The objective of this study was to examine the effects of arbuscular mycorrhizal fungi (AMF) and organic manure on the growth and element uptake of Trifolium repens and the soil chemical and ...biological characteristics in sterilized and natural soils. An increase in organic manure input reduced mycorrhizal colonization in the sterilized soil treatment. In comparison to the treatments in which AMF inoculation did not occur, AMF inoculation significantly promoted the biomass of shoots and roots in the sterilized soil at the 2.5 and 5% organic manure incorporation levels, whereas AMF had no effect on biomass in the natural soil. Regardless of soil type and whether AMF inoculation occurred, the N, P, K and Mg concentrations of the shoots and roots and the Cd concentration of the roots were increased by organic manure amendment. In comparison to the 2.5% organic manure amendment treatment, the addition of 5% organic manure to the sterilized soil decreased the N, P, K, Ca and Cd contents in the absence of AMF but had no negative effect on the N, P, K, Mg and Cd contents in the presence of AMF. The soil available P, pH, microbial biomass carbon, microbial biomass nitrogen, dissolved organic carbon, diethylenetriamine pentaacetic acid (DTPA)-extractable Cd and root Cd concentrations increased with increasing organic manure input. These results indicate that additive effects of AMF inoculation and organic manure on the growth and nutrient contents of Trifolium repens occurred in the Cd-contaminated sterilized soil but not in the natural soil, implying the limited function of nonnative AMF introduced into natural soils.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
White clover (WC) offers an alternative source of nitrogen (N) for pasture-based systems. Substituting energy- and carbon-intensive synthetic N fertilizers with N derived from biological fixation by ...WC has been highlighted as a promising environmental mitigation strategy through the omission of emissions, pollutants, and energy usage during the production and application of synthetic fertilizer. Therefore, the objective was to investigate the effect of the inclusion of WC in perennial ryegrass (PRG) swards on the environmental impact of pasture-based dairy systems. Cradle-to-farm gate life cycle assessment of 3 pasture-based dairy systems were conducted: (1) a PRG–WC sward receiving 150 kg of N/ha per year (CL150), (2) a PRG–WC sward receiving 250 kg of N/ha per year (CL250), and (3) a PRG-only sward receiving 250 kg of N/ha per year (GR250). A dairy environmental model was updated with country-specific N excretion equations and recently developed N2O, NH3, and NO3− emission factors. The environmental impact categories assessed were global warming potential, nonrenewable energy, acidification potential, and eutrophication potential (marine and freshwater). Impact categories were expressed using 2 functional units: per hectare and per metric tonne of fat- and protein-corrected milk. The GR250 system had the lowest milk production and highest global warming potential, nonrenewable energy, and acidification potential per tonne of fat- and protein-corrected milk for all systems. The CL250 system produced the most milk and had the highest environmental impact across all categories when expressed on an area basis. It also had the highest marine eutrophication potential for both functional units. The impact category freshwater eutrophication potential did not differ across the 3 systems. The CL150 system had the lowest environmental impact across all categories and functional units. This life cycle assessment study demonstrates that the substitution of synthetic N fertilizer with atmospheric N fixed by WC has potential to reduce the environmental impact of intensive pasture-based dairy systems in temperate regions, not only through improvement in animal performance but also through the reduction in total emissions and pollutants contributing to the environmental indicators assessed.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Soil salinity is an increasingly serious problem worldwide that reduces agricultural output potential. Selected beneficial soil bacteria can promote plant growth and augment tolerance to biotic and ...abiotic stresses. Bacillus subtilis strain GB03 has been shown to confer growth promotion and abiotic stress tolerance in the model plant Arabidopsis thaliana. Here we examined the effect of this beneficial soil bacterium on salt tolerance in the legume forage crop, white clover. Plants of white clover (Trifolium repens L. cultivar Huia) were grown from seeds with or without soil inoculation of the beneficial soil bacterium Bacillus subtilis GB03 supplemented with 0, 50, 100, or 150 mM NaCl water into soil. Growth parameters, chlorophyll content, malondialdehyde (MDA) content and osmotic potential were monitored during the growth cycle. Endogenous Na(+) and K(+) contents were determined at the time of harvest. White clover plants grown in GB03-inoculated soil were significantly larger than non-inoculated controls with respect to shoot height, root length, plant biomass, leaf area and chlorophyll content; leaf MDA content under saline condition and leaf osmotic potential under severe salinity condition (150 mM NaCl) were significantly decreased. Furthermore, GB03 significantly decreased shoot and root Na(+) accumulation and thereby improved K(+)/Na(+) ratio when GB03-inoculated plants were grown under elevated salt conditions. The results indicate that soil inoculation with GB03 promotes white clover growth under both non-saline and saline conditions by directly or indirectly regulating plant chlorophyll content, leaf osmotic potential, cell membrane integrity and ion accumulation.
