galeatus (Nematoda: Hoplolaimidae) for the EU. H. galeatus belongs to the order Rhabditida, subfamily Hoplolaiminae. This nematode is not reported from the EU and is not included in the EU Commission Implementing Regulation 2019/2072. It is widely distributed in the USA and is also reported from South America, Africa, Asia and Australia. The identity of H. galeatus is clearly defined and methods for its identification are available. H. galeatus is polyphagous and natural hosts include barley, wheat, rye, red and white clover, alfalfa, cabbage, pine, spruce, oak, apple, grapevine, as well as various ornamental plants and turf grasses. These hosts are grown over vast areas of the EU. The climate of the EU is suitable for the establishment of H. galeatus. Pathways of entry are host plants for planting except seeds, but also soil as a contaminant. Soil import to the EU is prohibited and special requirements apply to import of machinery for agricultural/forestry purposes from third countries. Impact of the nematode is best known for North American plant species. The nematode has been reported to damage cotton, maize, soybean, pine, oak and turfgrass. Many of the hosts represent a considerable economic and environmental value to the EU. Therefore, the Panel concludes that H. galeatus satisfies all the criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest. [ABSTRACT FROM AUTHOR]

Context: High-quality, summer-active pastures could improve meat production in south-eastern Australia by facilitating livestock finishing over summer, with legumes critical for enhancing the nutritive value of pasture mixes. Available legumes vary in their ability to withstand moisture stress and grazing. Aims: We aimed to identify legumes suitable for a summer–autumn finishing system. Methods: We tested pure swards of 12 cultivars across eight legume species in replicated small-plot experiments at Goulburn and Bombala, New South Wales, assessing productivity, persistence and warm-season nutritive characteristics over 2–3 years. Key results: Lucerne (Medicago sativa) was clearly the most productive species during summer and outperformed the clovers (Trifolium spp.) in terms of persistence and productivity throughout most of the experimental period at both sites, except during autumn 2021 after high rainfall in March. Caucasian clover (T. ambiguum) was also highly persistent at both sites. Talish clover (T. tumens) and strawberry clover (T. fragiferum) were more persistent than white clover (T. repens) and red clover (T. pratense). White clover recovered strongly under high rainfall after drought, whereas red clover established rapidly but showed less capacity for post-drought recovery. Hybrid Caucasian × white clover was the least productive legume. Alternative clover species sometimes had slightly lower values of nutritive characteristics than white clover; red clover sometimes had distinctly lower values. Conclusions: Lucerne performed best but several clovers were also productive, persistent and of high nutritive value over the summer–autumn period. Implications: Talish, Caucasian and strawberry clovers warrant further investigation for inclusion in summer-active pastures in south-eastern Australia. Profitability of livestock production over summer–autumn in the temperate environment of south-eastern Australia could be improved by including legumes in pastures that are persistent, productive and of high nutritive value. Evaluation of a range of legume species and cultivars showed that lucerne remained the most reliably productive warm-season option where soil conditions support it. Talish, Caucasian and strawberry clovers were persistent and of high nutritive value, warranting further evaluation in special-purpose warm-season pastures. [ABSTRACT FROM AUTHOR]

Legumes have underpinned transformational change on New Zealand sheep and beef farms over the last 30 years. This was through an emphasis on ewe nutrition based on lucerne or red clover dominant pastures, and increased use of subterranean and white clovers on uncultivatable hill country. Pre- and post-weaning lamb growth rates have increased, and enabled earlier slaughter of heavier lambs. The farm systems results include greater numbers of hoggets mated, higher lambing percentages and greater ewe efficiency (kg lamb weaned/kg ewe mated). Extension packages to support legume use have compared growth rates of resident and legume-based pastures, economic analyses of successful farms and management packages for the most appropriate legume in different environments. Over the same period, the dairy industry rapidly expanded in cow numbers and area onto flat irrigated land on the Canterbury Plains. The nitrogen deficiency of perennial ryegrass was overcome by a linear increase in nitrogen fertiliser use. Environmental concerns from this intensification has led to a legislated nitrogen cap of 190 kg/ha.year. This, coupled with a recent trebling in urea price, has returned attention to increasing the white clover content of these pastures. Nitrogen applications can be minimised by using diverse pastures sown with a legume, herb and <8 kg/ha of perennial ryegrass. Work on other legumes, including annuals and those with condensed tannins, has to date failed to increase their use in most pastoral settings, with the exception of the perennial lupin which is adapted to high-aluminium soils in the South Island High Country. Legumes have underpinned the New Zealand pastoral-based economy for over 100 years. White clover remains the most widely sown, but specialist red clover based pastures are providing high quality feed for fattening livestock in wetter regions of the country. In the drier east coast, lucerne management has been refined based on crop physiology principles. This has allowed earlier lambing, leading to transformational change on-farm. Subterranean clover remains the dominant annual legume used. Successful integration of legumes on-farm has required understanding of the genotype by environment by management interactions. [ABSTRACT FROM AUTHOR]

The experiment was to study nutritional composition of eight native leguminous forage in Xing'an league. The grey-correlation-analysis method was used to comprehensively evaluate eight types of potato family forage in rural areas. The results showed that the crude fat content of Medicago ruthenica L. Trautv. and Trifolium repens L. were significantly higher than that of other forage (P<0.05). The crude protein content of Trifolium pratense L. and Medicago ruthenica L. Trautv. were significantly higher than that of other forage (P<0.05). The acid detergent fiber content of Medicago lupulina L. and Astragalus laxmannii Jacquin were significantly lower than that of other forage (P<0.05). The content of neutral detergent fiber in Astragalus laxmannii Jacquin and Medicago lupulina L. were significantly lower than that of other forage (P<0.05). The crude ash content of Astragalus laxmannii Jacquin was significantly higher than that of other forage (P<0.05). The relative feeding value of Astragalus laxmannii Jacquin was significantly higher than that of other forage (P<0.05). The experiment indicates that nutritional quality of Vicia sepium L. and Melilotus albus Desr. are better, which have high development and utilization value in Xing'an league. [ABSTRACT FROM AUTHOR]

