Ryegrass Populations Research Articles

Stimulating dormancy release and emergence of annual ryegrass (Lolium rigidum) seeds using short-term hydrated storage in darkness

Experiments were performed to determine whether the dormancy release effect of hydrated storage in darkness (dark-stratification) is common amongst annual ryegrass populations and has the potential to occur under field conditions. Dormant seeds from all populations tested (22) became sensitive to light during dark-stratification, enabling them to germinate when subsequently exposed to light. Under controlled temperature (25/15°C), light (12-h photoperiod), and hydration (solidified agar-water) conditions, more seeds germinated by 28 days if the first 14 days were in darkness followed by exposure to light for 12 h per day than if they were exposed to light throughout or darkness throughout. Constraint over the conditions imposed during dark-stratification and germination was gradually reduced to investigate whether the dormancy release effect was diminished. Dark-stratification was effective in promoting germination when performed under natural diurnal temperatures, and burial in moist soil provided suitable conditions for dark-stratification to occur. The surface of moist soil, with natural diurnal temperatures and sunlight, was suitable for germination of dark-stratified seeds. Dark-stratification is a quick and effective means to enhance the sensitivity of dormant annual ryegrass seeds to light, enabling the majority of the population to germinate. However, large quantities of light are required to promote germination of dark-stratified seeds, so buried seeds must be moved to the soil surface to allow exposure to adequate light for germination.

Morphological and molecular characterization of italian ryegrass populations

Italian ryegrass is the most important temperate grass in Rio Grande do Sul, Brazil. Despite its overall importance, there are no breeding programs for this species in this State. The purpose of this study was to evaluate the existing variability within and between four Italian ryegrass populations, three from Rio Grande do Sul, Brazil, and one from Uruguay. The populations were characterized based on morphological traits such as: number of tillers, canopy diameter, heading date, total leaf area, number of leaves, area/leaf, leaf dry matter, stem dry matter, leaf/stem ratio and dry matter yield. Molecular variation was also characterized using RAPD markers. A large variability was found within the populations (98.41%), which limited the complete separation of populations. However, significant differences were found among populations for traits of great forage interest, such as the number of tillers, canopy and heading date, indicating that the present variability is suitable for the initiation of a breeding program.

Field assessment of thermal after‐ripening time for dormancy release prediction in Lolium rigidum seeds

SummaryDormancy release was studied in four populations of annual ryegrass (Lolium rigidum) seeds to determine whether loss of dormancy in the field can be predicted from temperature alone or whether seed water content (WC) must also be considered. Freshly matured seeds were after‐ripened at the northern and southern extremes of the Western Australian cereal cropping region and at constant 37°C. Seed WC was allowed to fluctuate with prevailing humidity, but full hydration was avoided by excluding rainfall. Dormancy was measured regularly during after‐ripening by germinating seeds with 12‐hourly light or in darkness. Germination was lower in darkness than in light/dark and dormancy release was slower when germination was tested in darkness. Seeds were consistently drier, and dormancy release was slower, during after‐ripening at 37°C than under field conditions. However, within each population, the rate of dormancy release in the field (north and south) in terms of thermal time was unaffected by after‐ripening site. While low seed WC slowed dormancy release in seeds held at 37°C, dormancy release in seeds after‐ripened under Western Australian field conditions was adequately described by thermal after‐ripening time, without the need to account for changes in WC elicited by fluctuating environmental humidity.

Analysis of the temperature effect on the components of plant digestibility in two populations of perennial ryegrass

