• Characteristaion of the initial generations of recombinant inbred lines in perennial ryegrass (lolium perenne L.) using molecular markers and cytogenetics

      Anhalt, Ulrike C. M.; Teagasc Walsh Fellowship Programme; National Development Plan (2009)
      In this study three early generations of a recombinant inbred line (RIL) breeding programme have been characterised by cytogenetic methods, metabolite profiling and a biomass quantitative trait loci (QTL) study. In situ hybridization was used to analyse recombination and the structure of the parental and the F1 genomes. A metabolite profiling study via GC/MS was conducted to characterise the metabolite activity of the parental inbred lines and the F1 hybrid. Total metabolites were compared across a growth season in a replicated field design. A genetic map was constructed with 75 nuclear DNA markers in the F2 generation, which consisted of 360 individuals. 63% of the markers in the F2 population did not fit the expected Mendelian ratios and showed significant (p<0.05) segregation distortion. Fresh weight, dry weight, and dry matter were analysed with an alpha lattice design throughout the greenhouse and field experiments. Additionally, leaf width was recorded in the greenhouse experiment. Heritablities ranged for the biomass related traits between 78 and 95%. This study provides an insight into the recombination of the parental and the F1 genotypes. Different levels of metabolite activities could be found among the two parental inbred lines across three harvest dates. The QTL study indicates the position of biomass QTL related traits. Major QTL with log of odds scores >3 were identified on linkage groups 2, 3, 4, 5 and 7. About 30% of overall variation could be explained. Increased biomass yield is still one of the most important traits in any Lolium perenne breeding programme. The present QTL study can be used for fine mapping of biomass yield related traits in L. perenne. In the long term biomass yield can be eventually monitored and predicted with marker assisted selection for some of the QTL identified in the present study.
    • An Immortalized Genetic Mapping Population for Perennial Ryegrass: A Resource for Phenotyping and Complex Trait Mapping

      Velmurugan, Janaki; Milbourne, Dan; Connolly, Vincent; Heslop-Harrison, J. S.; Anhalt, Ulrike C. M.; Lynch, M. B.; Barth, Susanne; Teagasc Walsh Fellowship Programme (Frontiers, 2018-05-31)
      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.
    • An Immortalized Genetic Mapping Population for Perennial Ryegrass: A Resource for Phenotyping and Complex Trait Mapping

      Velmurugan, Janaki; Milbourne, Dan; Connolly, Vincent; Heslop-Harrison, J. S.; Anhalt, Ulrike C. M.; Lynch, M. B.; Barth, Susanne; Teagasc Walsh Fellowship Programme; Teagasc (Frontiers, 2018-05-31)
      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.