Populations Of Perennial Ryegrass Research Articles

Locations of Perennial Ryegrass (Lolium perenne L.) Landraces in Central Anadolu Region of Türkiye and Some Soil Characteristics of the Locations

This study was aimed to determine the characteristics of the soils of perennial ryegrass genotypes found in the natural flora in the Central Anadolu Region in Türkiye. Within the scope of the study, perennial ryegrass populations were collected from the provinces of Ankara, Eskişehir, Afyon, Konya, Aksaray, Karaman and Mersin and Antalya provinces in 2006 and 2007. Plants were collected and soil samples were taken and analysed from 87 sites in Ankara (20), Eskişehir (9), Afyon (5), Konya (39), Aksaray (1), Karaman (5), Mersin (2) and Antalya (6). According to the analysis results, the pH varied between 6.35-8.50, EC 52-1395 µS/cm, organic matter 0.04-9.1 %, lime 1.2-64.2%, P2O5 4.28-128.7 mg/kg, K2O 24.7-1360. 8 mg/kg, Ca 820-4030 mg/kg, Mg 32.7-3569 mg/kg, Na 0.6-2016 mg/kg, Fe 2.78-67.50 mg/kg, Zn 0.10-8.97 mg/kg, Cu 0.21-9.43 and Mn 2.33-42.34 mg/kg. As a result of soil analyses, large differences were observed between the locations where plants were collected in terms of the characteristics examined. This change shows the potential of developing grass varieties that can grow in different soil properties with a good breeding study of perennial ryegrass populations.

Evaluation of the Relationship between Cultivar, Endophyte and Environment on the Expression of Persistence in Perennial Ryegrass Populations Using High-Throughput Phenotyping

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.

Effect of Lolium perenne population differences on shoot tissue nitrogen concentrations when grown on a peat soil

AbstractGrass nitrogen (N) concentrations of dairy grasslands are higher on peat soil than on mineral soils. This can lead to increased N losses to air and water from dairy farming systems on peat soils. Our hypothesis was that the use of low-N perennial ryegrass (Lolium perenne L.) genotypes could be a means to lower grass N concentrations, when grown on peat soils. Our objective was to determine whether perennial ryegrass populations with different shoot tissue N concentrations, recorded on a sandy soil, would show different shoot tissue N concentrations and N use efficiencies (NUE) or N uptake efficiencies (NUptE) when grown on a peat soil. First, a pot experiment lasting 62 days was carried out with nine diploid and seven tetraploid populations, followed by a field experiment with two diploid and two tetraploid populations and a control lasting 30 months. Both experiments had three N fertilization levels. In the pot experiment, populations explained 3% of the variation in shoot tissue N concentration among tetraploids, 5 and 7% of the variation in NUE among diploids and tetraploids and 12% of the variation in NUptE among diploids. In the field experiment, populations explained 44% of the variation in NUptE. A higher NUE coincided with lower shoot tissue N concentrations among tetraploid populations in the pot experiment. We conclude that there is potential to alter the shoot tissue N concentrations of perennial ryegrass grown on peat soil, via selection for shoot tissue N concentrations and NUE.

Physiological and transcriptional status of genetically diverse perennial ryegrass (Lolium perenne L.) populations under drought stress

AbstractThis study was designed to understand how drought stress impact on the physiological and molecular status of genetically diverse Lolium perenne L. (perennial ryegrass) populations. The sensitivities of the 36 perennial ryegrass populations were checked against PEG-6000-simulated drought stress by analysing physiological status. Afterward, drought-related gene expressions were evaluated under normal and stressed conditions. The drought stress significantly up-regulated the drought-related genes in P6, P10, P15 and P30, while no difference was observed in drought-sensitive ones. The results collectively showed that four populations (P6, P10, P15, P30) are tolerant to drought stress and activate drought-related genes. Genetic similarities of the populations were also monitored using microsatellite (SSR) markers. The ten SSR primers detected 38 alleles. Surprisingly, the drought-tolerant populations were classified into different groups by principal component (PCA) and cluster analysis, showing that they are genetically different. Moreover, four unique alleles were identified by two SSR primers (P-07, and P-08) in P6, P10 and P15. In addition, the drought-tolerant populations (P6, P10, P15, P30) can be used to develop new perennial ryegrass cultivars for areas having drought problems.

