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

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]

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]

Perennial ryegrass is an important turf and forage grass in temperate regions worldwide. Limited genetic gains have been made with current breeding strategies compared with other grass crops, such as rice and maize, which benefit from Filial 1 (F1) hybrid breeding. One of the largest constraints on hybrid breeding in ryegrass is self‐incompatibility preventing inbreeding, as homozygous parental lines are required to develop hybrids with maximal hybrid vigour. Obligate outcrossing in ryegrass has resulted in cultivars with high levels of heterozygosity, lacking trait uniformity across the population. A naturally occurring self‐fertile (SF) locus that overcomes the self‐incompatibility system has been identified in a European perennial ryegrass population. This study crossed the SF locus into an elite cultivar, producing a self‐compatible population that was inbred for several generations. Genotyping‐by‐sequencing (GBS) was used to assess the population structure and degree of inbreeding in the self‐compatible population. Phenotypic analysis indicated that increased homozygosity did not necessarily affect growth and performance. This study concludes that self‐compatible ryegrass is a promising tool for hybrid breeding and agronomic improvement of perennial ryegrass. [ABSTRACT FROM AUTHOR]

In summer 2018, brown stripe symptoms were observed on leaf blades of perennial ryegrass (Lolium perenne L.) used as a turfgrass in an athletic field in Saitama Prefecture, Japan. Bacterial ooze exuded from lesions, from which a pathogenic bacterium was isolated. Inoculation of blades of perennial ryegrass with the isolates reproduced the same symptoms. Pathogenicity and bacteriological properties of the isolates were almost identical to those of Acidovorax avenae, and the identification was supported by phylogenetic analysis. This is the first report of the brown stripe on perennial ryegrass caused by A. avenae (= A. avenae subsp. avenae) in Japan. [ABSTRACT FROM AUTHOR]

α-Ketoglutarate (AKG) is a key intermediate metabolite in the tricarboxylic acid cycle of respiration and a precursor for glutamate, playing important roles in regulating plant growth and stress tolerance. The objectives of this study were to examine effects of AKG on heat tolerance characterized by leaf senescence in a cool-season grass species by foliar application and to determine major metabolites and associated metabolic pathways regulated by AKG for its effects on heat tolerance. Perennial ryegrass (Lolium perenne L.) plants were exposed to heat stress (35/30 °C, day/night) or optimal temperature (25/20 °C, day/night, non-stress control) in controlled-environment growth chambers. The solution containing AKG (5 mM) was applied to leaves by spraying 7 d prior to the initiation of heat stress and every 7 d during the heat stress period. Exogenous application of AKG enhanced heat tolerance in perennial ryegrass, as manifested by significant increases in leaf chlorophyll content, photochemical efficiency, and membrane stability, as well as activities of antioxidant enzymes for H 2 O 2 scavenging in AKG-treated plants relative to untreated control plants exposed to heat stress. Metabolic profiling and pathway analysis demonstrated that exogenous AKG application enhanced metabolite accumulation in four major metabolic pathways, including antioxidant metabolism, amino acid metabolism, glycolysis and tricarboxylic acid cycle of respiration, and pyrimidine metabolism, contributing to AKG-improved heat tolerance in perennial ryegrass. • α-Ketoglutarate (AKG) enhanced heat tolerance in perennial ryegrass. • AKG suppressed heat-induced leaf senescence. • AKG regulated the metabolism of antioxidant, amino acids, pyrimidine, glycolysis, and TCA cycle of respiration. [ABSTRACT FROM AUTHOR]