AbstractFor the development of mechanistic models of herbage digestibility, quantitative insight into the effects of age, temperature and cultivar on digestibility characteristics of individual plant parts is needed. Towards that goal, glasshouse experiments were conducted at day/night temperatures of 13/8, 18/13 and 23/18 °C with vegetative and reproductive crops of two populations of perennial ryegrass (Lolium perenne L) selected for differences in leaf blade digestibility. Cell wall content (CWC) and true cell wall and organic matter digestibility (CWD and OMD) of vegetative and reproductive tillers were related to dimensions, mass, CWC and digestibility of separate plant parts. Compared with the vegetative tillers, the reproductive tillers had higher rates of leaf appearance, organic matter growth and CWD decline. Strikingly, for both tiller types, no direct effect of temperature on whole tiller CWD was observed, since temperature effects could be eliminated completely by relating CWD to development stage (DVS) expressed as number of leaves appearing on the main tiller. Temperature effects on CWD were restricted to its influence on tiller development rate only. The decline of CWD of individual plant parts with DVS in the reproductive tillers could be described with a negative exponential curve, which reached an asymptote that was higher for leaf blades (755 g kg−1) than for leaf sheaths (491 g kg−1) and stem internodes (230 g kg−1). However, all plant parts in both tiller types had the same fractional CWD decline rate of 0.395 per leaf appearance interval, independent of plant part insertion level, population or temperature. Differences between temperature treatments in OMD were caused by the higher CWC of plant parts at higher temperature, due to a stronger decline of the specific organic matter mass than of the specific cell wall mass of plant parts at increasing temperature. Differences in whole tiller OMD between populations were observed only for vegetative tillers and were also caused by differences in CWC. It is concluded that temperature increase accelerated both the tiller development rate and the rate of decline of CWD during aging to the same extent, whereas plant parts responded similarly in the fractional CWD decline pattern as a function of DVS. These trends offer unique possibilities for modelling grass digestibility under contrasting temperature regimes.© 2003 Society of Chemical Industry

Genetic diversity in Tunisian perennial ryegrass revealed by ISSR markers

The genetic diversity in Tunisian perennial ryegrass (Lolium perenne) was examined by the help of inter-simple sequence repeats (ISSR). Starting from eighteen accessions, a large number of polymorphic ISSR markers were currently generated using appropriate primers (a total of 136, which average of 12.6 polymorphic bands/primer). These markers were considered to estimate the genetic distance among accessions and to draw phylogenetic trees. Our data provide evidence of a high degree of genetic diversity in Tunisian ryegrass. In addition, both cultivars and wild types present a high degree of divergence suggesting a complex domestication process in this crop. Moreover, spontaneous populations of Tunisian ryegrass have been identified as important ecotypes that are suitable in selection programs to improve grasslands.

CHANGES IN ALLELE FREQUENCIES IN COLCHICINE-TREATED RYEGRASS POPULATIONS ASSESSED WITH RAPD MARKERS

shown that besides chromosomal duplication, ryegrass genotypes treated with colchicine present a significant amount of novel traits, suggesting that colchicine also has point mutagenic effects. Molecular approaches were used to evaluate diversity and relatedness in two annual ryegrass populations treated with colchicine. Colchicine treatment generated differences beyond the levels of inter and intrapopulational variability. Contrary to expectations, populations subjected to higher colchicine doses did not always contain more diversity than untreated populations. The use of Random Amplified Polymorfic DNA (RAPD) markers allowed the comparison of colchicine effects on different genome regions, which strongly support the findings at the morphological levels.

Investigating the mechanism of glyphosate resistance in rigid ryegrass (Lolium ridigum)

Glyphosate is a broad-spectrum herbicide that has been used extensively for more than 20 yr. The first glyphosate-resistant weed biotype appeared in 1996; it involved a rigid ryegrass population from Australia that exhibited an LD50 value approximately 10-fold higher than that of sensitive biotypes. We have characterized gene expression levels and glyphosate sensitivity of 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS), the target enzyme for glyphosate inhibition, in sensitive and resistant lines derived from this population. Restriction fragment length polymorphism analyses were also performed to examine the distribution of EPSPS gene variants and the gene copy number. A two- to threefold increase in basal EPSPS messenger RNA (mRNA) and enzyme activity levels was observed in the most resistant lines analyzed; however, differences among lines in the sensitivity of EPSPS to glyphosate were not apparent. Induction of EPSPS was observed within 48 h after application of 1.5 kg ae ha−1 of glyphosate. This was reflected in elevated levels of both EPSPS mRNA and enzyme activity. Similarly, 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase mRNA levels increased after glyphosate treatment; however, basal and induced transcript levels were comparable for sensitive and resistant lines in this case. The restriction fragment length polymorphism analyses showed no evidence for gene amplification or cosegregation of a specific EPSPS gene variant with glyphosate resistance. EPSPS expression in lines exhibiting an intermediate level of resistance was indistinguishable from that in glyphosate-sensitive lines, suggesting that the mechanism could, at least in part, be non–target-based.