Although traditional ryegrass pastures are still commonly used, they require intense management and input and do not perform well during the summer months. Multispecies systems have been recognised as more sustainable, needing less fertiliser and being tolerant to drought and heat. While data on monoculture system fat composition exist, information for multispecies is scarce. The present study compared the fatty acid (FA), carotenoid and fat-soluble composition of a multispecies system (consisting of perennial ryegrass, timothy, white clover, red clover, chicory and plantain) (MULTI) with two other conventional grazing systems (monoculture perennial ryegrass (PRG) and a binary mixture of perennial ryegrass and white clover (PRG+WC)) over the Irish late-summer grazing season of dairy cattle (July–September). The results showed that the three systems had similar levels of FAs (p > 0.05), with mean α-linolenic acid values of 20.00 mg/g in PRG, 18.51 mg/g in PRG+WC, and 17.90 mg/g in MULTI and mean linoleic acid values of 3.84 mg/g in PRG, 4.16 mg/g in PRG+WC, and 4.39 mg/g in MULTI. Fourfold and twofold increases in the concentrations of α-linolenic acid and linoleic acid, respectively, were observed throughout July–September. Variations in stearic acid concentrations were not significant (p > 0.05), and palmitic acid concentrations increased over time (p < 0.05). The average values of lutein (36.68 mg/kg in PRG, 31.26 mg/kg in PRG+WC, and 35.75 mg/kg in MULTI) and α-tocopherol (2.28 mg/kg in PRG, 1.49 mg/kg in PRG+WC, and 1.83 mg/kg in MULTI) were similar among the grazing systems (p > 0.05), and the average β-carotene value was 25% higher in monoculture ryegrass (140.74 mg/kg) than in the multispecies (102.51 mg/kg) (p < 0.05). Nonetheless, the levels of β-carotene found in the multispecies were still above the recommended intake required for ruminants. In conclusion, multispecies pastures could be included in dairy cattle production systems as a sustainable alternative to traditional pastures while also providing typically adequate sources of fats for animal diets. [ABSTRACT FROM AUTHOR]

This experiment investigated the variation in enteric methane production and associated gas exchange parameters, nutrient digestibility, rumen fermentation, and rumen microbiome when a range of silages based on different forage types (grass or clover), and different species within the 2 types, were fed as the sole feed to heifers. Three grass species (perennial ryegrass, festulolium, and tall fescue) and 2 clover species (red clover and white clover) were included. Perennial ryegrass was harvested at 2 maturity stages in the primary growth, white clover was harvested once in the primary growth, and 4 cuts of festulolium and tall fescue and 3 cuts of red clover were harvested during the growing season, giving 14 different silage batches in total. Sixteen Holstein heifers 16 to 21 mo old and 2 to 5 mo in pregnancy were fed the silages ad libitum as the sole feed in an incomplete crossover design. Each silage was fed to 4 heifers, except for the 2 perennial ryegrass silages, which were fed to 8 heifers; in total 64 observations. The CH 4 production was measured for 3 d in respiration chambers. Heifers fed clover silages had higher dry matter intake (DMI) compared with heifers fed grass silages, and heifers fed tall fescue silages had the numerically the lowest DMI. Compared with grass silages, feeding clover silages led to higher crude protein digestibility but lower neutral detergent fiber (NDF) digestibility. Rumen pH was higher in heifers fed clover silages compared with those fed grass silages. Based on composition analysis, the rumen microbiota of the heifers clustered clearly according to forage type and species. More specifically, 7 of the 34 dominating rumen bacterial genus-level groups showed higher relative abundances for the clover silages, whereas 7 genus-level groups showed higher abundances for the grass silages. Methane yield was higher for heifers fed grass silages than for those fed clover silages when methane production was related to dry matter and digestible organic matter intake, whereas the opposite was seen when related to NDF digestion. The gross energy lost as methane (CH 4 conversion factor, %) reduced from 7.5% to 6.7%, equivalent to an 11% reduction. The present study gives the outlines for choosing the optimal forage type and forage species with respect to nutrient digestibility and enteric methane emission in ruminants. [ABSTRACT FROM AUTHOR]

Mixed-species grassland containing legumes were suggested to increase yield compared to monocultures. Furthermore, some legumes were suggested to be able to sustain growth, even under drought conditions. The first aim of the current study was to measure if multispecies grassland with legumes is also more productive when their N input due to symbiotic N2 fixation is taken into account. Our second aim was to determine the benefit of grass–legume mixtures in terms of dry matter production under naturally occurring drought conditions. Mixed-species grasslands, consisting of monocultures and variable mixtures of (a) Trifolium pratense, (b) Trifolium. repens, (c) Lolium perenne, and (d) a mixture of drought-tolerant grasses (GSWT based), were assessed for their dry matter production over two years with contrasting weather patterns. The legume–grass seeding mixtures received either a fixed (180 kg N ha−1) or adapted N-fertilizer application (0–180 kg N ha−1), with the latter taking the assumed symbiotic N2 fixation by legumes into account. Mixed-species grassland showed improved yield compared to monocultures both in comparably humid and drought-affected years. The benefits of multispecies grass–legume mixtures were considerably more obvious under a fixed but still measurable under an adapted N-fertilizer regime. The species diversity effect appears to be significantly dependent on the additional N supply enabled by legumes' symbiotic N2-fixation. Legumes and drought-tolerant grasses yielded equally well under drought conditions, although legumes showed major advantages during moderate drought and humid conditions. White and red clover, although both legumes, differed significantly in their persistence under elevated-N and their dry matter production under low-N fertilizer application, but were equal in their tolerance towards drought. [ABSTRACT FROM AUTHOR]

