Microsatellite High ‐Resolution Melting (SSR‐HRM) to Track Olive Genotypes: From Field to Olive Oil

In this study, we report a fast, sensitive, and easy‐to‐use strategy for PDO olive varietal identification. To achieve thi s aim four different DNA extraction methods were tested and compared, based on initial volume, extraction time, the gDNA concentration, and quality ratios. The optimized DNA extraction protocol from extra virgin olive oils, based on CTAB–hexane–chloroform extraction, proved to be the most effect ive. High‐resolution melting (HRM) DNA assay was developed based on nuclear microsatellites (gSSR) and plastid DNA (cpDNA) aiming an accurate identification of the olive varieties present in the olive oil samples. After PCR reproducibility evaluation, six molecular markers: three SSRs and three cp DNAloci were chosen based on their discrimination power. The SSR ‐HRM strategy assays were designed to targetUDO99 ‐011,UDO99 ‐039, UDO99‐024, andssrOeUA ‐DCA16 loci. All SSR ‐PCR products generated from these primers were analyzed by capillary electrophoresis (CE) for HRM data validation. The SSR coupled with HRM melting curve analysis generated 14 HRM profiles sufficient to genotype all varieties, highlighting their potential use for varietal discrimination. Thelocus ssrOeUA ‐DCA16 generated a specific melting curve that allow a high ‐throughput discrimination of the Picual and Cobrançosa varieties in olive oil samples. Further, theUDO99 ‐024 was also tested by SSR ‐HRM assay in commercial olive oil samples with promising results. Conside...
Source: Journal of Food Science - Category: Food Science Authors: Tags: New Horizons in Food Research Source Type: research