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.     

Breeding two base populations of perennial ryegrass

Summary A program designed to develop two base populations of perennial ryegrass was initiated in 1985 in Galicia. Multitrait mass selection by a base index was practised in two genetic resources groups of wild Galician populations. Plants selected were intercrossed and the two resulting base populations (F and G) were evaluated in two locations at the spaced plant level and in plot trials with the aim of estimating the genetic parameters and the possibilities for future breeding. The two populations did not show significant differences from commercial cultivars. Production characters had low heritabilities (0.01–0.32) in both locations. Seasonal yields had medium to high family heritabilities (0.36–0.73). For the economic traits chosen, the expected genetic advances were important in both populations for summer and autumn growth in Mabegondo and spring and summer-autumn dry matter yield (1^st and 3^rd year) in Puebla de Brollón. Expected genetic advance for early spring growth was 0.62 in population F. In the autumn of 1991 equal numbers of plants, selected at each location by combined family and mass selection from each of these populations, were planted in two polycrosses with the aim of improving the genetic basis and adaptation of these populations.     

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. This revised version was published online in August 2006 with corrections to the Cover Date.     

多花黑麦草繁殖种与原种产量和品质的比较研究

本文采用随机区组试验设计,对多花黑麦草繁殖后种子与引进原种进行了产量和品质的比较研究.结果表明,繁殖种与原种No1和参照种Abundant在生育特性、鲜干草产量、营养品质和种子产量上均无显著差异.因此,多花黑麦草本地繁殖种子仍然保持了较好的产量和营养品质特性.     

Useful variation in in vitro digestibility within perennial ryegrass

In vitro dry matter digestibility (DMD) and dry matter yield (DMY) of herbage from plots sown with three selected perennial ryegrass varieties with similar heading dates (Ba11778, Talbot and Gator) under a range of harvesting frequencies (2, 3, 4 and 5-weekly harvests) were compared over two harvest years (1994 and 1995). Varieties varied in mean percentage DMD over all harvests and harvesting frequencies by 3.4 and 3.5 units in 1994 and 1995 respectively. Varietal ranking in mean DMD over all harvests was the same under each harvesting frequency in both years: Ba11778 being the highest, Gator the lowest and Talbot intermediate. In 1995, Gator had a higher proportion of unsown species in the herbage than both Ba11778 and Talbot but this was only a minor factor contributing to the differences among the varieties in mean DMD over all harvests. Although varieties consistently ranked in the same order of mean DMD over all harvesting frequencies during 13 5-week periods, the range among varieties varied from less than 1 unit to 8.6 units. This range was greatest in mid-season of each year when mean DMD over all varieties was lowest. The minimum mean DMD over all harvesting frequencies during both years was 75.5%, 70.9% and 66.8% for Ba11778, Talbot and Gator respectively. The most digestible variety Ba11778 also had higher annual DMY than the other two varieties in both harvest years and under all four harvesting frequencies. These results indicate that there is sufficient useful genetic variation for DMD within perennial ryegrass to justify breeding for the trait and its routine assessment in official variety trials.     

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.     

Continuous Plant Regeneration from Established Embryogenic Cell Suspension Cultures of Italian Ryegrass and Tall Fescue

The suitability of different protocols was compared for entire plant regeneration by somatic embryogenesis, of the forage plants Lolium multiflorum Lam. (Italian ryegrass) and Festuca arundinacea Schreb. (tall fescue). In the first protocol, miniature embryos were used as starting material, while mature seeds were retained in the other two. Whichever the considered protocol, undifferentiated calli were produced on Murashige and Skoog MS medium supplemented with 2,4-D. The calli were subcultured in the dark on solid MS agar medium, containing 5 mg/1 2,4-D (protocol 2) or on solid MS medium followed by transfer to a rotated liquid MS medium with 2 mg/1 2,4-D (protocol 1). In these conditions, induction of somatic embryogenesis occurred, and whole plants were regenerated during a limited lapse of time, upon transfer in the light, to MS medium supplemented with BAP but devoid of 2,4-D. The simultaneous elimination of 2,4-D and transfer to light appeared essential for full regeneration of the plants. Using this characteristic, an additional step was added to a new protocol (protocol 3) in which microcalli, cultured on liquid MS medium containing 5 mg/1 2,4-D, were transferred to the same medium with 2 mg/1 2,4-D, in the dark. In these conditions, the suspensions kept their embryogenic potential for months. In all cases, plantlets were successfully transferred into the soil. An evaluation of the somaclonal variation potential of the plants issued from each protocol is now underway.     