The yield and nutritional profile of grass and legume species in Kashmir Valley's rangelands are scantly reported. The study area in this paper included three types of sites (grazed, protected, and seed-sown) divided into three circles: northern, central, and southern Kashmir. From each circle, three districts and three villages per district were selected. Most sites showed higher aboveground biomass (AGB) compared to belowground biomass (BGB), which showed low to moderate effects on biomass. The comparison between northern, central, and southern Kashmir regions revealed that AGB (86.74, 78.62, and 75.22 t. ha−1), BGB (52.04, 51.16, and 50.99 t. ha−1), and total biomass yield (138.78, 129.78, and 126.21 t. ha−1) were the highest in central Kashmir region, followed by southern and northern Kashmir regions, respectively. More precisely, AGB and total biomass yield recorded the highest values in the protected sites of the central Kashmir region, whereas BGB scored the highest value in the protected sites of southern Kashmir region. The maximum yield (12.5 t. ha−1) recorded among prominent grasses was attributed to orchard grass, while the highest crude fiber and crude protein contents (34.2% and 10.4%, respectively), were observed for Agrostis grass. The maximum yield and crude fiber content (25.4 t. ha−1 and 22.7%, respectively), among prominent legumes were recorded for red clover. The highest crude protein content (33.2%) was attributed to white clover. Those findings concluded the successful management of Kashmir rangelands in protected sites, resulting in high biomass yields along with the considerable nutritional value of grasses and legumes. [ABSTRACT FROM AUTHOR]

Weeds are a major threat to agriculture and horticulture cropping systems that reduce yield. Weeds have a better ability to compete for resources compared to the main crops of various agro-ecosystems and act as a major impediment in reducing overall yield. They often act as energy drains in the managed agroecosystems. We studied weed infestation for five different agro-ecosystems in the part of Indian Western Himalayas represented by paddy, maize, mustard, apple and vegetable orchards. Systematic random sampling was done to record flowering phenology and diversity of weeds during the assessment period 2015–2020. We recorded 59 weed species, taxonomically distributed among 50 genera in 24 families. The Asteraceae family has the most species (15% species), followed by Poaceae (14% species), and Brassicaceae (12% species). The Therophytes were the dominant life form followed by Hemicryptophytes. The majority of the weeds were shown to be at their most blooming in the summer (predominantly from June to July). The Shannon index based diversity of weeds ranged from 2.307–3.325 for the different agro-ecosystems. The highest number of weeds was in the horticulture systems (apple > vegetable) followed by agriculture fields (maize > paddy > mustard). Agriculture and horticulture cropping systems were distinguished using indicator species analysis, which was supported by high and significant indicator values for a number of species. Persicaria hydropiper, Cynodon dactylon, Poa annua, Stellaria media, and Rorippa palustris had the highest indicator value in agriculture cropping systems, while Trifolium repens, Phleum pratense, and Trifolium pratense had the highest indicator value in horticulture cropping systems. We found that eleven weed species were unique to apple gardens followed by nine in maize, four in vegetables, two in mustard and one in paddy fields. Spatial turnover (βsim) and nestedness-resultant components (βsne) of species dissimilarity revealed dissimilarity lower than 50% among the five cropping systems. The study is expected to assist in formulating an appropriate management strategy for the control of weed infestation in the study region. [ABSTRACT FROM AUTHOR]

Red clover (Trifolium pratense L.), a key perennial pastoral species used globally, can strengthen pastural mixes to withstand increasingly disruptive weather patterns from climate change. Breeding selections can be refined for this purpose by obtaining an in-depth understanding of key functional traits. A replicated randomized complete block glasshouse pot trial was used to observe trait responses critical to plant performance under control (15% VMC), water deficit (5% VMC) and waterlogged conditions (50% VMC) in seven red clover populations and compared against white clover. Twelve morphological and physiological traits were identified as key contributors to the different plant coping mechanisms displayed. Under water deficit, the levels of all aboveground morphological traits decreased, highlighted by a 41% decrease in total dry matter and 50% decreases in both leaf number and leaf thickness compared to the control treatment. An increase in root to shoot ratio indicated a shift to prioritizing root maintenance by sacrificing shoot growth, a trait attributed to plant water deficit tolerance. Under waterlogging, a reduction in photosynthetic activity among red clover populations reduced several morphological traits including a 30% decrease in root dry mass and total dry matter, and a 34% decrease in leaf number. The importance of root morphology for waterlogging was highlighted with low performance of red clover: there was an 83% decrease in root dry mass compared to white clover which was able to maintain root dry mass and therefore plant performance. This study highlights the importance of germplasm evaluation across water stress extremes to identify traits for future breeding programs.

