Identification of closely related citrus cultivars with inter-simple sequence repeat markers

• Published in: Theoretical and applied genetics Vol. 95; no. 3; pp. 408 - 417
• Main Authors: Fang, D.Q, Roose, M.L
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer 01.08.1997; Berlin Springer Nature B.V
• Subjects: Biological and medical sciences; Citroncirus webberi; Citrus; Citrus fruits; Citrus limon; Citrus paradisi; Citrus reticulata; Citrus sinensis; Classical genetics, quantitative genetics, hybrids; cultivar identification; Cultivars; DNA fingerprinting; Fundamental and applied biological sciences. Psychology; Genetic aspects; Genetic markers; Genetics of eukaryotes. Biological and molecular evolution; Identification and classification; Nematoda; Plant genetics; Poncirus trifoliata; Pteridophyta, spermatophyta; repetitive sequences; restriction fragment length polymorphism; species differences; Vegetals; United States; Citrus; Genetic variability; Genetic marker; Citrus fruit; Dicotyledones; Microsatellite DNA; Angiospermae; Rutaceae; Genetic identification; Spermatophyta; Cultivar; Polymorphism
• ISSN: 0040-5752; 1432-2242
• DOI: 10.1007/s001220050577

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.