Varietal traceability of bread 'Pane Nero di Castelvetrano' by denaturing high pressure liquid chromatography analysis of single nucleotide polymorphisms

DNA-based traceability is a very powerful tool to allow the identification of genetic material along thefood production chain. In particular, it is able to verify the presence of a certain cultivar in order to meetthe quality claims of local and certified products, or to detect any adulteration or contamination. The aimof this work was to apply the Denaturing High Performance Liquid Chromatography (DHPLC) techniquefor setting up a Single Nucleotide Polymorphism (SNP)-based method to achieve the varietal traceabilityof the durum wheat cultivar‘Timilia’in the‘Pane Nero di Castelvetrano’(Castelvetrano Black Bread) SlowFood Presidium. The DHPLC technique was optimized using 20 SNPs (8 transitions and 12 transversions)for both the starting whole meals and the corresponding breads, giving comparable results representedby an amount of detectable Timilia cv. ranging from 2.5% to 60% or more, thus providing an efficient toolto trace such cultivar in bread claimed to be Pane Nero di Castelvetrano. The obtained results representthefirst application of DHPLC in thefield of food science, and will help to preserve the authenticity ofPane Nero di Castelvetrano.