Plants can affect N 2 O emissions by enhancing nitrogen (N) uptake and other below-ground interactions. However, the specific effect of the root systems of different plant species on the production and accumulation of N 2 O within the soil profile remain largely unknown. The aim of this study was to investigate how plant species from different functional groups, their productivity and root traits affect N 2 O emissions and N 2 O concentrations within the soil profile in a fertilised grassland. We conducted a field experiment with two grasses (Phleum pratense , Lolium perenne), two legumes (Trifolium repens , Trifolium pratense), two forbs (Cichorium intybus , Plantago lanceolata), and the six-species mixture in a fertilised grassland. The effects of these plant communities on N-cycling processes were then assessed through the measurement of above- and below-ground plant traits, plant productivity, soil nutrient availability, N 2 O emissions and its distribution in the soil profile. We found that C. intybus and P. pratense had the lowest N 2 O emissions from the soil, which was mainly related to higher root biomass. The six-species mixture also showed lower N 2 O emissions compared to L. perenne monoculture which was explained by complementary effects between the different plant species. We did not find a relationship between N 2 O emission and its concentration in the soil profile. Higher specific root length and root length density coincided with higher N 2 O concentrations at 10–20 and 20–30 cm soil depths. Since these two traits have been previously linked to reductions in N 2 O emissions emitted from the soil, our results show that the relationships between root traits and N 2 O emissions may not be reflected down in the soil profile. Overall, this study underscores the often-neglected importance of root traits for N-cycling and emphasises the need to better understand how root traits modify N 2 O consumption within the soil profile to design more sustainable grasslands. • C. intybus and P. pratense decreased N 2 O emissions due to higher root biomass. • Mixture lower N 2 O emissions compared to L. perenne due to complementary effects. • There is no relationship between N 2 O emission and its concentration in the soil profile. • Higher root traits coincided with higher N 2 O concentrations in the soil profile. [ABSTRACT FROM AUTHOR]

Since the most important deficiency of forage stands is the inability to maintain adequate legume participation in mixture, it is the purpose of this paper to examine persistence in red clover in Târgovişte Plain eco-climatic conditions, together with the factors that affect it. Six red clover cultivars (Napoca-Tetra, Dacia Tetra, Vesna – tetraploids; Flora, Roxana, Start – diploids) and one white clover diploid cultivar (Karina) were used in pure culture and in mixture (50:50) with hybrid ryegrass (Zefir – tetraploid) in a randomized block design with three replicates. Ground cover assessment in early spring was a suggestive indicator of the stand persistence to define the stability and sustainability boundaries of a reliable intensive system. In the beginning of the third year of cropping, ground cover was 54.33% for tetraploid cultivars (CV = 43.25%), and 67% for diploid cultivars (CV = 6.83%) in pure stands. Same ground cover average of 27% was established both for tetraploid cultivars (CV = 36.47%), and for diploid cultivars (CV = 16.97%) in mixtures with hybrid ryegrass.

【Objective】The flower color of plant was mainly determined by anthocyanins that were regulated by dihydroflavonol 4-reductase (DFR), a key regulatory enzyme in the biosynthetic pathway of anthocyanins. It was important for white clover breeding to explore TrDFR properties and its role in the formation of white clover flower color. 【Method】The petals from the wild-type and red-flowered white clover were used as materials to clone the TrDFR sequence, and the bioinformatics analysis and tissue expression analysis were carried out.【Result】The open reading frame of TrDFR sequence was 969 bp in length, encoding 322 amino acids. The molecular weight of DFR protein in white clover mutant was 36.59 kD, which showed an unstable hydrophilic non-secreted protein. The secondary structure was mainly α-helix and random coil, indicating the closest evolutionary relationship with red clover. Tissue expression analysis demonstrated that TrDFR gene was abundantly expressed in petals, petioles and pedicels of white clover, and the expression levels in stems and petals of red flower mutant were significantly higher than those in wild type (P<0.01). 【Conclusion】The gene TrDFR could be involved in the biosynthesis of anthocyanins in white clover petals, which provided a theoretical basis for further research on the functional mechanism of TrDFR in white clover. [ABSTRACT FROM AUTHOR]

The current study investigated the influence of mixture composition on the biomass yield and early establishment of legume–grass mixtures. The legume species alfalfa (Medicago sativa L. (A)), white clover (Trifolium repens L. (WC)), and red clover (Trifolium pratense L. (RC)) and grass species orchardgrass (Dactylis glomerata L. (O)), tall fescue (Festuca arundinacea Schreb. (TF)), and perennial ryegrass (Lolium perenne L. (PR)) were grown in monocultures and in different legume–grass mixtures. Legume–grass mixtures (M1: WC + O + TF; M2: A + O + TF; M3: A + WC + O + TF; M4: RC + WC + O + TF; M5: A + WC + O + TF + PR; M6: RC + WC + O + TF + PR; and M7: A + RC + WC + O + TF + PR) were sown in a legume–grass seeding ratio of 3:7. The results showed that M2 had the greatest two-year average biomass yield (12.92 t ha−1), which was significantly (p < 0.05), 4.7%, 5.4%, 15.8%, and 29.1% greater than that of WC monoculture, M7, M4, and M1, respectively. The grass biomass yield proportions of all mixtures significantly decreased, while legume biomass yield proportions significantly increased in the second year compared to the first year of establishment. The land-equivalent ratio values of M2 and M4 were greater than 1 in each cutting period. The competition rate of grasses gradually decreased with prolonged establishment time. Overall, the biomass yield, legume and grass biomass yield proportions, land equivalent ratio, and competition rate data highlighted that M2 is the best choice to achieve greater productivity and early establishment in southwest China. [ABSTRACT FROM AUTHOR]