Dry matter yield, herbage quality and persistency of equivalent populations of perennial ryegrass with and without reduced flowering

AbstractReducing the number of flowering tillers that perennial ryegrass produces would improve the feeding value of herbage and could also be used to reduce pollen flow to wild ryegrass populations. Equivalent populations with and without one or more genes that reduce flowering (normal and leafy populations, respectively) were compared over three harvest years (1999‐2001) in a field plot experiment with two cutting managements (silage and simulated grazing). In 1999, samples of dried herbage from each harvest were analysed for leaf lamina content, in vitro digestibility (DMD), crude protein (CP) and water‐soluble carbohydrate (WSC). The leafy population produced one‐fifth to one‐third as many reproductive tillers in May as did the normal population, depending on the management. Mean leaf content of the herbage over all harvests and managements was 63 g/kg higher in the leafy population but its mean DMD was only 7 g/kg higher. In August, September and October, when the leaf contents of the two populations were very similar, the mean DMD of the leafy population was 7 g/kg lower than that of the normal population. Mean annual dry matter yield over both managements of the leafy population was 6‐8% lower than that of the normal population but the leafy population was slightly more persistent.

Molecular genecology of temperature response in Lolium perenne: 2. association of AFLP markers with ecogeography.

Improved winter hardiness is an important breeding objective in the forage grass Lolium perenne. This is a complex trait with several components, including the ability to survive and grow at low temperature, to acclimate to cold, tolerate wind, snow cover and ice encasement. Marker-assisted selection has the potential to increase the efficiency of breeding for improved cold tolerance. Here we describe a genecological approach to identifying molecular markers that are associated with adaptation to low winter temperatures. AFLP was used to assess the genetic diversity in 29 wild populations of ryegrass (Lolium perenne) representing a pan-European temperature cline in terms of their geographical origin. A further 18 populations from a temperature cline in Bulgaria were also analysed. In addition, two varieties and five populations representing parents of mapping families currently in use at IGER were included in the analysis. Principal coordinate (PCoA) and cluster analyses of the molecular marker data showed that the Bulgarian altitude cline populations could be distinguished clearly from the other populations. Two regression analyses were carried out; one to identify AFLP markers that correlated in frequency with low mean January temperature of the geographical origin of the population, and another to identify AFLP markers correlating in frequency with the cold tolerance phenotype of the populations, as determined by LT50 values in freezing tests. In the first analysis six AFLP markers showed significant type II trends with mean January temperature, and in the second analysis 28 bands had a significant univariate relationship with the LT50 value of the accessions. In steps 2 and 3 of the stepwise analysis a further 4 and 5 bands, respectively, improved the fit significantly. The results of the two types of regression analysis are discussed in relation to ecogeography and cold tolerance phenotype of the populations.

Change of Ryegrass Seedling Root Fluorescence Expression during Three Generations of Seed Increase

Seedling root fluorescence (SRF) has been used since the early 1940s to discriminate Italian (annual) ryegrass (Lolium multiflorum Lam.) from perennial ryegrass (L perenne L.). Generally, roots of Italian ryegrass fluoresce under ultraviolet light, while those of perennial ryegrass do not fluoresce, but the trait has readily introgressed from Italian to perennial. Breeders document fluorescence levels of new ryegrass cultivars before they enter seed certification programs. The objective of this study was to ascertain if there was genetic change for the expression of SRF during generations of seed multiplication. Four ryegrass populations, constructed to have low, medium, and high SRF levels, were increased independently for three generations in the field at each of three Oregon locations (Aurora, Corvallis, and Madras). The SRF was measured on the initial population parents, and for each generation cycle at each location. There were significant differences in SRF among locations within populations and for seed production generation within locations. One population initially at 11% SRF increased to 36% across the three generations of seed multiplication at Corvallis, but decreased to 8 and 2% at the other two locations. The other three populations responded differently, showing large population × location and population × generation interactions for SRF expression. Location of increase and the seed production generation must be examined and carefully considered when describing fluorescence levels of cultivars for seed certification purposes. The large amount of variation associated with environmental influences indicates SRF is not a reliable characteristic to describe and predict ryegrass cultivar genetic purity.