Impact of ALS Herbicide-Resistant Perennial Ryegrass (Lolium perenne) Population on Growth Rate and Competitive Ability against Wheat

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.

Assessing inbreeding in perennial ryegrass (Lolium perenne) as a step towards F1 hybrid breeding

AbstractPerennial 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.

Epichloë seed transmission efficiency is influenced by plant defense response mechanisms.

Asexual Epichloë are endophytic fungi that form mutualistic symbioses with cool-season grasses, conferring to their hosts protection against biotic and abiotic stresses. Symbioses are maintained between grass generations as hyphae are vertically transmitted from parent to progeny plants through seed. However, endophyte transmission to the seed is an imperfect process where not all seeds become infected. The mechanisms underpinning the varying efficiencies of seed transmission are poorly understood. Host gene expression in response to Epichloë sp. LpTG-3 strain AR37 was examined within inflorescence primordia and ovaries of high and low endophyte transmission genotypes within a single population of perennial ryegrass. A genome-wide association study was conducted to identify population-level single nucleotide polymorphisms (SNPs) and associated genes correlated with vertical transmission efficiency. For low transmitters of AR37, upregulation of perennial ryegrass receptor-like kinases and resistance genes, typically associated with phytopathogen detection, comprised the largest group of differentially expressed genes (DEGs) in both inflorescence primordia and ovaries. DEGs involved in signaling and plant defense responses, such as cell wall modification, secondary metabolism, and reactive oxygen activities were also abundant. Transmission-associated SNPs were associated with genes for which gene ontology analysis identified "response to fungus" as the most significantly enriched term. Moreover, endophyte biomass as measured by quantitative PCR of Epichloë non-ribosomal peptide synthetase genes, was significantly lower in reproductive tissues of low-transmission hosts compared to high-transmission hosts. Endophyte seed-transmission efficiency appears to be influenced primarily by plant defense responses which reduce endophyte colonization of host reproductive tissues.

Divergent Selection for Seed Ability to Germinate at Extreme Temperatures in Perennial Ryegrass (Lolium perenne L.).

Various adaptive mechanisms can ensure that seedlings are established at the most favourable time and place. These mechanisms include seed dormancy i.e., incapacity to germinate in any environment without a specific environmental trigger and inhibition i.e., incapacity to germinate in an unfavourable environment (water availability, temperature: thermoinhibition and light). The objective of this research was to study in the temperate range for germination of forage and turf grass species perennial ryegrass, if the thermal requirements for germination are under genetic controlled and could be selectively bred. Two divergent selections of three cycles were realized on a natural population: one to select for the capacity to germinate at 10°C vs. the impossibility to germinate at 10°C, and one to select for the capacity to germinate at 32°C vs. the impossibility to germinate at 32°C. Seeds of all the lots obtained from the two divergent selections were then germinated at constant temperatures from 5 to 35°C to evaluate their germination ability. Concerning the positive selection, the first cycle of positive selection at 10°C was highly efficient with a very strong increase in the germination percentage. However, afterward no selection effect was observed during the next two cycles of positive selection. By contrast, the positive selection at 32°C was efficient during all cycles with a linear increase of the percentage of germination at 32°C. Concerning the negative selection, we observed only a large positive effect of the first cycle of selection at 10°C. These findings demonstrate that seed thermoinhibition at 10 and 32°C observed in a natural population of perennial ryegrass has a genetic basis and a single recessive gene seems to be involved at 10°C.