The purpose of the experiment is to determine the composition and energy value of fodder from monoculture and mixed grass stands of perennial ryegrass with legume fodder crops under mountain conditions. It was found that the concentration of the fiber components of the cell walls in the monoculture grass stands of perennial ryegrass was significantly higher compared to the mixed ones, which affected the forage mass digestibility. The highest dry matter digestibility was registered in the biomass of Lolium perenne L. + Trifolium repens L. mixture (642.54 g kg-1 DM) followed by Lolium perenne L. + Lotus corniculatus L. (636.28 g kg-1 DM) and Lolium perenne L. + Trifolium pratense L. (622.18 g kg-1 DM). The monoculture grass stands with red clover had the highest content of neutral detergent fibers (418.03 g kg-1 DM), whereas the bird's-foot-trefoil biomass had the highest concentration of acid detergent fibers (291.98 g kg-1 DM), acid-detergent lignin (70.72 g kg-1 DM) and cellulose (221.27 g kg-1 DM). The forage mass of pasture ryegrass registered lower values for all the investigated parameters compared to its mixtures with leguminous forage crops. The highest caloric content is the harvested grass stands of Lolium perenne L. with Lotus corniculatus L. (17.48 MJ/kg DM). Biomass also has the highest average value of energy useful for the normal course of physiological needs in the animal organism (7.45 MJ/kg DM), as well as in terms of the number of feed units for milk (0.68 in kg DM) and growth (0.62 in kg DM). [ABSTRACT FROM AUTHOR]

Many of the fruits that add diversity and nutrition to our diet are wholly or partially dependent upon flower‐visiting insects. For example, apples (Malus spp.) are self‐incompatible and therefore rely on insect pollinators for fruit development and seed production. Honey bees (Apis mellifera) are often migrated into these orchards when the apples are in bloom. While previous studies have focused on the impact of honey bees to fruit orchards, fewer studies have examined the reciprocal relationship of the orchards to honey bees, particularly if the bees are in the orchard for the entire foraging season, not just during bloom. Here we investigated the foraging dynamics of honey bees in apple orchards in Virginia for two full foraging seasons (April–October, 2018–2019). We decoded, mapped, and analyzed the waggle dances (n = 3710) made by returning foragers, which communicate the distance and direction from the hive to valuable resources, usually nectar or pollen. We found that bees foraged locally at <2 km throughout the season in both 2018 and 2019, with some long‐range recruitment of up to 11 km. Contrary to our expectations, apple blooms did not drive honey bee foraging. We determined in our calculations of percent (%) foraging that honey bees recruit more to the apple orchards after the bloom than during the bloom (29.4% vs. 18.6% in 2018 and 28.5% vs. 21.4% in 2019, respectively). Interestingly, honey bees recruited more to forests while the apples bloomed (36.9% and 25.7% in 2018 and 2019, respectively). Lastly, our odds ratio analysis, which includes a distance correction, indicates the honey bees were more than twice as likely to recruit to apple orchards in June, which is after the bloom, than in April or May, which is during the bloom. Our ground truthing revealed that post‐bloom apple orchards provided foraging opportunities on the growing understory of red and white clover (Trifolium spp.) and plantain (Plantago spp.). These data might therefore have important implications for best management practice decisions for bees located in fruit orchards. [ABSTRACT FROM AUTHOR]

The production and pest susceptibility of eight clover cultivars were compared over two years in the pasture at two contrasting sites: a rain-fed Waikato cattle-grazed pasture and an irrigated Canterbury dairy pasture. The cultivars were four white clovers (Trifolium repens L.) (Grasslands Kopu II, Grasslands Mainstay, SF Quest and Weka), two red clovers (T. pratense L.) (Grasslands Relish, Rubitas), a strawberry clover (T. fragiferum) (Palestine) and a white clover × Caucasian (T. ambiguum M. Bieb.) hybrid (Aberlasting). Grasslands Relish outperformed all other clovers and was associated with the fewest root-damaging nematodes. Both red clovers were less susceptible to insect pests than the white clovers, but Rubitas was vulnerable to root pathogens in Waikato. The poorest performing cultivar was Aberlasting, which was the most susceptible cultivar in all pest and disease measures. The white clovers and strawberry clover performed similarly across most measures. An assay using soils from each site showed that clover seedlings grown in pasteurised Waikato soil were almost ten times larger than those in the non-pasteurised soil whereas pasteurisation made no significant difference with the Canterbury soil. This study demonstrated that root health was a major factor in the contrasting site and cultivar differences in the early years of production. [ABSTRACT FROM AUTHOR]

More than 20 species of bumblebees are reported in South America, nine of them in the Eastern Cordillera of the Colombian Andes. They are known to be important pollinators, but their distribution has been decreasing due to increasing land use change; this has caused an impoverishment of Andean ecosystems. Identifying and protecting the floral resources used by wild bees is key in the management of wild areas to mitigate the impact of human activities in highly intervened areas such as cities or crops. Pollen analysis is an important tool to preserve floral resources used by bees, but it depends on having pollen catalogs that allow adequate taxonomic identifications. The objective of this study was to develop a pollen catalog of plants used by three of the most common Andean bumblebees, Bombus rubicundus, B. hortulanus and B. pauloensis. Sampling was conducted in six localities of the Bogotá savanna, covering an altitudinal gradient between 2600 and 3500 m a. s. l. Floral samples from all localities were prepared by acetolysis, and then analyzed. A reference catalog was digitized from the pollen extracted from the body of workers of each bumblebee species. Pollen grains were described according to their size and morphology. The similarity of the resources used by the different bumblebee species was compared using Jaccard's similarity and distance index. Pollen grains from 99 plants were isolated, identified and described, with Taraxacum officinale, Vallea stipularis, Senna multiglandulosa, Trifolium pratense and Trifolium repens being the pollen sources shared by all species. The most generalist species and the one that used the most exotic plants was B. pauloensis. An illustrated pollen catalog is provided with all the resources used by the bees. [ABSTRACT FROM AUTHOR]