White clover (Trifolium repens L.) belongs to the Fabaceae family legume and is cultivated in China for its medicinal properties and ornamental value. White clover is grown around the world for ...forage, turf , green manure and soil conservation purposes (Zhang el al. 2016). In October 2021, an investigation of a 1,000 m2 plant nursery in Lanzhou, China (36°06'N, 103°83'E) found that 80% of White clover plants were infected, and powdery mildew covered 95% of the leaf area. The disease had seriously destroyed the forage quality and reduced the ornamental value. Initially, thin, radial, irregular white colonies appeared on leaves and gradually spread to stems. The white colonies then expanded and thickened to cover upper surface of the leaf, and microscopic hyphae appeared on the bottom of the leaf. In severe cases, the infection resulted in dieback of the leaf. A small area of sporulating fungus was stripped off from the leaf surface with tape and mounted in sterile water for microscopic examination (Mukhtar et al. 2017). Conidiophores were cylindrical, consisting of a foot cell followed by three to four short cells, measuring 75 to 160 × 7 to 10 μm. Conidiophores had straight, cylindric foot cells ranging from 25 to 40 µm long. Singly produced conidia were hyaline and ranged in shape from oblong to cylindrical. Conidia lacked distinct fibrotic bodies and measured 30 to 45 × 15 to 25 μm in length. Long, unbranched germ tubes formed from the ends of the conidia and nipple-shaped appressoria developed on epiphytic mycelia. Based on these morphological characteristics, the pathogen was initially identified morphologically as Erysiphe polygoni (Braun and Cook 2012). To validate the identity, the internal transcribed spacer (ITS) region of the pathogen (SY77) rDNA was amplified by PCR and sequenced using the ITS1/ITS4 primers (White et al. 1990). The resulting sequences were registered to GenBank (GenBank Accession No.OM280998). The ITS sequence of the SY77 was 100% (640/640) identical to E. polygoni (LC009892) on Polygonum aviculare in the United Kingdom and 99% (638/640) identical to E. polygoni (MK685172) on Antigonon leptopus in Taiwan. MEGA 7.0 was used to conduct the neighbor-joining phylogenetic analysis using the ITS sequences from GenBank. The data indicated that the strain SY77 and E. polygoni clustered together on the same branch. Pathogenicity tests were conducted by gently pressing the infected leaves onto five healthy potted White clover plants, while five non-inoculated plants were used as controls (Michael et al. 2021). The plants were maintained in a growth chamber (25 ℃, 14 h light, and 10 h dark period, RH > 80%). After 10 days, the inoculated plants developed powdery mildew symptoms, whereas the control plants remained symptom-free. The fungus on the inoculated plants was re-isolated, re-identified, and confirmed as E. polygoni based on morphological observations and molecular identification. There is no previous report on E. polygoni causing powdery mildew on White clover in China. The powdery mildew caused by E. polygoni on Red clover has been reported in China and Bulgaria, respectively (Yuan el al.1991; Galina el al. 2017). To our knowledge, this is the first report of powdery mildew caused by E. polygoni on White clover in China. References: 1. Zheng, L., et al. 2018. Plant Dis. 102:628. 2. Mukhtar, G., et al. 2017. Plant Dis.101:1, 246. 3. Braun, U., and Cook, R. T. A. 2012. Taxonomic Manual of the Erysiphales (Powdery Mildews), CBS Biodiversity Series No.11. CBS, Utrecht. 4. Michael, R. F., et al. 2021.Plant Dis. First look.( doi.org/ 10.1094/PDIS-09-21-2060-PDN). 5. Yuan, Q. H., el al.1991. Pratacult Sci.05:59 (in Chinese). 6. Galina, N., et al, 2017. BIOTECHNOL Anim Husb.33.127.
•Similar yield ranking of white clover populations in monocultures and mixtures.•Relative performance of white clover population depended on site conditions.•The mixture benefit was higher at the ...fertile than at the more marginal site.•Including chicory into the mixtures increased the between-year yield stability.
