A Cost-Efficient and Simple Plant Oil DNA Extraction Protocol Optimized for DNA-Based Assessment of Product Authenticity

DNA-based assays offer precision in ascertaining the species/cultivar origin of agro-food products. Yet, obtaining DNA of sufficient quality and quantity is the main challenge while performing DNA-based food authentication analyses. The aim of the present work was to standardize a cost-efficient, easy-to-apply, yet effective plant oil DNA isolation protocol that allows reliable downstream PCR-based analyses. Because capillary electrophoresis (CE) separation of species/cultivar discriminating genomic fragments is a widely adopted approach in food genomics, a CE system was utilized in order to assess the performance of the proposed cetyl trimethyl ammonium bromide (CTAB)-based protocol. A plastid intergenic spacer and a nuclear olive gene were used as targets in order to evaluate the amplificability of DNA extracted with the CTAB-based protocol. The plastid barcode not only allowed assessing the reproducibility of PCR amplifications from the extracted oil DNA samples (olive, hazelnut, corn, rapeseed, cottonseed, and soybean oils) but also proved successful in discriminating all tested oil crop species based on amplified fragment length polymorphisms. Moreover, the barcode assay proved successful in correctly identifying the tested olive oil: cottonseed oil blends as admixtures of the two oil species. Thus, it was also feasible to demonstrate the potential of the barcode sequence as a discriminatory analyte to detect adulteration in plant oils. In addition, application of a CAPS (cleaved amplified polymorphic sequence) assay designed to genotype a nuclear SNP (single nucleotide polymorphism) marker resulted in the successful identification of the two single-cultivar olive oils included in the study. As a result of the present work, it was feasible to standardize a reliable and cost-efficient DNA extraction protocol that works well with both unrefined (olive and hazelnut) and refined (corn, rapeseed, cottonseed, and soybean) oils.