Grapevine (Vitis vinifera L.) SNP Detection through HRM Approach

In the last few years, single nucleotide polymorphisms (SNPs) have become the most popular genetic marker system in animals and plants. Their higher abundance combined with recent technological improvements makes SNP markers an excellent tool for grapevine genotyping. High resolution melting curve (HRM) is a recent advance technique for the detection of SNPs and INDELs in PCR amplicons and depends on DNA melting in the presence of saturating intercalating double-stranded DNA binding dyes. HMR presents several advantages: simple, rapid, high-throughput and low-cost technique. Anthocyanins represent a major group of the flavonoid family, with a crucial role in grapes once they are responsible for wine colour and its organoleptic properties. In Vitis vinifera L. several structural genes involved in anthocyanin biosynthesis have been identified. The aim of the study was to identify SNP markers in selected Portuguese red grapevine varieties concerning UFGT gene coding for UDP glucose–flavonoid 3-O-glucosyl-transferase. In silico analysis of UFGT gene sequence was performed and primer pairs were designed using Prime 3 software. Five red grapevine varieties, Tinto Cão, Tinta Roriz, Touriga Franca, Touriga Nacional and Cabernet Sauvignon, were sequenced and SNPs were annotated. An assay was designed to rapidly detect SNPs using HRM approach, allowing the differentiation of the grapevine varieties according to their sequence variants.

Acknowledgments: Portuguese Science Foundation (FCT) PTDC/AGR-ALI/117341/2010 and PhD grant SFRH/BD/44781/2008