Allelopathic and autotoxic interactions in selected populations of Loliumperenne grown in monoculture and mixed culture

Autotoxicity and allelopathy affect the respiration and yield of GL66 and GL72, two populations of perennial ryegrass (Lolium perenne L. cv. S23) that were originally selected for contrasting rates of mature-leaf dark respiration under conditions where allelopathic effects could not occur and autotoxic effects were minimal. The aim of this study was to further investigate growth and biomass allocation of these two populations in relation to their autotoxic and allelopathic properties. To this end, plants were subjected to two conditions (monoculture and mixed culture) and two treatments (growth in 'renewed' and 'replenished' nutrient solution, allowing for short- and long-term accumulation of allelochemicals, respectively). The fast-respiring population, GL66, showed a reduced total yield due to allelopathic effects only when long-term accumulation of allelochemicals was allowed (mixed culture, replenished). However, short-term accumulation (mixed culture, renewed) of allelochemicals was sufficient to affect allocation of biomass to leaf sheaths. The slow-respiring population, GL72, suffered from autotoxicity only when long-term accumulation was allowed (monoculture, replenished), and from allelopathy under both short- and long-term accumulation (mixed culture, either renewed or replenished). The predominant allelopathic and autotoxic effect was on dry matter percentage and dry weight of leaf sheaths. We conclude that the roots of both populations release one or more chemical compounds that primarily affect biomass allocation to leaf sheaths, both of the same and of the other population. Sensitivity to the putative inhibitor(s) released by the other population was greater than sensitivity to the inhibitor(s) released by a population's own roots.

Intraspecific phenotypic variation associated with nuclear DNA content in Lolium perenne L.

The variation in nuclear DNA content and its association with phenotypic traits were examined in 15 cultivated populations of perennial ryegrass (Lolium perenne L.), including herbage and turf populations. Nuclear DNA contents of 15 populations were measured by a flow cytometric method using DAPI (4′-6-diamidino-2-phenylindole) as a fluorochrome. DNA contents were also measured using PI (propidium iodide) for six populations that showed large differences in DAPI values to confirm the difference. There were significant differences in nuclear DNA contents among the populations for both dyes. Of the total variation in nuclear DNA content measured with DAPI, 29% was ascribed to the inter-population variation, 46% was ascribed to the intra-population variation, 24% was ascribed to the random error variation. Herbage populations tended to have larger DNA content than did turf populations. 2C DNA content was positively correlated with cell size, seed size and single leaf size, although significant correlations were mainly due to a small number of populations with large and small DNA contents. These results suggest that intraspecific variation in nuclear DNA content plays important roles in determining phenotypic differences between cultivated populations of L. perenne.

The role of fungal endophyte infection in the evolution of Lolium multiflorum resistance to diclofop‐methyl

Diclofop‐methyl resistance was evaluated in populations of Italian ryegrass (Lolium multiflorum Lam.) infected and uninfected with fungal endophyte (Neotyphodium). Survival was tested in susceptible populations using herbicide screening. The results served as inputs to a model for investigating the role of endophyte infection in the evolution of L. multiflorum resistance to herbicide. The tolerance of infected plants varied depending on the origin of the population and the herbicide dose. Only in some populations and at some diclofop‐methyl doses did plants infected with these endophytes have higher survivorship than endophyte‐free plants. The model demonstrated that endophytes might indeed play an important role in the evolution of herbicide‐resistant weeds, delaying the appearance of herbicide resistance.

Comparison of methods to estimate weed populations and their performance in yield loss description models

Accurate weed population estimation and yield loss prediction are important components of integrated weed management. Field experiments using Italian ryegrass as a weed in broccoli were conducted from 1994 to 1997 to compare weed density to other methods of weed population estimation, to evaluate the performance of weed population estimates in yield description models, and to study the affect of environmental variability on the predictive ability of models. A strong linear relationship was obtained between Italian ryegrass density and direct leaf area (r2 = 0.60 to 0.99). For Italian ryegrass, density and estimates of canopy from the optical device (crosswire device) had a hyperbolic relationship with high coefficients of determination (r2 > 0.72). Both direct leaf area and canopy estimates described broccoli yield as well as or better than Italian ryegrass density. The Li-Cor LAI-2000 Plant Canopy Analyzer (PCA) provided poor estimates of Italian ryegrass population (r2 from 0.00 to 0.63) that failed to describe broccoli yield. No relationship was observed between estimates of light interception through the plant canopy obtained with the Li-Cor LI-191-S Line Quantum Sensor (LQS) and either Italian ryegrass density or broccoli yield. The low performance of the PCA and lack of performance of the LQS were likely due to the smaller size of the plants and larger gaps in the plant canopy caused by wide bed spacing. At similar densities, Italian ryegrass competition with broccoli was stable from year to year. Under high Italian ryegrass density, water supply affected competition. This may limit construction of robust yield prediction models, especially in areas where water is mainly from rainfall.

