SCREENING RAPD PRIMERS FOR MOLECULAR TAXONOMY AND CULTIVAR FINGERPRINTING IN THE GENUS ACTINIDIA

Eighty ten-base long arbitrary primers were tested for PCR-based DNA a mplification of three species of the genus Actinidia (A. deliciosa the kiwifruit, A. chinensis, and A. kolomikta), with the aim of screening species-specific and genotype-specific markers. Of the 80 primers tes ted, 30 gave an average of 3.5 bands which were monomorphic within one or two species and absent in the remaining one(s), thus resulting in useful markers for taxonomic and phylogenetic purposes. None of the pr imers tested produced bands linked to sex. Twenty primers out of the t wenty-five selected from a preliminary screening showed high levels of polymorphism, producing two to eleven patterns each from the 13 kiwif ruit cultivars examined. We found the Stoffel fragment and the Tag pol ymerase were both suitable for RAPD analysis, the most noticeable diff erence being the smaller size of fragments (0.4-1.2 kb) produced by th e former in comparison to the latter (1.0-3.4 kb). We tested also thre e different annealing temperatures (35, 37, and 39 degrees C) and foun d the intermediate one best for number of amplified bands and reproduc ibility of results.