The aim of the study was to estimate the impact of temporal sward formation on soil organic carbon (SOC) variation and soil microbial biomass carbon (MBC) content in rhizosphere. The soil of experimental sites was Bathygleyic Dystric Glossic Retisol, where the soil acidity (pH) was 3.9 and 5.0, and four mixtures of legumes and grasses were cultivated. The mixtures were composed of one legume variety and two grass varieties -- timothy (Phleum pratense L.) 35% and meadow grass (Poa pratensis L.) 15%, 50% of each: 1) red clover (Trifolium pratense L.), 2) white clover (Trifolium repens L.), 3) hybrid clover (Trifolium hybridum L.), and 4) alfalfa (Medicago sativa L.). In the soil of different swards, SOC was more accumulated at 0-10 cm depth. The species composition of the sward did not have any significant influence on the studied indicators. In the soil of swards with pH 5.0, the SOC content in three years increased by 3.3%, SOC stock by 8.5%, and the C to N ratio was favourable for the SOC accumulation. In the naturally acidic soil (pH 3.9), the SOC content and SOC stock decreased by 12.0% and 3.3%, respectively. In the 4th year of sward formation, irrespective of the soil acidity, the SOC stock decreased, but it was similar as in the beginning of sward formation. Sward formation had a significant impact on the soil MBC. In the 2nd year of sward formation, the MBC content increased by 1.7 times in the soil with pH 5.0 and by 2 times in the naturally acidic soil compared to the year of sowing and remained relatively stable in the 3rd and the 4th year of sward formation irrespective of the soil acidity. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the growth dynamics based on the dry matter production of Lotus corniculatus L. variety 202700 and its morphological composition. Design/Methodology/Approach: The study was carried out in greenhouses conditions, from December 2020 to May 2021. The effect of indole butyric acid (IBA) growth promoter in solid Radix 1500 and liquid Radix T 3000 presentation (S+RSolid and S+RLiquid) and a control (Substrate) was evaluated. A completely randomized design with three replications was used, each with twenty pots as replications. 1300 pots were planted, of which 20 were taken monthly from each treatment (30, 60, 90, 120 and 150 days) for subsequent data recording. Results: S+RSolid was the one that presented the highest values followed by S+RLiquid and Control (S) respectively. Study Limitations/Implications: Destructive sampling of less than 30 days or greater than 150 days was not considered. Findings/Conclusions: The plants with the application of 1500 ppm of IBA registered the greatest response in terms of variables: number of leaves, number of stems, leaves (g), stems (g), roots (g), root height (cm), root volume (cm3), greenness index, leaf area (cm²/g) and plant height (cm), with respect to the rest of the treatments. [ABSTRACT FROM AUTHOR]

This review covers a comprehensive overview of the phytoconstituents and bioactivities reported to date for clover honeys produced from various Trifolium spp. against the backdrop of a more general discussion of the chemistry and bioactivity of these important agricultural species. While research into the phytochemical composition of various honeys and their associated bioactivities is growing, this review demonstrates that the literature to date has seen only a limited number of studies on clover honeys. Surprisingly, there appear to be no comparative data on the concentration of flavonoids in general or isoflavonoids specifically in different clover honeys, although the latter have been identified as a main group of bioactive compounds in red clover plants. Based on the findings of this review, the presence of phytoestrogenic isoflavonoids (e.g., formononetin, biochanin A, genistein, daidzein, glycitein) in clover plants and, by extension, in clover honeys should be further investigated, specifically of clover species outside the three popular perennial clovers (red, white and alsike clovers) to exploit new opportunities of potential benefit to both the pharmaceutical and apiculture industries. [ABSTRACT FROM AUTHOR]

A two-year field experiment was conducted to address the effects of mixture composition and legume-grass seeding ratio on the biomass yield and nutritional quality of legume–grass mixtures. Alfalfa (Medicago sativa L.), white clover (Trifolium repens L.), red clover (Trifolium pratense L.), orchardgrass (Dactylis glomerata L.), perennial ryegrass (Lolium perenne L.), and tall fescue (Festuca arundinacea Schreb.) were selected as plant materials. A total of seven legume–grass mixtures (A1: white clover, orchardgrass, and tall fescue; A2: alfalfa, orchardgrass, and tall fescue; B1: alfalfa, white clover, orchardgrass, and tall fescue; B2: red clover, white clover, orchardgrass, and tall fescue; C1: alfalfa, white clover, orchardgrass, tall fescue, and perennial ryegrass; C2: red clover, white clover, orchardgrass, tall fescue, and perennial ryegrass; and D: alfalfa, red clover, white clover, orchardgrass, tall fescue, and perennial ryegrass) were sown in two legume-grass seeding ratios (L:G) of 4:6 and 5:5. The results showed that A2 produced a higher two-year average biomass yield (14.20 t/ha) in L:G of 4:6 than that of other mixtures. The grasses biomass yield proportion decreased while legume biomass yield proportion increased with prolonged establishment time. A2 showed a higher crude protein yield (2.5 t/ha) in L:G of 4:6. C2 and A1 showed lower neutral detergent fiber (4.6 t/ha) and acid detergent fiber (2.8 t/ha) yields in L:G 5:5, while diverse mixtures showed higher water-soluble carbohydrate yields. Overall, A2 showed a relative feed value of 146.50 in L:G of 4:6, indicating that it has not only produced the higher biomass yield but also had a better nutritional quality. [ABSTRACT FROM AUTHOR]

