Morphological variation in Dutch perennial ryegrass (Lolium perenne L.) populations,in relation to environmental factors

Summary Twenty-one Dutch Lolium perenne populations, fifteen European populations and six L. perenne cultivars were compared for morphological variation. Dutch populations clearly differed from the European populations and the cultivars. Dutch populations generally had reduced plant length and smaller leaves. For other characters, e.g. date of ear emergence, the Dutch populations showed as much variation as the European populations and cultivars they were compared with in this trial. Correlations between morphology and environmental factors at the site of origin were significant in several cases but were generally weak, and dependent on the set of populations studied. In order to maximize the phenotypic differences between the collected populations, soil type and management type appeared to be the most important factors for the choice of collection sites in the Netherlands. To determine suitable collection sites all over Europe, other factors like precipitation, latitude, altitude and temperature factors were also important. For subsampling of the locations, variation in management type within the location determined whether or not phenotypic different samples could be collected. The extensive use of cultivars in the Netherlands does not seem to have prevented the formation of distinct populations. Therefore in situ conservation of grassland seems a good alternative for genetic conservation of L. perenne in the Netherlands.Also associated with the WAU Department of Plant Taxonomy, P.O. Box 8010, 6700 ED, Wageningen, The Netherlands     

Co-adaptation between neighbours? A case study with Lolium perenne genotypes

Summary Co-adaptation between neighbouring genotypes of Lolium perenne was examined in a pot experiment over an 18 month period. Competitive interactions in pure and mixed stands were examined using 9 genotype pairs (established as neighbours from electrophoresis data), 7 from a 10 year-old, and 2 from a 40+ year-old pasture. Combinations of each pair of neighbours grown together, and each grown with 5 randomly chosen alien (non-neighbour) genotypes sampled from the same pastures were also assessed. Seven harvests of green matter produced above 20 mm were made, and 50–135 kg N ha^-1 was administered after each harvest. Data for dry weight at each harvest for pure and mixed stands, and Relative Yield Totals (RYT) were obtained for neighbours and aliens in each treatment.In 6 of the 9 pairs of neighbours grown as pure stands, the yields of neighbours did not differ significantly at the final harvest. In 8 of the 9 pairs grown as mixtures of pairs of neighbours, the yield of one of the neighbours was significantly reduced and one neighbour had been eliminated. Over the last 5 harvests, yields from (alien + neighbour) mixtures had the greatest dry matter yields. No suppression of one component by the other was recorded for this (alien + neighbour) combination.Based upon RYT estimates, neighbours did not yield more when grown together than when grown separately with aliens. There was no relationship between original genotype size in their fields of origin and dry matter yield in this experiment.This experiment provides no evidence for he evolution of co-adaptation between surviving neighbours in the two pastures from which these individuals were sampled. The genotypic architecture of pasture would seem to be the product of random and/or competitive exclusion of individual genotypes, rather than the evolution of co-adapted groups of individuals through time. The consequences of extreme depletion of genetic variation in the first two years after establishment, the very low rates of seedling establishment in established pastures, and the impact of the grazing animal would suggest that there is little likelihood of or potential for, co-adaptation evolving between neighbours.     

Hybrid production in Lolium perenne based on incompatibility

Summary SI-hybrids were produced by crossing 30 partially inbred lines in 3 sets of 5×5 factorial crosses. Annual dry matter yields from 3 successive years and 2 locations were on average 9.5 T/ha for the hybrids compared to 8.9 T/ha for their parental lines, with the best hybrid yielding 10.3 T/ha. Heterosis was up to 20%. Problems and prospects of SI-hybrid production in grasses are discussed.This paper is dedicated to Prof. Dr. F.W. Schnell on the occasion of his 80th birthday.     

Testing perennial ryegrass (Lolium perenne L.) as spaced plants in swards

Summary A method of testing spaced plants in swards of another grass crop is described. The procedure provides information on sward performance at each stage of the breeding scheme. A large number of entries may be included, together with whatever cutting frequency.When used for perennial ryegrass, the method favours the selection of productive and persistent plants.     

