Identification of closely related citrus cultivars with inter-simple sequence repeat markers |
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Authors: | D Q Fang M L Roose |
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Institution: | (1) Department of Botany and Plant Sciences, University of California, Riverside, CA 92521, USA, US |
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Abstract: | Inter-simple sequence repeat (ISSR) markers generated by 22 primers were tested for their ability to distinguish among samples
from 94 trees of 68 citrus cultivars. Within each of the six cultivar groups studied, most of these cultivars are so closely
related that they are difficult to distinguish by other molecular-marker techniques. ISSR markers involve PCR amplification
of DNA using a single primer composed of a microsatellite sequence anchored at the 3′ or 5′ end by 2–4 arbitrary, often degenerate,
nucleotides. The amplification products were separated on non-denaturing polyacrylamide gels and detected by silver staining.
ISSR banding profiles were very repeatable on duplicate samples. Different citrus species had very different fingerprint patterns.
Within Citrus sinensis (L.) Osbeck and C. paradisi Macf., in which all cultivars have originated by the selection of mutants, ISSR markers distinguished 14 of 33 sweet orange
and 1 of 7 grapefruit cultivars. Five of six lemon cultivars were discriminated by ISSR markers. Many differences were found
among mandarin cultivars; however, all five satsuma cultivars analyzed had identical ISSR fingerprints. Four of five citrange
cultivars were distinguishable, but ‘Troyer’ and ‘Carrizo’ had identical ISSR fingerprints. ‘Kuharske Carrizo’ citrange, which
has better citrus nematode resistance than other ‘Carrizo’ citrange accessions, had unique ISSR fingerprints. Three ISSR markers
that differentiated certain sweet orange cultivars were hybridized to Southern blots of sweet orange DNA digested with different
restriction endonucleases. The sweet orange cultivars tested could be distinguished by these ISSR-derived RFLP markers. Moreover,
one ISSR marker unique to ‘Ruby’ blood orange was observed in its progeny trees.
Received: 9 September 1996 / Accepted: 4 April 1997 |
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Keywords: | Citrus Poncirus ISSR markers RFLPs Cultivar identification |
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