Due to complementary resource use of the components, grass-clover mixtures are usually higher yielding compared to unfertilized pure grass stands. The number and identity of species in a mixture are known as important factors for the mixture’s productivity. How intraspecific variability of component species affects the productivity of mixtures has received less attention. We established an experiment to investigate how and to what extent different populations of white clover (Trifolium repens L.) affect mixture performance. Eight novel populations of white clover and one variety of perennial ryegrass (Lolium perenne L.) and chicory (Cichorium intybus L.) were grown as monocultures and in two- and three-species mixtures at two sites differing in soil fertility. Aboveground herbage was cut twice in the establishing year and four times in each of the three following years. The accumulated dry matter yield was calculated as the sum of the four annual dry matter yields. There was no significant interaction between white clover population and crop stand (i.e., white clover monoculture or mixtures with partner species) on dry matter yield accumulated over four years; the white clover population that performed well in monoculture also did so in mixtures. At both sites, the binary mixtures of white clover and chicory produced significantly higher dry matter yields than the white clover monocultures and other mixtures. Inclusion of chicory in mixtures significantly increased the stability of yield production as well. Site conditions strongly affected the performance of the forage species mixtures. We found that the benefit of mixtures over monocultures was more related to the identity of species in mixture than to the white clover population.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
White clover (Trifolium repens L.) is an important perennial legume forage widely cultivated in China (Zhang et al. 2016). In April 2018, severe necrotic lesions on leaves were observed in a ...cultivated white clover field in Chongqing, China. Approximately 90% of plants in the field were affected. Leaf spots were amphigenous, dark-brown, elliptic to subcircular, with diameter ranging between 1 to 12 mm, well defined by brown margins and yellow halos. Severely infected leaves became withered and abscised. Stems and flowers were not affected by the disease. Symptomatic leaves were surface sterilized with 70% ethanol for 30 s followed by 0.1% HgCl2 treatment for 3 min, and rinsed in sterile water three times. Thereafter, tissue samples from margins of individual lesions were placed on potato dextrose agar (PDA) amended with 50 mg/L of chloramphenicol and incubated at 25℃ in the dark. An olivaceous gray fungal colony was consistently isolated (90.5% isolation frequency). After 15 days of incubation, subglobose, black pycnidia developed in the cultural medium. Conidia were hyaline, ellipsoid to oblong, nonseptate(n = 50), ranging from 4.0 to 7.5 μm long (5.6 ± 2.3µm) × 2.0 to 3.8 μm wide(2.8 ± 1.0 µm). On the basis of its morphological characteristics, the fungus was identified as Boeremia sp. (Aveskamp et al. 2010). To confirm the identity, the internal transcribed spacer region (ITS), large subunit ribosomal RNA (LSU), partial actin (ACT), RNA polymerase II second largest subunit (rpb2) and beta-tubulin (tub2) genes were amplified with primers ITS1/ITS4, LR0R/LR7, ACT-512F/ACT-783R, RPB2-5F2/fRPB2-7cR, and Btub2Fd/Btub4Rd, respectively, in eight representative isolates and sequenced(Aveskamp et al. 2009; Chen et al. 2015). BLAST results showed 100% identity of the ITS (506/506 nucleotides), LSU (966/966 nucleotides), ACT (244/244 nucleotides) and tub2 (297/297 nucleotides) sequences with those of B. exigua (KY419536, MK398746, EU880878, and MK514090) and 99.83% identity of those of the rpb2 (593/594 nucleotides) sequence with B. exigua (KT389572).Based on morphology and DNA sequence analysis, the associated fungus was identified as B. exigua. Representative sequences of one isolate (BT2-1) were deposited in GenBank (MN826339, MN836592, MT265217, MT265218 and MT265219). In a pathogenicity test, ten 2-month-old potted white clover plants were spray-inoculated with a spore and mycelial suspension (approximately 105 CFU/mL) and the control plants were inoculated with sterile distilled water. Plants were incubated in a greenhouse at 20 to 24°C under natural light and enclosed in plastic bags for the first 3 days to maintain high humidity. After 10 days, typical dark-brown lesions similar to those seen in naturally infected leaves developed on the inoculated leaves and not on the control plants. B. exigua was reisolated from the lesions, thus completing Koch's postulates. There is some evidence that B exigua is capable of invading seedling root tissue of white clover and causing necrotic lesions on roots (Skipp and Christensen,1982). However, to our knowledge, this is the first report of leaf spot on T. repens caused by B. exigua. This disease severely reduces forage quality and yield. Proper identification of the causal organism is essential in formulating management strategies.
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.