Genetic variation in leaf epidermal cell size and shape in Lolium perenne

Summary Six diploid perennial ryegrass populations, two diploid F₁ hybrid families and the autotetraploid cv. Tove were grown in a glasshouse and the mean length and mean width of epidermal cells on the abaxial leaf surface of young fully expanded leaves compared. The diploids varied in cell length by 26%, in cell width by 9% and in length: width ratio by 39%. The cells of cv. Tove were 25% longer and 12% wider than those of the highest ranking diploid and the length: width ratio 9% higher. The DNA contents per telophase root-tip cell nucleus of a large-celled and a small-celled diploid population were similar, suggesting that the variation in epidermal cell size among diploids was due to the action of specific genes and not to differences in genome size. The mean epidermal cell dimensions of four of the above populations growing as plots in a field trial employing three levels of fertiliser nitrogen and four harvesting frequencies were measured on three occasions. Varietal ranking in cell dimensions was consistent over the different managements and dates, and agreed well with the glasshouse results. The results indicate that measurements of leaf epidermal cell length and width can be useful aids both in breeding for herbage yield and in distinguishing varieties with similar flowering dates.     

Introduction of a self-compatible gene of Lolium temulentum L. to perennial ryegrass (Lolium perenne L.) for the purpose of the production of inbred lines of perennial ryegrass

Interspecific hybrids between self-compatible species, Lolium temulentum L. and self-incompatible species, L. perenne L. were obtained using embryo rescue. Two cycles of backcrossing of interspecific hybrids with L. perenne were carried out. A 1: 1 segregation ratio of self comatibility and incompatibility was observed in backcross generations. These segregation data confirmed that self-compatibility of L. temulentum was controlled by a single gene. It suggested that the self-compatible gene of L. temulentum could be introduced to self-incompatible plants of L. perenne through interspecific hybridization. It appears that utilization of a self-compatible gene of L. temulentum would be useful for production of inbred lines of L. perenne. The possibility of breeding procedures of perennial ryegrass using a self-compatible gene was discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Mitochondrial DNA analysis reveals cytoplasmic variation within a single cultivar of perennial ryegrass (Lolium perenne L.)

Summary The extent of mitochondrial DNA restriction fragment length polymorphisms among individual plant samples of perennial ryegrass was determined. A total of 72 plants from three cultivars Yorktown II, S23 and Riikka were surveyed using three restriction enzymes (BamHI,EcoRI andHindIII) and three mitochondrial gene probes (coxI, coxIII andnad9). Polymorphisms were noted within each of the two cultivars Yorktown II and S23, whereas in Riikka no variation was detected. It seems most likely that the mitochondrial genome diversity within the same cultivar has resulted from non-homogeneous ancestor cytoplasms. The hybridization-based assay employed is simple to perform, gives unambiguous results, and may thus be used in mass screening of perennial ryegrass populations for breeding purpose.     

Components of resistance to ryegrass mosaic virus in a clone of Lolium perenne and their strain-specificity

Summary The extreme resistance to ryegrass mosaic virus (RMV) of a clone of Lolium perenne L. was due to a combination of two distinct types of resistance: resistance to infection and resistance to multiplication and movement of virus within the plant. Resistance to infection was quantitatively inherited and highly effective against three strains of RMV, while resistance to multiplication and movement was controlled by two complementary recessive genes and was effective against only two of the three RMV strains.     

Genetic improvement of yield of nitrogen of Lolium perenne pastures

Summary Four varieties of Lolium perenne with similar flowering dates bred over a 30 year period were grown in an orthogonal plot trial with two contrasting harvesting frequencies and three levels of fertiliser (0.2, 0.4 and 0.6 t/ha of N annually). In the second harvest year, the nitrogen (N) content of herbage at each harvest was determined and the yields of N calculated. The newest variety (Ba 10761) gave a 20% higher mean annual yield of N than the oldest (S.321). The influence of variety in increasing the ratio of N recovered: N applied was as great or greater than the influence of harvesting frequency or level of applied N. There were small differences between varieties in N content (%N), N content being inversely related to N yield.     