The productivity of perennial ryegrass (Lolium perenne L.) is constrained by summer water deficit. We hypothesised that selection for desirable water relations traits might partially mitigate this constraint. Genotypes of a 1970s, and two recent ryegrass cultivars were grown in 55 cm tall pots for three months and exposed to moisture deficit for four weeks, then 66 plants were assessed for shoot (SDW) and root (RDW) dry weight, leaf elongation rate (LER), specific water use (SWU), leaf osmotic potential (OP), OP change, leaf relative water content (RWC), and gravimetric soil moisture. Among the 66 genotypes, SWU ranged from 935 ± 19 to 1754 ± 74 g H2O g–1 DM. In principal component (PC) analysis, PC1 indicated greater growth and lower SWU from greater soil water extraction as the common trait association among genotypes tested and ANOVA of PC scores indicated that recent cultivars outperformed the 1970s cultivar. PC2 indicated water saving by decreased LER and SDW, with increased RDW indicative of summer dormancy. PC3 associated more negative OP with improved SWU and RWC. This experiment demonstrates not only genotypic diversity for SWU but also diversity in trait associations determining SWU, which should be considered when investigating SWU, both in ecology and agronomy research. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the effects of cultivar, nitrogen (N) rate, and biostimulant application on the chemical composition of the aboveground biomass of perennial ryegrass (Lolium perenne L.). A small-area field experiment was established in the Agricultural Experiment Station in Tomaszkowo (53°42′40.8″ N 20°26′04.7″ E, north-eastern Poland). The experiment had a split-plot design with three replications, and the experimental variables were as follows: (i) perennial ryegrass cultivar: Bajka and Baronka, (ii) N fertilizer rate: 0, 120, 240 kg N ha−1, (iii) application of biostimulants: Blatt Boden-Foliar (BB-F) and Blatt Boden-Foliar + Blatt Boden-Multical (BB-F + BB-M). This study demonstrated that the tetraploid cultivar Baronka had a more desirable chemical composition than the diploid cultivar Bajka. The biomass of cv. Baronka had a higher content of CP, CF, and K, and it was characterized by higher leaf greenness (SPAD) values. Nitrogen fertilization considerably increased the content of CP, P, and K, and leaf greenness (SPAD) values in both cultivars, and the noted increase was higher when N was applied at 240 kg ha−1. The N fertilizer rate of 120 kg ha−1 led to a significant decrease in the average Ca content of plants, whereas the N fertilizer rate of 240 kg ha−1 had no significant effect on Ca concentration. The tested biostimulants significantly affected the chemical composition of perennial ryegrass biomass, and their influence was greater when they were applied in combination. The foliar application of Blatt Boden-Foliar and Blatt Boden-Multical increased the content of CP, P, and chlorophyll in perennial ryegrass leaves, whereas it decreased the accumulation of CF, K, and Ca in plants. The analyzed biostimulants had a positive effect on the chemical composition of perennial ryegrass biomass. This is an important practical consideration because high-quality green fodder for livestock can be produced while minimizing the use of mineral fertilizers and adverse environmental impacts. [ABSTRACT FROM AUTHOR]

Perennial ryegrass (Lolium perenne) is a widely used cool season turfgrass with outstanding turf quality and grazing tolerance. High temperature is the key factor restricting the distribution of perennial ryegrass in temperate and sub-tropic regions. In this study, we found that one HEAT SHCOK TRANSCRIPTION FACOTR (HSF) class A gene from perennial ryegrass, LpHSFA3 , was highly induced by heat stress. LpHSFA3 is localized in nucleus and functions as a transcription factor. Ectopic overexpression of LpHSFA3 in Arabidopsis improved thermotolerance and rescued heat sensitive deficiency of athsfa3 mutant. Overexpression of LpHSFA3 in perennial ryegrass enhanced heat tolerance and increased survival rate in summer season as evidenced by decreased EL and MDA, increased number of green leaves and total chlorophyll content. LpHSFA3 binds to the HSE region in LpHSFA2a promoter to constitutively activate the expression of LpHSFA2a and downstream heat stress responsive genes. Ectopic overexpression of LpHSFA2a consequently rescued thermal sensitivity of athsfa3 mutant and enhanced thermotolerance of athsfa2 mutant. Perennial ryegrass protoplasts with overexpression of LpHSFA3 and LpHSFA2a exhibited induction of similar subsets of heat responsive genes. These results indicated that transcription factor LpHSFA3 functions as positive regulator of LpHSFA2a to improve thermotolerance of perennial ryegrass, providing further evidence to understand the regulatory networks of plant heat stress response. • LpHSFA3 and LpHSFA2a were significantly induced by heat stresss. • LpHSFA3 improves plant heat tolerance and survival rate in summer season. • LpHSFA3 directly activates the expression of LpHSFA2a. • LpHSFA2a enhances plant heat tolerance. [ABSTRACT FROM AUTHOR]