High Levels of Herbicide Resistance in Rigid Ryegrass (Lolium rigidum) in the Wheat Belt of Western Australia1

A random survey of 264 cropping fields in the Western Australian wheat belt was conducted to determine the extent of rigid ryegrass resistance to commonly used acetolactate synthase- and acetyl-CoA carboxylase-inhibiting herbicides. Rigid ryegrass infestation density was assessed and seed samples collected and subsequently tested for resistance to diclofop-methyl, clethodim, chlorsulfuron, and sulfometuron. Of these randomly collected populations, 46% exhibited resistance to diclofop-methyl and 64% to chlorsulfuron, with 37% exhibiting multiple resistance to both herbicides. Only 28% of tested populations were classified as susceptible to both diclofop-methyl and chlorsulfuron, although all but one population were susceptible to clethodim. Large differences in the proportion of fields containing resistant populations were found between agronomic areas, reflecting different cropping and, therefore, herbicide use history. There was no significant association between resistance status and the density at which rigid ryegrass was present. Herbicide-resistant rigid ryegrass populations are now more common than susceptible populations across much of the Western Australian wheat belt.Nomenclature: Chlorsulfuron; clethodim; diclofop-methyl; sulfometuron; rigid ryegrass, Lolium rigidum Gaud. #3LOLRI.Additional index word: Weed density.Abbreviations: ACCase, acetyl-CoA carboxylase; ALS, acetolactate synthase.

Morphogenetic adaptation to defoliation and soil fertility in perennial ryegrass (Lolium perenne)

Morphogenetic adaptation, plasticity, and mortality of four perennial ryegrass populations, representing different selection histories, were compared in response to soil fertility and to defoliation severity. In a second experiment, morphogenetic adaptation and plasticity in response to defoliation frequency were studied in two divergent sub‐populations. Genetic variability was found between ryegrass populations for leaf appearance and elongation rates, leaf size, and tiller number. All populations exhibited plasticity in response to defoliation severity and soil fertility. There was no genetic variation for plasticity, the morphological response to the environment. Morphogenetic differences between populations remained constant across environments and resulted in differential mortality under the most severe defoliation. Both leaf length and rate of leaf appearance were positively correlated with greater mortality. The ryegrass ideotype for hill country is a compromise between yield and persistence.

AFLP analysis of genetic diversity within and between populations of perennial ryegrass (Lolium perenne L.)

Amplified fragment length polymorphism (AFLP) analysis has been used to measure genetic diversity in perennial ryegrass (Lolium perenne L.) and to relate intra- and interpopulation variation to breeding history. Cluster analysis of AFLP data from contrasting populations showed features consistent with the origins of these varieties. Significant differences in intrapopulation diversity were detected and partial separation of different cultivars was observed. Restricted base cultivars, derived from small numbers of foundation clones, were suitable for this type of study, allowing near complete discrimination of closely related cultivars. Analysis of bulked samples was based on the pooling of genomic DNA from 20 individuals from 6 selected populations. Cluster analysis of AFLP data from bulked samples produced a phenogram showing relationships consistent with the results of individual analysis. AFLP profiling provides an important tool for the detection and quantification of genetic variation in perennial ryegrass.

Leaf and root respiration of Lolium perenne populations selected for contrasting leaf respiration rates are affected by intra- and interpopulation interactions.