Aneuploids of perennial ryegrass (Lolium perenne)

Summary Aneuploid plants of perennial ryegrass (Lolium perenne L.) with 2n=15 to 30 chromosomes were obtained by crossing a near-triploid (2n=3x+1=22) with a diploid or on open-pollination with diploids and tetraploids. Aneuploids occurred with a frequency of 83% in near triploid × diploid progeny and 92% on open-pollination with diploid and tetraploid plants. Aneuploid plants with 15 to 18 chromosomes resembled diploids in morphology and those with 19 to 30 chromosomes were akin to tetraploids. Meiotic studies suggested that most aneuploid plants resulted from transmission of aneuploid egg cells (n=8 to 23). Aneuploid plants with 2n=27 to 30 chromosomes in the progeny of 22×14 cross originated from unreduced egg cells. Plants with 19 to 21 chromosomes were recovered only by immature seed culture. Aneuploid plants with 26 to 30 chromosomes and triploids (2n=21) had higher pollen fertility and bigger seeds than plants with 15 to 22 chromosomes.     

Colchicine-induced heritable variation in cell size and chloroplast number in the leaf cells of inbred ryegrasses (Lolium perenne,L. multiflorum)

Summary A 3 hr period of treatment with a 0.2% aqueous solution of colchicine was given to one week old seedlings of 3 inbred lines of perennial ryegrass (Lolium perenne) and 4 lines of Italian ryegrass (L. multiflorum). The surviving plants developed as mixoploids and were self-pollinated. In the following generation (CT1) the seeds were grown without further treatment, and were screened cytologically at the seedling stage to separate out diploids from tetraploids. Only the diploids were kept, and in this CT1 generation comparisons were then made between controls (2x) and colchicine-treated diploids (C2x) for leaf mesophyll cell size and chloroplast number variation. Heritable differences were found such that the cell plan areas and/or the numbers of chloroplasts per cell were significantly greater in the C2x compared with the 2x treatment in 3 and 5 out of the 7 lines. In each case there was enhancement of chloroplast numbers independently of the variation in cell size.     

Effect of date of sampling and plant density upon size and digestibility of reproductive tillers of Lolium perenne L.

Summary Evaluation of forage germplasm digestibility is an important factor in screening programmes. The major problems involve the relevance of estimations made in spaced plants to plot performance, the effect of maturity, leaf: stem ratio and the choice of plant organ to be analysed. The use of stems of reproductive tiller proved successful in Lolium multiflorum. In the present experiment the effect of dates of sampling (date 1=28/6; date 2=5/7; date 3=12/7/83) and of plant densities (spaced plants and plots) on the digestibility of reproductive tillers of L. perenne cv. Marta 23 days after ear emergence was evaluated. After harvesting, the following characters were recorded on each tiller: tiller length and diameter, and spike and flag leaf length. Leaf blades and sheaths were then removed, the tillers dried and milled individually, and the in vitro digestibility assessed. The data were analysed by Analysis of Variance, Principal Components Analysis and Numerical Classification followed by a study of how the variables contributed to the derived group structure. Results: I-Plots: digestibility values were 63.4, 60.3 and 56.6% (P<0.01), respectively, for the sampling dates 1, 2 and 3. The Cramer Value (0.41) shows some correlation between tiller size and digestibility. Tillers from date 3 tend to be smaller than the others but the separation is not very clear. II-Spaced plants: digestibility values were 63.2, 60.0 and 60.8%, respectively, for dates 1, 2 and 3. Analysis of variance showed differences (P<0.01) between dates and also between genotypes. The Cramer Value (0.23) shows less correlation between tiller size and digestibility: however, tillers from dates 1 and 3 are clearly separated on the basis of size. Tillers from date 3 being smaller, date 2 tillers are located in between those from dates 1 and 3. III-Plots and single plants combined: Tillers from plots are larger than tillers from spaced plants in sampling dates 2 and 3. The Cramer Value for digestibility (0.14) is the lowest one. The results indicate differences in size and digestibility of reproductive tillers of L. perenne when contrasting situations are being compared, which suggest influence of environmental conditions upon the characters evaluated in the experiment. The differences are sufficiently high so that the use of this technique, when populations of L. perenne with wide variation in flowering dates are being evaluated, is not recommended.     

