Search

Herbicide promotes physiological and biochemical changes even in tolerant species. The objective of this study was to evaluate the interference of the herbicides clodinafop-propargyl and 2,4-D in the antioxidant system of wheat, and iodosulfuron-methyl in wheat and ryegrass. Two studies, divided into three experiments, were conducted in an entirely randomized design in a greenhouse and phytotron. The first study tested herbicides iodosulfuron-methyl, clodinafop-propargyl, 2,4-D, and a control without application in wheat. The second, divided into two experiments with wheat and ryegrass, tested iodosulfuron-methyl doses (0, 1.75, 3.5, and 7.0 g a.i. ha-1). The variables analyzed were the concentrations of chlorophylls a, b, carotenoids, hydrogen peroxide (H2O2), lipid peroxidation (TBARS), electrolyte leakage, and antioxidant system activity. The herbicide treatments iodosulfuron-methyl, clodinafop-propargyl, and 2,4-D decreased the concentrations of chlorophylls and carotenoids in wheat plants. The herbicides promoted oxidative stress with increased H2O2 and TBARS concentrations. Increasing the dose of iodosulfuron-methyl provided a reduction in the activity of the enzymatic antioxidant system in wheat and ryegrass. [ABSTRACT FROM AUTHOR]

Background: Seed retention is the basic prerequisite for seed harvest. However, only little breeding progress has been achieved for this trait in the major forage grasses. The aim of this study was to evaluate the potential of plant genetic resources of the important fodder grasses Festuca pratensis Huds. and Lolium perenne L. as source for seed retention in the breeding process. Furthermore, the morphology of the abscission zone, where shattering occurs, was studied on the cell tissue level in different developmental stages of contrasting accessions. Results: 150 and 286 accessions of Festuca pratensis and Lolium perenne were screened for seed retention, respectively. Contrasting accessions were selected to be tested in a second year. We found a great variation in seed retention in Festuca pratensis and Lolium perenne, ranging from 13 to 71% (average: 35%) and 12 to 94% (average: 49%), respectively, in the first year. Seed retention was generally lower in the second year. Cultivars were within the accessions with highest seed retention in Festuca pratensis, but had lower seed retention than ecotypes in Lolium perenne. Field-shattered seeds had a lower thousand grain weight than retained seeds. Cell layers of the abscission zone appeared already in early seed stages and were nested within each other in accessions with high seed retention, while there were two to three superimposed layers in accessions with low seed retention. Conclusions: Plant genetic resources of Lolium perenne might be a valuable source for breeding varieties with high seed retention. However, simultaneous selection for high seed weight is necessary for developing successful commercial cultivars. [ABSTRACT FROM AUTHOR]

One of the largest isotopic datasets of the ancient Eastern Mediterranean region is evaluated, based on plants (n = 410), animals (n = 210) and humans (n = 16) from Tell Tweini (Syria). Diachronic analysis of plant and faunal specimens from four main periods of occupation: Early Bronze Age (2600–2000 BC), Middle Bronze Age (2000–1600 BC), Late Bronze Age (1600–1200 BC) and Iron Age (1200–333 BC) were investigated. Mean Δ13C results from seven plant species reveal emmer and free threshing wheat, olives, bitter vetch, rye grass and barley were adequately or well-watered during all periods of occupation. The grape Δ13C results suggest excellent growing conditions and particular care for its cultivation. The δ15N results indicate that especially the emmer and free threshing wheats received some manure inputs throughout the occupation sequence, while these were likely further increased during the Iron Age, encompassing also the olive groves and grape vineyards. Generally, domestic animals (cattle, sheep, goats) had C3 terrestrial diets and were kept together in similar environments. However, some animals consumed significant amounts of marine or C4 plants, possibly from disturbed habitats due to land use pressure or salt tolerant grasses and shrubs from wetland environments, which were recorded in the direct vicinity of the site. Middle Bronze Age humans consumed a C3 terrestrial diet with no measurable input from C4, freshwater or marine protein sources. Interestingly, the human diet was relatively low in animal protein and appears comparable to what is considered today a typical Mediterranean diet consisting of bread (wheat/barley), olives, grapes, pulses, dairy products and small amounts of meat. The combined isotopic analysis of plants, animals and humans from Tell Tweini represents unbroken links in the food chain which create unparalleled opportunities to enhance our current understanding of environmental conditions, climate change and lifeways in past populations from the Eastern Mediterranean. [ABSTRACT FROM AUTHOR]

Food allergies mediated by specific IgE (sIgE) have a significant socioeconomic impact on society. Evaluating the IgE cross-reactivity between allergens from different allergen sources can enable the better management of these potentially life-threatening adverse reactions to food proteins and enhance food safety. A novel banana fruit allergen, S-adenosyl-L-homocysteine hydrolase (SAHH), has been recently identified and its recombinant homolog was heterologously overproduced in E. coli. In this study, we performed a search in the NCBI (National Center for Biotechnology Information) for SAHH homologs in ryegrass, latex, and kiwifruit, all of which are commonly associated with pollen-latex-fruit syndrome. In addition, Western immunoblot analysis was utilized to identify the cross-reactive IgE to banana SAHH in the sera of patients with a latex allergy, kiwifruit allergy, and ryegrass allergy. ClustalOmega analysis showed more than 92% amino acid sequence identity among the banana SAHH homologs in ryegrass, latex, and kiwifruit. In addition to five B-cell epitopes, in silico analysis predicted eleven T-cell epitopes in banana SAHH, seventeen in kiwifruit SAHH, twelve in ryegrass SAHH, and eight in latex SAHH, which were related to the seven-allele HLA reference set (HLA-DRB1*03:01, HLA-DRB1*07:01, HLA-DRB1*15:01, HLA-DRB3*01:01, HLA-DRB3*02:02, HLA-DRB4*01:01, HLA-DRB5*01:01). Four T-cell epitopes were identical in banana and kiwifruit SAHH (positions 328, 278, 142, 341), as well as banana and ryegrass SAHH (positions 278, 142, 96, and 341). All four SAHHs shared two T-cell epitopes (positions 278 and 341). In line with the high amino acid sequence identity and B-cell epitope homology among the analyzed proteins, the cross-reactive IgE to banana SAHH was detected in three of three latex-allergic patients, five of six ryegrass-allergic patients, and two of three kiwifruit-allergic patients. Although banana SAHH has only been studied in a small group of allergic individuals, it is a novel cross-reactive food allergen that should be considered when testing for pollen-latex-fruit syndrome. [ABSTRACT FROM AUTHOR]

Background: With the accumulating omics data, an efficient and time-saving transient assay to express target genes is desired. Mesophyll protoplasts, maintaining most stress-physiological responses and cellular activities as intact plants, offer an alternative transient assay to study target genes' effects on heat and oxidative stress responses. Results: In this study, a perennial ryegrass (Loliumperenne L.) mesophyll protoplast-based assay was established to effectively over- or down-regulate target genes. The relative expression levels of the target genes could be quantified using RT-qPCR, and the effects of heat and H2O2-induced oxidative stress on protoplasts' viability could be quantitatively measured. The practicality of the assay was demonstrated by identifying the potential thermos-sensor genes LpTT3.1/LpTT3.2 in ryegrass that over-expressing these genes significantly altered protoplasts' viability rates after heat stress. Conclusion: This protoplast-based rapid stress regulatory gene identification assay was briefed as 'PRIDA' that will complement the stable genetic transformation studies to rapidly identify candidate stress-regulatory genes in perennial ryegrass and other grass species. [ABSTRACT FROM AUTHOR]

Biofertilizers with phosphate-solubilizing microorganism (PSM) inoculations have been suggested to diminish the limitation of phosphorus (P) deficiency in plants. However, their applications in agriculture are restricted due to the inconstant effects of various PSMs. Proper carriers for the inoculations may overcome this shortcoming and improve PSMs' effectiveness. The objective of this study was to investigate whether straw compost, a type of organic material, can act as a carrier for improving the efficiencies of phosphate-solubilizing bacteria and fungi named Acinetobacter sp. and Aspergillus niger, respectively, in soils. We monitored the growth and cellular physiological responses of one type of model plants, named perennial ryegrass (Lolium perenne L.), under four soil treatments, including non-fertilization, PSM inoculation alone, straw compost addition alone, and the combined applications of both PSMs and straw compost. We found the combined treatments significantly improved the growth by 14.7% for shoot height and 79.7% for shoot weight, respectively, on average. P and potassium (K) uptakes of ryegrass were also increased by 102.5% and 65.3%, respectively, after the application of both PSMs and straw compost. Furthermore, physiological properties, such as photosynthetic efficiency and P-transportation capacity, of ryegrass were also significantly improved under combined treatments when compared to other treatments, regardless of the types of PSM included. The piecewise structural equation model further indicated that PSM inoculation and straw compost input are synergistically contributing to the nutrient uptake of ryegrass through many direct and indirect ways. We propose that straw compost is a good carrier material for PSMs' survival and would improve their plant growth promotion ability in soil. Our results provide valuable insights into the exploitation and utilization of P-biofertilizers in agriculture. [ABSTRACT FROM AUTHOR]

