Articles
TOWARDS A LINKAGE MAP IN KIWIFRUIT (ACTINIDIA CHINENSIS PLANCH.) BASED ON MICROSATELLITES AND SATURATED WITH AFLP MARKERS
Article number
498_8
Pages
79 – 84
Language
Abstract
We have isolated and sequenced 263 microsatellites (or SSR, Simple Sequence Repeats) containing clones from two small insert libraries of A. chinensis enriched for (AC/GT)n and (AG/CT)n repeats respectively.
The polymorphism of twenty microsatellites was evaluated in ten genotypes of A. chinensis. All microsatellites tested were polymorphic, showing 9 to 17 alleles per locus. About 20% of the primers generated banding patterns consistent with the amplification of 2 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. We have also assayed the transportability of microsatellites from A. chinensis to other Actinidia species and have found that microsatellite flanking regions are extensively conserved across species, as 75% of primer pairs gave successful amplifications in all 8 species assayed (A. arguta, A. rufa, A. polygama, A. chrysantha, A. callosa, A. hemsleyana, A. eriantha, A. deliciosa). An inter-specific cross between A. chinensis and A. callosa was made in order to produce a linkage map, using the pseudo-testcross mapping strategy.
We describe a preliminary linkage map for A. chinensis based on 61 SSR and saturated with 110 AFLP markers, produced using MseI/EcoRI restriction enzymes and 15 primer combinations.
The polymorphism of twenty microsatellites was evaluated in ten genotypes of A. chinensis. All microsatellites tested were polymorphic, showing 9 to 17 alleles per locus. About 20% of the primers generated banding patterns consistent with the amplification of 2 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. We have also assayed the transportability of microsatellites from A. chinensis to other Actinidia species and have found that microsatellite flanking regions are extensively conserved across species, as 75% of primer pairs gave successful amplifications in all 8 species assayed (A. arguta, A. rufa, A. polygama, A. chrysantha, A. callosa, A. hemsleyana, A. eriantha, A. deliciosa). An inter-specific cross between A. chinensis and A. callosa was made in order to produce a linkage map, using the pseudo-testcross mapping strategy.
We describe a preliminary linkage map for A. chinensis based on 61 SSR and saturated with 110 AFLP markers, produced using MseI/EcoRI restriction enzymes and 15 primer combinations.
Publication
Authors
R. Testolin, W.-G. Huang, G. Cipriani
Keywords
Simple Sequence Repeat, molecular markers, linkage map, fingerprinting
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