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RAPD MARKERS TO GENETIC MAPPING AND SELECTION OF CITRUS HYBRIDS

Molecular markers have many applications in plant breeding, enabling some types of genetic analyses. The aim of this work was to establish RAPD markers to be used to genetic mapping studies and selection of hybrids between 'Cravo' tangerine (Citrus reticulata Blanco) and 'Pêra' orange (C. sinensis (L.) Osbeck). DNA of the parents and six hybrids F1 was isolated from the leaves. The amplification reactions were performed in volumes of 13 µL, composed by GIBCO BRL 1x buffer, 1,54 mM MgCl2, 0,2 mM of each dNTP, 15 ng of each primer, 1,5 unit of Taq DNA Polymerase and 15 ng of genomic DNA. These reactions were carried out in thermocyclers programmed for 36 cycles of 1 min at 92ºC, 1 min at 36ºC, 2 min at 72ºC and 10 min of extension at 72ºC. It were evaluated random decamer primers of the kits A, AB, AT, AV, B, C, D, E, G, H, M, N, P, Q, R e U from Operon. One hundred thirteen primers were selected as polymorphics, with number of markers varying from 1 to 6 per primer. These primers amplified 201 (23,13%) polymorphic fragments, with application in genetic mapping and selection of hybrids. The frequency of primers with 1, 2, 3, 4, 5 and 6 polymorphic fragments was 49,5%, 33,6%, 9,7%, 4,4%, 1,8% e 1,0%, respectively.

'Pêra' sweet orange; 'Cravo' tangerine; hybrids; molecular makers


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