Forage legumes such as white and red clover improve the nutritional quality of grazed pasture but can cause bloat. Their rapid fermentation in the rumen also contributes to environmental issues through higher methane emissions and urinary nitrogen losses. Condensed tannins are known to reduce urinary nitrogen and methane production from grazing animals, reduce bloat, reduce internal parasite burden, and improve animal productivity. Several forage legumes including birdsfoot trefoil and sainfoin do have good levels of condensed tannins but unfortunately these species fail to persist in intensively grazed pasture systems. Conventional breeding approaches including mutagenesis and phenotypic selection have failed to deliver condensed tannins in legumes, such as white clover, red clover or lucerne, that do persist under grazing. A recent advance using a molecular biology approach has identified a transcription factor or master switch that can 'turn on' the condensed tannin pathway present in white clover allowing biologically significant levels of condensed tannin expression in leaf tissue. In vitro tests have demonstrated that the condensed tannins produced in white clover leaves can bind protein at a pH 6.5, as found in the rumen, and then release them at pH 2.5, the pH in the abomasum, before entering the small intestine for amino acid absorption. Additional tests have demonstrated that these condensed tannins can reduce methane production by up to 15.7% in the first 6 h of incubation. The journey to this point, and the challenges ahead to deliver white clover cultivars with condensed tannin expression, is described. [ABSTRACT FROM AUTHOR]

This article presents the data obtained from a field experiment in which grass–clover leys were fertilized with increasing N rates applied in either mineral N fertilizer and/or cattle slurry forms. The leys were composed of a 2–species mixture of white clover (Trifolium repens L.) and ryegrass (Lolium perenne L.) and a 4–species mixture of white clover, red clover (Trifolium pratense L.), festulolium (Festulolium braunii) and ryegrass. In total, eighty fields were established at two farm sites in the western part of Denmark on sandy soils and monitored for two herbage seasons (2018–2019). Dry matter yield, botanical composition, N concentration and the proportion of N derived from the atmosphere using the 15 N dilution method were recorded in the harvestable biomass after each cut. Furthermore, the specific growth, N uptake and quantitative biological N fixation of the species were determined.The dataset can be used to establish the N balance, to calculate the optimal economic fertilization rate based on grass-clover composition and to predict N leaching and residual effect.The data presented were thoroughly used and discussed in the research article “Contrasting effects of slurry and mineral fertilizer on N2 fixation in grass-clover mixtures”.

Background: The nitrogen (N) response in grass-clover is influenced by quantity and timing of N-fertilizer, but less is known about the effect of fertilizer type (animal manure vs. mineral N) and N form (ammonium vs. nitrate). Method: A field trial was established in a mixture of white clover (Trifolium repens L.), red clover (Trifolium pratense L.) and perennial ryegrass (Lolium. perenne L.) with mineral N and slurry. DM yield in grass and clover and N2-fixation activity (%Ndfa) was determined in four cuts during the season. Results: Fertilization resulted in a peak in soil mineral N within 1-2 weeks of application and decreased to background rate prior to plant harvest. No differences in white clover N2-fixation activity were found between fertilizer types within each N level. Conclusions: The experiment showed that mineral N form does not affect the regulation of N2-fixation activity. The fertilizer N level was the only factor affecting white clover N2-fixation activity. Further, the results demonstrated an N threshold below which annual %Ndfa was always high (~90%). This was controlled by the companion grass' ability to deplete the soil mineral N pool. When N fertilization exceeded this threshold (above 180 kg N ha-1 yr-1 in our case) annual %Ndfa significantly decreased as did qBNF in the harvested biomass. [ABSTRACT FROM AUTHOR]

Desiccation with diquat about one week before seed harvest has been common practise in Norwegian clover seed production. However, after withdrawal of diquat in 2020, clover seed growers no longer have desiccators available. In 2019 and 2020, six field trials in red clover and two field trials in white clover were carried out to evaluate alternative chemical products at different rates and at two different spraying dates, either early at 50% mature seed heads and / or late at 65% mature seed heads. Products included, either for one or two years, was Spotlight Plus (carfentrazonethyl), Beloukha (pelargonic acid), Glypper (glyphosate), Gozai (Pyraflufen-ethyl), Harmonix LeafActive (acetic acid), Harmonix FoliaPlus (pelargonic acid), Flurostar (fluroxypyr) and Saltex (sodium chloride) and liquid urea-based fertilizers. In addition, swathing was examined as an alternative in two red clover trials in 2020. While none of the tested chemicals were superior to diquat, the most promising alternatives were Harmonix FoliaPlus and Harmonix LeafActive in red clover or Harmonix FoliaPlus in white clover. Although usually less effective than these products, Beloukha also had an acceptable desiccation effect, especially when sprayed early and late. Swathing before harvest, using finger bar cutters, was an effective drying method under favourable weather conditions. [ABSTRACT FROM AUTHOR]