Autumn closing date (CD) in intensive pasture‐based systems influences spring herbage availability. Little is known about the impact of earlier closing of swards in autumn on sward structure. A two‐year experiment was undertaken to investigate the impact of five CD on herbage production, leaf extension rate (LER), leaf senescence rates (LSR), sward free leaf lamina (FLL) content and the production and survival of tillers over winter and into spring. The five CD ranges were October 1–15 (CD1), October 16–30 (CD2), October 31––November 14 (CD3), November 15–29 (CD4), November 30–December 14 (CD5) and sward measurements were made on four measurement dates (MD): December 11 (MD1), January 6 (MD2), January 27 (MD3) and prior to spring defoliation (MD4). Thirty perennial ryegrass tillers were measured until the first defoliation in spring (average 1 March). Herbage mass was greatest in early CD and tillers had greater LER and LSR (4.6 and 4.5 mm tiller−1 day−1 respectively) compared to swards with a later CD (3.2 and 2.3 mm tiller−1 day−1 respectively). Early CD had greater parent and daughter tiller mortality than later CD (parent tiller mortality 1.1 and 0.3 ± 0.19 tillers; daughter tiller mortality 1.1 and 0.3 ± 0.27 tillers respectively). Early CD also had greater LER and LSR in daughter tillers (2.6 and 1.5 mm tiller−1 day−1, respectively) compared to late (1.5 and 0.5 mm tiller−1 day−1, respectively). Earlier closing of sward increased herbage mass in spring, however, there were negative implications, such as increased LSR and tiller mortality. [ABSTRACT FROM AUTHOR]

The aim of this study was to determine and compare the sensitivity of new high yield Bulgarian varieties of perennial ryegrass (Lolium perenne L.) - Harmoniya, Tetrany and Tetramis to glyphosate in laboratory conditions. The effect of three doses of glyphosate - 360, 720 and 1440 g a.i./ha applied in the growth stage tillering (BBCH - 22-23) on perennial ryegrass on the dynamics of plant growth and formed fresh biomass in g per plant was studied. Genotypic differences were observed between the studied varieties of perennial ryegrass to glyphosate. With a relatively high sensitivity to glyphosate conditionally can be determined diploid variety Harmoniya in which the depression coefficients (B) on the dynamics of growth and accumulation of fresh biomass are in the range from -5.7 to - 4.7, while for tetraploids Tetrany (B) (average) is in the range of -3.3 to -3.9 and for Tetramis (B) (average) is within limits from -3.2 to -3.9, they have relatively high tolerance to glyphosate and can be used as components in future programs. [ABSTRACT FROM AUTHOR]

Despite the progress made in DNA sequencing over the last decade, reconstructing telomere-to-telomere genome assemblies of large and repeat-rich eukaryotic genomes is still difficult. More accurate basecalls or longer reads could address this issue, but no current sequencing platform can provide both simultaneously. Perennial ryegrass (Lolium perenne L.) is an example of an important species for which the lack of a reference genome assembly hindered a swift adoption of genomics-based methods into breeding programs. To fill this gap, we optimized the Oxford Nanopore Technologies' sequencing protocol, obtaining sequencing reads with an N50 of 62 kb—a very high value for a plant sample. The assembly of such reads produced a highly complete (2.3 of 2.7 Gb), correct (QV 45), and contiguous (contig N50 and N90 11.74 and 3.34 Mb, respectively) genome assembly. We show how read length was key in determining the assembly contiguity. Sequence annotation revealed the dominance of transposable elements and repeated sequences (81.6% of the assembly) and identified 38,868 protein coding genes. Almost 90% of the bases could be anchored to seven pseudomolecules, providing the first high-quality haploid reference assembly for perennial ryegrass. This protocol will enable producing longer Oxford Nanopore Technology reads for more plant samples and ushering forage grasses into modern genomics-assisted breeding programs. [ABSTRACT FROM AUTHOR]