Forage potential of winter‐hardy perennial ryegrass populations in monoculture and binary alfalfa mixture

AbstractPerennial ryegrass (Lolium perenne L.) is a cool‐season grass valued for its grazing characteristics and nutritive value. MSP3984 and MSP4029, breeding populations selected for winter‐hardiness, were evaluated for biomass and nutritive value in monoculture and grass–alfalfa mixtures (Medicago sativa L.). Plots were sampled at conventional hay harvest intervals at three Minnesota locations for two production years, 2017 and 2018. Persistence was evaluated in the spring of the 3rd year, 2019. MSP3984 and MSP4029 were compared to commercial cultivars of perennial ryegrass (Lolium perenne L.), meadow fescue (Festuca pratensis Huds.), tall fescue (F. arundinacea Schreb.), and orchardgrass (Dactylis glomerata L.). Location and year × location effects largely influenced grass monoculture yields (p < .001) and botanical composition of mixtures (p < .001). In general, perennial ryegrass monocultures yielded less and were less persistent (p < .05) compared to the other cool‐season grasses. Biomass, nutritive value, and persistence did not differ among MSP3984, MSP4029, and the perennial ryegrass check ‘Remington’. However, MSP4029 was less competitive in grass–alfalfa mixtures than ‘Remington’ (p = .01). Perennial ryegrasses had lower neutral detergent fiber (NDF) concentrations than meadow fescue (p = .001) and orchardgrass (p < .001). Perennial ryegrasses also had lower acid detergent fiber (ADF) values on average than the other grasses, but cultivar effects were influenced by a year × location interaction (p = .02). This research reinforces (a) the importance of evaluating cool‐season forage grasses across latitudinal and precipitation gradients, (b) the value of grass–legume mixtures for yield stability compared to monocultures across locations and production years, and (c) the sustained need to improve forage production and persistence of perennial ryegrass for cold environments.

Management options to recover perennial ryegrass populations and productivity in run-out pastures

In parts of the upper North Island, farmers frequently report perennial ryegrass (Lolium perenne L.) pastures failing within 3 or 4 years post-sowing. This appears to be related to interactions between several factors: climatic (drier, hotter summers), biotic (insect pest), soil (texture, water-holding capacity) and grazing management factors that vary spatially and temporally. The efficacy of three management interventions for recovering ryegrass populations and production in runout pasture was assessed in an experiment initiated in 2018/19 in central Waikato. Treatments were a long-spring rotation (LSR), a longer grazing deferral during late spring and summer (pasture deferral, PD) and under-sowing with perennial ryegrass (US), each applied to pastures of four ryegrass cultivars. In the year after the treatments were implemented, the yield of ryegrass in PD was 2.4 t DM/ha greater than for the control (7-year-old pasture), and ryegrass tiller populations initially doubled but later declined. Yields in US and LSR were intermediate but not significantly different from the control. Ground score changes responded more positively to PD compared with the other treatments. Relative to the baseline prior to initiation of the study, tiller populations increased for PD and US but declined for control and LSR. There were no interactions between management treatment and cultivar for any of the variables measured. Pasture deferral shows promise as an intervention for recovering failing ryegrass pastures through natural reseeding. However, the longevity of the benefits observed here has yet to be determined.

Improving Accuracy of Herbage Yield Predictions in Perennial Ryegrass with UAV-Based Structural and Spectral Data Fusion and Machine Learning

High-throughput field phenotyping using close remote sensing platforms and sensors for non-destructive assessment of plant traits can support the objective evaluation of yield predictions of large breeding trials. The main objective of this study was to examine the potential of unmanned aerial vehicle (UAV)-based structural and spectral features and their combination in herbage yield predictions across diploid and tetraploid varieties and breeding populations of perennial ryegrass (Lolium perenne L.). Canopy structural (i.e., canopy height) and spectral (i.e., vegetation indices) information were derived from data gathered with two sensors: a consumer-grade RGB and a 10-band multispectral (MS) camera system, which were compared in the analysis. A total of 468 field plots comprising 115 diploid and 112 tetraploid varieties and populations were considered in this study. A modelling framework established to predict dry matter yield (DMY), was used to test three machine learning algorithms, including Partial Least Squares Regression (PLSR), Random Forest (RF), and Support Vector Machines (SVM). The results of the nested cross-validation revealed: (a) the fusion of structural and spectral features achieved better DMY estimates as compared to models fitted with structural or spectral data only, irrespective of the sensor, ploidy level or machine learning algorithm applied; (b) models built with MS-based predictor variables, despite their lower spatial resolution, slightly outperformed the RGB-based models, as lower mean relative root mean square error (rRMSE) values were delivered; and (c) on average, the RF technique reported the best model performances among tested algorithms, regardless of the dataset used. The approach introduced in this study can provide accurate yield estimates (up to an RMSE = 308 kg ha−1) and useful information for breeders and practical farm-scale applications.

