Durum what cultivar traceability in PDO Altamura bread by analysis of DNA microsatellites

Altamura bread is an Italian baking product that obtained the European mark of protected designation of origin (PDO). The varietal requirements of the official production protocol of this bread require it to be prepared from the durum wheat cultivars Appulo, Duilio, Arcangelo and Simeto (single or in combination, accounting for minimum 80%), and eventually other cultivars diffused in the production area. The aim of this work was to set up a microsatellite-based method for verifying the presence of the four required durum wheat cultivars in PDO Altamura bread, also in the presence of other cultivars up to 20%. Ten microsatellites were tested and the combination of the amplification profiles of four of them, characterised by high polymorphism and simple electrophoretic patterns, enabled to distinguish and identify breads from all the possible combinations of the cultivars required for PDO mark. The obtained amplicons were all in the range of molecular weight between 115 and 272 bp, and were analysed by capillary electrophoresis. The contribution of the single cultivars was detectable in the amplification profiles, enabling to verify their presence. The analysis was also effective in the case of additional cultivars.