The premixtures flufenacet plus diflufenican and flufenacet plus diflufenican plus metribuzin are two herbicides recently registered in Greece for weed control in bread wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) with application early post-emergence to the crop (1st–3rd leaf growth stage). To evaluate the selectivity of these new herbicides, pot experiments were conducted by applying flufenacet plus diflufenican at 240 + 120 g ai ha−1 and flufenacet plus diflufenican plus metribuzin at 119.7 + 119.7 + 44.8 g ai ha−1 to bread wheat and barley, at 1st (BBCH-11), 2nd (BBCH-12) and 3rd (BBCH-13) leaf growth stage. The efficacy of the herbicides at the above-mentioned rates in pre-emergence application was also tested on three ALS/ACCase herbicide-resistant populations of Lolium rigidum Gaudin in comparison with the pre-emergence herbicides prosulfocarb and chlorotoluron plus diflufenican at 3200 g ai ha−1 and 1380 + 92 g ai ha−1, respectively. The results revealed decreased selectivity of both premixtures when applied at BBCH-11 for both winter cereals, with flufenacet plus diflufenican being less selective compared to flufenacet plus diflufenican plus metribuzin. Both herbicides highly controlled the three herbicide-resistant L. rigidum populations. The results indicated that both premixtures are effective chemical options for the management of herbicide resistant L. rigidum. To ensure crop safety and optimize efficacy, application at BBCH-12 is recommended. [ABSTRACT FROM AUTHOR]

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

Heavy metal contamination is an urgent ecological governance problem in mining areas. In order to seek for a green and environmentally friendly reagent with better plant restoration effect to solve the problem of low efficiency in plant restoration in heavy metal pollution soil. In this study, we evaluated the effects of three biodegradable chelating agents, namely citric acid (CA), fulvic acid (FA) and polyaspartic acid (PASP), on the physicochemical properties of copper tailings, growth of ryegrass (Lolium perenne L.) and heavy metal accumulation therein. The results showed that the chelating agent application improved the physicochemical properties of copper tailings, increased the biomass of ryegrass and enriched more Cu and Cd in copper tailings. In the control group, the main existing forms of Cu and Cd were oxidizable state, followed by residual, weak acid soluble and reducible states. After the CA, FA or PASP application, Cu and Cd were converted from the residual and oxidizable states to the reducible and weak acid soluble states, whose bioavailability in copper tailings were thus enhanced. Besides, the chelating agent incorporation improved the Cu and Cd extraction efficiencies of ryegrass from copper tailings, as manifested by increased root and stem contents of Cu and Cd by 30.29–103.42%, 11.43–74.29%, 2.98–110.98% and 11.11–111.11%, respectively, in comparison with the control group. In the presence of multiple heavy metals, CA, FA or PASP showed selectivity regarding the ryegrass extraction of heavy metals from copper tailings. PCA analysis revealed that the CA-4 and PASP-7 treatment had great remediation potentials against Cu and Cd in copper tailings, respectively, as manifested by increases in Cu and Cd contents in ryegrass by 90.98% and 74.29% compared to the CK group. [ABSTRACT FROM AUTHOR]

The co-allocation of photovoltaic arrays with crops presents a promising strategy to mitigate the conflict between photovoltaics and agricultural land. However, there is a notable lack of quantitative research on the impact of agrivoltaic system on land quality in fragile areas. In this study, peanuts (Arachis hypogaea) and ryegrass (Lolium perenne) were cultivated in photovoltaic array in the dry–hot valley of southwest China, with an off-site native land serving as the control. Sixteen soil physicochemical and biochemical parameters were measured in the gap and under-panel and control area. Results demonstrated that the agrivoltaic system significantly enhanced soil moisture, organic carbon, nitrogen–phosphorus–potassium nutrients, microbial biomass, and urease activity. It also led to varying degrees of increase in soil pH and electrical conductivity, along with reduced soil sucrase and phosphatase activity. In comparison to the control, the agrivoltaic system notably improved soil quality and multifunctionality. Specially, gap cultivation had a more pronounced positive impact on soil quality than under-panel cultivation, and the cultivation of peanuts had a greater effect on soil quality and multifunctionality improvement than ryegrass. This study provides fundamental data to support the improvement of land quality in photovoltaic developed regions, and to alleviate the conflict between photovoltaics and agricultural land. [ABSTRACT FROM AUTHOR]

A standardized preparation process is proposed in this study for achieving optimal strength and vegetative properties in vegetated concrete, using Yunnan red soil as a growth substrate for plants. The porosity of vegetated concrete is a crucial factor influencing plant growth, while compressive strength is a significant mechanical property. To assess the strength and porosity of vegetated concrete, different design porosities (22%, 24%, 26%, 28%) and cement-to-aggregate ratios (4, 5, 6, 7) were utilized in the preparation of vegetated concrete samples. The shell-making and static-pressure-molding methods were optimized for specimen preparation. Analyzing the stress–strain full curve characteristics of vegetation-type concrete under different influencing factors, an in-depth investigation into its failure mechanism was conducted. It was determined that the design porosity and cement content significantly impact the concrete's performance, particularly in terms of 30-day compressive strength and effective porosity. Furthermore, an increase in the fly ash ratio led to an increase in porosity and a decrease in compressive strength, providing a certain guidance for optimizing concrete performance. Comparative analysis through vegetation experiments revealed that black rye grass exhibited favorable growth adaptability compared to other grass species. [ABSTRACT FROM AUTHOR]

Crop irrigation requirements are usually estimated based on crop evapotranspiration (ETc) as determined by the reference evapotranspiration (ETo) and crop coefficient (Kc). There is a lack of knowledge on the irrigation requirements of tropical forage crops in Brazil, contrasting with the increasing use of irrigation in pastures. The effort of this study was to investigate what would be the water needs of tropical forages in Southern Brazil, based on a robust experimental database. The study was carried out in São Paulo State-Brazil using different forages species and their combinations [Guinea grass (GG); Guinea grass + black oat + ryegrass (GOR); Bermuda grass (BG), and Bermuda + black oat + ryegrass (BOR)]. The experimental fields were fully irrigated, and the Kc values were derived from ETc measurements on lysimeters; ETo was estimated using daily data from a nearby weather station and the standard FAO56 parameterization. Mean daily ETc values for GG, GOR, BG and BOR were 4.1, 2.9, 3.6, and 3.4 mm, respectively, and respective mean Kc values were 0.99, 0.90, 1.0, and 0.94. Average Kc values for all plots decreased as ETo increased, producing a negative Kc-ETo relationship, mainly when ETo reached values greater than 5 mm d−1. This was most likely due to internal plant stomatal resistance to vapor release from the leaves diffusing to the atmosphere at high ETo. So, the time-based Kc curves described by FAO 56 manual should be adjusted for the analyzed crops considering different ranges of ETo to improve the required irrigation depth. [ABSTRACT FROM AUTHOR]

Perennial ryegrass (Lolium perenne L.) is one of the most widely used species for sports fields in temperate climates because of its high wear tolerance. However, wear tolerance of cultivars may vary according to local environmental conditions and turfgrass management. In this study, we evaluated the wear tolerance of six perennial ryegrass cultivars (Adagio, Apple SGL, Equate, Firebird, Principal 2, Tetradark) under two fertility treatments (100 or 200 kg N-ha21-yr21) over 2 years. The field trial was performed at the Experimental Agricultural Farm at the University of Padova in northeastern Italy in a silty loam soil. Plots were arranged in a randomized complete block with three replications and subjected to three traffic events per week using a sports field wear simulator. Turfgrass quality (TQ), percent green cover (PGC), and normalized difference vegetation index (NDVI) were recorded every 2 weeks and averaged over each month. Although perennial ryegrass cultivars responded differently to wear stress, the higher nitrogen (N) rate positively affected the TQ of them all. 'Tetradrak' and 'Equate' had the lowest TQ, especially during the active growing seasons (spring and autumn). However, 'Tetradark' was particularly negatively affected during the cool fall months. The impact of a higher N fertilization rate on PGC and NDVI appeared to be more pronounced in spring than in fall. Furthermore, slight differences among cultivars and treatments were observed in summer and winter when temperatures were a limiting growth factor. [ABSTRACT FROM AUTHOR]

Green manuring is an established strategy for soil nutrient enrichment; however, its effects on the soil microbiome remain unclear. In this study, we assessed the impact of green manuring on soil microbial diversity, functional profiles, and enzymatic activities in a pot experiment involving tobacco rotation with three types of green manure: smooth vetch, ryegrass, and radish. Our data revealed distinct temporal shifts in bacterial communities under diverse tillage while fungi communities showed obvious clustering between fallow and green manuring. Moreover, green manure-sensitive bacterial and fungal taxa constitute a greater proportion of total sequences than their fallow-sensitive counterparts, indicating a more pronounced influence of green manuring on microbial communities. Functionally, green manuring augmented the relative abundance of N-cycle-related genes and was strongly associated with elevated N-acetyl-glucosaminidase activity. Notably, a subset of green manure-sensitive microbes exhibited significant antagonistic interactions with the fungal pathogens Fusarium spp. and Fusarium solani, and synergistic interactions with the beneficial bacteria Pseudomonas spp. Collectively, these findings suggest that green manuring promotes an optimized soil environment characterized by efficient nutrient cycling and diminished pathogenicity, thereby providing a scientific basis for targeted microbiota manipulation in agricultural systems. [ABSTRACT FROM AUTHOR]

