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ISHS Acta Horticulturae 546: International Symposium on Molecular Markers for Characterizing Genotypes and Identifying Cultivars in Horticulture

MATHEMATICAL ANALYSIS OF RAPD DATA TO ESTABLISH RELIABILITY OF VARIETAL ASSIGNMENT IN VEGETATIVELY PROPAGATED SPECIES

Author:   J. Ibañez
Keywords:   Vitis vinifera, grapevine, similarity, probability-of-coincidence, legal- protection, patented-variety, breeders'-rights, intellectual-property
Abstract:
An unknown grapevine sample (US) and a seedless grape variety (V) were compared by RAPDs analysis using 122 different decamer oligonucleotides. No differences were found between them after studying 1741 bands. In order to determine the reliability of the assignment of US to V, two mathematical analysis have been developed. In the first one, individual bands are used as comparison unit. The probability of finding at least one non-shared band between V and US can be estimated as P= 1- SN, being S a similarity coefficient, and N the number of bands on the assumption of independence of loci. In the example, the calculated similarity between V and US is higher than 0.9974 with a P=0.9999, while the highest similarity found among a set of 45 seedless grape varieties was 0.9250. For the second approach, the complete RAPDs pattern obtained with each primer for V was compared with those obtained for the other 44 seedless grape varieties, and the number of non-differentiated pairs was established for each of the 10 primers used. Then, the probability of coincidence was calculated for these 10 primers (2.3×10-13) and for the 122 used between US and V (7.7× 10-155). Both mathematical analyses clearly demonstrate that US belongs to the variety, and these analysis are applicable to other asexually propagated plants.

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