• Genome-wide identification and comprehensive analysis of NAC gene family. • LpNAC genes expression analysis under various abiotic stress. • Candidate function gene identification based on phylogenetic analysis. NAC is one of the largest family of plant-specific transcription factors, and it plays important roles in plant development and stress responses. The study identified 72 LpNACs genes from the perennial ryegrass genome database. Gene length, MW and pI of NAC family transcription factors varied, but the gene structure and motifs were relatively conserved in bioinformatics analysis. Phylogenetic analyses of perennial ryegrass, rice and Arabidopsis were performed to study the evolutionary and functional relationships in various species. The expression of LpNAC genes that respond to various abiotic stresses including high salinity, ABA, high temperature, polyethylene glycol (PEG) and heavy metal was comprehensively analyzed. The present study provides a basic understanding of the NAC gene family in perennial ryegrass for further abiotic stress studies and improvements in breeding. [ABSTRACT FROM AUTHOR]

The plant growth regulator trinexapac-ethyl (TE) is widely used to enhance ryegrass seed yields by reducing lodging (i.e. delaying collapse of the crop). However, lodging data are usually a single evaluation related to TE rate for a particular date of measurement. In eight field trials with varying rates of TE, weekly lodging assessments were made from full head emergence and days to 50% lodging were determined. In all trials, dynamic assessment of TE treatment showed there was a strong positive correlation (R 2 = 0.82) between the number of days to 50% lodging and seed yield. Nil TE reached 50% lodging in 7 days compared with 33 days for TE treatments of 400 g ha-1. On average, one days' delay in reaching 50% lodging increased seed yield by 24 kg ha-1 (response range 16-33 kg ha-1). The data indicate that TE rate per se is less important than the rate required to delay the date that 50% lodging is reached. The seed yield response was linear for delayed lodging from full head emergence to harvest, suggesting that the highest yields are achieved if the crop is not more than 50% lodged at harvest. [ABSTRACT FROM AUTHOR]

The effects of one cycle of selection for adventitious root system shape (i.e. percentage of total root mass in successive depth increments) were investigated in progeny of four perennial ryegrass pools with contrasting rooting patterns: (1) low surface (0-10 cm) root mass and roots to 1 m; (2) high surface root mass and roots to 1 m; (3) high root mass 10-20 cm and roots to 1 m; (4) high surface root mass, shallow rooting. Ten half-sib families were selected from each of the root type progenies, and five seeds from each family were sown and raised as stock plants. Tiller cuttings of each plant were planted into individual 1 m deep root screening tubes of sand and irrigated with nutrient solution After 115 days, the shoots were cut off, the sand/root column cut into 10 cm increments and the roots washed free of sand. The shoots and root samples were oven-dried and weighed. The four root shape progeny groups did not differ significantly for shoot or root dry weight (DW), or root/shoot DW ratio. Root type 2 (high surface root mass and roots to 1 m) progeny had a significantly higher percentage of total root mass between 0 and 10 cm than did root type 1 (low surface root mass and roots to 1 m) but none of the other differences were significant for percentage roots 0-10 cm. The root type 2 progeny had a lower percentage of total root DW between 10 and 20 cm than all the other root types, none of which differed and had a significantly lower percentage of root DW 20-30 cm than types 1 and 4 but did not differ at this depth from type 3 progeny. The probability of root type 4 (high surface root mass, shallow rooting) plants having roots at 1 m depth was 0.67; this was significantly lower than for the other selections, none of which varied significantly. These results indicate that it should be possible to change root system shape in perennial ryegrass using conventional breeding techniques. [ABSTRACT FROM AUTHOR]

Root and shoot dry weights (DW), and root DW depth distribution were determined for wild type perennial ryegrass, breeding lines and cultivars. The plants were grown in 1 m deep tubes of sand culture in a glasshouse experiment. There were significant differences in shoot DW among accessions in the wild types but not within the bred material, and the wild types had more accessions with low shoot DW. Root DW varied significantly among accessions in both the bred material and the wild types. There were significant differences in root/shoot DW ratios among accessions, and between pools in the breeding lines, but no differences within the wild types. Root/ shoot ratios in the wild types had a much narrower and generally higher range of values than those in the bred lines. The percentage of root DW in the top 10 cm of sand did not vary significantly in the bred lines but it did in the wild types. Variation was recorded in root DW depth distribution at both the accession and genotype levels. We concluded that selection for merit based on shoot performance does not necessarily result in large root systems, and better plant performance might result from an increase in root system size in perennial ryegrass. [ABSTRACT FROM AUTHOR]

