Development and characterisation of simple sequence repeat (SSR) markers for white clover (Trifolium repens L.) |
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Authors: | R Kölliker E S Jones M C Drayton M P Dupal J W Forster |
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Institution: | Plant Biotechnology Centre, Agriculture Victoria, La Trobe University, Bundoora, Victoria 3083, Australia e-mail: john.forster@nre.vic.gov.au Tel.: +61-3-9479 2995, Fax: +61-3-9479 3618, Present address: R. K?lliker, Swiss Federal Research Station for Agroecology and Agriculture, Reckenholzstrasse 191, 8046 Zurich, Switzerland, CH
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Abstract: | Highly informative molecular markers, such as simple sequence repeats (SSRs), can greatly accelerate breeding programs. The
aim of this study was to develop and characterise a comprehensive set of SSR markers for white clover (Trifolium repens L.), which can be used to tag genes and quantitative trait loci controlling traits of agronomic interest. Sequence analysis
of 1123 clones from genomic libraries enriched for (CA)
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repeats yielded 793 clones containing SSR loci. The majority of SSRs consisted of perfect dinucleotide repeats, only 7% being
trinucleotide repeats. After exclusion of redundant sequences and SSR loci with less than 25 bp of flanking sequence, 397
potentially useful SSRs remained. Primer pairs were designed for 117 SSR loci and PCR products in the expected size range
were amplified from 101 loci. These markers were highly polymorphic, 88% detecting polymorphism across seven white clover
genotypes with an average allele number of 4.8. Four primer pairs were tested in an F2 population revealing Mendelian segregation. Successful cross-species amplification was achieved in at least one out of eight
legume species for 46 of 54 primer pairs. The rate of successful amplification was significantly higher for Trifolium species when compared to species of other genera. The markers developed in this study not only provide valuable tools for
molecular breeding of white clover but may also have applications in related taxa.
Received: 3 April 2000 / Accepted: 12 May 2000 |
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Keywords: | Simple sequence repeats SSR-enriched libraries Molecular marker Trifolium repens Cross-species amplification |
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