Microsatellite DNA in Actinidia chinensis: isolation, characterisation, and homology in related species |
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Authors: | W-G Huang G Cipriani M Morgante R Testolin |
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Institution: | (1) Dipartimento di Produzione Vegetale e Tecnologie Agrarie, University of Udine Via delle Scienze 208, 33100 Udine, Italy Tel.: +39 432 558632 Fax: +39 432 558603 E-mail: raffaele.testolin@dpvta.uniud.it, IT |
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Abstract: | We have isolated and sequenced 263 microsatellite-containing clones from two small insert libraries of Actinidia chinensis enriched for (AC/GT) and (AG/CT) repeats, respectively. Primer pairs were designed for 203 microsatellite loci and successfully
amplified from both plasmid and A. chinensis genomic DNA. In this paper we report the sequences of 40 primer pairs for which we have demonstrated Mendelian segregation
in the progeny from controlled crosses. The polymorphism of ten microsatellites of each type was evaluated in four diploid
and six tetraploid genotypes of A. chinensis. All microsatellites proved to be polymorphic, the number of alleles per locus detected in polyacrylamide sequencing gels
ranging from 9 to 17. The high degree of polymorphism in Actinidia renders these markers useful either for mapping in A. chinensis or for fingerprinting cultivars of both domesticated kiwifruit species (A. chinensis and A. deliciosa). While most primer pairs produced single amplification products, about 20% generated banding patterns consistent with the
amplification of two different loci. This supports the hypothesis that diploid species of Actinidia (2n=2x=58) are polyploid in origin with a basic chromosome number x=14/15 and that chromosome duplication may have occurred during the evolution of the genus. Finally, we have assayed the cross-species
transportability of primer pairs designed from A. chinensis sequences and have found extensive cross-species amplification within the genus Actinidia; 75% of primer pairs gave successful amplification in the eight species assayed (A. arguta, A. rufa, A. polygama, A. chrysantha, A. callosa, A. hemsleyana, A. eriantha, and A. deliciosa), which are representative of the four sections into which the genus is currently split.
Received: 14 February 1998 / Accepted: 26 May 1998 |
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Keywords: | Simple sequence repeat (SSR) Microsatellites Molecular markers Genetics Kiwifruit |
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