To grow or survive: Which are the strategies of a perennial grass to face severe seasonal stress?

Abstract More severe seasonal stresses resulting from climate change affect the survival of perennial plant species. The growth–survival trade‐off exemplified in dormant species is a key issue to understand adaptation. As the validity of this trade‐off has yet to be tested in non‐dormant species, it was assessed by exploring the intraspecific variability of strategies to face drought and frost within perennial ryegrass. Three common gardens compared 385 European perennial ryegrass populations along a latitudinal environmental gradient over 3‐years. Persistence, productivity and physiological traits were recorded under contrasting seasonal environments. Decoupling plant responses, that is, growth under favourable summers/winters and plant survival under harsh summers/winters, showed a general trade‐off between growth potential and dehydration survival. Three groups of perennial ryegrass populations were identified according to their contrasting strategies: (a) year‐round productive but stress sensitive populations from wet areas; (b) drought‐tolerant populations with low summer growth potential from drought‐prone areas and (c) frost‐tolerant populations with low winter growth potential from frost‐prone areas. Overall, the populations surviving drought best were more resource conservative, whereas populations of the other groups were more resource acquisitive. However, such overall functional patterns were less meaningful than seasonal variations of resource acquisition potentials. The predicted potential biogeographical distribution of these groups suggests shifts of areas of suitability under climate change over the next decades in Europe. Dehydration escape and dehydration tolerance through reduction of growth potential in summer may become the strategies best adapted to an increasingly large area of Europe. The large intraspecific variability of phenological adaptations within perennial ryegrass reveals that the seasonal modulation of growth potential is crucial to plant adaptation under severe chronic abiotic stresses. The global plant economics spectrum cannot account for contrasting seasonal trade‐offs, which points out the importance of integrating phenological traits as key components of plant strategies. The identification of the trade‐off between growth potential and frost or drought stress survival in this non‐dormant species provides key knowledge to understand the future regional distribution of this major species for grassland ecosystem services. A free Plain Language Summary can be found within the Supporting Information of this article.

A new genetic locus for self-compatibility in the outcrossing grass species perennial ryegrass (Lolium perenne).

BackgroundSelf-incompatibility (SI) is a physiological mechanism that many flowering plants employ to prevent self-fertilization and maintain heterozygosity. In the grass family this is known to be controlled by a two locus (S-Z) system; however, the SI system is intrinsically leaky. Modifier genes of both the S and Z loci and a further locus, T, are known to override SI leading to self-fertilization and self-seed production. This has implications for the ecological and evolutionary success as well as the commercial breeding of grasses. Here we report a study where the genetic control of self-compatibility (SC) was determined from the results of self-pollinating an F2 population of perennial ryegrass from two independently derived inbred lines produced by single-seed descent.Methods In vitro self-pollinations of 73 fertile plants were analysed. A genetic association analysis was made with a panel of 1863 single-nucleotide polymorphism (SNP) markers, generated through genotype-by-sequencing methodology. Markers were placed on a recombination map of seven linkage groups (LGs) created using Joinmap v.5. The seed set on self- and open-pollinated inflorescences was determined on 143 plants, including the 73 plants analysed for self-pollination response.Key ResultsSelf-pollinations revealed a bimodal distribution of percentage SC with peaks at 50 and 100 %. A single quantitative trait locus (QTL) was identified with peak association for marker 6S14665z17875_11873 that mapped to LG 6. Peak position was associated with maximum marker segregation distortion. The self-compatible plants were equally fecund after self- and open pollination.ConclusionsThis is the first report in the Poaceae family of an SC locus located on LG 6. This new SC QTL discovery, as well as indicating the complex nature of the pollen–stigma recognition process and its evolutionary significance, provides an additional source of SC for breeding perennial ryegrass.