Precise information about carbohydrates and proteins in relation to their utilization in the rumen is useful for the breeding purposes of perennial ryegrass cultivars used for animal nutrition. The objective of the current study was to evaluate 20 diploid perennial ryegrass accessions from the intermediary heading stage. The ruminal kinetics of different carbohydrate and protein fractions of grasses and legumes are important for forage breeding programs. The Cornell Net Carbohydrate and Protein System (CNCPS) was used to provide such information. Accession-based variation can be observed after considering dynamic degradation processes. Variation among the 20 accessions was observed. Ruminally digested (RDC) and undigested (UDC) carbohydrate and ruminally degraded (RDP) and undegraded (UDP) protein contents, total carbohydrate and total protein contents, and carbohydrate and protein fractions for the first cut and annual averages show significant differences. Although the variation was smaller for the protein fractions, the calculated usable protein content (uCP, sum of UDP and amount of synthesized microbial protein in the rumen) in the small intestine is mainly influenced by decreasing the neutral detergent fiber (NDF) and increasing the sugar content among cultivars. Carbohydrate and protein fractionation is suitable for characterizing perennial grass accessions as it uses parameters relevant to ruminant nutrition, allowing a step forward in forage plant breeding for forage quality. To conclude, using parameters related to ruminal degradation kinetics should favor the selection of accessions with higher amounts of ruminally digested carbohydrates (RDC). The selection of accessions based on protein quality (proportion of UDP) is less favorable for achieving a higher usable protein content. [ABSTRACT FROM AUTHOR]

Autumn grazing management aims to accumulate herbage for defoliation prior to a decrease in growth rates for the extension of the grazing season. The current study investigated the impact of building different target herbage masses (THMs) in autumn and imposing one of three different defoliation dates (DDs) between mid-October and late November on light transmitted to the base of the sward, free leaf lamina (FLL), leaf stage and internode elongation. Four THMs (low, medium, high and very high) and three DDs (DD1—15 October, DD2—7 November and DD3—21 November) were assigned to a 4 × 3 split plot design over two years. Light transmitted to the base of the sward was greatest in the low THM and decreased in all other THMs. Internode elongation increased in tillers in the medium to the very high THMs. Defoliation of the medium, high and, in particular, the very high THMs earlier in autumn reduced the effect of decreased light transmission on internode elongation. This study highlights that, as light transmitted to the base of the sward decreases, internode elongation increases, and this could negatively impact sward structure. [ABSTRACT FROM AUTHOR]

The relationship between phosphorus (P) availability and water restriction was explored in this study, focusing on its impact on phosphorus use efficiency (PUE) and water use efficiency (WUE) in various bromegrass (Bromus spp.) genotypes. Under controlled conditions, five bromegrass genotypes, as well as one ryegrass (Lolium perenne) cultivar, were compared by subjecting them to two P levels and two watering regimes. It was determined that combining water and phosphorus limitations led to reduced plant productivity. Initially, the ryegrass outperformed the bromegrass, but this result declined over time, while bromegrass exhibited consistent stability. Notably, under P and water stress, enhanced root development was observed in bromegrass compared to that in ryegrass. Distinct patterns of PUE and WUE allowed for the categorization of bromegrass genotypes into three groups. Genotype 3457 emerged as the most efficient, scoring 20 out of 24, while Pro 94-49 A achieved a score of only 10 out of 24. This study suggests that the drought resilience of bromegrass may be linked to increased root growth during the early vegetative stages, which potentially facilitates improved P acquisition. However, further validation through long-term field experiments is needed. The insights from this study are potentially valuable for use in shaping plant breeding programs by revealing the plant adaptation mechanisms for both P and water absorption. [ABSTRACT FROM AUTHOR]

Background: Commercial cultivars of perennial ryegrass infected with selected Epichloë fungal endophytes are highly desirable in certain pastures as the resulting mutualistic association has the capacity to confer agronomic benefits (such as invertebrate pest deterrence) largely due to fungal produced secondary metabolites (e.g., alkaloids). In this study, we investigated T2 segregating populations derived from two independent transformation events expressing diacylglycerol acyltransferase (DGAT) and cysteine oleosin (CO) genes designed to increase foliar lipid and biomass accumulation. These populations were either infected with Epichloë festucae var. lolii strain AR1 or Epichloë sp. LpTG-3 strain AR37 to examine relationships between the introduced trait and the endophytic association. Here we report on experiments designed to investigate if expression of the DGAT + CO trait in foliar tissues of perennial ryegrass could negatively impact the grass-endophyte association and vice versa. Both endophyte and plant characters were measured under controlled environment and field conditions. Results: Expected relative increases in total fatty acids of 17–58% accrued as a result of DGAT + CO expression with no significant difference between the endophyte-infected and non-infected progeny. Hyphal growth in association with DGAT + CO expression appeared normal when compared to control plants in a growth chamber. There was no significant difference in mycelial biomass for both strains AR1 and AR37, however, Epichloë-derived alkaloid concentrations were significantly lower on some occasions in the DGAT + CO plants compared to the corresponding null-segregant progenies, although these remained within the reported range for bioactivity. Conclusions: These results suggest that the mutualistic association formed between perennial ryegrass and selected Epichloë strains does not influence expression of the host DGAT + CO technology, but that endophyte performance may be reduced under some circumstances. Further investigation will now be required to determine the preferred genetic backgrounds for introgression of the DGAT + CO trait in combination with selected endophyte strains, as grass host genetics is a major determinant to the success of the grass-endophyte association in this species. [ABSTRACT FROM AUTHOR]

Ryegrass mottle virus (RGMoV; genus: Sobemovirus) is a single-stranded positive RNA virus with a 30 nm viral particle size. It exhibits T = 3 symmetry with 180 coat protein (CP) subunits forming a viral structure. The RGMoV genome comprises five open reading frames that encode P1, Px, a membrane-anchored 3C-like serine protease, a viral genome-linked protein, P16, an RNA-dependent RNA polymerase, and CP. The RGMoV genome size varies, ranging from 4175 nt (MW411579.1) to 4253 nt (MW411579.1) in the deposited sequences. An earlier deposited RGMoV complete genome sequence of 4212 nt length (EF091714.1) was used to develop an infectious complementary DNA (icDNA) construct for in vitro gRNA transcription from the T7 promoter. However, viral infection was not induced when the transcribed gRNA was introduced into oat plants, indicating the potential absence of certain sequences in either the 5' or 3' untranslated regions (UTR) or both. The complete sequence of the 3' UTR was determined through 3' end RACE, while the 5' UTR was identified using high-throughput sequencing (HTS)-RNA-Seq to resolve the potential absences. Only the icDNA vector containing the newly identified UTR sequences proved infectious, resulting in typical viral infection symptoms and subsequent propagation of progeny viruses, exhibiting the ability to cause repeated infections in oat plants after at least one passage. The successful generation of icDNA highlighted the synergistic potential of utilizing both methods when a single approach failed. Furthermore, this study demonstrated the reliability of HTS as a method for determining the complete genome sequence of viral genomes. [ABSTRACT FROM AUTHOR]

Estimating the role of specific processes in the spread of alien species necessitates the determination of introduction pathways and source populations of invaded areas. Alien grasses in the genus Lolium that have extensively invaded Japan provide a unique opportunity to estimate the expansion process through direct comparison between source and naturalised populations because the introduction pathways, contaminants in grain commodities and commercial cultivars for fodder crops or revegetation materials are well‐known. Therefore, by directly comparing source and naturalised populations, we estimated the introduction pathways and whether adaptative evolution occurred in Lolium species on sandy coasts in Japan.Lolium individuals sampled from naturalised populations in croplands, seaports, and sandy coasts were compared with those from two introduction sources for morphological and genetic variations based on a genome‐wide single nucleotide polymorphism analysis and a common garden experiment. Furthermore, we conducted a reciprocal transplant experiment between cropland and sandy coast.Populations naturalised in croplands were closely related to the cultivars, whereas those naturalised in seaports and sandy coasts were associated with contaminants. These results indicate that the cropland and sandy coast populations are derived from cultivars and contaminants, respectively. In addition, asymmetric gene flow from cropland populations to sandy coast populations was observed. The reciprocal transplant experiment clearly demonstrated the home site advantage; populations derived from croplands yielded higher floret numbers than those derived from other habitats at the cropland site; sandy coast populations had higher survival rates than those from croplands at the coastal site. Port populations exhibited a similar tendency as sandy coast populations, indicating that contaminants may be originally adapted to salty and dry environments, such as that in sandy coasts. The flowering phenology in the sandy coast populations evolved in the late flowering; therefore, late flowering alleles may have been transferred from cropland populations to sandy coast populations.Synthesis. We demonstrated that two congeneric species with different ecological characteristics were introduced through multiple introduction pathways and spread across different habitats. A direct comparison between source and naturalised populations can considerably elucidate the patterns and processes of biological invasions. [ABSTRACT FROM AUTHOR]