Potential for improving adaptation ofLolium perenne L. to continental climates in Norway

Summary Breeding of perennial ryegrass has been conducted in Norway for more than 30 years. The little progress achieved so far can, most probably, be explained by a restricted genetic variation within our indigeneous plant material. In order to increase the variation in the Norwegian ryegrass germplasm, we have tested populations of diverse origin and adaptations under contrasting climatic conditions in Norway. Data is presented for winter survival and dry matter yield obtained in two experiments, one in a dense stand with 20 populations of Norwegian and Russian origin, and one as a spaced plant experiment with 26 populations of Norwegian, Russian and Swiss origin. In both cases commercial foreign cultivars and breeding populations were included.The results show that the commercial varieties were superior when grown in dense stand. The Norwegian material showed, however, a significant better adaptation at the continental location, measured as plant cover after three years. There was considerable variation between populations in all characters. In the spaced plant experiment, the Norwegian diploid breeding populations were the highest yielding. The commercial cultivars also performed well. Winter survival was generally good in this experiment, and only small differences between populations could be detected. Winterhardy and productive populations of different origin and contrasting adaptations have been selected, and breeding populations constructed. Surprisingly enough, Swiss Alp populations, presumably adapted to long lasting snow-cover, do not show any better adaptation to the continental climates in Norway than indigeneous ryegrass populations.     

Assessment of perennial ryegrass (Lolium perenne L.) for breeding. II. Components of winter hardiness

Summary When breeding for good winter hardiness a broad range of adaptations must be considered. Such a multitrait approach requires some understanding of physiological and genetical relationships between relevant traits.Fourteen winter related characters were measured on field grown plants from 86 perennial ryegrass accessions originating from a wide range of climates. Genetic components of variance and covariance were extracted from the data and a principal components analysis carried out. The first three independent components accounted for 80% of the variance represented by the genetic correlations and were broadly identified as winter growth, winter hardiness and early spring activity factors. None of the accessions exhibited a combination of extreme winter growth and winter hardiness although several with excellent winter hardiness showed better than average winter growth. Amounts of water soluble carbohydrate maintained through the winter appeared important for good survival.It was concluded that a selection index approach may prove useful in future breeding programmes as further information concerning physiological relationships becomes available.     

Seed yield related to crop development and to yield components in nine cultivars of perennial ryegrass (Lolium perenne L.)

Summary We had previously found differences for seed yield among nine perennial ryegrass cultivars which were not associated with variation for seed weight. To detect the physiological basis of these genetic differences for seed yield, growth analyses were carried out. We related crop development and components of seed yield to seed yield during three years on clay and sandy soil. No significant differences occurred among cultivars for accumulation and partitioning of dry matter or the pattern of tiller production. Seed yield of the cultivars was not associated with ear number or total dry matter yield of the seed crop. Seed yield was more correlated with the number of seeds per unit area than with seed weight. The number of seeds as calculated after harvest from seed yield and seed weight was much lower than the number of seeds as estimated prior to harvest from seed yield components. The number of spikelets differed significantly among the cultivars, but the ranking was different from that for seed yield. The physiological basis of the genetic differences for seed yield is not clear. Implications for breeding perennial ryegrass are discussed.     