Multiple herbicide–resistant Lolium spp. is prevalent in wheat production in Texas Blacklands

AbstractField surveys were conducted across the Blacklands region of Texas during 2016 and 2017 to document the distribution of herbicide-resistant Lolium spp. infesting winter wheat production fields in the region. A total of 68 populations (64 Italian ryegrass, four perennial ryegrass) were evaluated in a greenhouse for sensitivity to herbicides of three different modes of action: an acetolactate synthase (ALS) inhibitor (mesosulfuron-methyl), two acetyl-coenzyme-A carboxylase (ACCase) inhibitors (diclofop-methyl and pinoxaden), and a 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) inhibitor (glyphosate). Herbicides were applied at twice the label-recommended rates for mesosulfuron-methyl (29 g ai ha−1), diclofop-methyl (750 g ai ha−1), and pinoxaden (118 g ai ha−1); and at the recommended rate for glyphosate (868 g ae ha−1). The herbicide screenings were followed by dose-response assays of the most-resistant ryegrass population for each herbicide at eight rates (0.5, 1, 2, 4, 8, 16, 32, and 64×), compared with a susceptible population at six rates (0.0625, 0.125, 0.25, 0.5, 1, and 2×). The initial screening and dose-response experiments were conducted in a completely randomized design with three replications and two experimental runs. Survivors (<80% injury) were characterized as highly resistant (0% to 20% injury) or moderately resistant (21% to 79%). Results showed that 97%, 92%, 39%, and 3% of the Italian ryegrass populations had survivors to diclofop-methyl, mesosulfuron-methyl, pinoxaden, and glyphosate treatments, respectively. Of the four perennial ryegrass populations, three were resistant to diclofop-methyl and mesosulfuron-methyl, and one was resistant to pinoxaden as well. Perennial ryegrass populations did not exhibit any resistance to glyphosate. Dose-response assays revealed 37-, 196-, and 23-fold resistance in Italian ryegrass to mesosulfuron-methyl, diclofop-methyl, and pinoxaden, respectively, compared with a susceptible standard. One Italian ryegrass population exhibited three-way multiple resistance to ACCase-, ALS-, and EPSPS-inhibitors. The proliferation of multiple herbicide–resistant ryegrass is a challenge to sustainable wheat production in Texas Blacklands and warrants diversified management strategies.

Differential gene expression and gene ontologies associated with increasing water-stress in leaf and root transcriptomes of perennial ryegrass (Lolium perenne).

Perennial ryegrass (Lolium perenne) is a forage and amenity grass species widely cultivated in temperate regions worldwide. As such, perennial ryegrass populations are exposed to a range of environmental conditions and stresses on a seasonal basis and from year to year. One source of potential stress is limitation on water availability. The ability of these perennial grasses to be able to withstand and recover after periods of water limitation or drought can be a key component of grassland performance. Thus, we were interested in looking at changes in patterns of gene expression associated with increasing water stress. Clones of a single genotype of perennial ryegrass were grown under non-flowering growth room conditions in vermiculite supplemented with nutrient solution. Leaf and root tissue was sampled at 4 times in quadruplicate relating to estimated water contents of 35%, 15%, 5% and 1%. RNA was extracted and RNAseq used to generate transcriptome profiles at each sampling point. Transcriptomes were assembled using the published reference genome sequence and differential gene expression analysed using 3 different programmes, DESeq2, edgeR and limma (with the voom transformation), individually and in combination, deriving Early, Middle and Late stage comparisons. Identified differentially expressed genes were then associated with enriched GO terms using BLAST2GO. For the leaf, up-regulated differentially expressed genes were strongly associated with GO terms only during the Early stage and the majority of GO terms were associated with only down-regulated genes at the Middle or Late stages. For the roots, few differentially expressed genes were identified at either Early or Middle stages. Only one replicate at 1% estimated water content produced high quality data for the root, however, this indicated a high level of differential expression. Again the majority of enriched GO terms were associated with down-regulated genes. The performance of the different analysis programmes and the annotations associated with identified differentially expressed genes is discussed.