Studies with dairy cows receiving mixed ration and grazing annual pastures, which have low tiller density, are scarce. This study examined the effect of partial replacement of a total mixed ration (TMR) with annual temperate pasture. Treatments were ad libitum TMR (TMR100), 75% ad libitum TMR + ryegrass (Lolium multiflorum 'Maximus') (pTMR75), and 50% ad libitum TMR + ryegrass (pTMR50). Twelve multiparous Holstein and F1 Jersey x Holstein cows were divided into six groups (experimental units), taking account of milk production (26.6 ± 4.55 kg/day), days-in-milk (129 ± 50.8) and body weight (546 ± 30.6 kg). Treatments were compared using two 3 x 3 Latin square arangements, comprising three 21-day periods (measurements during final 5 days). Herbage DM intake increased from 4.8 kg/day with pTMR75 to 6.7 kg/day with pTMR50. Total DM intake decreased from 19.4 kg/day (TMR100), to 18.1 and 15.8 kg/day (pTMR75 and pTMR50, respectively). Milk production, energy corrected milk production (ECM), and milk fat content were similar between treatments, averaging 25.6, 28.4 kg/day and 44.6 g/ kg, respectively. Ryegrass pastures were able to replace up to 50% of TMR offered to mid lactation dairy cows with no adverse effects on ECM production. [ABSTRACT FROM AUTHOR]

Bioconversion of plant material with white-rot fungi has demonstrated improvements in in vitro digestibility, however, effects on rumen fermentation parameters and methane (CH4) production, when included as a dietary ingredient, are not clear. The aim of this study was to assess changes in rumen fermentation parameters and in vitro CH4 production by dietary inclusion of ryegrass-fescue hay biodegraded with P. ostreatus (H-PO) during 14 days of solid fermentation. The experiment used 2 diets with contrasting acid detergent fibre (aADFom) content which were added with 0, 3 and 6% H-PO. Substrates were incubated 3 times with sampling at 24 h. Results were analysed by ANOVA, and compared with Tukey's test and polynomial contrasts. Inclusion of H-PO at concentrations of 3 and 6% resulted in increased in vitro dry matter (DM) disappearance (IVDMD) by 10 and 12% respectively, whereas high concentrations of aADFom reduced IVDMD by 10% as compared to low aADFom. Methane production (mg/g DM incubated) and yield (mg/g DM degraded) were reduced by increasing aADFom and P. ostreatus biodegraded hay concentrations in the diets (p < 0.001) whereas concentrations of total short-chain fatty acids were not affected by the inclusion of biodegraded hay (p = 0.080) or aADFom concentration (p = 0.076). Increasing concentrations of biodegraded hay resulted in increased acetate (p < 0.001), and reduced propionate proportions (p = 0.021) but did not affect butyrate (p = 0.233). In conclusion the use of biodegraded hay as a pre-treated ingredient improved the utilisation of low and high fibre diets by increasing the in vitro disappearance, and decreasing CH4 emissions, without increasing the propionate proportion. The use of hay biodegraded with P. ostreatus (H-PO) improves the digestibility of ruminant diets. The use of H-PO decreases methane, and increases acetate production in in vitro conditions. The use of H-PO as an ingredient in ruminant diets could improve the sustainability of livestock systems. [ABSTRACT FROM AUTHOR]

The essential oil (EO) of Thymbra capitata has been demonstrated to possess herbicidal activity and could be used as an alternative to synthetic herbicides with reduced persistence in soil and new mode of action. Nevertheless, it is necessary to determine the adequate doses for its use, the proper way for its application and the best phenological stage of weeds and crops in which the EO should be applied to obtain maximum efficacy against weeds without compromising crop production. In this work, T. capitata EO was tested at three different concentrations against weeds grown from a citrus orchard soil seedbank untreated with herbicides and against three important weed species grown in substrate to determine the efficacy of the concentrations on different weed species. All experiments were carried out under greenhouse conditions. To find out the best way for applying the EO, it was applied by irrigation and by spraying on the targeted weeds, and to verify the influence of timing, it was tested on Lolium rigidum at two different phenological stages and on wheat at a later phenological stage than weeds. The highest concentration tested (12 µL·mL−1) showed the best performance to control weeds. The more effective mode of application was by spraying on dicotyledons and by irrigation on monocotyledons at the earliest phenological stage. T. capitata EO was phytotoxic for wheat. More trials in different crops are needed to determine the best conditions for its use. [ABSTRACT FROM AUTHOR]

The widespread practice of using high doses of nitrogen to increase unit area yield in annual ryegrass introduces ecological and economic problems. This research was carried out over two years and aimed to determine the potential forage yield and quality of annual ryegrass by applying manure, humic acid, and low doses of nitrogen fertilizer, within the framework of sustainable forage crop cultivation. The study was carried out from 2020 to 2022 with three replications according to the randomized block split plots experimental design. According to the study results, the highest values were achieved with combinations that included manure application, and even higher values were observed when manure was applied along with low doses of humic acid and nitrogen. The M20 + H20 + N100 treatment yielded the highest fresh yield, dry matter, and crude protein. The treatments with manure, humic acid, and nitrogen fertilizer had varying effects on the ADF and NDF content of annual ryegrass, resulting in fluctuating values. In conclusion, 20 t manure, 20 L humic acid, and 100 kg ha−1 N application can be suggested for sustainable and higher dry matter production with good quality for annual ryegrass cultivation under semi-humid climatic conditions. The results obtained from this research hold promise for sustainable agricultural practices. [ABSTRACT FROM AUTHOR]

Epichloë festucae uses a siderophore‐mediated system to acquire iron, which is important to maintain endophyte–grass symbioses. Here we investigate the roles of the alternative iron acquisition system, reductive iron assimilation (RIA), via disruption of the fetC gene, which encodes a multicopper ferroxidase, either alone (i.e., ΔfetC) or in combination with disruption of the gene sidA, which encodes a siderophore biosynthesis enzyme (i.e., ΔfetC/ΔsidA). The phenotypic characteristics of these mutants were compared to ΔsidA and wild‐type (WT) strains during growth under axenic culture conditions (in culture) and in symbiosis with the host grass, perennial ryegrass (in planta). Under iron deficiency, the colony growth rate of ΔfetC was slightly slower than that of WT, while the growth of ΔsidA and ΔfetC/ΔsidA mutants was severely suppressed. Siderophore analyses indicated that ΔfetC mutants hyperaccumulate ferriepichloënin A (FEA) at low iron concentrations and ferricrocin and FEA at higher iron concentrations. When compared to WT, all mutant strains displayed hyperbranching hyphal structures and a reduced ratio of Epichloë DNA to total DNA in planta. Furthermore, host colonization and vertical transmission through infection of the host seed were significantly reduced in the ΔfetC/ΔsidA mutants, confirming that high‐affinity iron uptake is a critical process for Epichloë transmission. Thus, RIA and siderophore iron uptake are complementary systems required for the maintenance of iron metabolism, fungal growth, and symbiosis between E. festucae and perennial ryegrass. [ABSTRACT FROM AUTHOR]

This study was carried out to determine the quality of sugar beet pulp (Beta vulgaris L.), lenox (Brassica rapa L.), and ryegrass (Lolium multiflorium cv. caramba) silages used in feeding dairy cattle, in comparison to corn silage. Whole corn was harvested at the dough stage, lenox was harvested at the end of flowering, and ryegrass was harvested at the beginning of spike. No additives were added to the corn silage, but 5% molasses was added to the sugar beet pulp, ryegrass, and lenox silages that have low levels of easily degradable sugar. Additionally, 5% sainfoin was added to the sugar beet pulp silage to increase its dry matter level to 20%. All silages used in the study were classified as "very good" according to the Fleig scoring system. Four Holstein dairy cattle at a mean age of 3 years old, at the 3rd month of lactation, and with milk yields close to each other were used in the feeding experiment. In this study, the 4 × 4 Latin square trial design was applied, and each period of the experiment consisted of 21 days, 14 days for exercise and 7 days for sample collection. The amount of feed to be consumed by the animals according to the periods was calculated based on the National Research Council (NRC) standards considering their yield levels. In the experiment, milk yield was found to be similar in the silage groups, but milk fat levels were higher in the group consuming corn silage than the other groups. The rumen fluid and blood serum values of the animals were within the reference ranges. As a result, it was determined that these silages were competitive with corn silage in the feeding of dairy cattle as long as 5% molasses would be added to wet the beet pulp, lenox and ryegrass silages, whereas it was concluded that the digestibility and energy content of the ryegrass silage was lower than those of the other silages. [ABSTRACT FROM AUTHOR]

