Development of sex-linked PCR markers for gender identification in Actinidia |
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Authors: | G P Gill C F Harvey R C Gardner L G Fraser |
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Institution: | (1) The Horticulture and Food Research Institute of New Zealand Ltd., Mt Albert Research Centre, Private Bag 92169, Auckland, New Zealand, NZ;(2) Plant Molecular Biology, School of Biological Sciences, University of Auckland, Auckland, New Zealand, NZ |
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Abstract: | Two sex-linked random amplified polymorphic DNA (RAPD) markers identified from Actinidia chinensis were converted into sequence-characterised amplified regions (SCARs) for the large-scale screening of Actinidia breeding populations. Initial SCAR primers converted one RAPD (SmX) into a dominant marker, but the other (SmY), which was
potentially more useful because of its linkage to the male determining ‘Y’ locus, failed to retain polymorphism. This difficulty
was overcome by cloning and sequencing the alternate ‘allele’ from female plants, and then designing ‘allele’-specific primers
that utilised nucleotide differences between the sexes. Using a quick squash-blot method of DNA extraction, the SCAR primers
were tested in 120 A. chinensis plants to determine their gender. The system is now in use for large-scale screening of seedling populations in the Actinidia breeding programme. The sex-linked SCAR primers also functioned with plants from some other geographically separate accessions
of A. chinensis and with plants in the closely related polyploid species A. deliciosa, but did not amplify a sex-linked band in more distantly related species of Actinidia.
Received: 27 December 1997 / Accepted: 5 March 1998 |
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Keywords: | Actinidia RAPD SCAR Sex-linked markers Marker-assisted selection |
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