Microsatellite markers isolated in olive (Olea europaea L.) are suitable for individual fingerprinting and reveal polymorphism within ancient cultivars

Abstract

We have isolated and sequenced 52 microsatellites or simple sequence repeats (SSRs) from nearly 60 positive clones obtained from two 'Frantoio' olive genomic libraries enriched in (AC/GT) and (AG/CT) repeats, respectively. The repeat-containing fragments obtained from genomic DNA restricted with Tsp509I were separated using a biotinylated probe bound to streptavidin-coated paramagnetic beads. Fragments were then cloned into lambda ZAPII vector and sequenced. Thirty of the 36 primer pairs which gave correct re-amplification in the source genome were used to assay the polymorphism of 12 olive cultivars, namely four well-known cultivars ('Coratina', 'Frantoio', 'Leccino', 'Pendolino') and eight ancient cultivars grown locally near Lake Garda ('Casaliva', 'Favarol', 'Fort', 'Grignan', 'Less', 'Raza', 'Rossanel', 'Trep'). The local cultivars were each re- presented by two to four long-lived individuals. The analysis was carried out using (33)P-labelled primers and 6% polyacrylamide sequencing gels. All except two microsatellites showed polymorphism, the number of alleles varying from 1 to 5. The average genetic diversity ( H) was 0.55. The power of discrimination ( PD) was 0.60. All cultivars, including the local ones, were easily separated from each other. Variations in the SSR pattern were observed among individual plants of the same cultivar in four out of the eight local cultivars analysed. Several primer pairs (17%) amplified more than one locus.

Genetic diversity of Brazilian and introduced olive germplasms based on microsatellite markers

“…Synonymy in olives is a frequently reported problem (Carriero et al, 2002;Cipriani et al, 2002;Chafari et al, 2008;Khadari et al, 2008;Poljuha et al, 2008;Baldoni et al, 2009;Bracci et al, 2011). Indeed, the lack of an international organization responsible for standardizing names and cultivars and the limited centralization and availability of information about this species, which is associated with the global diffusion of olive plantlets and seedlings, makes the occurrence of synonymy and homonymy even more common now.…”

Traceability of Origin and Authenticity of Olive Oil

“…Many authors have reported on SSR development in olive and several of them are currently available for DNA analysis (Cipriani et al 2002;De la Rosa et al 2002;Rabiei & Tahmasebi Enferadi 2009;Sabino Gil et al 2006;Sefc et al 2000). This technique benefits from the use of microsatellites which are short stretches (1-6-bp long) of DNA, tandemly repeated several times.…”

“…The power of discrimination (PD) [21] of microsatellite primer pairs are also calculated as reported by Cipriani et al (2002), where the allele frequency of the He formula is replaced by the genotype frequency. (Alba et al, 2009) A research carried on the use of SSRs as a tool to identify the genetic background of olive oil which was involved the analysis of DNA sequences using a panel of seven simple sequence repeats (SSRs) to provide genotype-specific allelic profiles (Rabiei et al, 2010).…”

Fruit Germplasm Characterization: Genomics Approaches for the Valorisation of Genetic Diversity

“…Once these markers have been ascertained, the data can be readily shared among laboratories. Since not all microsatellites are identical Rallo et al, 2000;Sefc et al, 2000;Carriero et al, 2002;Cipriani et al, 2002;Muzzalupo et al, 2006Muzzalupo et al, , 2009a, however, successful utilization of known microsatellite markers requires prior information regarding the characteristics of a particular genetic locus ).…”

“…Many SSRs have been developed in olive and applied with success (Sefc et al, 2000;Carriero et al, 2002;Cipriani et al, 2002;De la Rosa et al, 2003;Sabino Gil et al, 2006). All these characteristics make them ideal markers for applications in analysis of intravariety variability issues (Cipriani et al, 2002;Lopes et al, 2004;Muzzalupo et al, 2009bMuzzalupo et al, , 2010, linkage mapping (Wu et al, 2004) and for characterizing olive germplasm resources (Belaj et al, 2004;Montemurro et al, 2007;Muzzalupo and Perri, 2009). Sarri et al, (2006) confirmed the power of SSR markers in the identification of 118 varieties from different Mediterranean countries to study the genetic diversities of olive varieties.…”