Assessment of perennial ryegrass (Lolium perenne L.) for breeding. I Freezing tolerance

Summary Assessments of freezing tolerance, estimated in terms of temperature at which 50% mortality occurs (=LT₅₀), were obtained for 86 Lolium perenne accessions using a glycol bath test on naturally hardened field plants. Significant correlations were obtained between LT₅₀ and both heading dates of accessions and minimum temperature of the coldest month near their sites of origin. The effects of temperature and reproductive development on the hardening/dehardening of plants and their growth after freezing are discussed.     

Association of agronomic traits with isozyme loci in perennial ryegrass (Lolium perenne L.)

Summary Inbred lines possessing different genotypes at three isozyme loci, PGI-2, GOT-3 and ACP-1, were produced by two generations of selfing from two contrasting cultivars of perennial ryegrass. Four separate F₂ populations were obtained by selfing or crossing F₁ plants from crosses between cultivar inbred lines. F₂ plants were scored for genotype at the three isozyme loci and for eight agronomic traits. The three isozyme loci showed independent segregation confirming that they belonged to separate linkage groups. A consistent association was observed between water soluble carbohydrate content and genotype at the PGI-2 locus. Some association was also detected between genotype at the ACP-1 locus and heading date. The relevance of these observations to ryegrass breeding programmes is discussed.     

Selection for yield in Lolium perenne. II. performance of spaced plant selections under competitive conditions

Summary The performance of six lines of perennial ryegrass (Lolium perenne L.) which had been selected for three generations for production under spaced plant conditions were assessed under increasing levels of competition, namely spaced plants, drills, miniplots and large sown plots. Successful selection for yield under spaced plant conditions was not accompanied by comparable production patterns in the alternative managements. The extreme situation occurred in swards where a significant decline in production was encountered. Negative or no relationships were found in the establishment year between spaced plant production and yield in the alternative conditions whereas in the first harvest year positive correlations occurred between spaced plants, drills and miniplots and only the latter with plots. These varying patterns of performance are considered in relation to the prediction of sward behaviour and the appropriate environment for selection.INIA. CRIDA 01, La Coruna, Spain.     

Spaced-plant traits related to seed yield in plots of perennial ryegrass (Lolium perenne L.)

Summary Seed yield in perennial ryegrass is low and selection criteria for high seed production are lacking. Indirect selection in spaced plants would be efficient, but spaced-plant traits need to be identified that correlate with seed yield in drilled plots. Spaced plants were studied of nine perennial ryegrass cultivars with a different seed production when grown in plots. Genotypic variation within cultivars for seed yield components and other traits was assessed in four environments in replicated clonal experiments. Extensive genotypic variation for most traits was present within each cultivar. Based on cultivar means of 25 or 50 genotypes, significant differences among cultivars were found for most traits. Cultivar-year interactions were found for most traits, but no significant cultivar-location interactions were generally found. Spaced-plant traits in general showed poor correlation to corresponding traits in drilled plots. Cultivars with contrasting seed yield in plots could not be distinguished on the basis of their spaced-plant traits and therefore no criteria for indirect selection for seed production in drilled plots could be identified in spaced plants.     

Variation in digestibility in nine populations of perennial ryegrass (Lolium perenne) and its genetic basis

Summary Thirty five individuals from each of nine perennial ryegrass populations were sampled from set-stocked and zero-grazed swards and from an old permanent pasture. The swards were 6, 18, 30 months and 11 years old when sampled. The permanent pasture was at least 40 years old.Dry matter digestibility (DMD) and organic matter digestibility (OMD) were determined for green leaf material harvested 4 days after ear emergence. Significant differences were found between the populations, but genotypic differences were found only for OMD. The largest proportion of the total variation was due to differences between blocks. In the zero-grazed populations, DMD and OMD declined significantly with increasing sward age. This was not found for the set-stocked swards. Two sets of pair crosses, from which mid-parent, progeny regressions could be determined, were carried out.The regressions of offspring on mid-parent for set-stocked and zero-grazed swards were used to assess the presence of additive genetic variation and to estimate narrow sense heritability of DMD. The coefficients of correlation, 0.122 and 0.226 respectively, were non-significant, indicating an absence of additive genetic variation.