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.
ABSTRACT
In this study polyphenolic compounds extracted from olive fruits of five registered cultivars were analyzed. A solid-liquid extraction (LSE) procedure with Extrelut cartridge (diatomaceous earth) using different eluents was developed to obtain polyphenolic compounds. HPLC-DAD and HPLC-MS methods were applied for the quali-quantitative analysis of each fraction obtained from LSE. The results of this work show that the LSE procedure with diatomaceous earth cartridge supplies a rapid and reproducible fractioning method able to obtain a quantitative recovery of all compounds and to collect fractions directly analyzed by HPLC. A comparison among different cultivars shows significant quantitative differences in some polyphenols, such as verbascoside, anthocyanic compounds, and oleuropein derivatives.
Subject(s)
Flavonoids , Phenols/analysis , Polymers/analysis , Vegetables/chemistry , Italy , Polyphenols , Seeds/chemistry , Species SpecificityABSTRACT
A modification of the Asakawa-Matsushita iodometric assay method for the determination of the content of lipid hydroperoxides was developed which permits the simultaneous processing of many samples of high lipid content. The method has the advantages of simplicity as well as good reproducibility, so it is not necessary to process standards with each determination. Our technique exceeds the sensitivity attained with other spectrophotometric determinations reported in the literature. The method requires the total elimination of water from the samples, and this was accomplished using an azeotropic mixture of ethanol:water of 96:4. The results obtained with liposomes indicate that the method is applicable to biological material limited to small volume samples, ranging 5-50 microliters. We want to emphasize that this method permits the study of the peroxidation process as function of time.