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

Simple Summary: New Zealand has a temperate climate, and dairy farming relies on ryegrass pasture-based systems, normally with a winter-calving system. Although there is adequate pasture growth in some regions during winter, dairy farmers use autumn-saved pasture to sustain cows for one to two months before and after calving. During the transition period at calving, dairy cows have major changes in both physiology and nutritional requirements. The knowledge of ryegrass cultivars would enable the most appropriate selection to benefit cows pre- and post-calving. However, studies on Winter ryegrass are scarce. In this study, we measured nine ryegrass cultivars, including perennial, hybrid, and Italian types grown in late autumn and winter and harvested after seven or nine weeks of regrowth. The crude protein concentration was similar across types, but perennial ryegrasses contained the most fibre. Italian ryegrasses had the greatest proportion of soluble nutrients. The degradation rate of insoluble (fibrous) nutrients was the highest in Italian and the lowest in perennial grasses. Ryegrasses grown during winter had lower crude protein and fibre and higher non-fibre carbohydrate concentrations than that grown in late autumn. In conclusion, this study′s insights into the characteristics of nine ryegrass cultivars, grown in different seasons, offer valuable guidance for selecting the most suitable options to support the nutritional requirements of dairy cows during the transition period around calving. During winter and early spring, pasture supply is usually lower than the demand in New Zealand dairy farming systems and thus the 'autumn saved pastures' (stockpiling) are introduced to fill the gap. This study aimed to investigate the chemical, physical, and degradation characteristics of ryegrass pastures, the predominant forage in New Zealand, grown in autumn and winter. To serve as 'autumn saved pasture', nine ryegrass cultivars, comprising three types (three perennial, three hybrid, and three Italian), were grown in late autumn (Autumn) and early winter (Winter) and harvested after 7 and 9 weeks of regrowth, respectively. The experiment had two experimental factors: ryegrass type (or cultivar) and harvest season. These experimental factors were in a randomised block design with the forage plot as the experimental unit. The degradation characteristics were assessed in the rumen of fistulated cows using the in situ incubation technique. Perennial ryegrass had a greater neutral detergent fibre (NDF) concentration (468 g/kg dry matter (DM)) than the hybrid (435 g/kg DM) or Italian (414 g/kg DM) ryegrasses. Italian ryegrasses had the greatest soluble fraction of DM (64.2% vs. 46.7% and 40.7%) and the greatest degradation rate of an insoluble but degradable fraction of DM (0.221 vs. 0.189 and 0.145/h) than the hybrid and perennial ryegrasses. Compared with the Winter ryegrass, the Autumn ryegrass had a greater crude protein concentration (246 vs. 208 g/kg DM) and a greater NDF concentration (486 vs. 392 g/kg DM) but a lower calculated soluble carbohydrate concentration (152 vs. 263 g/kg DM). It is concluded that there are notable variations among the cultivars, highlighting distinctions in parameters, such as NDF concentration, soluble fractions, degradation rates, and nutrient content among the perennial, hybrid, and Italian ryegrasses, as well as the seasonal variations observed between autumn and winter growth. These findings will not only facilitate enhanced nutrition for dairy cows as they undergo the transition phase but also have practical implications for future research and dairy cow nutrition. [ABSTRACT FROM AUTHOR]

This study investigated the effects of acacia (extract of Acacia mearnsii) and sainfoin (Onobrychis viciifolia) as condensed tannin (CT)-rich sources on ruminal and manure methane (CH 4) emissions in comparison with non-CT silages characterized by different contents of the cell wall and water-soluble carbohydrates. In a 3 × 6 incomplete Latin square design, 30 Holstein cows (63 ± 23 d in milk; mean ± SD; 33.8 ± 7.6 kg of milk per day, body weight 642 ± 81 kg) were provided with ad libitum access to 1 of 6 total mixed rations comprising 790 g of silage and 210 g of concentrate per kilogram of dry matter (DM). The silages were either rich in sainfoin [neutral detergent fiber (NDF): 349 g/kg of DM], perennial ryegrass (NDF: 420 g/kg of DM), or red clover (NDF: 357 g/kg of DM). Each silage was supplemented with 20 g/kg (of total diet DM) of acacia or straw meal. Feed intake and milk yield were recorded daily. Milk composition and ruminal fluid characteristics and microbiota were analyzed. The individual ruminal CH 4 production was determined using the GreenFeed system, and CH 4 emissions from the manure of cows fed the same diets were measured in a parallel experiment over 30 d at 25°C using a dynamic flux chamber. The CT sources did not reduce CH 4 yield or emission intensity. Acacia reduced milk production (from 26.3 to 23.2 kg/d) and DM intake (from 19.7 to 16.7 kg/d) when supplemented with ryegrass, and both CT sources reduced the milk protein content and yield. Acacia supplementation and ryegrass silage reduced the ruminal acetate:propionate ratio. Furthermore, during acacia treatment, the abundance of Methanobrevibacter archaea tended to be lower and that of Thermoplasmata was higher. Acacia reduced the CH 4 emissions from manure for the ryegrass group by 17% but not for the sainfoin and clover groups. Feeding sainfoin silage resulted in the lowest manure-derived CH 4 emissions (−47% compared with ryegrass). In conclusion, acacia reduced ruminal CH 4 production by 10%, but not emission intensity, and the mitigation effect of sainfoin depended on the silage to which it was compared. Because mitigation was partially associated with animal productivity losses, careful evaluation is required before the implementation of tanniferous feeds in farm practice. [Display omitted] [ABSTRACT FROM AUTHOR]

The Quick Wash (QW) treatment extracts phosphorus (P) from manure and municipal sludge (MS), producing an organic acidified by-product with adequate nitrogen (N):P ratio to meet crop N requirements. Yet, data on crop response to N using QW by-products are lacking. We evaluated the response of annual ryegrass (Lolium multiflorum Lam.) and potential N leaching in sandy soil to N applications using raw wastes, their corresponding QW by-products, and ammonium sulfate (AMS) fertilizer. Treatments included a control (no amendment added), raw and acid-washed chicken litter, dairy and swine manure, MS, and AMS at 100, 200, and 400 kg N ha−1. We found no significant differences in annual ryegrass yield and N uptake between the raw and acidified organic QW by-products. However, ryegrass produced 4–30% more biomass with AMS than organic amendments. The total residual soil inorganic N under AMS treatments ranged between 6.3 and 67.9 mg pot−1 and accounted for 5–17% of the total N applied, but it was <1% for all the organic amendments. We found no differences in soil N leaching between raw and acid-washed forms of each organic soil amendment. Our results indicated that acidified organic QW by-products can improve environmental quality by substantially reducing the amount of applied P with no penalties for crop yield losses compared to raw manure and MS. [ABSTRACT FROM AUTHOR]

Background: Drought resistance is a complex characteristic closely related to the severity and duration of stress. Perennial ryegrass (Lolium perenne L.) has no distinct drought tolerance but often encounters drought stress seasonally. Although the response of perennial ryegrass to either extreme or moderate drought stress has been investigated, a comprehensive understanding of perennial ryegrass response to both conditions of drought stress is currently lacking. Results: In this study, we investigated the genetic variation in drought resistance in 18 perennial ryegrass varieties under both extreme and moderate drought conditions. The performance of these varieties exhibited obvious diversity, and the survival of perennial ryegrass under severe stress was not equal to good growth under moderate drought stress. 'Sopin', with superior performance under both stress conditions, was the best-performing variety. Transcriptome, physiological, and molecular analyses revealed that 'Sopin' adapted to drought stress through multiple sophisticated mechanisms. Under stress conditions, starch and sugar metabolic enzymes were highly expressed, while CslA was expressed at low levels in 'Sopin', promoting starch degradation and soluble sugar accumulation. The expression and activity of superoxide dismutase were significantly higher in 'Sopin', while the activity of peroxidase was lower, allowing for 'Sopin' to maintain a better balance between maintaining ROS signal transduction and alleviating oxidative damage. Furthermore, drought stress-related transcriptional and posttranscriptional regulatory mechanisms, including the upregulation of transcription factors, kinases, and E3 ubiquitin ligases, facilitate abscisic acid and stress signal transduction. Conclusion: Our study provides insights into the resistance of perennial ryegrass to both extreme and moderate droughts and the underlying mechanisms by which perennial ryegrass adapts to drought conditions. [ABSTRACT FROM AUTHOR]

In current paper a comparative study of bioethanol yields from ryegrass biomass by different type of pretreatment and enzyme hydrolysis is done. Acidic and alkali hydrolysis was used as a pretreatment of this feedstock. Cellulase blend 10 FPU g-1 dry matter was used for enzyme hydrolysis. The pre-treatment was conducted by autoclaving for 20 minutes at 121°C. Best results were obtained by pretreatment with 1 % HCl and 1 % NaOH. Hydrolysate obtained by pretreatment with hydrochloric acid contained 33.6 mg mL-1 reducing sugars, and that obtained by pretreatment with base - 29.0 mg mL-1. Both hydrolysates were found to be suitable substrates for bioethanol generation, where 5.55 g ethanol/100 g dry matter was produced from grass hydrolysate obtained by acidic hydrolysis followed by enzyme hydrolysis and from alkali pretreated hydrolysate-4.73 g ethanol/100 g dry matter. The results demonstrated that ryegrass biomass is suitable feedstock for bioethanol production. [ABSTRACT FROM AUTHOR]

