High Resolution Melting (HRM) applied to wine authenticity

Abstract

Wine authenticity methods are in increasing demand mainly in Denomination of Origin designations. The DNA-based methodologies are a reliable means of tracking food/wine varietal composition. The main aim of this work was the study of High Resolution Melting (HRM) application as a screening method for must and wine authenticity. Three sample types (leaf, must and wine) were used to validate the three developed HRM assays (Vv1-705bp; Vv2-375bp; and Vv3-119bp). The Vv1 HRM assay was only successful when applied to leaf and must samples. The Vv2 HRM assay successfully amplified all sample types, allowing genotype discrimination based on melting temperature values. The smallest amplicon, Vv3, produced a coincident melting curve shape in all sample types (leaf and wine) with corresponding genotypes. This study presents sensitive, rapid and efficient HRM assays applied for the first time to wine samples suitable for wine authenticity purposes.

Study of Commercially Available Lobelia chinensis Products Using Bar-HRM Technology

“…HRM technology has been utilized for discriminating cultivated varieties, medicinal plant species, herbal tea, wine authenticity (Jung et al, 2010; Osathanunkul et al, 2015, 2016; Costa et al, 2016; Li et al, 2016; Song et al, 2016; Pereira et al, 2017). However, a reliable HRM approach to identify herbal medicines on Chinese herb market is still lacks.…”

Microsatellite High‐Resolution Melting (SSR‐HRM) to Track Olive Genotypes: From Field to Olive Oil

“…The development of methodologies that require minimum manipulation is mandatory when dealing with samples that are being submitted for authenticity testing, diminishing substantially the identification errors. Therefore, the HRM technology may be a way to achieve that goal once it allows the genotyping of the varieties in a closed-tube reaction, without further analysis, as long as the variants have been previously identified (Distefano, Caruso, La Malfa, Gentile, & Wu, 2012;Pereira et al, 2017;Simko, 2016). Among the SSR loci previously selected, the UDO99-011 and UDO99-024 loci were the most discriminatory markers based on number of repeat motifs (Cipriani, Marrazzo, Marconi, Cimato, & Testolin, 2002).…”

“…The use of such an assay allows to quickly genotype the olive varieties based on allelic variants, avoiding the use of individually labelled primers, probes, and post-PCR handling, as required for SSRs analysed in a fragment analyser (Simko, 2016;Villano et al, 2017). HRM analysis based on SSR markers has been widely applied for fingerprinting in several species, such as grapevine and olive (Mackay, Wright, & Bonfiglioli, 2008;Pereira et al, 2017), cherry (Ganopoulos et al, 2011), lentils (Bosmali, Ganopoulos, Madesis, & Tsaftaris, 2012), among others, saving time and resources, once the reaction is carried out in a closed-tube.…”

“…In addition, several plastidial genes, such as the rpoC1, rbcL, and matK have also been proposed as barcode regions (Chase et al, 2005;Kress, Wurdack, Zimmer, Weigt, & Janzen, 2005). These characteristics make these molecular markers excellent for olive oil authenticity as they are based on small amplicons that are considered as the best targets when dealing with highly degraded DNA, such as present in olive oil (Fernandes, Costa, Oliveira, & Mafra, 2017;Hoseinpour, Foroughi, Nomanpour, & Nasab, 2017;Montemurro et al, 2015;Pasqualone et al, 2007Pasqualone et al, , 2014Pereira et al, 2017;Vietina, Agrimonti, & Marmiroli, 2013;Vietina, Agrimonti, Marmiroli, Bonas, & Marmiroli, 2011).…”

“…This characteristic grants the HRM technique a high-resolution power allowing the detection of single-base variants, small insertions or deletions and SNPs (Druml & Cichna-Markl, 2014). HRM has been used in the analysis of processed foods (Pereira et al, 2017;Villano et al, 2017), including olive oils (Ganopoulos, Bazakos, Madesis, Kalaitzis, & Tsaftaris, 2013) being regarded as an informative, easy, and low-cost method able to greatly reduce operating time in authentication processes.…”