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Characterisation of microsatellites from Actinidia chinensis

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Abstract

We have identified a set of informative microsatellite markers for genome analysis in kiwifruit and related Actinidia species. A small-insert genomic library was constructed from Actinidia chinensis DNA, and screened for microsatellites. About 1.2% of the total colonies hybridised to a (GA)8 probe, 0.4% to (GT)8, and 0.1% to a mixture of three different trinucleotide repeat probes, (CAA)5, (GAA)5 and (CTA)5. From the DNA sequences of 35 hybridising clones, 18 primer pairs were designed, and used to amplify genomic DNA from 38 individual plants, representing 30 different accessions of ten Actinidia species. The banding patterns for most of the dinucleotide repeats showed a high degree of polymorphism in the diploid and tetraploid A. chinensis, and in the hexaploid A. deliciosa (kiwifruit). Heterozygosity levels of up to 100% were found among eight diploid accessions of A. chinensis examined, and the number of different-sized bands among all the species varied from 3 to 36 for each microsatellite. One simple CT microsatellite gave 21 bands with sizes suggesting that the number of repeats ranged from 9 to 37. The highest number of bands (36) and the largest size variation (>100 bp) were observed with a complex microsatellite harbouring four different repeat motifs. The majority of primer pairs amplified bands from most of the ten Actinidia species tested. The most polymorphic primer pairs were used successfully to fingerprint a range of closely related varieties of kiwifruit (A. deliciosa).

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Abbreviations

PCR:

polymerase chain reaction

RFLP:

restriction fragment length polymorphism

VNTR:

variable number of tandem repeats

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Authors and Affiliations

  1. Centre for Gene Technology, School of Biological Sciences, University of Auckland, Private Bag, 92019, Auckland, New Zealand

    Kurt Weising, Raymond W. M. Fung, D. Jeannette Keeling & Richard C. Gardner

  2. Plant Molecular Biology Group, Biozentrum, Johann Wolfgang Goethe University, Marie-Curie-Strasse 9, D-60439, Frankfurt, Germany

    Kurt Weising

  3. Horticulture and Food Research Institute of New Zealand Ltd, Private Bag 92169, Mt Albert, Auckland, New Zealand

    Ross G. Atkinson

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  1. Kurt Weising
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  4. Ross G. Atkinson
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  5. Richard C. Gardner
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Weising, K., Fung, R.W.M., Keeling, D.J. et al. Characterisation of microsatellites from Actinidia chinensis . Mol Breeding 2, 117–131 (1996). https://doi.org/10.1007/BF00441427

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