Theoretical and Applied Genetics

, Volume 104, Issue 2, pp 223–228

Microsatellite markers isolated in olive (Olea europaea L.) are suitable for individual fingerprinting and reveal polymorphism within ancient cultivars

  • G. Cipriani
  • M. T. Marrazzo
  • R. Marconi
  • A. Cimato
  • R. Testolin

DOI: 10.1007/s001220100685

Cite this article as:
Cipriani, G., Marrazzo, M., Marconi, R. et al. Theor Appl Genet (2002) 104: 223. doi:10.1007/s001220100685

Abstract  

We have isolated and sequenced 52 microsatellites or simple sequence repeats (SSRs) from nearly 60 positive clones obtained from two ’Frantoio’ olive genomic libraries enriched in (AC/GT) and (AG/CT) repeats, respectively. The repeat-containing fragments obtained from genomic DNA restricted with Tsp509I were separated using a biotinylated probe bound to streptavidin-coated paramagnetic beads. Fragments were then cloned into lambda ZAPII vector and sequenced. Thirty of the 36 primer pairs which gave correct re-amplification in the source genome were used to assay the polymorphism of 12 olive cultivars, namely four well-known cultivars (’Coratina’, ’Frantoio’, ’Leccino’, ’Pendolino’) and eight ancient cultivars grown locally near Lake Garda (’Casaliva’, ’Favarol’, ’Fort’, ’Grignan’, ’Less’, ’Raza’, ’Rossanel’, ’Trep’). The local cultivars were each re- presented by two to four long-lived individuals. The analysis was carried out using 33P-labelled primers and 6% polyacrylamide sequencing gels. All except two microsatellites showed polymorphism, the number of alleles varying from 1 to 5. The average genetic diversity (H) was 0.55. The power of discrimination (PD) was 0.60. All cultivars, including the local ones, were easily separated from each other. Variations in the SSR pattern were observed among individual plants of the same cultivar in four out of the eight local cultivars analysed. Several primer pairs (17%) amplified more than one locus.

Keywords SSRSimple sequence repeatMolecular markersDNA polymorphismGenetics

Copyright information

© Springer-Verlag Berlin Heidelberg 2002

Authors and Affiliations

  • G. Cipriani
    • 1
  • M. T. Marrazzo
    • 1
  • R. Marconi
    • 1
  • A. Cimato
    • 2
  • R. Testolin
    • 1
  1. 1.Dipartimento di Produzione vegetale e tecnologie agrarie, University of Udine, Via delle Scienze 208, 33100 Udine, Italy e-mail: testolin@dpvta.uniud.it Fax: +39-0432-558603IT
  2. 2.C.N.R. – Istituto sulla Propagazione delle Specie legnose, Scandicci, Firenze, ItalyIT