Theoretical and Applied Genetics

, Volume 102, Issue 2, pp 405–415

Development and characterisation of simple sequence repeat (SSR) markers for perennial ryegrass (Lolium perenne L.)

  • E. S. Jones
  • M. P. Dupal
  • R. Kölliker
  • M. C. Drayton
  • J. W. Forster
Original Article

DOI: 10.1007/s001220051661

Cite this article as:
Jones, E., Dupal, M., Kölliker, R. et al. Theor Appl Genet (2001) 102: 405. doi:10.1007/s001220051661

Abstract 

Enrichment methods were optimised in order to isolate large numbers of simple sequence repeat (SSR) markers for perennial ryegrass (Lolium perenne L.), with the aim of developing a comprehensive set of loci for trait mapping and cultivar identification. Two libraries were constructed showing greater than 50% enrichment for a variety of SSR-motif types. Sequence characterisation of 1853 clones identified 859 SSR-containing clones, of which 718 were unique. Truncation of flanking sequences limited potential primer design to 366 clones. One-hundred selected SSR primer pairs were evaluated for amplification and genetic polymorphism across a panel of diverse genotypes. The efficiency of amplification was 81%. A relatively high level of SSR polymorphism was detected (67%), with a range of 2–7 alleles per locus. Mendelian segregation of alleles detected by selected SSR-locus primer pairs was demonstrated in the F1 progeny of a pair cross. Cross-species amplification was detected in a number of related pasture and turfgrass species, with high levels of transfer to other Lolium species and members of the related genus Festuca. The identity of putative SSR ortholoci in these related species was confirmed by DNA sequence analysis. These loci constitute a valuable resource of ideal markers for the molecular breeding of ryegrasses and fescues.

Keywords Simple sequence repeats SSR-enriched libraries Molecular marker Lolium perenne Cross-species amplification 

Copyright information

© Springer-Verlag Berlin Heidelberg 2001

Authors and Affiliations

  • E. S. Jones
    • 1
  • M. P. Dupal
    • 1
  • R. Kölliker
    • 1
  • M. C. Drayton
    • 1
  • J. W. Forster
    • 1
  1. 1.Plant Biotechnology Centre, Agriculture Victoria, La Trobe University, Bundoora, Victoria 3083, and CRC for Molecular Plant Breeding, Australia e-mail: john.forster@nre.vic.gov.au Tel.: +61 3 9479 2995, Fax: +61 3 9479 3618, Present address: R. Kölliker, Swiss Federal Research Station for Agroecology and Agriculture, Reckenholzstrasse 191, 8046 Zürich, SwitzerlandCH

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