Abstract
The Tubulin Based Polymorphism (TBP) method was used to genotype olive cultivars of different origin and to produce short-size cultivar-specific molecular probes. Both the first and the second intron of the members of the olive β-tubulin gene family were exploited as sources of DNA polymorphism. Compared with the data obtained with the use of a set of 11 SSR markers selected from an Olea europea L. database, TBP is shown to provide similar, if not better, information about the polymorphic content of the olive genomes, releasing, at the same time, a simple and discriminatory DNA barcode specific for any of the analyzed cultivars. Such a barcode is the source for the preparation of variety specific molecular probes.
Highlights
Native to the Mediterranean basin, olive (Olea europaea L.) represents one of the most ancient and precious agricultural trees for both cultural and economic reasons
The Tubulin Based Polymorphism (TBP) method, based on the detection of intron length polymorphisms present in the members of the β-tubulin gene family, was applied to 15 different olive varieties, mainly of Greek origin provided by the Agricultural University of Athens within the context of an exchange Erasmus program
The tubulin identity of the amplified fragments was verified by nucleotide sequencing and such information was further exploited for the preparation of a specific molecular probe
Summary
Native to the Mediterranean basin, olive (Olea europaea L.) represents one of the most ancient and precious agricultural trees for both cultural and economic reasons. The actual richness and diffusion of olive germplasms reflects on one side its longevity and scarce breeding improvement, and on the other a way of propagation that, largely based on cutting and grafting, has promoted a vast and multidirectional dispersal of the plant genetic material. A new approach that made use of 10 standard simple sequence repeats (SSRs) and 9 olive expressed tag sequences (OLEST) based on tri-nucleotidic EST-SSR, has been proposed as a reference combinatorial method of analysis that could possibly overcome the well known limit of the olive genotyping based on dinucleotide SSR motifs [9] Because of all these uncertainties, the laboriousness of the proposed methods and the lack of agreement on commonly shared molecular markers, the feasibility of the TBP (Tubulin-Based-Polymorphism) approach to the genotyping and classification of olive germplasm has been assessed. Data with reference to DNA barcoding, phylogenetic relationships and the production of varieties-specific molecular probes are reported
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