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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]

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]

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]

The type-B response regulator (ARR) family members represent the elements of two-component regulatory systems and play crucial roles in the cytokinin signaling, plant growth, and stress responses. The objective of this study was to characterize type-B ARR genes from perennial ryegrass, and to investigate their roles in regulating to salt tolerance. Twelve type-B ARRs , namely LpARR1 - 12 , were identified in perennial ryegrass. They were clustered into three subgroups and were unevenly distributed on chromosomes. The stress responsive elements (SREs) and hormone responsive elements (HREs) were identified in LpARRs promoters and the transcription of LpARRs genes were responsive to various abiotic stress (heat, salt, and osmotic) and hormonal (cytokinin, ABA, and ethylene) treatments. LpARR10 was strong induced by salt stress, therefore it was selected for function analysis. Overexpressing LpARR10 in Arabidopsis led to a significant improvement in salt tolerance. The LpARR10 -overexpressing (LpARR10 -OE) lines exhibited lower levels of Na+ content compared to WT plants, and the expression levels of SOS1 and SOS3 were significantly higher in the LpARR10 -OE lines. Results of yeast auto-transactivation and plant cell transcriptional activity assays shows LpARR10 is a nucleus localized transcriptional activator and directly bind and transactivated the promoter of SOS1 and SOS3. Together, LpARR10 enhances salt tolerance may via trans-activating SOS1 / SOS3 in Arabidopsis. Knowledge gained in this study will facilitate further functional analysis of LpARRs for molecular breeding of ryegrass and other related grass species. • Overexpressing LpARR10 resulted in a reduction in Na+ content compared to WT plants. • The LpARR10 -OE lines exhibited significantly higher expression levels of SOS1 and SOS3. • LpARR10 acts as a transcriptional activator and positively regulates salt tolerance by up-regulating the expressions of SOS1 and SOS3. [ABSTRACT FROM AUTHOR]