Developing new tools for pasture plant breeding

The rate of genetic gain represented in the Forage Value Index of perennial ryegrass (Lolium perenne) is a major factor underpinning sustained profitability in pastoral farming. Effective new technologies for trait data acquisition and parent plant selection are used in many animal and crop improvement programmes to lift the rate of gain, but have yet to be developed and integrated in forage breeding. For forage improvement, hypotheses tested were: a) genomic selection (GS) offers a viable breeding strategy, and b) key enabling technologies for non-destructive, high-throughput phenotyping (HTP) in the field will improve trait data acquisition. To evaluate GS, extensive molecular marker and phenotypic datasets in structured populations of perennial ryegrass were developed. Phenotypic data for seasonal dry matter yield (DMY), the core trait in the Dairy NZ Forage Value Index, were obtained replicated field trials. Data on heading date (HD) as a useful trait to assess the efficacy of GS for simply inherited traits, were also collected. Genomic prediction models were developed for seasonal DMY and HD. Application of GS for HD was effective in selecting for both early and late heading, with movement of up to 7 days in a single generation of selection. The HTP research used iterative development of computational methods supporting a repeatable, non-invasive means of accurately and rapidly measuring DMY of perennial ryegrass in single row plots. These findings demonstrate effective genetic prediction and phenotyping approaches which may enable breeders to lift the rate of genetic gain in perennial ryegrass.

Genetic Loci Governing Androgenic Capacity in Perennial Ryegrass (Lolium perenne L.).

Immature pollen can be induced to switch developmental pathways from gametogenesis to embryogenesis and subsequently regenerate into homozygous, diploid plants. Such androgenic production of doubled haploids is particularly useful for species where inbreeding is hampered by effective self-incompatibility systems. Therefore, increasing the generally low androgenic capacity of perennial ryegrass (Lolium perenne L.) germplasm would enable the efficient production of homozygous plant material, so that a more effective exploitation of heterosis through hybrid breeding schemes can be realized. Here, we present the results of a genome-wide association study in a heterozygous, multiparental population of perennial ryegrass (n = 391) segregating for androgenic capacity. Genotyping-by-sequencing was used to interrogate gene- dense genomic regions and revealed over 1,100 polymorphic sites. Between one and 10 quantitative trait loci (QTL) were identified for anther response, embryo and total plant production, green and albino plant production and regeneration. Most traits were under polygenic control, although a major QTL on linkage group 5 was associated with green plant regeneration. Distinct genetic factors seem to affect green and albino plant recovery. Two intriguing candidate genes, encoding chromatin binding domains of the developmental phase transition regulator, Polycomb Repressive Complex 2, were identified. Our results shed the first light on the molecular mechanisms behind perennial ryegrass microspore embryogenesis and enable marker-assisted introgression of androgenic capacity into recalcitrant germplasm of this forage crop of global significance.

An Immortalized Genetic Mapping Population for Perennial Ryegrass: A Resource for Phenotyping and Complex Trait Mapping.