Perennial grasses have got greater attention in the past decade due to their potential to be productive over several years and their positive impacts on the environment. A great resource in this regard are the native species, which are well adapted to the specific conditions of the environment, but are often underestimated and irrationally used. The objects of study in the present research were 8 grasses (Festuca rubra, Lolium perenne, Dactilis glomerata, Arrhenatherum elatius, Festuca arundinacea, Briza maxima, Trisetum flavescens, Agrostis alba), typical for the Middle Balkan Mountains region, in order to assess their productivity and stability. The two-factor analysis of variance in terms of dry mass yield revealed significant influence of the factors of species, environment, and their interaction. During the 9-year experimental period, the highest yield was demonstrated by F. rubra (5.75 t ha-1), followed by F. arundinacea (5.54 t ha-1) and A. elatius (5.43 t ha-1). The stability parameters, calculated by the methods of regression analysis defined as stable the species of A. elatius and A. alba, and those, calculated by variance and non-parametric analyzes - F. rubra, D. glomerata and F. arundinacea. Differences in the stability assessment of the individual parameters are due to the fact that the different methods are based on different concepts of stability. The GGE biplot analysis, which integratedly assesses the species for their productivity and stability, identified F. arundinacea and A. elatius as the most favorable combining of these indicators. Trisetum flavescens was also characterized by good stability and average productivity. These species are suitable for cultivation in a wide range of environmental conditions. [ABSTRACT FROM AUTHOR]

Background: Genomic selection has the potential to accelerate genetic gain in perennial ryegrass breeding, provided complex traits such as forage yield can be predicted with sufficient accuracy. Methods: In this study, we compared modelling approaches and feature selection strategies to evaluate the accuracy of genomic prediction models for seasonal forage yield production. Results: Overall, model selection had limited impact on predictive ability when using the full data set. For a baseline genomic best linear unbiased prediction model, the highest mean predictive accuracy was obtained for spring grazing (0.78), summer grazing (0.62) and second cut silage (0.56). In terms of feature selection strategies, using uncorrelated single‐nucleotide polymorphisms (SNPs) had no impact on predictive ability, allowing for a potential decrease of the data set dimensions. With a genome‐wide association study, we found a significant SNP marker for spring grazing, located in the genic region annotated as coding for an enzyme responsible for fucosylation of xyloglucans—major components of the plant cell wall. We also presented an approach to increase interpretability of genomic prediction models with the use of Gene Ontology enrichment analysis. Conclusions: Approaches for feature selection will be relevant in development of low‐cost genotyping platforms in support of routine and cost‐effective implementation of genomic selection. [ABSTRACT FROM AUTHOR]

Perennial ryegrass (Lolium perenne L.) is a commonly grown pasture species in temperate agriculture, mainly serving as a primary energy source for dairy cows. However, its limited persistence often leads to missed production potential and early resowing, especially in countries that experience summer drought, e.g., Australia and New Zealand. Therefore, understanding the factors influencing perennial ryegrass pasture persistence is crucial for sustainable land management and climate resilience in pasture-based animal production systems. Significant gaps in knowledge exist regarding the factors influencing pasture persistence, as the number of conducted studies in this area remains limited. This study aimed to investigate the factors influencing the expression of persistence in perennial ryegrass populations using airborne and ground-based sensors. A field experiment was conducted in the southwest region of Victoria, Australia, involving ten commercial perennial ryegrass cultivar–endophyte combinations in two different populations. Persistence was evaluated using sensor-based and conventional pasture measurements over two consecutive autumns. The results revealed significant fixed effects of cultivar, endophyte, and environment and their interactions on persistence traits of perennial ryegrass. Cultivars Alto, Samson, and One50 exhibited high levels of persistence when infected with novel endophyte strains. Furthermore, prolonged environmental stresses were found to drive directional selection within pasture populations. The findings emphasise the importance of selecting appropriate cultivar–endophyte combinations and early detection of signs of poor persistence to optimise sward longevity and financial returns from pasture-based animal production systems. This study fills a knowledge gap regarding the factors influencing pasture persistence and provides valuable insights for sustainable pasture management strategies. [ABSTRACT FROM AUTHOR]

Fly ash (FA) is promising for environmental remediation, but how to modify the FA with high remediation efficiency through an environmentally friendly and low-cost modification method is scare. A modified FA (MFA) was prepared through a one-step hydrothermal modification with Ca(OH)2 and KH2PO4. Results indicated that irregular agglomerates occurred on the surface of the MFA and that the specific surface area increased by 1.94 times compared to that of FA. Compared to FA, glassy compositions in MFA were destroyed and amorphous Si/Al and alkaline aluminosilicate gels were formed. The soil application of 0.2–0.6% MFA significantly increased soil pH by 0.23–0.86 units compared to FA and decreased available lead (Pb) and copper (Cu) by 25–97.1% and 13.5–75%, respectively. MFA significantly decreased exchangeable Pb and Cu by 12.5–32% and 11.4–35.2%, respectively, compared to FA. This may be due to the high pH and specific surface area of MFA, which promoted to the formation of amorphous Si/Al, metal–phosphate precipitation, and complexation with functional groups. In addition, MFA slightly increased the biomass of shoots and roots and decreased the uptake of Pb and Cu by ryegrass. This study provides a new modification method for the utilization of FA in the heavy metal-contaminated soils. [ABSTRACT FROM AUTHOR]

Simple Summary: Production of small ruminants, specifically hair sheep, has increased in recent years and has resulted in an increased presence of hair sheep in feedlots for finishing. In 2017, the United States banned the use of antibiotic growth promoters. This ban has resulted in an interest in non-medicated feed additives, such as yeast, to aid in growth promotion. While hair lambs tend to take longer to reach an adequate finishing weight, the utilization of a yeast fermentation product may aid as a natural growth promoter in the feedlot. Additionally, interest in grass-based finishing systems has grown, but there are limited data on using stored cool-season forages as the base of the diet. Based on the results of this experiment, the inclusion of a yeast fermentation product can improve the feedlot performance and carcass texture of hair lambs in a feedlot with an annual ryegrass-hay-based diet. Hair sheep production has increased in recent years, which has resulted in an increased presence in feedlots. Additionally, grass-based finishing systems for ruminant animal production have increased. Data are limited for finishing hair lambs on diets based on cool-season hay. The objective was to evaluate a Saccharomyces cerevisiae fermentation product (SCFP) on the feedlot performance and carcass characteristics of Katahdin lambs offered an annual ryegrass (Lolium multiflorum)-hay-based diet. Twenty-four Katahdin lambs (21.5 ± 2.5 kg BW) were assigned to either the control (CON) or the yeast-supplemented group (SCFP) in a completely randomized design. Lambs were offered a 14% crude protein total mixed ration diet based on annual ryegrass hay. The SCFP group also received the yeast supplement in their diet. Lambs in the SCFP group had a higher final body weight (p < 0.01) and ADG (p = 0.01). Less maximum and total energy were required to cut SCFP lamb meat compared to CON lamb meat (p < 0.03). Results from this study indicated that SCFP supplementation may prove to be beneficial in hair lamb finishing diets. Future research will need to specifically evaluate the use of these products in hair lambs with a larger sample size. [ABSTRACT FROM AUTHOR]

Extended waterlogging (WL) conditions in heavily manured soils can change soil phosphorus (P) dynamics. We assessed the effects of soil moisture regimes (field capacity (FC) and WL) and P rates on (i) dry matter (DM) yield and P offtake of ryegrass, (ii) changes in soil Fe3+, Fe2+, and soil P, and (iii) risk of P leaching. The treatments were tested in a four-month greenhouse experiment using intact soil columns and annual ryegrass (Lolium multiflorum). The DM yield and P offtake were lower under WL compared with FC. The concentration of Fe3+ was 1984 mg kg−1 (0–30 cm) under FC, but 1213 mg kg−1 at 0–5 cm and 2024 mg kg−1 at 25–30 cm depth under WL. The concentration of Fe2+ was 244 mg kg−1 (0–30 cm) under FC, but 2897 at 0–5 cm and 687 mg kg−1 at 25–30 cm under WL. The water extractable P (Pw) was 12.7 mg kg−1 at 0–5 cm and 9.5 mg kg−1 at 25–30 cm under FC, but 8.6 mg kg−1 at 0–5 cm and 10.5 mg kg−1 at 25–30 cm under WL. The P saturation index (PSI) was 27.2% at 0–5 cm and 17.4% at 25–30 cm under FC, but averaged 11.9% at 0–30 cm under WL. We can conclude that extended WL associated with flooding creates reducing conditions in the soil, thus decreasing the concentration of Fe3+, but increasing the concentrations of Fe2+ and the solubility of P which can exacerbate the risk of P loss with runoff and leaching. [ABSTRACT FROM AUTHOR]