• Toxicity threshold for ryegrass staggers is lowered the longer perennial ryegrass is fed. • Sub-chronic (>60 day) threshold for lolitrem B is 1554 µg/kg in Angus beef cattle. • Lolitrem B accumulates in fat tissue in a dose- and time-responsive manner. Lolitrem B is a potent tremorgen responsible for the syndrome known as "ryegrass staggers" in livestock consuming perennial ryegrass (Lolium perenne) infected with the endophyte fungus Epichloë festucae var. lolii. We evaluated clinical signs in cattle continuously fed endophyte-infected perennial ryegrass straw at four concentrations of lolitrem B (LB)/ergovaline (EV) (groups: I-2256/638, II-1554/373, III-1012/259 or IV-<300/0 µg/kg LB/EV, n =5-6 steers/group) over 64 days to establish a sub-chronic threshold of toxicity for this mycotoxin; animals were supplemented with 1 kg grain mix/day. Most animals in groups I and II developed clinical signs consistent with ryegrass staggers by day 64; thus, it is recommended that a threshold level of 1554 µg/kg LB should not be exceeded for extended feeding regimens using endophyte-infected perennial ryegrass feed material alone. This toxicity threshold is lower than that observed with shorter feeding periods, i.e. 1800–2000 µg/kg LB when fed for less than 30 days. No clinical signs associated with ergovaline toxicosis (i.e. decreased weekly weight gain) were observed. As adipose tissue appears to be the main compartment where LB is deposited, the concentration of LB and its metabolic derivatives in fat samples was determined at 37 and 64 days. Lolitrem B and three previously defined metabolites bioaccumulated in a dose- and time-dependent manner throughout the feeding experiment in fat tissue, at levels similar to those found in past research. Ratios of metabolites to LB were consistent across time points and dose groups, indicating that the enzyme(s) responsible for LB metabolism were not saturated. In summary, LB toxicity thresholds are dependent upon specificities of the dietary program including the duration feed material is fed, and should be considered when formulating safe livestock feeding regimens. [Display omitted] [ABSTRACT FROM AUTHOR]

The aim of this study was to determine the effects of cultivar, nitrogen (N) rate and harvest time on the content of water-soluble carbohydrates (WSC), crude protein (CP) and the WSC:CP ratio in the aboveground biomass of perennial ryegrass (Lolium perenne L.). A small-area field experiment was conducted in the Agricultural Experiment Station in Tomaszkowo (53°42′40.8″ N 20°26′04.7″ E, north-eastern Poland). Data were presented for three years of full utilization (2013–2015). The experimental factors were as follows: (i) perennial ryegrass cultivar: Bajka and Baronka, (ii) N rate (kg ha−1): 0, 120, and 240, and (iii) harvest time: 8.00–10.00 a.m., 12.00–2.00 p.m., and 4.00–6.00 p.m. The tetraploid cultivar Baronka had higher WSC content and lower CP content on a dry matter (DM) basis than the diploid cultivar Bajka (by approx. 3% on average). An increase in N fertilizer rate to 240 kg ha−1 contributed to a decrease in WSC concentrations by 23% in cv. Bajka and by 19% in cv. Baronka, and to an increase in the CP content of aboveground biomass by 32% and 23%, respectively. Both cultivars accumulated significantly higher WSC amounts when biomass was harvested at noon and in the afternoon than in the morning. The CP content of biomass was also affected by harvest time, and it was higher in cv. Bajka when harvesting was carried out at 12.00–2.00 p.m. and 4.00–6.00 p.m., and in cv. Baronka when harvesting was carried out at 4.00–6.00 p.m. The WSC:CP ratio in the biomass of the analyzed cultivars was within the optimal range for ruminants, and it was higher in cv. Baronka (1:1.11). Nitrogen applied at 240 kg ha−1 had a negative influence on the WSC:CP ratio, decreasing its value by around 36% (relative to the rate of 120 kg N ha−1) and around 35% (relative to the unfertilized treatment). In both perennial ryegrass cultivars, biomass harvested in the morning was characterized by the lowest WSC:CP ratio. [ABSTRACT FROM AUTHOR]