To address the lack of a truly portable, universal reference mapping population for perennial ryegrass, we have been developing a recombinant inbred line (RIL) mapping population of perennial ryegrass derived via single seed descent from a well-characterized F2 mapping population based on genetically distinct inbred parents in which the natural self-incompatibility (SI) system of perennial ryegrass has been overcome. We examined whether it is possible to create a genotyping by sequencing (GBS) based genetic linkage map in a small population of the F6 generation of this population. We used 41 F6 genotypes for GBS with PstI/MspI-based libraries. We successfully developed a genetic linkage map comprising 6074 SNP markers, placing a further 22080 presence and absence variation (PAV) markers on the map. We examined the resulting genetic map for general and RIL specific features. Overall segregation distortion levels were similar to those experienced in the F2 generation, but segregation distortion was reduced on linkage group 6 and increased on linkage group 7. Residual heterozygosity in the F6 generation was observed at a level of 5.4%. There was a high proportion of chromosomes (30%) exhibiting the intact haplotype of the original inbred parents of the F1 genotype from which the population is derived, pointing to a tendency for chromosomes to assort without recombining. This could affect the applicability of these lines and might make them more suitable for situations where repressed recombination is an advantage. Inter- and intra-chromosomal linkage disequilibrium (LD) analysis suggested that the map order was robust. We conclude that this RIL population, and subsequent F7 and F8 generations will be useful for genetic analysis and phenotyping of agronomic and biological important traits in perennial ryegrass.

Genomic prediction of crown rust resistance in Lolium perenne

BackgroundGenomic selection (GS) can accelerate genetic gains in breeding programmes by reducing the time it takes to complete a cycle of selection. Puccinia coronata f. sp lolli (crown rust) is one of the most widespread diseases of perennial ryegrass and can lead to reductions in yield, persistency and nutritional value. Here, we used a large perennial ryegrass population to assess the accuracy of using genome wide markers to predict crown rust resistance and to investigate the factors affecting predictive ability.ResultsUsing these data, predictive ability for crown rust resistance in the complete population reached a maximum of 0.52. Much of the predictive ability resulted from the ability of markers to capture genetic relationships among families within the training set, and reducing the marker density had little impact on predictive ability. Using permutation based variable importance measure and genome wide association studies (GWAS) to identify and rank markers enabled the identification of a small subset of SNPs that could achieve predictive abilities close to those achieved using the complete marker set.ConclusionUsing a GWAS to identify and rank markers enabled a small panel of markers to be identified that could achieve higher predictive ability than the same number of randomly selected markers, and predictive abilities close to those achieved with the entire marker set. This was particularly evident in a sub-population characterised by having on-average higher genome-wide linkage disequilibirum (LD). Higher predictive abilities with selected markers over random markers suggests they are in LD with QTL. Accuracy due to genetic relationships will decay rapidly over generations whereas accuracy due to LD will persist, which is advantageous for practical breeding applications.

Low pyrrolizidine alkaloid levels in perennial ryegrass is associated with the absence of a homospermidine synthase gene

BackgroundPyrrolizidine alkaloids (PAs) are a class of secondary metabolites that function as feeding deterrents in a range of different plant species. In perennial ryegrass (Lolium perenne L.) the only PAs that have been identified are the thesinine-rhamnoside group, which displays significant genetic variation. Homospermidine synthase (HSS) has evolved from deoxyhypusine synthase (DHS) and catalyses the first step in the PA pathway, making it a key candidate for the investigation of genes influencing observed PA trait variation.ResultsDuring PCR amplification and sequence analysis of DHS we identified two putative HSS genes in perennial ryegrass. One of the genes (LpHSS1) was absent in some perennial ryegrass plants. Thesinine-rhamnoside levels were measured using liquid chromatography coupled with mass spectrometry in a diverse association mapping population, consisting of 693 plants free of fungal endophytic symbionts. Association tests that accounted for population structure identified a significant association of absence of the LpHSS1 gene with lower levels of thesinine-rhamnoside PAs. HSS-like gene sequences were identified for other grass species of the Poaceae, including tall fescue, wheat, maize and sorghum.ConclusionHSS is situated at the crucial first step in the PA pathway making it an important candidate gene for investigation of involvement in PA phenotypic variation. In this study, PA level in perennial ryegrass was strongly associated with the presence or absence of the LpHSS1 gene. A genetic marker, developed for the presence/absence of LpHSS1, may be used for marker-assisted breeding to either lower or increase PAs in breeding populations of perennial or Italian ryegrass to investigate a potential role in the deterrence of herbivore pests. The presence of HSS-like genes in several other Poaceae species suggests that PA biosynthesis may occur in plant family members beyond perennial ryegrass and tall fescue and identifies a potential route for manipulating PA levels.