Heat stress can hinder the growth of perennial ryegrass (Lolium perenne L.). Methyl jasmonate (MeJA) applied exogenously can increase heat stress tolerance in plants; however, the regulatory mechanisms involved in heat tolerance mediated by MeJA are poorly understood in perennial ryegrass. Here, the microRNA (miRNA) expression profiles of perennial ryegrass were assessed to elucidate the regulatory pathways associated with heat tolerance induced by MeJA. Plants were subjected to four treatments, namely, control (CK), MeJA pre-treatment (T), heat stress treatment (H), and MeJA pre-treatment + heat stress (TH). According to the results, 102 miRNAs were up-regulated in all treatments, with 20, 27, and 33 miRNAs being up-regulated in the T, H, and TH treatment groups, respectively. The co-expression network analysis between the deferentially expressed miRNAs and their corresponding target genes showed that 20 miRNAs modulated 51 potential target genes. Notably, the miRNAs that targeted genes related to with regards to heat tolerance were driven by MeJA, and they were involved in four pathways: novel-m0258-5p mediated signal transduction, novel-m0350-5p mediated protein homeostasis, miR397-z, miR5658-z, and novel-m0008-5p involved in cell wall component, and miR1144-z and miR5185-z dominated chlorophyll degradation. Overall, the findings of this research paved the way for more research into the heat tolerance mechanism in perennial ryegrass and provided a theoretical foundation for developing cultivars with enhanced heat tolerance. [ABSTRACT FROM AUTHOR]

The efficient preservation of protein in silage for livestock feed is dependent on the rate and extent of proteolysis. Previous research on fresh forage indicated enhanced protein stability in certain Festulolium (ryegrass × fescue hybrids) cultivars compared to ryegrass. This is the first report of an experiment to test the hypothesis that a Lolium perenne × Festuca arundinacea var glaucescens cultivar had reduced proteolysis compared to perennial ryegrass (L. perenne) during the ensiling process. Forages were harvested in May (Cut 2) and August (Cut 4), wilted for 24 h and ensiled in laboratory‐scale silos. Silage was destructively sampled at 0 h, 9 h, 24 h, 48 h, 72 h, 14 days and 90 days post‐ensiling, and dry matter (DM), pH and chemical composition were determined. At Cut 2, there was no difference in crude protein between treatments but ryegrass had higher soluble nitrogen (SN) (P < 0.001) and grass × time interactions (p = 0.03) indicated higher rates of proteolysis. By Cut 4, Festulolium had (5.5% units) higher CP than ryegrass (p < 0.001) but SN did not differ. Ammonia‐N did not differ between silages in either cut. DM differences (11.8% units) between treatments in Cut 4 (v.2.2% in Cut 2) may have masked effects on proteolysis, highlighting the importance of management on silage quality. This was despite higher WSC in ryegrass in both cuts (p < 0.001), with grass × time interactions (Cut 2; p = 0.03) showing slower WSC decline in ryegrass in Cut 4 (p < 0.001). Silage pH values did not differ between grasses in either cut, but grass × time interactions (p < 0.001) showed a slower decline in both ryegrass cuts, resulting in higher (p < 0.05) pH at 24 h and 72 h for Cuts 2 and 4, respectively. Overall, the hypothesis for an enhanced protein stability in Festulolium when ensiled as ruminant feed was evidenced by lower SN but not ammonia‐N in an early‐cut silage with a comparable DM to ryegrass. An experiment tested the hypothesis that a Festulolium had reduced proteolysis compared to perennial ryegrass when ensiled. The benefits of Festulolium silage as ruminant feed were evidenced by lower soluble nitrogen but not ammonia‐N in an early‐cut silage with a comparable DM to ryegrass. [ABSTRACT FROM AUTHOR]

Intercropping perennial legumes with trees can reduce Nitrogen (N) losses, due to the high amount of N accumulated in stable forms in the soil and permanent soil cover during the whole year. Although N cycling improvement in mature agroforestry systems (AFS) was well documented, there is a lack of knowledge regarding systems in transition to AF. In this work, we studied the association of two perennial forage crops, namely ryegrass (Lolium multiflorum Lam.) and sulla (Hedysarum coronarium L.), with 1-year old poplars, to evaluate: (i) the agronomic performance of sulla and ryegrass with vs. without intercropped poplar trees; (ii) the N-fixing ability of sulla in association with trees; (iii) the N transfer effect from sulla and growth promoting effect on poplar; and finally (iv) the nitrate leaching reduction due to the presence of poplar trees associated to forage crops. The layout was arranged in a two-factor randomized complete blocks design (RCB) with three replicates. The first factor tested (crop species) implied two different swards, namely sulla and ryegrass. The second factor (cropping system) included two different systems: PAST i.e., a pastoral system without trees, and SIPAST, i.e., a silvo-pastoral system with one poplar tree row beside the sward. Sulla resulted more productive than ryegrass when associated with trees (+35%). No clear trend was observed about the tree influence on N-fixation in sulla, but the amount of N fixed resulted higher in in sulla grown in the SIPAST near the trees (+35%). Poplar plants, even in the first year after planting, resulted effective yet in reducing the nitrate flux from the crops towards ditches. Further investigations are needed to study other swards in young AF and better understand the N dynamics; in particular, it could be worth to assess the nutrient flux in the soil solution. [ABSTRACT FROM AUTHOR]

Simple Summary: A Festulolium grass is an intergeneric hybrid of fescue (Festuca L.) and ryegrass (Lolium L.) that combines the nutritional qualities and weather resistance of each. Its high nutritional value and resistance to extreme weather conditions are important considerations in the era of global climate change. Festulolium has been used as a forage crop for many years, but little information is available on the levels of biologically active compounds (β-carotene and tocopherols) in Festulolium-based diets and in the meat of animals fed such diets. The present study assessed the contents of β-carotene, retinol and tocopherols and the proximate chemical composition of meat from young rams receiving diets based on silage made from different Festulolium cultivars (Becva, Felopa, Paulita), as compared with meat from rams fed a diet based on hybrid ryegrass. The meat of animals fed Festulolium silage-based diets contained more tocopherols and retinol than the meat of sheep receiving a hybrid ryegrass-based diet. Meat from rams fed diets based on silage made from Festulolium cvs. Felopa and Paulita was characterized by the highest concentrations of tocopherols and retinol. The results of this study indicate that Festulolium silage can be a desirable component of ram diets, but its efficacy may vary across cultivars. This study analyzed the chemical composition and the β-carotene, tocopherol and retinol content of meat (musculus longissimus lumborum, musculus gluteus maximus) from young rams fed different silage-based diets. The animals were divided into four groups depending on the type of silage in the feed ration (hybrid ryegrass, Festulolium cv. Becva, Festulolium cv. Felopa, Festulolium cv. Paulita). The concentrations of the examined lipophilic compounds were measured via high-performance liquid chromatography (HPLC). Animals fed a diet based on ryegrass silage had the highest intakes of α-, β-, γ- and δ-tocopherol and total tocopherols (p < 0.01). Muscles of rams fed Festulolium silage-based diets contained more tocopherols (p < 0.01) and retinol (p < 0.01 in m. g. maximus; p = 0.053 in m. l. lumborum) than the meat of animals receiving hybrid ryegrass-based diets. The highest amounts of tocopherols (p < 0.01) and retinol (p < 0.01 in m. g. maximus; p = 0.053 in m. l. lumborum) were found in meat from rams fed rations based on silage made from Festulolium cvs. Felopa and Paulita. The analysis revealed a positive correlation between the intake of tocopherols and the α-tocopherol content of meat (r = 0.99), as well as between the β-carotene intake and the retinol content of meat (r = 0.92 in m. l. lumborum; r = 0.91 in m. g. maximus). [ABSTRACT FROM AUTHOR]

Systematic measurement of pasture biomass (kg DM/ha) is crucial for optimising pasture utilisation and increasing dairy farm profitability. On-farm pasture monitoring can be conducted using various sensors, but calibrations are necessary to convert the measured variable into pasture biomass. In this study, we conducted three experiments in New South Wales (Australia) to evaluate the use of the rising plate meter (RPM), pasture reader (PR), unmanned aerial vehicles (UAV) and satellites as pasture monitoring tools. We tested various calibration methods that can improve the accuracy of the estimations and be implemented more easily on-farm. The results indicate that UAV and satellite-derived reflectance indices (e.g., Normalised Difference Vegetation Index) can be indirectly calibrated with height measurements obtained from an RPM or PR. Height measurements can be then converted into pasture biomass ideally by conducting site-specific sporadic calibrations cuts. For satellites, using the average of the entire paddock, root mean square error (RMSE) = 226 kg DM/ha for kikuyu (Pennisetum clandestinum Hochst. ex Chiov) and 347 kg DM/ha for ryegrass (Lolium multiflorum L.) is as effective as but easier than matching NDVI pixels with height measurement using a Global Navigation Satellite System (RMSE = 227 kg DM/ha for kikuyu and 406 kg DM/ha for ryegrass). For situations where no satellite images are available for the same date, the average of all images available within a range of up to four days from the day ground measurements were taken could be used (RMSE = 225 kg DM/ha for kikuyu and 402 kg DM/ha for ryegrass). These methodologies aim to develop more practical and easier-to-implement calibrations to improve the accuracy of the predictive models in commercial farms. However, more research is still needed to test these hypotheses under extended periods, locations, and pasture species. [ABSTRACT FROM AUTHOR]