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IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Increased emissions of N2O, a potent greenhouse gas (GHG), from agricultural soils is a major concern for the sustainability of grassland agriculture. Emissions of N2O are closely associated with the ...rates and forms of N fertilisers applied as well as prevailing weather and soil conditions. Evidence suggests that multispecies swards require less fertiliser N input, and may cycle N differently, thus reducing N loss to the environment. This study used a restricted simplex-centroid experimental design to investigate N2O emissions and soil N cycling following application of urea-N (40 kg N ha−1) to eight experimental swards (7.8 m2) with differing proportions of three plant functional groups (grass, legume, herb) represented by perennial ryegrass (PRG, Lolium perenne), white clover (WC, Trifolium repens) and ribwort plantain (PLAN, Plantago lanceolata), respectively. Swards were maintained under two contrasting soil moisture conditions to examine the balance between nitrification and denitrification. Two N2O peaks coincided with fertiliser application and heavy rainfall events; 13.4 and 17.7 g N2O-N ha−1 day−1 (ambient soil moisture) and 39.8 and 86.9 g N2O-N ha−1 day−1 (wet soil moisture). Overall, cumulative N2O emissions post-fertiliser application were higher under wet soil conditions. Increasing legume (WC) proportions from 0% to 60% in multispecies swards resulted in model predicted N2O emissions increasing from 22.3 to 96.2 g N2O-N ha−1 (ambient soil conditions) and from 59.0 to 219.3 g N2O-N ha−1 (wet soil conditions), after a uniform N application rate. Soil N dynamics support denitrification as the dominant source of N2O especially under wet soil conditions. Significant interactions of PRG or WC with PLAN on soil mineral N concentrations indicated that multispecies swards containing PLAN potentially inhibit nitrification and could be a useful mitigation strategy for N loss to the environment from grassland agriculture.
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•Measurement of N2O emissions and N cycling from varying sward compositions.•Post N application (40 kg N ha−1) N2O loss increased with white clover proportion.•N2O emissions from PRG were 2.5 fold higher in wet soil (WFPS > 60%) compared to ambient.•Soil N dynamics suggest denitrification as dominant N2O source when WFPS > 60%.•Plantago lanceolata (forage herb) potentially regulates N cycling pathways.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Urban ecosystems are an increasingly dominant feature of terrestrial landscapes. While evidence that species can adapt to urban environments is accumulating, the mechanisms through which urbanization ...imposes natural selection on populations are poorly understood. The identification of adaptive phenotypic changes (i.e. clines) along urbanization gradients would facilitate our understanding of the selective factors driving adaptation in cities. Here, we test for phenotypic clines in urban ecosystems by sampling the frequency of a Mendelian-inherited trait—cyanogenesis—in white clover (Trifolium repens L.) populations along urbanization gradients in four cities. Cyanogenesis protects plants from herbivores, but reduces tolerance to freezing temperatures. We found that the frequency of cyanogenic plants within populations decreased towards the urban centre in three of four cities. A field experiment indicated that spatial variation in herbivory is unlikely to explain these clines. Rather, colder minimum winter ground temperatures in urban areas compared with non-urban areas, caused by reduced snow cover in cities, may select against cyanogenesis. In the city with no cline, high snow cover might protect plants from freezing damage in the city centre. Our study suggests that populations are adapting to urbanization gradients, but regional climatic patterns may ultimately determine whether adaptation occurs.
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BFBNIB, NMLJ, NUK, PNG, SAZU, UL, UM, UPUK
Allelopathy is a unique phenomenon in nature that refers to direct or indirect, positive or negative, impact of one plant (including microorganisms, insects, and herbivores) on another plant through ...chemical secretions (allelochemicals). Ambrosia artemisiifolia (ragweed) is a well-known invasive plant possessing allelochemicals that have been found to inhibit the growth and development of plants in their immediate environment. This study aimed to determine the allelopathic impact of the aqueous leaf extract isolated from ragweed leaves (concentrated extract, 1:2, 1:4, and 1:8) on seed germination and early seedling growth of barley (Hordeum vulgare L.) and white clover (Trifolium repens L.). The examined aqueous extract of ragweed had a higher inhibitory effect on the germination of white clover seeds compared to barley. The germination of barley seeds was the lowest at the concentrated leaf extract (up to 70%), while the same process in barley seeds was not determined at this concentration. In the other treatments, germination was significantly lower in both examined species compared to the control. Shoot and root growth of both researched species was inhibited in all treatments compared to the control. The obtained results indicate that the aqueous ragweed extract significantly inhibits the growth of roots and shoots of both tested species, which increases the need for more intensive research on this topic.
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