Disturbed Mendelian segregations at isozyme marker loci in early backcrosses of Lolium multiflorum × Festuca pratensis hybrids to L. multiflorum

Summary The segregation of interspecific recombinant Festuca pratensis (Fp) alleles, introgressed into the germolasm of Lolium multiflorum (Lm), at four loci (PGI/2, AcP/2, GOT/3 and BAP) is described. Heterozygous (Lm/Fp) plants were backcrossed to L. multiflorum (2n=2×=14) and subsequent BC2 Lm/Fp sibling genotypes intercrossed.In crosses between BC1 heterozygous plants (Lm/Fp) used as males and L. multiflorum, there was a reduced transmission of the F. pratensis (Donor Parent) derived alleles in the populations with PGI/2 and AcP/2 marker loci compared to the reciprocal cross but the reduction was not significantly different in those with GOT/3 and BAP markers.Two PGI/2 marked BC2 half-sib families in particular exhibited a more extreme deficiency of Fp/Fp progeny plants than anticipated from the BC2 segregations indicating possible linkage to zygotic lethals. Deficiencies of F. pratensis alleles were, in most cases, less marked in BC2 half-sib families indicating that a further round of recombination had reduced the size of the introgressed chromosome segment or that deleterious linkages had been broken. A tendency towards heterozygote advantage was found in one BAP marked halfsib family.The significance for forage grass breeding of reduced transmission rates of Donor Parent alleles in early back-cross generations especially through the male gametes is discussed.     

Anther Culture Response in Perennial Ryegrass (Lolium perenne L.)

20 diploid clones from 7 varieties, and 10 tetraploid clones from 3 varieties of Lolium perenne were tested in replicated anther culture experiments. Embryos or calluses, were obtained from all clones, and plants were regenerated from all clones except one. The total yield of plants (albino and green plants) ranged from 0 to 61 plants per 100 cultured anthers among genotypes, and there was a general tendency for tetraploic genotypes to be more responsive. Viable green plants were obtained from 5 diploid and 7 tetraploid clones representing 2 and 3 varieties, respectively. Their origin from reduced pollen was confirmed by a haploid chromosome number in some regenerants and by homozygosity for isozyme loci in spontaneously chromosome doubled plants produced from heterozygous diploid donor plants. A large number of the plants were successfully established in the soil. For most donor genotypes, green plants were rare exceptions, but two diploic clones consistently produced 2.3 and 3.8 green plants per 100 cultured anthers, respectively. Estimates of variance components from replicates with greenhouse and field-grown donor plants showed that genotypes accounted for about 73 per cent of the total variation in yield of embryos/calluses, while only 14—15 per cent of the total variation was due to replicates. Hence at present, emphasis should be placed on die selection of high-response genotypes in material of high agronomic potential.     

Factorial regression analysis of genotype by environment interaction in ryegrass populations, using both isozyme and climatic data as covariates

A sample of 58 natural ryegrass populations (Lolium perenne L.) from the French and Spanish oceanic coast was evaluated for three seasonal growth traits (i.e. spring, autumn and winter vigour traits) in 5 evaluation sites (three in France and two in Spain). This sample showed a high level of genotype (population) × environment (evaluation sites) interactions for the three agronomic traits. A factor regression analysis, using both isozyme frequencies of populations and climatic factors of evaluation sites as covariates, was carried out on a sub-sample of 30 populations in order to explain these interactions. This method succeeded in explaining most population × location interactions by the product of two covariates. For instance, for spring vigour trait, 72.8% of the interaction term could be explained by the use of two covariates: the PGI2-20 allelic frequency of populations and the minimum temperature of the coldest month of evaluation sites. This study shows the advantage of such a method for plant breeders who need to identify promising ryegrass populations for their breeding objectives. A number of genetic and evolutionary hypotheses are also discussed. This revised version was published online in July 2006 with corrections to the Cover Date.