Background: In the Western Highlands of Cameroon (WHC), information on the nutritional value of fodder species consumed by ruminants is very limited. Methods: Through interviews with farmers and monitoring of animals on the range, information was obtained on the types of fodder resources consumed by the ruminants. Samples of each forage species were collected in 15 districts, mixed, chopped, and dried in a ventilated oven at 60°C, and then ground for chemical composition analysis. Results: Twenty‐two forage species were identified. Among these species, Vernonia amygdalina (29.43% ± 0.45% dry matter [DM]) and Pennisetum clandestinum (87.21% ± 1.33% DM) were, respectively, the highest in protein and neutral detergent fiber contents. Manihot esculenta was one of the most energy‐rich forages in terms of forage unit for lactation and forage unit for meat production. Hierarchical ascending classification revealed three main groups of forages, respectively, rich in crude fiber (Group 1), protein (Group 2), and energy (Group 3). Conclusions: The present study identified 22 forage species browsed by ruminants in WHC. Further studies should be carried out to determine the antinutritional factors and to evaluate their nutrient value using in vitro or in situ digestibility techniques. [ABSTRACT FROM AUTHOR]

Background: Tall fescue (TF) and tall wheatgrass (TW) are the main temperate perennial grasses used for livestock grazing in the Pampas region of Argentina. In this region, pastures based on these grasses show low tiller density and the presence of bare soil in winter. Methods: An experiment was performed in pots kept outdoors. Plants of TF and TW were cut every 550°C day and 700°C day, respectively. In each species, reproductive tillers were removed at different times: (i) at each cut, (ii) when they reached the flag leaf stage, or (iii) when they reached full bloom. Tiller appearance, tiller death, and type of bud (basal or axillary) of the tillers present were determined. Results: Tiller appearance decreased from July to April. Minimum values were 0.2 and 1.1 tillers 100 tillers−1 day−1 in TF and TW, respectively. In TF, tiller death began in September–October and increased from 0.2 to 0.8 tillers 100 tillers−1 day−1 in March–April. In TW, tiller death began in October–November and increased from 0.6 to 0.8–1.0 tillers 100 tillers−1 day−1 from February to April. Conclusions: Defoliation of reproductive tillers affected the tiller density of TF plants in spring and in the following fall. In TW, the treatments did not modify the tiller density of the plants. [ABSTRACT FROM AUTHOR]

Three perennial ryegrass (Lolium perenne) populations (R1, R2, and R3) with suspected resistance (R) to acetolactate synthase (ALS) or acetyl-CoA carboxylase (ACCase) herbicides were collected from wheat (Triticum aestivum) fields in northwestern Greece to study the underlying mechanisms of resistance and their impact on growth rate and competitive ability against wheat. Preemergence and postemergence plant dose–response assays showed that the R1 population was cross-resistant to the ALS inhibitors chlorsulfuron, mesosulfuron + iodosulfuron, and pyroxsulam, but susceptible (S) to imazamox. However, all populations were susceptible to the ACCase inhibitors clodinafop-propargyl, clethodim, diclofop-methyl, and pinoxaden. The analysis of the ALS gene sequence revealed a substitution of Pro197 by His or Leu in the ALS enzyme in L. perenne, which is reported for the first time in this weed and indicates a potential mechanism of target site-mediated resistance. The R1 population grown in the absence or presence of wheat competition displayed similar aboveground biomass and tiller number trends, and therefore similar estimated growth rates. In addition, the aboveground biomass of wheat was similarly reduced by both the R1 and S populations, supporting the evidence of their similar competitive ability against wheat. In general, these findings indicate that there is no clear evidence for the fitness advantage of R1 over the S population. [ABSTRACT FROM AUTHOR]

Soil microorganisms play a vital role in increasing the availability of phosphorus (P) for plants through mineralization of organic P and solubilization of precipitated P compounds. In this two-year study, we analyzed several P-solubilizing microorganisms (PSMs) of the genus Bacillus and their consortiums for the ability to release soluble P from apatite concentrates of various grinding degrees using ryegrass (Lolium multiflorum Lam.) as a model plant. The effects were accessed by analyzing plant growth and nutrient assimilation. The greatest effect on root system development and plant biomass accumulation (dry weight) was observed for the apatite concentrate of standard grinding in combination with Bacillus megaterium BI14 and Bacillus subtilis BI2 and Bacillus velezensis BS89 strains. Although the introduction of apatite concentrates led to an increase in the content of total strontium in soil, the levels of strontium did not exceed the maximum allowable concentration, and the accumulation of mobile strontium by plants was unchanged; importantly, the use of tested PSMs led to a decrease in the strontium content in the green biomass of ryegrass. Our results indicate that biologized apatite concentrates in combination with PSMs represent promising fertilizers that can provide a source of soluble P to be readily assimilated by plants. [ABSTRACT FROM AUTHOR]

The current guidelines of waste management are aimed at the recovery and recycling of biowaste while respecting the protection of human health and the environment. The recent European legislation on fertilizers provides for the use of digestates derived from the organic fraction of municipal solid waste (OFMSW). The objectives of this study were to verify the fertilizing effect of three types of OFMSW digestates on the ryegrass culture comparing mineral fertilization and to evaluate the nitrogen lost to leaching in soil diversely fertilized following simulated rainfall. The ryegrass was grown in pots. The soil was fertilized with mineral fertilizer or OFMSW digestates. For each treatment, five mowing procedures were performed on the crop, and the ryegrass biomass production and nitrogen concentration were determined from the ryegrass samples. During the experiment, six rains were simulated, and the leached nitrogen was analyzed. The results showed that: (i) the fertilizing effect of OFMSW digestates on nitrogen nutrition of ryegrass was similar to ammonium sulphate fertilization, confirming the agronomic validity of these by-products to reduce the use of synthetic fertilizers; (ii) soil fertilization with OFMSW digestates had a positive effect limiting nitrogen loss due to leaching compared to mineral fertilization, highlighting the soil-improving properties of these by-products, in particular of the composted digestate. [ABSTRACT FROM AUTHOR]

The automotive upholstery industry, which processes bovine leathers, has struggled with vast amounts of solid waste, of which the majority are fractions, such as shavings and splits and offcuts, both containing chromium (Cr) and free of Cr. In this work, a novel leather waste-to-fertiliser approach has been tested: four lightly processed (incubation with effective microorganisms) and three aggressively processed (using pyrolysis or hydrolysis processes) tanned leather waste fractions were used. They were applied as organic nitrogen-based fertilisers in two separate glasshouse experiments, consisting of four subsequent harvests, under spring–autumn conditions in northern Poland. The Cr stressing effect caused by bovine shavings containing Cr and splits and offcuts containing Cr stimulated an increase in ryegrass growth after 30 days, providing twice the dry matter yield than bovine shavings free of Cr and splits and offcuts free of Cr, then (after 90 and 120 days) it inhibited growth. The Monod kinetics model was developed using the best fitting procedure (R2 > 0.94) for the efficiency of internal N use in ryegrass for the lightly processed fractions of tanned leather-waste fractions and each harvest. In the case of the processed leather waste-fractions, the relation was quasi linear, evidencing less growth inhibition and no over-fertilisation effects. The acid hydrolysate of bovine shavings containing Cr showed the best performance, reaching a relative agronomic effectiveness (RAE, N utilization-based) equal to 89%, 95%, 19%, and 38% for 30, 60, 90, and 120 days, respectively, and 71% in total. For the lightly processed fractions, it was lower than 12%. The future perspective should focus on steps including scaling up the tests to field conditions; transportation, storage, and application methods; effectiveness for other crops; usage in crop rotation systems; environmental impact assessment; production carbon footprint; commercial potential; and optimizing the production processes. [ABSTRACT FROM AUTHOR]

Summary: Some interspecific plant hybrids show unequal transmission of chromosomes from parental genomes to the successive generations. It has been suggested that this is due to a differential behavior of parental chromosomes during meiosis. However, underlying mechanism is unknown.We analyzed chromosome composition of the F2 generation of Festuca × Lolium hybrids and reciprocal backcrosses to elucidate effects of male and female meiosis on the shift in parental genome composition. We studied male meiosis, including the attachment of chromosomes to the karyokinetic spindle and gene expression profiling of the kinetochore genes.We found that Lolium and Festuca homoeologues were transmitted differently to the F2 generation. Female meiosis led to the replacement of Festuca chromosomes by their Lolium counterparts. In male meiosis, Festuca univalents were attached less frequently to microtubules than Lolium univalents, lagged in divisions and formed micronuclei, which were subsequently eliminated.Genome sequence analysis revealed a number of non‐synonymous mutations between copies of the kinetochore genes from Festuca and Lolium genomes. Furthermore, we found that outer kinetochore proteins NDC80 and NNF1 were exclusively expressed from the Lolium allele. We hypothesize that silencing of Festuca alleles results in improper attachment of Festuca chromosomes to karyokinetic spindle and subsequently their gradual elimination. [ABSTRACT FROM AUTHOR]