The natural abundance technique is used to estimate the amount of N fixed by a legume, but the B value of this particular species has to be known first. This value, however, varies with the rhizobium strains and climatic conditions. To assess this variation range for forage legumes growing under Uruguayan conditions, seeds of Lotus corniculatus L., Trifolium repens L. and Trifolium pratense L. were sown in pots containing vermiculite-sand and inoculated with a commercial or a promissory rhizobium strain, and grown in a greenhouse during the winter and spring, or in a growth chamber that simulated summer temperatures. At harvest, B values were determined by analyzing δ15N in shoots, and their proportion of N derived from air (%Ndfa) were also estimated. All B values were within the reported ranges. The rhizobium strain affected B only in L. corniculatus, resulting in a 0.55‰ shift between the commercial and the research strains. The growing seasonal effect, however, was important across the three species, with B values becoming more negatives as the temperature and the isotopic fractionation increased. However, the impact of these B value changes on the associated %Ndfa values were not always agronomically relevant, but depended on the particular legume–rhizobia combination. [ABSTRACT FROM AUTHOR]

1. The aim of this study was to evaluate the production of equol (4ʹ,7-isoflavandiol; a bacterial polyphenol metabolite which is an isoflavandiol oestrogen metabolised from daidzein from plants) enriched eggs from free-range hens fed different pasture species. Four species were tested: red clover, white clover, ryegrass and chicory. 2. The study was conducted from June to September 2017 on eight free range, outdoor areas, each containing fifteen laying hens and sown with a single pasture species 3. Precursors of equol (daidzein, formononetin) were analysed every fortnight from the fresh pasture cover in each area, as well as equol and daidzein levels in eggs. 4. Daidzein and formononetin concentrations in the fresh pasture samples differed significantly according to species (P < 0.001), whereby red clover had the highest concentrations of daidzein and formononetin (85 and 996 µg/g DM, respectively). 5. Equol concentration in eggs differed according to pasture species (P < 0.001). Equol concentrations reached about 1,200 ng/g DM in eggs from hens with access to red clover. These eggs can represent a valuable source of equol in the human diet. [ABSTRACT FROM AUTHOR]

A field experiment was conducted at the Research Institute of Mountain Stockbreeding and Agriculture (Bulgaria) to establish the impact of annual organic fertilizing on the bioproductive characteristics of a degraded grassland with a predominant share of grasses in mountain conditions. A significant effect on the biological productivity of the at all levels of fertilizing was found. The excess in the values of the indicator compared to the control was from 49.1% to 98.8%. Desired changes in the individual groups and species of the botanical composition of the pastures were observed. Lotus corniculatus, Vicia sativa, Trifolium hybridum (from legumes) and Festuca arundinaceae Scherb., Festuca rubra, Anthoxanthum odoratum, Agrostis capillaris (grasses) showed the highest resistance and adaptability to mountain conditions. The elimination of Trifolium repens, Trifolium aureum Pollich., Vicia villosa Roth. (second year), Vicia cracca, Medicago lupulina (third year) and Trifolium pratense (fourth year) from the composition of the grass mass affected the fodder and nutritional value of the grasslands. The grasslands treated with 1000 and 4000 kg/da dose of manure have the highest concentration of crude protein, respectively 14.39 and 14.05%. The values of the indicator exceeded the control by 20.0% and 17.2%, respectively. With a slight difference in protein content were grasslands treated with 2000 kg/da (12.03%) and 3000 kg/da (12.52%). The excess compared to the control variants was 0.3% и 4.4%, respectively. The highest fiber (44.37%) is the biomass with an imported dose of 3000 kg/da. The fertilizing variants registered with 2.3% to 11.4% higher content of crude ash compared to the control. [ABSTRACT FROM AUTHOR]

Toxic effect of Origanum vulgare subsp. hirtum (Link) Ietsw. essential oil was assayed on weeds (Lolium perenne L. Trifolium pratense L. and Trifolium repens L.), potato plants (Solanum tuberosum L.) and Myzus persicae Sulz. The GC-MS analysis of the essential oil revealed carvacrol as the main component. The essential oil solutions completely inhibited the seed germination of the target weeds at concentration 2 µL/mL in Petri dishes. In contrast, in soil, this effect was achieved at a concentration of 5 µL/mL for studied dicot and at 10 µL/mL for the monocot species. Total destruction of target weeds at post-emergence stage after spraying with solution of the essential oil in concentrate 10 µL/mL was found. Mature potato plants were poorly affected by the essential oil. A completely toxic effect on M. persicae was established by applying a solution with a concentration of 3 µL/mL of the essential oil. The experimental data indicate that the essential oil of the species has great potential, as an alternative to synthetic biocides, for the production of eco-friendly formulations for pest control. [ABSTRACT FROM AUTHOR]

Background: Nitrogen (N) plays an important role in dairy systems, both in production costs and as an indicator of environmental impact. Objective: The objective was to investigate the effects on milk production and nitrogen excretion by dairy cows fed cut pastures either of perennial ryegrass-red clover (PRG-RCL) or annual ryegrass-white clover (ARG-WCL), complemented with ground maize straw and of commercial concentrate. Methodology: Six dairy cows were used in a double cross-over on-farm experiment with 14-day experimental periods. Milk yield and composition, live weight, body condition score, chemical composition of feeds and N content in faeces, urine, milk and blood were recorded over the last four days of each experimental period. Results: No differences were found (P > 0.05) for milk yield nor composition. Cows on ARG-WCL had a higher N intake and N balance than those on PRG-RCL (P < 0.05); also, there were higher levels of blood urea nitrogen (BUN) and urine urea nitrogen (UUN) when cows were fed ARG-WCL than PRG-RCL (P < 0.05). Implications: This work allows us to know the amount of nitrogen excreted in milk production in small-scale dairy systems. Conclusions: It is concluded that PRG-RCL may be a feeding strategy to reduce N excretion and achieve higher efficiency in N use for small-scale dairy systems.