Autotoxicity and allelopathy affect the yield of GL66 and GL72, two populations of perennial ryegrass (Lolium perenne L. cv. S23) that were originally selected for contrasting rates of mature leaf dark respiration, but under conditions where allelopathic effects could not occur and autotoxic effects were minimal. To investigate if the respiration properties are also affected, the populations were subjected to two conditions (monoculture and mixed culture) and two treatments (grown in `renewed', i.e. solution completely replaced every week, and `replenished', i.e. nutrients added to the solution every week, to compensate for the nutrients that had been absorbed from the solution). When plants were grown in mixed culture with a `renewed' nutrient solution, the rate of mature leaf respiration in the absence of inhibitors was higher in GL66 than in GL72. In GL66, this rate was unaffected by condition or treatment, whereas in GL72 it increased in the `replenished' cultures, due to autotoxic and allelopathic effects. In contrast with GL66, for GL72 the effect of an inhibitor of the alternative path (salicylhydroxamic acid, SHAM) differed between conditions and treatments. Effects of an inhibitor of the cytochrome path (KCN) were affected by treatment in both populations. The increase in the rate of root respiration of GL66 due to autotoxic and allelopathic effects was accompanied by a higher rate of the SHAM-resistant component, i.e. maximum activity of the cytochrome path. Roots of GL72 showed a higher control respiration rate and a higher rate of the SHAM-resistant component when long-term accumulation of root exudates was allowed to occur in mixed cultures (allelopathy). We conclude that allelochemicals can modify both leaf and root respiration and that there is no consistent correlation between yield and respiration for GL66 and GL72 in the present study. This shows that selection for low rates of mature leaf respiration is not an appropriate method to select for high-yielding cultivars in perennial ryegrass.

Effect of drought on the growth of Lolium perenne genotypes with and without fungal endophytes

Abstract Grass leaves are often inhabited by fungal endophytes that can enhance host growth. In some forage species, endophytes improve host resistance to, and recovery from, drought. Our objective was to determine if the growth of genotypes of Lolium perenne L. was improved by endophytes during recovery from drought. Thirteen infected genotypes were cloned into ramets. Half were treated with a systemic fungicide to eliminate the endophyte (E−); half were untreated and retained high endophyte levels (E+). In a glasshouse, half of all E− and E+ ramets were watered regularly, whilst half were exposed to a 2 week drought on two occasions, each followed by a 3 week recovery period. After the first drought and recovery period, endophytes significantly reduced tiller production in the drought‐stressed group. After the second drought and recovery period, effects of drought on live leaf area and dry mass were highly dependent on host genotype, but not endophytes. The mean tiller mass of E+ ramets after drought was significantly less than that of watered E+ ramets, but this was not true in E− ramets. For six genotypes there was greater mass allocation to storage in the tiller bases of E− ramets after drought. This perennial ryegrass population showed marked genotypic variation in the ability to recover from drought stress, but endophytes played little or no role in this ability. For some host genotypes there may be a metabolic cost of harbouring endophytes during environmentally stressful conditions.

Incidence of ryegrass endophyte (Neotyphodium lolii) and diversity of associated alkaloid concentrations among naturalised populations of perennial ryegrass (Lolium perenne L.)

The mutualistic association between perennial ryegrass (Lolium perenne L.) and the ryegrass endophyte Neotyphodium lolii (Latch, Christensen and Samuels) Glenn, Bacon and Hanlin, previously assigned to the genus Acremonium, is known to have a major impact on pasture and animal production. The perennial ryegrass ecotypes Victorian and Kangaroo Valley are the most widely sown varieties of grass in Australian pasture. The incidence of ryegrass endophyte (N. lolii) in these ecotypes, and the production of alkaloids associated with theryegrass endophyte, are largely unexplored. Fifty-six populations of Victorian and 45 of Kangaroo Valley were sampled from old pasture within the recognised zones of naturalisation of both ecotypes. All of the 101 populations examined were infected with ryegrass endophyte. The mean ryegrass endophyte (N. lolii) frequency for plants within the populations of the Victorian and Kangaroo Valley ecotypes was 88% and 93%, respectively. Alkaloid concentrations were determined from infected plants for 39 Kangaroo Valley populations and, in a separate study, for 24 Victorian populations. Ergovaline concentration was above the critical level of 0.4 mg/kg (above which clinical symptoms are observed) for 10% of the Kangaroo Valley populations and 17% of the Victorian populations. For lolitrem B, all Victorian populations were below the critical level of 1.8 mg/kg, but 28% of the Kangaroo Valley populations had mean concentrations above the critical level, 1.8 mg/kg. Concentrations for individual plants were <5 mg/kg for ergovaline, <6 mg/kg for lolitrem B, and <48 mg/kg for peramine. Within each ecotype examined, significant variation (P < 0.05) in the mean concentrations of ergovaline, lolitrem B, and peramine was found between the populations of naturalised perennial ryegrass. Mean concentrations for some of the various naturalised populations were higher than that observed in some of the control cultivars. Low ergovaline and lolitrem B concentrations observed for individual genotypes indicate scope for selecting grass endophyte combinations characterised by optimum production of alkaloids.