Abstract
DNA authentication of wines is a process of verifying their authenticity by genetic identification of the main plant component. The sample preparation of experimental and commercial wines was carried out by precipitation of wine debris by centrifugation with preliminary exposure with precipitators and co-precipitators, including developed macro- and micro-volume methods applicable to white or red wines, using polyvinylpyrrolidone as a co-precipitator. Addition of 2-mercaptoethanol and proteinase K to the lysing solution made it possible to adapt the technology for DNA extraction from the precipitated wine debris. The additionally tested technique of DNA extraction from wine debris by dimethyl sulfoxide (DMSO) lysis had fewer stages and, consequently, a lower risk of contamination. The results of further testing of one of the designed primer pairs (UFGT-F1 and UFGT-R1) in conjunction with the tested methods of wine material sample preparation and nucleic acid extraction, showed the advantage in the given set of oligonucleotides over previously used ones in terms of sensitivity, specificity and reproducibility. The developing strategy for genetic identification of grape varieties and DNA authentication of wines produced from them based on direct sequencing of polymerase chain reaction (PCR) products is implemented by interpreting the detected polymorphic positions of variable Vitis vinifera L. UFGT gene locus with distribution and split into 13 UFGT gene-associated groups.
Highlights
DNA authentication of wines—a technological process of verification of their authenticity by genetic identification of the main plant component—technical varieties of grapes, molecular genetic analysis of residual nucleic acids of Vitis vinifera L., extracted from precipitated wine debris formed at the stage of sample preparation of biomaterial under research [3]
The results of preliminary studies indicated that the selected conditions for sample preparation, DNA extraction, and polymerase chain reaction (PCR) were applicable for amplification of the analyzed gene locus directly to the raw material, but not to the wine material produced from it
Taking into account the potential and limitations of tested methods, a number of proposed techniques expanding the arsenal of methodological tools, increased the rapidity of sample preparation of wine materials and wines and extraction of residual grapevine nucleic acids, enhanced the sensitivity, specificity and reproducibility of PCR, as well as provided identification of the UFGT gene-associated groups by sequencing
Summary
The search for objective criteria for assessing the authenticity and the place of origin of food products with the possibility of traceability of the entire life cycle “from field to counter” is a strategically important task that can only be achieved by multidisciplinary science-intensive approaches [1].The development of genomic, post-genomic and bioinformatic technologies increase the potential of molecular genetic approaches to DNA authentication of products and raw material for their production, unfolding perspectives of pilot design/elaboration in the quality management system with monitoring of counterfeit products, including wine [2].DNA authentication of wines—a technological process of verification of their authenticity by genetic identification of the main plant component—technical varieties of grapes, molecular genetic analysis of residual nucleic acids of Vitis vinifera L., extracted from precipitated wine debris formed at the stage of sample preparation of biomaterial under research [3].Various methods of sample preparation of wines based on centrifugation of debris precipitation with an additional use of precipitants and co-precipitants are known in the art.F. The development of genomic, post-genomic and bioinformatic technologies increase the potential of molecular genetic approaches to DNA authentication of products and raw material for their production, unfolding perspectives of pilot design/elaboration in the quality management system with monitoring of counterfeit products, including wine [2]. DNA authentication of wines—a technological process of verification of their authenticity by genetic identification of the main plant component—technical varieties of grapes, molecular genetic analysis of residual nucleic acids of Vitis vinifera L., extracted from precipitated wine debris formed at the stage of sample preparation of biomaterial under research [3]. Martinelli (2006) [4] performed sample preparation of commercial wines by precipitation of wine debris by centrifugation with preliminary exposure of the wine in a mixture with NaCl or isopropanol or sodium acetate in a freezer (−20 ◦ C) for
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